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      Antioxidant, Antimicrobial and Cytotoxic Properties as Well as the Phenolic Content of the Extract from Hancornia speciosa Gomes

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          Abstract

          Hancornia speciosa Gomes (Apocynaceae) is a fruit tree, popularly known as mangabeira, and it is widely distributed throughout Brazil. Several parts of the plant are used in folk medicine, and the leaf and bark extracts have anti-inflammatory, antihypertensive, antidiabetic, and antimicrobial properties. In this study, we investigated the chemical composition of the ethanolic extract of Hancornia speciosa leaves (EEHS) and its antioxidant, antimicrobial, and cytotoxic activities as well as the mechanisms involved in cell death. The chemical compounds were identified by liquid chromatography coupled to mass spectrometry (LC-MS/MS). The antioxidant activity of the EEHS was investigated using the method that involves the scavenging of 2,2-diphenyl-1-picrylhydrazyl free radicals as well as the inhibition of oxidative hemolysis and lipid peroxidation induced by 2,2’-azobis (2-amidinopropane) in human erythrocytes. The antimicrobial activity was determined by calculating the minimum inhibitory concentration, minimum bactericidal concentration, minimum fungicidal concentration, and zone of inhibition. Kasumi-1 leukemic cells were used to assess the cytotoxic activity and mechanisms involved in cell death promoted by the EEHS. The chemical compounds identified were quinic acid, chlorogenic acid, catechin, rutin, isoquercitrin, kaempferol-rutinoside, and catechin-pentoside. The EEHS demonstrated antioxidant activity via the sequestration of free radicals, inhibition of hemolysis, and inhibition of lipid peroxidation in human erythrocytes incubated with an oxidizing agent. The antimicrobial activity was observed against American Type Culture Collection (ATCC) and hospital strains of bacteria and fungi, filamentous fungi and dermatophytes. The cytotoxic activity of the EEHS was induced by apoptosis, reduction of the mitochondrial membrane potential, and activation of cathepsins. Together, these results indicate the presence of phenolic compounds and flavonoids in the EEHS and that their antioxidant, antimicrobial, and cytotoxic activities in acute myeloid leukemia cells are mediated by apoptosis.

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          Attributable mortality of nosocomial candidemia, revisited.

          We reexamined the attributable mortality of nosocomial candidemia 15 years after a retrospective cohort study performed at our hospital demonstrated an attributable mortality of 38%. For all episodes of nosocomial candidemia between 1 July 1997 and 30 June 2001, we matched control patients with case patients by age, sex, date of hospital admission, underlying disease(s), length of time at risk, and surgical procedure(s). We analyzed 108 matched pairs. There were no statistically significant differences in age, sex, underlying disease(s), time at risk, surgical procedure, or vital signs at admission between cases and controls. The crude mortality among case patients was 61% (66 of 108 patients), compared with 12% (13 of 108) among control patients, for an attributable mortality of 49% (95% CI, 38%-60%). Nosocomial candidemia is still associated with an extremely high crude and attributable mortality--much higher than that expected from underlying disease alone.
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            Oxidative damage in multiple sclerosis lesions

            Multiple sclerosis is a chronic inflammatory disease of the central nervous system, associated with demyelination and neurodegeneration. The mechanisms of tissue injury are currently poorly understood, but recent data suggest that mitochondrial injury may play an important role in this process. Since mitochondrial injury can be triggered by reactive oxygen and nitric oxide species, we analysed by immunocytochemistry the presence and cellular location of oxidized lipids and oxidized DNA in lesions and in normal-appearing white matter of 30 patients with multiple sclerosis and 24 control patients without neurological disease or brain lesions. As reported before in biochemical studies, oxidized lipids and DNA were highly enriched in active multiple sclerosis plaques, predominantly in areas that are defined as initial or ‘prephagocytic’ lesions. Oxidized DNA was mainly seen in oligodendrocyte nuclei, which in part showed signs of apoptosis. In addition, a small number of reactive astrocytes revealed nuclear expression of 8-hydroxy-d-guanosine. Similarly, lipid peroxidation-derived structures (malondialdehyde and oxidized phospholipid epitopes) were seen in the cytoplasm of oligodendrocytes and some astrocytes. In addition, oxidized phospholipids were massively accumulated in a fraction of axonal spheroids with disturbed fast axonal transport as well as in neurons within grey matter lesions. Neurons stained for oxidized phospholipids frequently revealed signs of degeneration with fragmentation of their dendritic processes. The extent of lipid and DNA oxidation correlated significantly with inflammation, determined by the number of CD3 positive T cells and human leucocyte antigen-D expressing macrophages and microglia in the lesions. Our data suggest profound oxidative injury of oligodendrocytes and neurons to be associated with active demyelination and axonal or neuronal injury in multiple sclerosis.
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              Caspase-independent cell death.

              Caspase activation has been frequently viewed as synonymous with apoptotic cell death; however, caspases can also contribute to processes that do not culminate in cell demise. Moreover, inhibition of caspases can have cytoprotective effects. In a number of different models, caspase inhibition does not maintain cellular viability and instead shifts the morphology of death from apoptosis to nonapoptotic pathways. Here, we explore the contribution of caspases to cell death, either as upstream signals or as downstream effectors contributing to apoptotic morphology, as well as alternative strategies for cell death inhibition. Such alternative strategies may either target catabolic hydrolases or be aimed at preventing mitochondrial membrane permeabilization and its upstream triggers.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, CA USA )
                1932-6203
                1 December 2016
                2016
                : 11
                : 12
                : e0167531
                Affiliations
                [1 ]School of Environmental and Biological Science, Federal University of Grande Dourados, Rodovia Dourados Ithaum, Dourados, MS, Brazil
                [2 ]Department of Biochemistry, Federal University of São Paulo, São Paulo, SP, Brazil
                [3 ]Center for Interdisciplinary Research Biochemistry, University of Mogi das Cruzes, Av. Dr. Cândido Xavier de Almeida Souza, Mogi das Cruzes, SP, Brazil
                [4 ]Federal University of Mato Grosso do Sul, Center of Biological and Health Sciences, Laboratory of Natural Products and Mass Spectrometry, Campo Grande, MS, Brazil
                [5 ]Agricultural College of Bragança, Polytechnic Institute of Bragança, Campus Santa Apolónia, Bragança, Portugal and Centre of Molecular and Environmental Biology, University of Minho, Campus de Gualtar, Braga, Portugal
                Institute of medical research and medicinal plant studies, CAMEROON
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                • Conceptualization: UPS JFC HFVT EJPG CAC LME KPS ELS.

                • Data curation: EJPG CAC LME KPS ELS.

                • Formal analysis: UPS JFC HFVT EJPG CAC LME KPS ELS.

                • Funding acquisition: EJPG CAC LME KPS ELS.

                • Investigation: UPS JFC HFVT EJPG CAC LME KPS ELS.

                • Methodology: UPS JFC HFVT EJPG CAC LME KPS ELS.

                • Project administration: ELS.

                • Resources: EJPG CAC LME KPS ELS.

                • Supervision: EJPG CAC LME KPS ELS.

                • Validation: UPS JFC HFVT EJPG CAC LME KPS ELS.

                • Visualization: UPS KPS ELS.

                • Writing – original draft: UPS JFC EJPG CAC LME KPS ELS.

                • Writing – review & editing: UPS JFC EJPG CAC LME KPS ELS.

                Author information
                http://orcid.org/0000-0002-6557-7914
                Article
                PONE-D-16-35638
                10.1371/journal.pone.0167531
                5132198
                27907185
                2a0cbc5a-0a33-4996-8e35-0067caecd9f9
                © 2016 Santos et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 5 September 2016
                : 9 November 2016
                Page count
                Figures: 7, Tables: 3, Pages: 19
                Funding
                Funded by: 1. Foundation to Support to Fundação de Apoio ao Desenvolvimento do Ensino, Ciência e Tecnologia do Estado de Mato Grosso do Sul (FUNDECT)—http://fundect.ledes.net/
                Funded by: 2. Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)—http://www.capes.gov.br/
                Funded by: 3. Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) – http://www.cnpq.br/
                Funded by: 4. PRODERÂ, (24.073 – Â, Portugal)
                Award Recipient :
                This work was supported by grants from Fundação de Apoio ao Desenvolvimento do Ensino, Ciência e Tecnologia do Estado de Mato Grosso do Sul (FUNDECT, Brazil), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Brazil) and PRODER, (24.073 – Â, Portugal). E.L.S.; E.J.P.G e C.A.L.C., were recipient of felowship from CNPq, Brazil. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Cell Biology
                Cell Processes
                Cell Death
                Apoptosis
                Biology and Life Sciences
                Biochemistry
                Antioxidants
                Medicine and Health Sciences
                Pharmacology
                Drugs
                Antimicrobials
                Biology and Life Sciences
                Microbiology
                Microbial Control
                Antimicrobials
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                Cell Biology
                Cellular Types
                Animal Cells
                Blood Cells
                Red Blood Cells
                Physical Sciences
                Chemistry
                Chemical Compounds
                Phenols
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                Physiology
                Electrophysiology
                Membrane Potential
                Medicine and Health Sciences
                Physiology
                Electrophysiology
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                Biology and Life Sciences
                Organisms
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                Candida Albicans
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                Candida Albicans
                Medicine and Health Sciences
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                Candida Albicans
                Biology and Life Sciences
                Mycology
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                Research and Analysis Methods
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                Yeast and Fungal Models
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                Biology and Life Sciences
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