Metodologia para o gerenciamento integrado saudável dos resíduos sólidos da bacia da estrada nova do município de Belém/PA

O presente trabalho tem o objetivo de criar uma metodologia para o Gerenciamento Integrado dos Resíduos Sólidos (GIRS) que associa a prática pertinente ao tema, com programas que envolvam a comunidade, no sentido de manter o meio ambiente limpo e saudável, tendo como cenário o município de Belém, especificamente a Bacia Hidrográfica da Estrada Nova (BHEN). Objetiva também identificar porquê, apesar da BHEN possuir coleta de resíduos e serviços de limpeza realizada pela Secretaria Municipal de Saneamento (SESAN), a mesma permanece, constantemente, suja, principalmente, de lixo e entulho lançados nas vias públicas e canais de drenagem dessa bacia. A pesquisa de campo consistiu de entrevistas com os principais atores desse trabalho, a comunidade da BHEN. Inova no município um modelo de programas de participação da população com o nome de Cidadania e Participação Ativa da Comunidade (CIPAC) propondo 20 programas: Mascote da educação ambiental; Boteco em boteco; Eu amo minha cidade; Alô comunidade; TV SOS “Meio ambiente”; Rádio “Desperta comunidade; Coral e teatro “Reciclar”; Educa móvel; Coleta seletiva nas escolas; O meio ambiente pede carona; Centro de memória; A escola do lixo; Conhecer o lixo; Comunidade nota 10; Futuro verde; Coleta seletiva “porta a porta” Implantação dos LEVs; Criação das unidades básicas ecológicas; Criação da central de reciclagem de entulho; Criação das unidades de triagem de materiais recicláveis; Criação das cooperativas de catadores e carroceiros. Faz uma previsão de investimentos para implantação e manutenção desses programas assim como o retorno do investimento aplicado com a implantação. Como resultado, apresenta um novo modelo, baseado na prática, como sustentação para o estabelecimento de uma política municipal, de acordo com a Lei da Política Nacional de Resíduos Sólidos (PNRS) que tramita no Congresso Nacional. Também foram identificados parâmetros capazes de identificar a inadequação do processo atual de coleta de lixo e dos serviços de limpeza nessa bacia. Esses resultados alcançados permitem concluir que grande parte da população da BHEN não está preparada para aderir a um programa de Gestão de Resíduos Sólidos (GRS), que tenha como ponto de partida o GIRS. O grau de escolaridade e o nível de conhecimento da comunidade não representam obstáculos para isso, mas sim a falta de programas que envolvam a sua participação, a coleta de resíduos e serviços de limpeza urbana corretamente prestados, pois, atualmente, na pesquisa de campo realizada, foram visivelmente reprovados. Finalmente, ainda conclui que somente com a implantação de um GIRS, com apoio do CIPAC o meio ambiente seria consideravelmente beneficiado, mas não resolveria os graves problemas ambientais da BHEN, será necessária a melhoria de todos os sistemas de infra-estrutura urbana nessa importante bacia, para um efeito realmente mais saudável. É importante destacar que, no momento atual, as propostas apresentadas por este trabalho são consideradas bastante oportunas, pois com o inicio da implantação do Programa de Recuperação Urbano-Ambiental da Bacia da Estrada Nova (PROMABEM) pela Administração Municipal financiado pelo Banco Interamericano de Desenvolvimento (BID), os problemas sanitários e ambientais dessa importante bacia têm grande probabilidade de ser resolvidos.

Pseudechis guttatus venom proteome: Insights into evolution and toxin clustering

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Proteomic analysis of Porphyromonas gingivalis exposed to nicotine and cotinine

Cogo K, de Andrade A, Labate CA, Bergamaschi CC, Berto LA, Franco GCN, Goncalves RB, Groppo FC. Proteomic analysis of Porphyromonas gingivalis exposed to nicotine and cotinine. J Periodont Res 2012; 47: 766775. (c) 2012 John Wiley & Sons A/S Background and Objective: Smokers are more predisposed than nonsmokers to infection with Porphyromonas gingivalis, one of the most important pathogens involved in the onset and development of periodontitis. It has also been observed that tobacco, and tobacco derivatives such as nicotine and cotinine, can induce modifications to P. gingivalis virulence. However, the effect of the major compounds derived from cigarettes on expression of protein by P.gingivalis is poorly understood. Therefore, this study aimed to evaluate and compare the effects of nicotine and cotinine on the P.gingivalis proteomic profile. Material and Methods: Total proteins of P gingivalis exposed to nicotine and cotinine were extracted and separated by two-dimensional electrophoresis. Proteins differentially expressed were successfully identified through liquid chromatography-mass spectrometry and primary sequence databases using MASCOT search engine, and gene ontology was carried out using DAVID tools. Results: Of the approximately 410 protein spots that were reproducibly detected on each gel, 23 were differentially expressed in at least one of the treatments. A particular increase was seen in proteins involved in metabolism, virulence and acquisition of peptides, protein synthesis and folding, transcription and oxidative stress. Few proteins showed significant decreases in expression; those that did are involved in cell envelope biosynthesis and proteolysis and also in metabolism. Conclusion: Our results characterized the changes in the proteome of P.gingivalis following exposure to nicotine and cotinine, suggesting that these substances may modulate, with minor changes, protein expression. The present study is, in part, a step toward understanding the potential smokepathogen interaction that may occur in smokers with periodontitis.

Proteomic profiling of the rat hypothalamus

Background: The hypothalamus plays a pivotal role in numerous mechanisms highly relevant to the maintenance of body homeostasis, such as the control of food intake and energy expenditure. Impairment of these mechanisms has been associated with the metabolic disturbances involved in the pathogenesis of obesity. Since rodent species constitute important models for metabolism studies and the rat hypothalamus is poorly characterized by proteomic strategies, we performed experiments aimed at constructing a two-dimensional gel electrophoresis (2-DE) profile of rat hypothalamus proteins. Results: As a first step, we established the best conditions for tissue collection and protein extraction, quantification and separation. The extraction buffer composition selected for proteome characterization of rat hypothalamus was urea 7 M, thiourea 2 M, CHAPS 4%, Triton X-100 0.5%, followed by a precipitation step with chloroform/methanol. Two-dimensional (2-D) gels of hypothalamic extracts from four-month-old rats were analyzed; the protein spots were digested and identified by using tandem mass spectrometry and database query using the protein search engine MASCOT. Eighty-six hypothalamic proteins were identified, the majority of which were classified as participating in metabolic processes, consistent with the finding of a large number of proteins with catalytic activity. Genes encoding proteins identified in this study have been related to obesity development. Conclusion: The present results indicate that the 2-DE technique will be useful for nutritional studies focusing on hypothalamic proteins. The data presented herein will serve as a reference database for studies testing the effects of dietary manipulations on hypothalamic proteome. We trust that these experiments will lead to important knowledge on protein targets of nutritional variables potentially able to affect the complex central nervous system control of energy homeostasis.

Studio della trasduzione del Segnale Nucleare Inositide-Dipendente: identificazione di eEF1A2 come nuovo Fosfosubstrato di PKC βI

Introduction Phospholipase Cb1 (PLC-β1) is a key player in the regulation of nuclear inositol lipid signaling and of a wide range of cellular functions, such as proliferation and differentiation (1,2,3). PLCb1 signaling depends on the cleavage of phosphatidylinositol 4,5-bisphosphate and the formation of the second messengers diacylglycerol and Inositol tris-phosphate which activate canonical protein kinase C (cPKC) isoforms. Here we describe a proteomic approach to find out a potential effector of nuclear PLC-b1 dependent signaling during insulin stimulated myogenic differentiation. Methods Nuclear lysates obtained from insulin induced C2C12 myoblasts were immunoprecipitated with anti-phospho-substrate cPKC antibody. Proteins, stained with Comassie blue, were excised, digested and subsequently analysed in LC-MS/MS. For peptide sequence searching, the mass spectra were processed and analyzed using the Mascot MS/MS ion search program with the NCBI database. Western blotting, GST-pull down and co-immunoprecipitation were performed to study the interaction between eEF1A2 and cPKCs. Site direct mutagenesis was performed to confirm the phosphorylated motif recognized by the antibody. Immunofluorescence analysis, GFP-tagged eEF1A2 vector and subcellular fractionation were performed to study nuclear localization and relative distribution of eEF1A2. Results We have previously shown that PLC-β1 is greatly increased at the nuclear level during insulin-induced myoblasts differentiation and that this nuclear localization is essential for induction of differentiation. Thus, nuclear proteins of insulin stimulated C2C12 myoblasts, were immunoprecipitated with an anti-phospho-substrate cPKC antibody. After Electrophoretic gel separation of proteins immunoprecipitated, several molecules were identified by LC-MS/MS. Among these most relevant and unexpected was eukaryotic elongation factor 1 alpha 2 (eEF1A2). We found that eEF1A2 is phosphorylated by PKCb1 and that these two molecules coimmunolocalized at the nucleolar level. eEF1A2 could be phosphorylated in many sites among which both threonine and serine residues. By site direct mutagenesis we demonstrated that it is the serine residue of the motif recognized by the antibody that is specifically phosphorylated by PKCb1. The silencing of PLCb1 gives rise to a reduction of expression and phosphorylation levels of eEF1A2 indicating this molecule as a target of nuclear PLCb1 regulatory network during myoblasts differentiation.

Elevated albumin in retinas of monkeys with experimental glaucoma.

PURPOSE: To establish the identity of a prominent protein, approximately 70 kDa, that is markedly increased in the retina of monkeys with experimental glaucoma compared with the fellow control retina, the relationship to glaucoma severity, and its localization in the retina. METHODS: Retinal extracts were subjected to 2-D gel electrophoresis to identify differentially expressed proteins. Purified peptides from the abundant 70 kDa protein were analyzed and identified by liquid chromatography/mass spectrometry/mass spectrometry (LC/MS/MS) separation, and collision-induced dissociation sequencing. Protein identity was performed on MASCOT (Matrix Science, Boston, MA) and confirmed by Western blot. The relationship between the increase in this protein and glaucoma severity was investigated by regression analyses. Protein localization in retina was evaluated by immunohistochemistry with confocal imaging. RESULTS: The abundant protein was identified as Macaca mulatta serum albumin precursor (67 kDa) from eight non-overlapping proteolytic fragments, and the identity was confirmed by Western blot. The average increase in retinal albumin content was 2.3 fold (P = 0.015). In glaucoma eyes, albumin was localized to some neurons of the inner nuclear layer, in the inner plexiform layer, and along the vitreal surface, but it was only found in blood vessels in control retinas. CONCLUSIONS: Albumin is the abundant protein found in the glaucomatous monkey retinas. The increased albumin is primarily localized to the inner retina where oxidative damage associated with experimental glaucoma is known to be prominent. Since albumin is a major antioxidant, the increase of albumin in the retinas of eyes with experimental glaucoma may serve to protect the retina against oxidative damage.

University of Michigan Football Team, 1901

Top Row: Wesley Winkler, Ralph Drake, Charles Rothschild, Norman Sterry, Kenneth Robinson, Alfred Chadwick, Weldon Fix, Ross Kidston, Les Barkenbus, Harrison Weeks, David Dunlap, George Gregory, Harold Baker, Cecil Gooding, George Sadler

Third Row: student mngr. H.C. Crafts, Edward Dickey(?), David Beardsley, William Snushall, ? Smith, George Davison, Kennedy Potter, ? Clark, Charles Van Valkenberg, Neil Snow, Joseph Horgan, James Forrest, William Foote, Webb Sadler, ? Hayes

Second Row: trainer Keene Fitzpatrick, ? Smith, Samuel Sackett, Everett Sweeley, Herbert Graver, Walter Shaw, Hugh White, Albert Herrnstein, Curtis Redden, Dan McGugin, Ebin Wilson, Bruce Shorts, Arthur Redner, Temple Owens

Front Row: Ralph Husson, Albert Preussman, Arthur Urquhart, Willie Heston, Benjamin Southworth, coach Fielding Yost, Charles Crane, Jerome (mascot), John Lewis, Frank Doty, Frank Belknap

1912 University of Michigan Baseball Team

Top Row: Leo? Kenyon, student mngr. Earl Good, ? Warehin, Thomas Wheat, Roy Baribeau

3rd Row:Francis Scully, Donald Duncanson, captain Elmer D Mitchell, coach Branch Rickey, Cecil Corbin, Goodloe Rogers, Charles Hippler, ? Ward

2nd Row:Francis Scully, Donald Duncanson, captain Elmer D Mitchell, coach Branch Rickey, Cecil Corbin, Goodloe Rogers, Charles Hippler, ? Ward

Front Row: ? Creamer (mascot), Emery Munson

1950 Ice Hockey Team

1913 University of Michigan Baseball Team

Top Row: student mngr McCarthy, assistant coach Campbell, coach Branch Rickey, ? Stewart, George Sisler, Roy Baribeau, ? Black, Wilbur Davidson, Russell Baer, captain Joseph Bell

Middle Row: Donald Duncanson, John Cory, John Lavan, Ernest Hughitt, Miller Pontius, Frank Sheehy, Cregar Quaintance, Perry Howard

Front Row: Goodloe Rogers, Sayres(?), Roy Baker, Edward Webber, Edmond McQueen, (Seward Cramer-mascot)

Performance of five different electrospray ionisation sources in conjunction with rapid monolithic column liquid chromatography and fast MS/MS scanning

The performances of five different ESI sources coupled to a polystyrene-divinylbenzene monolithic column were compared in a series of LC-ESI-MS/MS analyses of Escherichia coli outer membrane proteins. The sources selected for comparison included two different modifications of the standard electrospray source, a commercial low-flow sprayer, a stainless steel nanospray needle and a coated glass Picotip. Respective performances were judged on sensitivity and the number and reproducibility of significant protein identifications obtained through the analysis of multiple identical samples. Data quality varied between that of a ground silica capillary, with 160 total protein identifications, the lowest number of high quality peptide hits obtained (3012), and generally peaks of lower intensity; and a stainless steel nanospray needle, which resulted in increased precursor ion abundance, the highest-quality peptide fragmentation spectra (5414) and greatest number of total protein identifications (259) exhibiting the highest MASCOT scores (average increase in score of 27.5% per identified protein). The data presented show that, despite increased variability in comparative ion intensity, the stainless steel nanospray needle provides the highest overall sensitivity. However, the resulting data were less reproducible in terms of proteins identified in complex mixtures -- arguably due to an increased number of high intensity precursor ion candidates.

Mapping nitro-tyrosine modifications in fibrinogen by mass spectrometry as a biomarker for inflammatory disease

There is a growing awareness that inflammatory diseases have an oxidative pathology, which can result in specific oxidation of amino acids within proteins. It is known that patients with inflammatory disease have higher levels of plasma protein nitro-tyrosine than healthy controls. Fibrinogen is an abundant plasma protein, highly susceptible to such oxidative modifications, and is therefore a potential marker for oxidative protein damage. The aim of this study was to map tyrosine nitration in fibrinogen under oxidative conditions and identify susceptible residues. Fibrinogen was oxidised with 0.25mM and 1mM SIN-1, a peroxynitrite generator, and methionine was used to quench excess oxidant in the samples. The carbonyl assay was used to confirm oxidation in the samples. The carbonyl levels were 2.3, 8.72 and 11.5nmol/mg protein in 0, 0.25mM and 1mM SIN-1 samples respectively. The samples were run on a SDS-PAGE gel and tryptically digested before analysis by HPLC MS-MS. All 3 chains of fibrinogen were observed for all treatment conditions. The overall sequence coverage for fibrinogen determined by Mascot was between 60-75%. The oxidised samples showed increases in oxidative modifications in both alpha and beta chains, commonly methionine sulfoxide and tyrosine nitration, correlating with increasing SIN-1 treatment. Tyrosines that were most susceptible were Tyr135 (tryptic peptide YLQEIYNSNNQK) and Tyr277 (tryptic peptide GGSTSYGTGSETESPR), but several other nitrated tyrosines were also identified with high confidence. Identification of these susceptible peptides will allow design of sequences-specific biomarkers of oxidative and nitrative damage to plasma protein in inflammatory conditions.

Identification of protein targets of nevirapine reactive metabolites using click chemistry and mass spectrometry-based differential proteomics

Abstract : Adverse drug reactions (ADRs) are undesirable effects caused after administration of a single dose or prolonged administration of drug or result from the combination of two or more drugs. Idiosyncratic drug reaction (IDR) is an adverse reaction that does not occur in most patients treated with a drug and does not involve the therapeutic effect of the drug. IDRs are unpredictable and often life-threatening. Idiosyncratic reaction is dependent on drug chemical characteristics or individual immunological response. IDRs are a major problem for drug development because they are usually not detected during clinical trials. In this study we focused on IDRs of Nevirapine (NVP), which is a non-nucleoside reverse transcriptase inhibitor used for the treatment of Human Immunodeficiency Virus (HIV) infections. The use of NVP is limited by a relatively high incidence of skin rash. NVP also causes a rash in female Brown Norway (BN) rats, which we use as animal model for this study. Our hypothesis is that idiosyncratic skin reactions associated with NVP treatment are due to post-translational modifications of proteins (e.g., glutathionylation) detectable by MS. The main objective of this study was to identify the proteins that are targeted by a reactive metabolite of Nevirapine in the skin. The specific objectives derived from the general objective were as follow: 1) To implement the click chemistry approach to detect proteins modified by a reactive NVP-Alkyne (NVP-ALK) metabolite. The purpose of using NVP-ALK was to couple it with Biotin using cycloaddition Click Chemistry reaction. 2) To detect protein modification using Western blotting and Mass Spectrometry techniques, which is important to understand the mechanism of NVP induced toxicity. 3) To identify the proteins using MASCOT search engine for protein identification, by comparing obtained spectrum from Mass Spectrometry with theoretical spectrum to find a matching peptide sequence. 4) To test if the drug or drug metabolites can cause harmful effects, as the induction of oxidative stress in cells (via protein glutathionylation). Oxidative stress causes cell damage that mediates signals, which likely induces the immune response. The results showed that Nevirapine is metabolized to a reactive metabolite, which causes protein modification. The extracted protein from the treated BN rats matched 10% of keratin, which implies that keratin was the protein targeted by the NVP-ALK.

Personalidade da marca

Dissertação para obtenção do grau de Mestre em Design de Comunicação, apresentada na Universidade de Lisboa - Faculdade de Arquitetura.

Espectáculo de identidades nacionales. Presentación de Colombia en las Exposiciones Internacionales de 1992, 1998, 2010

El presente documento analiza como el Estado colombiano ha querido crear una identidad nacional en tres Exposiciones Internacionales a partir de representaciones elaboradas con un discurso entre político, comercial y cultural, generando imágenes que no siempre concuerdan con la realidad.