Phytochemical profile and some biological activities of three Centaurea species from Turkey

Purpose: To characterise the phytochemical profile of whole plants of Centaurea balsamita, C. depressa and C. lycopifolia with LC-ESI-MS/MS, and as well as their antioxidant, anticholinesterase and antimicrobial activities. Methods: Organic and aqueous extracts of the three Centaurea species were evaluated for DPPH free radical, ABTS cation radical scavenging and cupric reducing antioxidant capacity (CUPRAC). Acetyland butyryl-cholinesterase enzyme inhibition abilities of the extracts using petroleum ether, acetone, methanol and water were studied to determine anticholinesterase activity, while antimicrobial activity was determined by disc diffusion method using appropriate antimicrobial standards and organisms. The phytochemical components of the methanol extracts were assessed by LC-MS/MS. Results: The methanol extract of C. balsamita exhibited much higher DPPH free and ABTS cation radicals scavenging activities (with IC50 of 62.65 ± 0.97 and 24.21 ± 0.70 mg/ml, respectively) than the other extracts. The petroleum ether extracts of the plant species exhibited moderate inhibitory activity against butyrylcholinesterase enzymes while the acetone extract of C. balsamita showed good antifungal activity against Candida albicans. Quinic acid (17513 ± 813 μg/g, 63874 ± 3066 μg/g and 108234 ± 5195 μg/g) was the major compound found in the methanol extracts of C. balsamita, C. depressa and C. Lycopifolia, respectively. Conclusion: These results indicate quinic acid is the major compound in the three plant species and that Centaurea balsamita has significant antioxidant, anticholinesterase and antimicrobial properties. Further studies to identify the compounds in the extracts responsible for the activities are required.

Chemical Reaction Dynamics of the Liquid/Vapor Interface Studied by Mass Spectrometry

This thesis presents investigations of chemical reactions occurring at the liquid/vapor interface studied using novel sampling methodologies coupled with detection by mass spectrometry. Chapters 2 and 3 utilize the recently developed technique of field-induced droplet ionization mass spectrometry (FIDI-MS), in which the application of a strong electric field to a pendant microliter droplet results in the ejection of highly charged progeny droplets from the liquid surface. In Chapter 2, this method is employed to study the base-catalyzed dissociation of a surfactant molecule at the liquid/vapor interface upon uptake of ammonia from the gas phase. This process is observed to occur without significant modulation of the bulk solution pH, suggesting a transient increase in surface pH following the uptake of gaseous ammonia. Chapter 3 presents real-time studies of the oxidation of the model tropospheric organic compound glycolaldehyde by photodissociation of iron (III) oxalate complexes. The oxidation products of glycolaldehyde formed in this process are identified, and experiments in a deoxygenated environment identify the role of oxygen in the oxidation pathway and in the regeneration of iron (III) following photo-initiated reduction. Chapter 4 explores alternative methods for the study of heterogeneous reaction processes by mass spectrometric sampling from liquid surfaces. Bursting bubble ionization (BBI) and interfacial sampling with an acoustic transducer (ISAT) generate nanoliter droplets from a liquid surface that can be sampled via the atmospheric pressure interface of a mass spectrometer. Experiments on the oxidation of oleic acid by ozone using ISAT are also presented. Chapters 5 and 6 detail mechanistic studies and applications of free-radical-initiated peptide sequencing (FRIPS), a technique employing gas-phase free radical chemistry to the sequencing of peptides and proteins by mass spectrometry. Chapter 5 presents experimental and theoretical studies on the anomalous mechanism of dissociation observed in the presence of serine and threonine residues in peptides. Chapter 6 demonstrates the combination of FRIPS with ion mobility-mass spectrometry (IM-MS) for the separation of isomeric peptides.

Evaluation of sperm retrieval rate with bilateral testicular sperm extraction in infertile patients with azoospermia

Background: About 10% to 15% of infertile men have azoospermia, which could be obstructive or non-obstructive. Diagnostic biopsy from the testis and recently testicular sperm extraction (TESE) are the most precise investigations in these patients. Testicular biopsy can be done unilaterally or bilaterally. The worth of unilateral or bilateral testicular biopsy in men with azoospermia is controversial. Objective: To evaluate the necessity of bilateral diagnostic biopsy from the testis in new era of diagnosis and treatment of male infertility. Materials and Methods: In this retrospective study, we reviewed the results of testis biopsy in 419 azoospermic men, referred to Yazd Research and Clinical Center for Infertility from 2009-2013. Patients with known obstructive azoospermia were excluded from the study. Results: In totally, 254 infertile men (60.6%) were underwent unilateral TESE, which in 175 patients (88.4%) sperm were extracted from their testes successfully. Bilateral testis biopsy was done in 165 patients (39.4%) which in 37 patients (22.4%), sperm were found in their testes tissues. Conclusion: Due to the low probability of positive bilateral TESE results especially when we can’t found sperm in the first side, we recommend that physicians re-evaluate the risk and benefit of this procedure in era of newer and more precise technique of sperm retrieval like micro TESE.

Caractérisation de l’activité transcriptionnelle antivirale et immunorégulatrice dépendante de STAT2 et IRF9, mais indépendante de STAT1, induite par la costimulation par TNF

Les cellules épithéliales pulmonaires constituent la première ligne de défense face aux virus respiratoires via la sécrétion de mucus, de peptides, de cytokines et chimiokines qui déterminent l'élimination ou la progression de l’infection. Les principales cytokines antivirales produites par les cellules épithéliales alvéolaires (AEC) sont les interférons (IFN) type I (α/β) et III (λ). La liaison d’IFNβ à son récepteur induit une voie antivirale bien caractérisée qui aboutit à l’activation du complexe ISGF3 (STAT1, STAT2 et IRF9) qui permet la transcription de multiples gènes codant pour des protéines à activité antivirale et immunorégulatrice. Il a récemment été démontré que la costimulation des cellules épithéliales pulmonaires par l’IFNβ et le Tumor Necrosis Factor α (TNFα), également produit lors d’une infection, synergisent pour induire un état antiviral tardif distinct. D’autre part, il a été montré que la synergie entre le TNFα et l'IFNβ induit une voie de signalisation impliquant STAT2 et IRF9, mais indépendante de STAT1 permettant l’expression du gène DUOX2. Notre but est de déterminer l’importance de cette nouvelle voie de signalisation induite par la costimulation du TNFα+IFNβ, impliquant STAT2 et IRF9 indépendamment de STAT1 dans la régulation d’un programme transcriptionnel antiviral et immunorégulateur tardif. Notre premier objectif est de déterminer si des gènes antiviraux et immunorégulateurs qui sont induits par la costimulation par TNFα+IFNβ sont dépendants de la voie STAT2/IRF9, indépendamment de STAT1. En utilisant la technique de qRT-PCR, nous avons identifié 3 gènes immunorégulateurs, CXCL10, IDO et APOBEC3G, induits de manière synergique en réponse à TNFα+IFNβ dans les cellules A549, un modèle de cellules épithéliales pulmonaires. Afin de confirmer que ces gènes sont induits indépendamment de STAT1, nous avons validé leur expression dans la lignée cellulaire U3A déficiente en STAT1. Par l'utilisation d'ARN interférants (ARNi) dirigés contre STAT2 et IRF9, nous avons confirmé que l’induction de ces gènes est dépendante de STAT2 et IRF9. Finalement, l’analyse de l’activité du promoteur de CXCL10 en réponse à TNFα+IFNβ par des essais rapporteurs luciférases a permis de montrer que la régulation se fait au niveau transcriptionnel. Notre deuxième objectif, est de déterminer si STAT6 pourrait remplacer STAT1 dans la voie de signalisation induite par TNFα+IFNβ. En effet, STAT6 est un inducteur connu de l’expression de DUOX2 en réponse à IL4+IL13. Contrairement à notre hypothèse, l’inhibition de STAT6 par ARNi augmente l’expression de DUOX2 en réponse à TNFα+IFNβ suggérant que STAT6 est un régulateur négatif. Nos résultats ont permis de comprendre de manière plus détaillée les mécanismes mis en place dans le développement d’une réponse antivirale. D’autre part, l’étude de l’effet de l’IFNβ et du TNFα est également pertinente pour les maladies chroniques inflammatoires et autoimmunes. De plus, nos résultats illustrent un nouveau paradigme concernant les mécanismes de signalisation cellulaire impliqués dans la synergie entre deux cytokines qui pourrait être applicable à des combinaisons de cytokines autres que TNFα+IFNβ.

ANALYSIS OF POPULUS TREMULOIDES CLONAL VARIATION AND DELINEATION IN THE OTTAWA NATIONAL FOREST

The technique of delineating Populus tremuloides (Michx.) clonal colonies based on morphology and phenology has been utilized in many studies and forestry applications since the 1950s. Recently, the availability and robustness of molecular markers has challenged the validity of such approaches for accurate clonal identification. However, genetically sampling an entire stand is largely impractical or impossible. For that reason, it is often necessary to delineate putative genet boundaries for a more selective approach when genetically analyzing a clonal population. Here I re-evaluated the usefulness of phenotypic delineation by: (1) genetically identifying clonal colonies using nuclear microsatellite markers, (2) assessing phenotypic inter- and intraclonal agreement, and (3) determining the accuracy of visible characters to correctly assign ramets to their respective genets. The long-term soil productivity study plot 28 was chosen for analysis and is located in the Ottawa National Forest, MI (46° 37'60.0" N, 89° 12'42.7" W). In total, 32 genets were identified from 181 stems using seven microsatellite markers. The average genet size was 5.5 ramets and six of the largest were selected for phenotypic analyses. Phenotypic analyses included budbreak timing, DBH, bark thickness, bark color or brightness, leaf senescence, leaf serrations, and leaf length ratio. All phenotypic characters, except for DBH, were useful for the analysis of inter- and intraclonal variation and phenotypic delineation. Generally, phenotypic expression was related to genotype with multiple response permutation procedure (MRPP) intraclonal distance values ranging from 0.148 and 0.427 and an observed MRPP delta value=0.221 when the expected delta=0.5. The phenotypic traits, though, overlapped significantly among some clones. When stems were assigned into phenotypic groups, six phenotypic groups were identified with each group containing a dominant genotype or clonal colony. All phenotypic groups contained stems from at least two clonal colonies and no clonal colony was entirely contained within one phenotypic group. These results demonstrate that phenotype varies with genotype and stand clonality can be determined using phenotypic characters, but phenotypic delineation is less precise. I therefore recommend that some genetic identification follow any phenotypic delineation. The amount of genetic identification required for clonal confirmation is likely to vary based on stand and environmental conditions. Further analysis, however, is needed to test these findings in other forest stands and populations.

The Discursive Construction of Japanese Identity and its Haunting Others

This dissertation examined the formation of Japanese identity politics after World War II. Since World War II, Japan has had to deal with a contradictory image of its national self. On the one hand, as a nation responsible for colonizing fellow Asian countries in the 1930s and 1940s, Japan has struggled with an image/identity as a regional aggressor. On the other hand, having faced the harsh realities of defeat after the war, Japan has seen itself depicted as a victim. By employing the technique of discourse analysis as a way to study identity formation through official foreign policy documents and news media narratives, this study reconceptualized Japanese foreign policy as a set of discursive practices that attempt to produce renewed images of Japan’s national self. The dissertation employed case studies to analyze two key sites of Japanese postwar identity formation: (1) the case of Okinawa, an island/territory integral to postwar relations between Japan and the United States and marked by a series of US military rapes of native Okinawan girls; and (2) the case of comfort women in Japan and East Asia, which has led to Japan being blamed for its wartime sexual enslavement of Asian women. These case studies found that it was through coping with the haunting ghost of its wartime past that Japan sought to produce “postwar Japan” as an identity distinct from “wartime imperial Japan” or from “defeated, emasculated Japan” and, thus, hoped to emerge as a “reborn” moral and pacifist nation. The research showed that Japan struggled to invent a new self in a way that mobilized gendered dichotomies and, furthermore, created “others” who were not just spatially located (the United States, Asian neighboring nations) but also temporally marked (“old Japan”). The dissertation concluded that Japanese foreign policy is an ongoing struggle to define the Japanese national self vis-à-vis both spatial and historical “others,” and that, consequently, postwar Japan has always been haunted by its past self, no matter how much Japan’s foreign policy discourses were trying to make this past self into a distant or forgotten other.

Caractérisation de l’activité transcriptionnelle antivirale et immunorégulatrice dépendante de STAT2 et IRF9, mais indépendante de STAT1, induite par la costimulation par TNF

Les cellules épithéliales pulmonaires constituent la première ligne de défense face aux virus respiratoires via la sécrétion de mucus, de peptides, de cytokines et chimiokines qui déterminent l'élimination ou la progression de l’infection. Les principales cytokines antivirales produites par les cellules épithéliales alvéolaires (AEC) sont les interférons (IFN) type I (α/β) et III (λ). La liaison d’IFNβ à son récepteur induit une voie antivirale bien caractérisée qui aboutit à l’activation du complexe ISGF3 (STAT1, STAT2 et IRF9) qui permet la transcription de multiples gènes codant pour des protéines à activité antivirale et immunorégulatrice. Il a récemment été démontré que la costimulation des cellules épithéliales pulmonaires par l’IFNβ et le Tumor Necrosis Factor α (TNFα), également produit lors d’une infection, synergisent pour induire un état antiviral tardif distinct. D’autre part, il a été montré que la synergie entre le TNFα et l'IFNβ induit une voie de signalisation impliquant STAT2 et IRF9, mais indépendante de STAT1 permettant l’expression du gène DUOX2. Notre but est de déterminer l’importance de cette nouvelle voie de signalisation induite par la costimulation du TNFα+IFNβ, impliquant STAT2 et IRF9 indépendamment de STAT1 dans la régulation d’un programme transcriptionnel antiviral et immunorégulateur tardif. Notre premier objectif est de déterminer si des gènes antiviraux et immunorégulateurs qui sont induits par la costimulation par TNFα+IFNβ sont dépendants de la voie STAT2/IRF9, indépendamment de STAT1. En utilisant la technique de qRT-PCR, nous avons identifié 3 gènes immunorégulateurs, CXCL10, IDO et APOBEC3G, induits de manière synergique en réponse à TNFα+IFNβ dans les cellules A549, un modèle de cellules épithéliales pulmonaires. Afin de confirmer que ces gènes sont induits indépendamment de STAT1, nous avons validé leur expression dans la lignée cellulaire U3A déficiente en STAT1. Par l'utilisation d'ARN interférants (ARNi) dirigés contre STAT2 et IRF9, nous avons confirmé que l’induction de ces gènes est dépendante de STAT2 et IRF9. Finalement, l’analyse de l’activité du promoteur de CXCL10 en réponse à TNFα+IFNβ par des essais rapporteurs luciférases a permis de montrer que la régulation se fait au niveau transcriptionnel. Notre deuxième objectif, est de déterminer si STAT6 pourrait remplacer STAT1 dans la voie de signalisation induite par TNFα+IFNβ. En effet, STAT6 est un inducteur connu de l’expression de DUOX2 en réponse à IL4+IL13. Contrairement à notre hypothèse, l’inhibition de STAT6 par ARNi augmente l’expression de DUOX2 en réponse à TNFα+IFNβ suggérant que STAT6 est un régulateur négatif. Nos résultats ont permis de comprendre de manière plus détaillée les mécanismes mis en place dans le développement d’une réponse antivirale. D’autre part, l’étude de l’effet de l’IFNβ et du TNFα est également pertinente pour les maladies chroniques inflammatoires et autoimmunes. De plus, nos résultats illustrent un nouveau paradigme concernant les mécanismes de signalisation cellulaire impliqués dans la synergie entre deux cytokines qui pourrait être applicable à des combinaisons de cytokines autres que TNFα+IFNβ.

Favorability conditions in the adoption of agile method practices for software development in a public banking

The main purpose of this paper is to propose and test a model to assess the degree of conditions favorability in the adoption of agile methods to develop software where traditional methods predominate. In order to achieve this aim, a survey was applied on software developers of a Brazilian public retail bank. Two different statistical techniques were used in order to assess the quantitative data from the closed questions in the survey. The first, exploratory factorial analysis validated the structure of perspectives related to the agile model of the proposed assessment. The second, frequency distribution analysis to categorize the answers. Qualitative data from the survey opened question were analyzed with the technique of qualitative thematic content analysis. As a result, the paper proposes a model to assess the degree of favorability conditions in the adoption of Agile practices within the context of the proposed study.

Gobierno empresarial: etnografía de un proyecto de cultura organizacional en la Cía. Aceros Ltda

La presente investigación analiza las dinámicas y relaciones sociales de la implementación de un estudio de cultura organizacional en un escenario etnográfico concreto. El propósito del presente trabajo, es analizar el estudio de la cultura organizacional como una técnica de gobierno con unas prácticas específicas, negociadas, consensuadas y establecidas en el marco de la participación de los empleados en el estudio realizado por la empresa.

Subjective Aspects of People with Chronic Ulcers: a Qualitative Study in Brazil and Portugal

Introduction: The experience built in the process of living with chronic ulcer is marked by changes such as the inability to work, to relate socially and causes the need to adapt to a routine care. Understanding this network of meanings is important to develop techniques of individual or collective care. Objective: To understand impregnated subjectivities in the everyday experiences of Brazilian and Portuguese patients with chronic ulcers, in the light of the Oral History of life. Method: Comparative study with a qualitative approach, using Oral History as method and technique. The network of collaborators, formed by males and females, aged 39-82 years, was structured into two groups, one consisting of 06 people in Natal/RN, Brazil and another composed of 10 people in Évora/Portugal. In both groups, the narratives were collected through open questions, which were recorded, transcribed and analyzed by the technique of thematic content analysis. Results: Through the analysis, three themes were revealed: Social repercussion in Brazil and in Portugal; Trajectory of theinjured person; and Coping mechanisms. Conclusion: Changes were observed in social life, leading contributors to isolation, in addition to confronting the stigma experienced. There were also identified points as the reinterpretation of chronic wound carrier about their disease and coping strategies of their chroniccondition.

Manifestations of Stigma and Prejudice Informed by Treated Lepers

Introduction: The segregation of people affected by leprosy in lepercolonies as well as presenting itself ineffective, caused irreversible and irreparable consequences in the lives of individuals who came to carry the marks of stigma and prejudice surrounding the disease. Objective: To identify traits related to the stigma and prejudice in the content of lepers' 12 speeches, separated compulsorily in Saint Francis of Assisi Colony in the city of Natal, located in the state of Rio Grande do Norte. Method: Descriptive study focusing on the oral history of life. The narratives were collected in April 2010 through a recorded interview and analyzed by means of the technique of Thematic Content Analyses. Results: The stigma and the prejudice crystallized in our culture had caused and still cause huge suffering and pain in the participants, who grew up and several aged in biological terms, living with a stigmatizing disease and its complications, sometimes disabling ones. In social and psychological terms, they were discriminated, rejected and even expelled from the familiar conviviality and the original social nucleus. Conclusion: In this manner, one notices the need of these people recover their attachments, values and self-esteem, share feelings and build relationships so as to be integrated in the real world.

“Losanga” decorated imitations of italic late republican black gloss tableware from South-Western Iberia: A multi-analytical/microchemical characterization

The micro-chemical/mineralogical composition of samples of grey-paste imitations of Italic Late Republican black gloss tableware displaying a particular kind of lozenge-shaped decoration (“Losanga pottery”) from Portuguese and Spanish archaeological sites in SW Iberia has been analysed by BSEM + EDS, μXRD, Powder XRD, Portable XRF and μRaman spectroscopy. “Losanga” decorated ceramics have been found throughout the Western Mediterranean. Most of the sherds display a green-brown to greyish-black engobe at the surface resembling the gloss found in Attic pottery from Classical Greece. The overall chemical, mineralogical and fossiliferous homogeneities of the ceramic paste show common features (low K-feldspar/plagioclase ratio, high Ca content, abundance of well-preserved fragments of foraminifera microfossils) that indicate low firing conditions in the kiln ranging from 650 to 900 °C. With respect to the ceramic body, analytical results confirm an enrichment in the surface gloss layer of iron, potassium and aluminium and a depletion in silicon and calcium; the very fine grain size of the surface coating suggests elutriation of iron oxide-rich clays as confirmed by the presence of magnetite, maghemite and goethite in μ-XRD scan. Chemical and mineralogical data also suggest that the firing process was performed in a 600–850 °C temperature range, adopting the well-known technique of alternating oxidizing and reducing firing conditions largely employed at the time. The analytical results, while compatible with the archaeological hypothesis of a common provenance of the raw materials for pottery production from the Guadalquivir valley workshops cannot be considered conclusive due to the similarity in the geological substrate in the two SW Iberian regions under study.

SOCIAL REPRESENTATIONS OF BASICS LIFE SUPPORT

Victims of cardiac arrest need immediate Basic Life Support, in order to preserve as much as possible, the flow of blood to the brain and heart and other vital organs, it is essential to gain time pending differentiated help, performing simple acts and practical (BLS) to save lives. Learn how to perform RPC is an interactive process that requires knowledge and skills, but at the same time an act of solidarity, social responsibility, civic consciousness, and a duty of citizenship. Because no one revives alone, it requires a coordinated work of a team, all citizens must join forces in a single goal: Save Lives, the massification of the BLS (RPC, 2014). We conducted an exploratory study that aimed to identify the social representations of basic life support in the general population. We used the technique of free association of words through a short questionnaire, we obtained a sample of 45 participants. The results show that participants were mostly female and 27 that fashion of age was in the age group 40 to 59 years. With regard to social representations, we find an organized structure follows the core: help, help to revive, and save is giving life, are in fact structural and consensual elements in basic life support. In more peripheral elements we find extremely important elements, which can be worked in a way so that the core is more efficient such as to act coordinately as a team in face of an accident, it can thus be successful in practice. The social representation of basic life support does not differ from that referred in the literature on the subject, but it is common knowledge that these skills can only be acquired if they are systematically trained, because they obey an algorithm that if it is not settled theoretical and instrumentally it is not effective in practice.

Effect of phytoremediation with Atriplex nummularia in irrigated soils with saline waste..

In the semi-arid region of northeastern Brazil, the Atriplex plant genus has been efficient in removing salts from soils irrigated with saline wastewater. However, this removal might not be significant compared with the amount of salts added to the soil by the wastewater irrigation. Considering this aspect, the aim of this work was to evaluate the effectiveness of Atriplex nummularia Lindl plants in the remediation of a soil submitted to saline wastewater irrigation. Despite the known inhibition effect of saline wastewater on soil enzyme activity, the cultivation of Atriplex nummularia Lindl maintained the treated soil enzyme activity levels similar to the ones found in natural soils.

Kinetic and mechanistic studies on unconventional antioxidant activities of natural compounds

In chapter one, the autoxidation kinetics of natural oil substrates, including, triglyceric sunflower oil, olive oil, terpenic squalene, and p-cymene were calibrated through differential oximetry methods. Calibration allows their use as reference oxidizable substrates for further studies, e.g. for quantitative testing of antioxidants under biomimetic settings. Several essential oils samples, of different botanical species or different productions of same species were studied for their antioxidant activity in inhibited autoxidation kinetics. Their antioxidant activities were matched with their composition analyzed by GC-MS. In chapter two, the molecular mechanism of the synergy between the common phenolic antioxidants such as tocopherol and catechols with widespread essential component gamma-terpinene was studied through lipid oxidation kinetics. Wherein, gamma-terpinene was able to disclose the key intermediacy HOO·, which acted as a reducing agent regenerating the phenolic antioxidant. This counterintuitive role of HOO· radicals was further investigated in detail and allowed to rationalize for the first time the purported antioxidant behavior of PDA melanin nanoparticles. It will also open to a deeper understanding of the redox biology of quinones. Regarding melanin, its role is broadly important in living organisms and its control, including its inhibition, is of great importance with several relevant applications ranging from food preservation to control of human skin pigmentation. In chapter three, an oximetry method combined with the traditional UV-Vis spectroscopy was developed to study the tyrosinase inhibition kinetics, which allowed identifying Glabridin (from G. glabra, L.), as one of the most effective natural tyrosinase inhibitors.