Análise do polimorfismo de inversões cromossômicas em populações de Drosophila polymorpha do sul da ilha de Santa Catarina - Florianópolis

TCC(graduação) - Universidade Federal de Santa Catarina. Centro de Ciências Biológicas. Biologia.O grupo cardini do subgênero Drosophila apresenta quesitos muito interessantes principalmente no que se refere à distribuição, pigmentação e padrões evolutivos. Sua distribuição abrange a região Neotropical e, apesar de sua alta representatividade nesta região, ainda há poucos estudos sobre o grupo. A espécie D. polymorpha pertencente a este grupo tem recebido recentemente grande atenção dos pesquisadores devido aos seus polimorfismos: pigmentação e inversão cromossômica paracêntrica. No Brasil, estudos apontam sua maior abundância na região Sul e em levantamentos taxonômicos na região de Florianópolis, tem sido uma das espécies de drosofilídeos mais abundante. Os polimorfismos de inversão cromossômica em espécies de Drosophila constituem um dos sistemas mais estudados em evolução e dados recentes apontam D. polymorpha como uma das espécies mais polimórficas do grupo cardini. O presente estudo teve como objetivo analisar o polimorfismo de inversões cromossômicas em D. polymorpha na Caiera da Barra do Sul, área de Mata Atlântica do extremo sul da região insular de Florianópolis. As 6 isolinhagens cultivadas em laboratório foram obtidos a partir da captura com rede sobre iscas de banana fermentada deixadas no campo por no mínimo três dias. As lâminas com os cromossomos politênicos foram obtidas a partir das larvas de terceiro estágio destas linhagens. Duas inversões, XA e IIRE sendo esta última descrita pela primeira vez na literatura. O estudo contribui no complemento do mapa cromossômico da espécie, com informações citogenéticas agregadas a aspectos ecológicos do grupo bem como para futuras pesquisas sobre sua história evolutiva

Innate Immunity to Ebola virus

Ebola virus is a negative sense RNA virus and belongs to the family of Filoviridae. It can cause severe disease including hemorrhagic fever and multiorgan failure. Initial Ebola virus replication occurs in dendritic cells and macrophages, the sentinel cells of our immune system. However, infected dendritic cells fail to orchestrate an effective immune response. Virulence is partly associated to Ebola virus protein VP35, an interferon-antagonist which effectively counteracts the activation of RNA receptors from the RIG-I-like receptor pathway family. Furthermore, potential editing of viral RNA could have pro-viral effects, avoiding interferon induction. ADAR1 is an RNA editing enzyme, which modifies double-stranded RNA by adenosine-to-inosine editing, essential for differentiation between self and foreign RNA. This crucial negative regulator of the interferon response is expressed in two isoforms: the interferoninducible p150 present in both the cytoplasm and nucleus, and the constitutively expressed p110, which is restricted to the nucleus. Potential adenosine-to-inosine editing of Ebola virus genomes was shown recently in different approaches and in samples from Ebola virus disease survivors. Therefore, the early immune response to Ebola virus seems to be crucial for disease outcome. The aim of this thesis was to determine and analyze innate sensors and regulators of the innate signaling pathways relevant for Ebola virus infections. The goal was to research innate sensors of Ebola virus transcription and replication competent virus-like particles as well as to analyze sensing of Ebola virus RNA in cell-based assays and to investigate a link to altered innate sensing depending on the presence or absence of ADAR1 isoforms. The transcription and replication competent virus-like particle system allows life cycle modeling of Ebola virus under biosafety level-1 conditions. Innate sensing was measured by monitoring expression of the direct IRF3-target gene ISG54. Two different VP35 mutants were analyzed regarding their interferon antagonistic function as well as their function in replication and transcription. Production of transcription and replication competent virus-like particles including the VP35 mutants in HEK 293T cells, leads to a strong interferon-β response compared to wildtype VP35, suggesting that mutant VP35 proteins lost their antagonistic activity compared to wildtype VP35. Nevertheless, infection of cells with mutant VP35 virus-like particles does not lead to an immune response. To further investigate the lack of innate response to particles infection, the immunostimulatory potential of naked Ebola virus RNAs isolated from particles was assessed in a quantitative assay using monocyte-derived dendritic cells and monocytederived macrophages as a model for highly immunocompetent cells. Upon transfection of viral nucleic acids into immunocompetent cells, high sensing inductions are observed, suggesting that viral RNA components are sensed. To identify the particular innate pathways that are triggered by Ebola RNA, THP-1 knock-out cell lines deficient for key molecules of RNA and DNA sensing pathways were exposed to Ebola virus RNA. As expected, THP-1 cells deficient in the key molecule of the RNA sensing pathway lose the ability to trigger an immune response upon stimulation with Ebola virus RNA, suggesting members of the RIG-I like receptor family as initial sensors. Indeed, in gene knock-down experiments sensing of Ebola virus RNA was abrogated upon knock-down of RIG-I. Furthermore, ADAR1 knock-out HEK 293T cells, as well as knock-out cells stably expressing ADAR1p150, catalytically inactive ADAR1p150in, and the ADAR1p110 isoform were generated. Ebola virus particles were produced in respective cells or wildtype cells, followed by Ebola virus RNA extraction. Target cells were transfected with respective Ebola virus RNA and innate sensing was measured by monitoring the expression of the IRF-3 target gene ISG54 as well as by Western Blots for IRF-3 activation. Here, it was shown that Ebola virus RNA extracted from particles produced in wildtype cells induce an IRF-3-dependent response after transfection in primary myeloid cells. Interestingly, Ebola virus RNA produced in ADAR1 knock-out cells induce a higher immune response after transfection in A549 cells than RNA produced in wildtype cells. This suggests ADAR1 as a negative regulator for sensing. In addition, the innate response to particle-associated RNA stemming from cells overexpressing ADAR1p150 is strongly diminished in comparison to RNA stemming from ADAR1 knock-out cells or cells overexpressing the catalytically inactive form of p150 or the nuclear isoform p110. This suggests that strong RNA editing activity by the active interferon-stimulated p150, but not p110 influences the capacity for Ebola virus RNA sensing. In conclusion, this work leads to a better understanding of Ebola virus-host interactions and established ADAR1 as a pro-viral factor during Ebola virus infection and as a negative regulator of innate sensing of Ebola virus RNA. A better understanding of the first interactions between Ebola virus and innate regulators can help to advance therapeutic strategies

On Hegel, Women, and the Foundation of Ethical Life: Why Gender Doesn’t Belong in the Family.

Feminist philosophers are right to criticize Hegel’s prejudices against women. In many of his works, Hegel reduces women to their physiology as means of explaining why they occupy a subordinate role in nature and in society. Such treatment seems arbitrary at best, for the gendering of roles disrupts Hegel’s dialectical approach to spirit without any meaningful gain. Despite this defect in Hegel’s work, what is positive in Hegelian social and political philosophy remains intact. In this paper I argue that the sexist claims that Hegel makes about women are irrelevant to his theory of the family in the Philosophy of Right. Therein, Hegel outlines three components that are necessary for the completion of the family: marriage, property and assets, and the raising of children. Hegel also includes a description of the different roles occupied by family members and divides these roles along gender lines. Given the three components that are essential to the family, I argue that there is no necessary basis for familial roles to be divided by gender

What is a Family? Considerations on Purpose, Biology, and Sociality

There are many different interpretations of what the family should be – its desired member composition, its primary purpose, and its cultural significance – and many different examples of what families actually look like across the globe. I examine the most paradigmatic conceptions of the family that are based upon the supposed primary purpose that the family serves for its members and for the state. I then suggest that we ought to reconceptualize how we understand and define the family in an effort to move away from these paradigmatic conceptions. This approach requires that we examine the way(s) in which the family has been defined descriptively – that is, how families have been defined historically – in an effort to determine what a normative theory of the family might look like. The goal of this inquiry is to define a family in terms of what it ought to be – a goal that moves our understanding of the family to a new conceptual landscape. I then present my own account of familial relations that aims to capture a normative understanding of the unique primary purpose that the family serves for its members

Cardiovascular effects of lead and mercury and their mixtures in rats

Cardiovascular diseases are the major cause of death worldwide. It is a group of diseases, which affect the heart, the vasculature and the brain. Lifestyle and metabolic risk factors are major contributors to cardiovascular ill-health. In addition to these risk factors, a growing number of scientific studies show that some environmental pollutants, e.g. lead and mercury, can adversely affect cardiovascular health. Despite the increasing amount of knowledge from human and animal studies, cardiovascular effects of lead, mercury species or their mixtures are not well understood. It is also unknown if safe exposure thresholds for these metals exist or the underlying mechanisms of action for the elicitation of cardiovascular toxicity. The first set of studies had the objectives to elucidate the range of effects of single exposure to lead, inorganic mercury or methylmercury on the cardiovascular system. Therefore, male Wistar rats were exposed to a broad range of doses of lead, inorganic mercury or methylmercury for four weeks through the drinking water. Cardiovascular health of the rats was assessed by measuring the blood pressure and the cardiac electrical activity after four weeks of exposure, while the heart function and blood flow in the carotid artery was measured at baseline and at the end of the exposure duration. The study showed that all three metals differ in their effects on the cardiovascular system. Lead showed bi-phasic dose-response curves for several cardiovascular end-points. No cardiovascular effects were observed for inorganic mercury, while methylmercury showed linear dose-response curves. Based on these results, safe levels of exposure for lead and methylmercury were derived. The second study applied the same experimental design as the previous study in order to investigate the cardiovascular effects of combined exposures to lead, inorganic mercury and methylmercury. The mixture ratios were based on reference and exposure values published in the scientific literature. The adverse cardiovascular effects, which were observed for single exposures were reversed for the mixtures indicating antagonism. In contrast to single exposures, mixtures negatively affected the electrical activity of the heart (synergism), which could lead to arrhythmias and heart failure. The third set of studies focused on the exploration of oxidative stress, kidney function and damage, and global DNA methylation as potential mechanisms of action for the development of elevated blood pressure. Results for lead showed an increase in oxidative stress but not mercury. While only lead was associated with kidney damage, only inorganic mercury was related to altered global DNA methylation. Methylmercury appears to elevate blood pressure through a not investigated mechanism. Therefore, oxidative stress and kidney damage seem to be associated with elevated blood pressure but not global DNA methylation. Overall, the research presented in this thesis shows that lead, inorganic mercury and methylmercury and their mixtures have the ability to adversely affect the cardiovascular system. However, each metal affected the cardiovascular system differently and surprisingly, mixtures showed antagonism or synergism depending on the examined end-point, which was reflected in the results of the mechanistic study. As health problems of the cardiovascular system, e.g. hypertension, occur mainly in the adult population and in particular the elderly, cardiovascular effects should be considered as an important end-point for this age group in addition to neurodevelopmental effects in children

The history of neuromyelitis optica

The discovery of a novel serum autoantibody (termed NMO-IgG or AQP4-Ab) in a subset of patients in 2004 has revived interest in neuromyelitis optica (NMO). While the history of classical multiple sclerosis has been extensively studied, only little is known about the history of NMO. In the present article, we provide a comprehensive review of the early history of this rare but intriguing syndrome. We trace the origins of the concept of NMO in the 19th century medical literature and follow its evolution throughout the 20th and into the 21st century. Finally, we discuss recent proposals to revise the concept of NMO and explain why there is indeed a need for a more systematic and descriptive nomenclature

‘Medusa-head ataxia’: the expanding spectrum of Purkinje cell antibodies in autoimmune cerebellar ataxia. Part 1: Anti-mGluR1, anti-Homer-3, anti-Sj/ITPR1 and anti-CARP VIII

Serological testing for anti-neural autoantibodies is important in patients presenting with idiopathic cerebellar ataxia, since these autoantibodies may indicate cancer, determine treatment and predict prognosis. While some of them target nuclear antigens present in all or most CNS neurons (e.g. anti-Hu, anti-Ri), others more specifically target antigens present in the cytoplasm or plasma membrane of Purkinje cells (PC). In this series of articles, we provide a detailed review of the clinical and paraclinical features, oncological, therapeutic and prognostic implications, pathogenetic relevance, and differential laboratory diagnosis of the 12 most common PC autoantibodies (often referred to as ‘Medusa-head antibodies’ due to their characteristic somatodendritic binding pattern when tested by immunohistochemistry). To assist immunologists and neurologists in diagnosing these disorders, typical high-resolution immunohistochemical images of all 12 reactivities are presented, diagnostic pitfalls discussed and all currently available assays reviewed. Of note, most of these antibodies target antigens involved in the mGluR1/calcium pathway essential for PC function and survival. Many of the antigens also play a role in spinocerebellar ataxia. Part 1 focuses on anti-metabotropic glutamate receptor 1-, anti-Homer protein homolog 3-, anti-Sj/inositol 1,4,5-trisphosphate receptor- and anti-carbonic anhydrase-related protein VIII-associated autoimmune cerebellar ataxia (ACA); part 2 covers anti-protein kinase C gamma-, anti-glutamate receptor delta-2-, anti-Ca/RhoGTPase-activating protein 26- and anti-voltage-gated calcium channel-associated ACA; and part 3 reviews the current knowledge on anti-Tr/delta notch-like epidermal growth factor-related receptor-, anti-Nb/AP3B2-, anti-Yo/cerebellar degeneration-related protein 2- and Purkinje cell antibody 2-associated ACA, discusses differential diagnostic aspects and provides a summary and outlook

‘Medusa head ataxia’: the expanding spectrum of Purkinje cell antibodies in autoimmune cerebellar ataxia. Part 2: Anti-PKC-gamma, anti-GluR-delta2, anti-Ca/ARHGAP26 and anti-VGCC

Serological testing for anti-neural autoantibodies is important in patients presenting with idiopathic cerebellar ataxia, since these autoantibodies may indicate cancer, determine treatment and predict prognosis. While some of them target nuclear antigens present in all or most CNS neurons (e.g. anti-Hu, anti-Ri), others more specifically target antigens present in the cytoplasm or plasma membrane of Purkinje cells (PC). In this series of articles, we provide a detailed review of the clinical and paraclinical features, oncological, therapeutic and prognostic implications, pathogenetic relevance, and differential laboratory diagnosis of the 12 most common PC autoantibodies (often referred to as ‘Medusa head antibodies’ due their characteristic somatodendritic binding pattern when tested by immunohistochemistry). To assist immunologists and neurologists in diagnosing these disorders, typical high-resolution immunohistochemical images of all 12 reactivities are presented, diagnostic pitfalls discussed and all currently available assays reviewed. Of note, most of these antibodies target antigens involved in the mGluR1/calcium pathway essential for PC function and survival. Many of the antigens also play a role in spinocerebellar ataxia. Part 1 focuses on anti-metabotropic glutamate receptor 1-, anti-Homer protein homolog 3-, anti-Sj/inositol 1,4,5-trisphosphate receptor- and anti-carbonic anhydrase-related protein VIII-associated autoimmune cerebellar ataxia (ACA); part 2 covers anti-protein kinase C gamma-, anti-glutamate receptor delta-2-, anti-Ca/RhoGTPase-activating protein 26- and anti-voltage-gated calcium channel-associated ACA; and part 3 reviews the current knowledge on anti-Tr/delta notch-like epidermal growth factor-related receptor-, anti-Nb/AP3B2-, anti-Yo/cerebellar degeneration-related protein 2- and Purkinje cell antibody 2-associated ACA, discusses differential diagnostic aspects, and provides a summary and outlook