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Fluorescent probes for the detection of reactive oxygen species in human spermatozoa

Asian Journal of Andrology 2020年第5期22卷 465-471页

作者： sara escada-rebelo Francisca G Mora Ana P Sousa Teresa Almeida-Santos Artur Paiva Joao Ramalho-Santos phD Programme in Experimental Biology and Biomedicine CNC-Center for Neuroscience and Cell BiologyUniversity of CoimbraCoimbra 3004-504Portugal Biology of Reproduction and Stem Cell Group CNC-Center for Neuroscience and Cell BiologyUniversity of CoimbraCoimbra 3004-504Portugal IIIUC-Institute for Interdisciplinary Research Casa Costa AlemaoUniversity of CoimbraCoimbra 3030-789Portugal Reproductive Medicine Unit University Hospitals of CoimbraCoimbra 3004-561Portugal Faculty of Medicine University of CoimbraCoimbra 3000-370Portugal Clinical Pathology Unit University Hospitals of CoimbraCoimbra 3004-561Portugal Department of Life Sciences University of CoimbraCoimbra 3000-456Portugal

Reactive oxygen species(ROS)production is a by-product of mitochondrial activity and is necessary for the acquisition of the capacitated state,a requirement for functional ***,an increase in oxidative stress,due to an abnormal production of ROS,has been shown to be related to loss of sperm function,highlighting the importance of an accurate detection of sperm ROS,given the specific nature of this *** this work,we tested a variety of commercially available fluorescent probes to detect ROS and reactive nitrogen species(RNS)in human sperm,to define their *** both flow cytometry(FC)and fluorescence microscopy(FM),we confirmed that MitoSOX™Red and dihydroethidium(DHE)detect superoxide anion(as determined using antimycin A as a positive control),while DAF-2A detects reactive nitrogen species(namely,nitric oxide).For the first time,we also report that RedoxSensor™Red CC-1,CellROX®Orange Reagent,and MitoPYl seem to be mostly sensitive to hydrogen peroxide,but not ***,mean fluorescence intensity(and not percentage of labeled cells)is the main parameter that can be reproducibly monitored using this type of methodology.

关键词： flow cytometry fluorescent probes human spermatozoa oxidative stress reactive oxygen species

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Trophic niche changes associated with the eradication of invasive mammals in an insular lizard: an assessment using isotopes

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Current Zoology 2022年第2期68卷 211-219页

作者： sara F.NUNES Mario MOTA-FERREIRA Marta SAMPAIO Joana ANDRADE Nuno OLIVEIRA Rui rebelo Ricardo ROCHA Faculty of Sciences Centre for EcologyEvolution and Environmental ChangesUniversity of LisbonLisbon1749-016Portugal CIBI0-lnBI0 Research Center in Biodiversity and Genetic ResourcesUniversity of PortoVairao4485-661Portugal CIBI0-lnBI0 Research Center in Biodiversity and Genetic ResourcesInstitute of AgronomyUniversity of LisbonLisbon1349-017Portugal Sociedade Portuguesa Para o Estudo das Aves Av.Columbano Bordalo Pinheiro873°AndarLisboa1070-062Portugal

Invasive species are a major threat to island biodiversity,and their eradications have substantially contributed to the conservation of island ***,the consequences of eradications on the trophic ecology of native taxa are largely ***,we used the eradication of invasive black rats Rattus rattus and European rabbits Oryctolagus cuniculus from the Berlenga Island,in the western coast of Portugal,as a whole-ecosystem experiment to investigate the effects of the eradication of invasive mammals on the trophic niche and body dimensions of the island-restricted Berlenga wall lizard Podarcis carbonelli berlengensis over a 2-year *** results suggest an expansion of the isotopic niche and an intensification of the sexual dimorphism of the lizard following mammal ***,we found considerable variability in isotopic niche across the island and detected evidence of sex-specific and season-modulated nutritional requirements of this threatened *** findings support that the eradication of 2 of the planets most problematic invasive vertebrates led to changes in the lizard trophic niche and sexual dimorphism in just 2 *** suggests that the ecological pressuresfor example,prey availability and habitat structureto which lizards are exposed have substantially changed *** study emphasizes the scientific value of island eradications as experiments to address a wide range of ecological questions and adds to the increasing body of evidence supporting substantial conservation gains associated with these restoration interventions.

关键词： food-webs invasive species eradication Oryctolagus cuniculus Podarcis carbonelli berlengensis Rattus rattus

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Phosphoprotein Phosphatase 1 Isoforms Alpha and Gamma Respond Differently to Prodigiosin Treatment and Present Alternative Kinase Targets in Melanoma Cells

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Journal of Biophysical Chemistry 2014年第2期5卷 67-77页

作者： Margarida Fardilha Joao Figueiredo Margarita Espona-Fiedler Juliana Felgueiras Luis Korrodi-Gregorio sara L.C.Esteves Sandra rebelo Odete A.B.da Cruz Silva Edgar da Cruz e Silva Ricardo Perez-Tomas Laboratory of Signal Transduction Centre for Cell BiologyBiology Department and Health Sciences DepartmentUniversity of AveiroAveiroPortugal Department of Pathology and Experimental Therapeutics Cancer Cell Biology Research GroupUniversity of BarcelonaBarcelonaSpain Laboratory of Neuroscience Centre for Cell BiologyBiology Department and Health Sciences DepartmentUniversity of AveiroAveiroPortugal

Reversible protein phosphorylation is a central regulatory mechanism of cell function. Deregulation of the balanced actions of protein kinases and phosphatases has been frequently associated with several pathological conditions, including cancer. Many studies have already addressed the role of protein kinases misregulation in cancer. However, much less is known about protein phosphatases influence. Phosphoprotein Phosphatase 1 (PPP1) is one of the major serine/threonine protein phosphatases who has three catalytic isoforms: PPP1CA, PPP1CB, and PPP1CC. Its function is achieved by binding to regulatory subunits, known as PPP1-interacting proteins (PIPs), which may prefer a catalytic isoform. Also, some inhibitors/enhancers may exhibit isoform specificity. Here we show that, prodigiosin (PG), a molecule with anticancer properties, promotes the formation of PPP1CA-AKT complex and not of PPP1CC-MAPK complex. Both, AKT and MAPK, are well-known PIPs from two pathways that crosstalk and regulate melanoma cells survival. In addition, the analysis performed using surface plasmon resonance (SPR) technology indicates that PPP1 interacts with obatoclax (OBX), a drug that belongs to the same family of PG. Overall, these results suggest that PG might, at least in part, act through PPP1C/PIPs. Also, this study is pioneer in demonstrating PPP1 isoform-specific modulation by small molecules.

关键词： Phosphoprotein Phosphatase 1 Catalytic Subunit Surface Plasmon Resonance Mitogen-Activated Protein Kinase V-Akt Murine Thymoma Viral Oncogene Glycogen Synthase Kinase 3

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