Effect of C5-Methylation of Cytosine on the UV-Induced Reactivity of Duplex DNA: Conformational and Electronic Factors

International audienceC5-methylation of cytosines is strongly correlated with UV-induced mutations detected in skin cancers. Mutational hot-spots appearing at TCG sites are due to the formation of pyrimidine cyclobutane dimers (CPDs). The present study, performedfor the model DNA duplex (TCGTA)3·(TACGA)3 and the constitutive single strands, examines the factors underlying the effect of C5-methylation on pyrimidine dimerization at TCG sites. This effect is quantified for the first time by quantum yields ϕ.They were determined following irradiation at 255, 267, and 282 nm and subsequent photoproduct analysis using HPLC coupled to mass spectrometry. C5-methylation leads to an increase of the CPD quantum yield up to 80% with concomitant decrease of that of pyrimidine(6−4) pyrimidone adducts (64PPs) by at least a factor of 3. The obtained ϕ values cannot be explained only by the change of the cytosine absorption spectrum upon C5-methylation. The conformational and electronic factors that may affect the dimerization reaction are discussed in light of results obtained by fluorescence spectroscopy,molecular dynamics simulations, and quantum mechanical calculations. Thus, it appears that the presence of an extra methyl on cytosine affects the sugar puckering, thereby enhancing conformations of the TC step that are prone to CPD formation but less favorable to 64PPs. In addition, C5-methylation diminishes the amplitude of conformational motions in duplexes; in the resulting stiffer structure, ππ* excitations may be transferred from initially populated exciton states to reactive pyrimidines giving rise to CPDs

Evaluación de la estabilidad en el tiempo del contenido proteico en variedades de arveja

El objetivo del presente trabajo ha sido evaluar la estabilidad del contenido proteico de las variedades desarrolladas por INTA y la Facultad de Ciencias Agrarias de la Universidad Nacional de Rosario y las disponibles en el mercado, contemplando tanto el comportamiento en la producción primaria como la calidad industrial de los granos producidos a la hora de producir aislados proteicos.EEA OliverosFil: Accoroni, Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Oliveros. Laboratorio de Agroindustria; ArgentinaFil: Magnano, Luciana. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Oliveros; ArgentinaFil: Espósito, María Andrea. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Oliveros; Argentin

Excited State Pathways Leading to Formation of Adenine Dimers

International audienceThe reaction intermediate in the path leading to UV-induced formation ofadenine dimers A=A and AA* is identified for the first time quantum mechanically, usingPCM/TD-DFT calculations on (dA)2 (dA: 2′deoxyadenosine). In parallel, its fingerprint isdetected in the absorption spectra recorded on the millisecond time-scale for the singlestrand (dA)20 (dA: 2′deoxyadenosine)

Chapter The offshore environmental impact by Sarno river in Naples bay (South-West Italy)

The goal of this work has been to establish the influence of the Sarno river on the present sedimentation in the Naples bay continental shelf by evaluating organic matter contribution and pollution. Sediments samples were collected, by van Veen grab, in 71 stations located offshore the Sarno river between Vesuvian and Sorrento Peninsula coasts. The characteristics of the surface sediments were analysed to highlight spatial trends in the (i) granulometry (grain-size); (ii) total nitrogen, organic carbon and total phosphorus; (iii) metal content (Hg, Cd, Pb, As, Cr, Cu, Ni, Zn, Fe and Mn)

Identification of a Sorbicillinoid-Producing Aspergillus Strain with Antimicrobial Activity Against Staphylococcus aureus: a New Polyextremophilic Marine Fungus from Barents Sea

The exploration of poorly studied areas of Earth can highly increase the possibility to discover novel bioactive compounds. In this study, the cultivable fraction of fungi and bacteria from Barents Sea sediments has been studied to mine new bioactive molecules with antibacterial activity against a panel of human pathogens. We isolated diverse strains of psychrophilic and halophilic bacteria and fungi from a collection of nine samples from sea sediment. Following a full bioassay-guided approach, we isolated a new promising polyextremophilic marine fungus strain 8Na, identified as Aspergillusprotuberus MUT 3638, possessing the potential to produce antimicrobial agents. This fungus, isolated from cold seawater, was able to grow in a wide range of salinity, pH and temperatures. The growth conditions were optimised and scaled to fermentation, and its produced extract was subjected to chemical analysis. The active component was identified as bisvertinolone, a member of sorbicillonoid family that was found to display significant activity against Staphylococcus aureus with a minimum inhibitory concentration (MIC) of 30 μg/mL. © 2018, Springer Science+Business Media, LLC, part of Springer Nature

Human neutrophil migration and activation by BJcuL, a galactose binding lectin purified from Bothrops jararacussu venom

<p>Abstract</p> <p>Background</p> <p>Neutrophil migration to an inflamed site constitutes the first line of the innate immune response against invading microorganisms. Given the crucial role of endogenous lectins in neutrophil mobilization and activation, lectins from exogenous sources have often been considered as putative modulators of leukocyte function. Lectins purified from snake venom have been described as galactoside ligands that induce erythrocyte agglutination and platelet aggregation. This study evaluated human neutrophil migration and activation by C-type lectin BJcuL purified from <it>Bothrops jararacussu </it>venom.</p> <p>Results</p> <p>Utilizing fluorescence microscopy, we observed that biotinylated-BJcuL was evenly distributed on the neutrophil surface, selectively inhibited by D-galactose. Lectin was able to induce modification in the neutrophil morphology in a spherical shape for a polarized observed by optical microscopy and exposure to BJcuL in a Boyden chamber assay resulted in cell migration. After 30 minutes of incubation with BJcuL we found enhanced neutrophil functions, such as respiratory burst, zymozan phagocytosis and an increase in lissosomal volume. In addition, BJcuL delays late apoptosis neutrophils.</p> <p>Conclusion</p> <p>These results demonstrate that BJcuL can be implicated in a wide variety of immunological functions including first-line defense against pathogens, cell trafficking and induction of the innate immune response since lectin was capable of inducing potent neutrophil activation.</p

Cytocompatibility of poly(p-dioxanone)/ poly(hydroxybutic) (PPD/PHB) blends to cartilage tissue engineering

In order of seek strategies to improve the interaction between bioreabsorbable polymer materials and cellular growth, this work aimed at evaluating in vitro the influence of PPD/PHB blends on cell adhesion and fibrochondrocytes growth. Fibrochondrocytes cells were obtained by primary extraction from enzymatic digestion methods. The PPD/PHB blends were prepared by casting with 100/0, 60/40 and 50/50 compositions, and were characterized by scanning electron microscopy (SEM). After 6, 48, 120 and 168 hours in culture, ultrastructural observations showed changes in cell morphology, suggesting that the fibrochondrocytes can respond to substrate modifications, changing their phenotypic profile. The MTT analyses showed that the blends did not present cytotoxicity and allowed fibrochondrocytes adhesion and proliferation on the membranes in all compositions. The colorimetric Sirius Red test revealed the capability of extracellular matrix synthesis on the blends, from which one can conclude that the PPD/PHB blends are not cytotoxic and can be indicated for cell culture.Buscando estratégias que repercutam na melhoria da interação entre materiais poliméricos biorreabsorvíveis e o crescimento celular, o presente estudo in vitro teve como objetivo estudar a influência de blendas de PPD/PHB na adesão celular e crescimento de fibrocondrócitos obtidos a partir de cultura primária. As blendas de PPD/PHB foram preparadas pelo método de evaporação de solvente nas composições 100/0, 60/40 e 50/50 e caracterizadas por microscopia eletrônica de varredura (MEV). Observações ultra-estruturais mostraram alterações na morfologia celular, sugerindo que os fibrocondrócitos podem responder a alterações no substrato alterando seu perfil fenotípico. As análises com MTT demonstraram que as blendas não apresentaram citotoxicidade e permitiram a adesão e proliferação dos fibrocondrócitos sobre os substratos em todas as suas composições. O ensaio colorimétrico com Sirius Red evidenciou a capacidade de manutenção da síntese de matriz extracelular colágena sobre as amostras, concluindo-se que as blendas de PPD/PHB podem ser indicadas para o cultivo celular.383388Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Expression of aryl hydrocarbon receptor (AHR) and AHR-interacting protein in pituitary adenomas: pathological and clinical implications.

peer reviewedaudience: researcher, professionalGermline mutations of the aryl hydrocarbon receptor (AHR)-interacting protein (AIP) gene confer a predisposition to pituitary adenomas (PA), usually in the setting of familial isolated PA. To provide further insights into the possible role of AIP in pituitary tumour pathogenesis, the expression of AIP and AHR was determined by real-time RT-PCR and/or immunohistochemistry (IHC) in a large series of PA (n=103), including 17 with AIP mutations (AIP(mut)). Variable levels of AIP and AHR transcripts were detected in all PA, with a low AHR expression (P<0.0001 versus AIP). Cytoplasmic AIP and AHR were detected by IHC in 84.0 and 38.6% of PA respectively, and significantly correlated with each other (P=0.006). Nuclear AHR was detected in a minority of PA (19.7%). The highest AIP expression was observed in somatotrophinomas and non-secreting (NS) PA, and multivariate analysis in somatotrophinomas showed a significantly lower AIP immunostaining in invasive versus non-invasive cases (P=0.019). AIP expression was commonly low in other secreting PA. AIP immunostaining was abolished in a minority of AIP(mut) PA, with a frequent loss of cytoplasmic AHR and no evidence of nuclear AHR. In contrast, AIP overexpression in a subset of NS PA could be accompanied by nuclear AHR immunopositivity. We conclude that down-regulation of AIP and AHR may be involved in the aggressiveness of somatotrophinomas. Overall, IHC is a poorly sensitive tool for the screening of AIP mutations. Data obtained on AHR expression suggest that AHR signalling may be differentially affected according to PA phenotype

Nationwide genetic analysis of more than 600 families with inherited eye diseases in Argentina

This study corresponds to the first large-scale genetic analysis of inherited eye diseases (IED) in Argentina and describes the comprehensive genetic profile of a large cohort of patients. Medical records of 22 ophthalmology and genetics services throughout 13 Argentinian provinces were analyzed retrospectively. Patients with a clinical diagnosis of an ophthalmic genetic disease and a history of genetic testing were included. Medical, ophthalmological and family history was collected. A total of 773 patients from 637 families were included, with 98% having inherited retinal disease. The most common phenotype was retinitis pigmentosa (RP, 62%). Causative variants were detected in 379 (59%) patients. USH2A, RPGR, and ABCA4 were the most common disease-associated genes. USH2A was the most frequent gene associated with RP, RDH12 early-onset severe retinal dystrophy, ABCA4 Stargardt disease, PROM1 cone-rod dystrophy, and BEST1 macular dystrophy. The most frequent variants were RPGR c.1345 C > T, p.(Arg449*) and USH2A c.15089 C > A, p.(Ser5030*). The study revealed 156/448 (35%) previously unreported pathogenic/likely pathogenic variants and 8 possible founder mutations. We present the genetic landscape of IED in Argentina and the largest cohort in South America. This data will serve as a reference for future genetic studies, aid diagnosis, inform counseling, and assist in addressing the largely unmet need for clinical trials to be conducted in the region