O encontro dos desencontros : a população em situação de rua e os edifícios abandonados no centro de Curitiba

Orientadora: Profa. Dra. Daniele Regina PontesDissertação (mestrado) - Universidade Federal do Paraná, Setor de Tecnologia, Programa de Pós-Graduação em Planejamento Urbano. Defesa : Curitiba, 20/10/2021Inclui referências: p. 108-113Resumo: Num cenário de crise econômica, observa-se a ampliação do déficit habitacional e o aumento de pessoas em situação de rua nas capitais do Brasil. Em Curitiba, não é diferente. A cidade possui histórico excludente e de estigmatização dessa população, que em março de 2021 já contava com 2832 pessoas e que aumenta diante do contexto instável. A crítica à cidade modelo está assentada no silenciar e na insuficiência das políticas públicas adotadas, tanto no campo assistencial, com a não adequação dos usuários aos equipamentos disponibilizados, habitacional com a oferta ínfima de unidades habitacionais, e urbanísticas com inações administrativas para o cumprimento das funções sociais da cidade e da propriedade. Considerando a contradição que existe observando que a área de maior localização de pessoas em situação de rua coincide com o bairro de maior concentração de unidades habitacionais ociosas, esta pesquisa busca verificar as relações existentes entre estes dois temas. Para tanto, foram realizados dois levantamentos para a verificação de concentrações de pessoas em situação de rua, sendo um in loco e o segundo com dados da Central 156. Também foi realizado levantamento quanto as edificações ociosas no Centro de Curitiba, com espacialização e tabulação das metragens estimadas. Concluiu-se não ser possível afirmar que a ocorrência de concentrações de edifícios não utilizados ou subutilizados e pessoas em situação de rua são causa-efeito, no entanto, verificou-se que o Centro é dividido em duas partes, onde a porção com maior caráter público, com praças e equipamentos, também possui menor valor venal, maior concentração de edifícios ociosos e maior concentração de pessoas em situação de rua. Além disso, verificou-se permissividade por parte do município de Curitiba para o descumprimento da função social da propriedade, não apenas ao não aplicar instrumentos que oneram os proprietários de imóveis ociosos, mas ao solicitar que estes edifícios sejam lacrados para evitar o uso por pessoas em situação de rua. Portanto, verifica-se que a questão da população em situação de rua não é inerente apenas ao setor de assistência social, como é realizado, mas muito permeia aspectos do Planejamento Urbano.Abstract: In a financial crisis situation, the housing deficit and the homelessness increase seemingly over Brazilian large cities. Curitiba is not an exception. There were 2832 homeless people in March, 2021, number that keeps escalating, coming from a history of prejudice and stigmatization toward this population. The status of an International Model City is often questioned, considering the history of social silencing and lack of effective public policies that address people fighting through life without a roof. Furthermore, there is a noticeable inertia of the government to apply regulations that guarantee the social purpose of the city and the property, as well as a difficulty to officially discuss numbers and formal features of the provided public equipment. By examining this subject, there is a drastic contradiction that affects Curitiba's downtown: the biggest concentration of homeless people overlaps the area with the largest amount of vacant residential properties, thus, the following Research is aimed at verifying the association of these two subjects. Two extensive surveys were conducted to check on the groups of homeless people, the first one being a field research based on technical observation and the second conducted through the municipality's given data. On top of that, a different field survey was lead to estimate how many vacant unities currently exist in Curitiba's downtown, providing the results in square metres. After a comprehensive analysis of the gathered material, it is not possible to state that the concentration of vacant residential units and the large number of homeless people in the same area are to be considered "cause-effect", nonetheless it was found that the city's downtown has two distinct typologies, of which the one with the largest area of public spaces, also with the lesser economic value, is the one with both the largest amount of vacant buildings and homeless people. Additionally, the municipal government is far too lenient with the non observance of social purpose of the property, not only by ignoring the regulations that were created to guarantee that landlords consistently give use to units, but also by requiring vacant buildings to be kept safely closed in order to keep them from being used by the homeless population. As a final observation, the homelessness subject is not only inherently connected to social assistance programs, but also deeply rooted to aspects of Urban Planning

Stabilization of Hfq-mediated translational repression by the co-repressor Crc in Pseudomonas aeruginosa.

In Pseudomonas aeruginosa the RNA chaperone Hfq and the catabolite repression control protein (Crc) govern translation of numerous transcripts during carbon catabolite repression. Here, Crc was shown to enhance Hfq-mediated translational repression of several mRNAs. We have developed a single-molecule fluorescence assay to quantitatively assess the cooperation of Hfq and Crc to form a repressive complex on a RNA, encompassing the translation initiation region and the proximal coding sequence of the P. aeruginosa amiE gene. The presence of Crc did not change the amiE RNA-Hfq interaction lifetimes, whereas it changed the equilibrium towards more stable repressive complexes. This observation is in accord with Cryo-EM analyses, which showed an increased compactness of the repressive Hfq/Crc/RNA assemblies. These biophysical studies revealed how Crc protein kinetically stabilizes Hfq/RNA complexes, and how the two proteins together fold a large segment of the mRNA into a more compact translationally repressive structure. In fact, the presence of Crc resulted in stronger translational repression in vitro and in a significantly reduced half-life of the target amiE mRNA in vivo. Although Hfq is well-known to act with small regulatory RNAs, this study shows how Hfq can collaborate with another protein to down-regulate translation of mRNAs that become targets for the degradative machinery

New functions for an archaeal translation factor

La caratterizzazione del processo di sintesi proteica nei tre domini del mondo dei viventi è una sfida di importanza fondamentale che comporta, tra i vari aspetti, una piena comprensione della funzione dei fattori di traduzione. L'apparato traduzionale archeo condivide diverse caratteristiche con la controparte eucariota, pertanto lo studio del processo di traduzione negli Archaea potrebbe aiutare a chiarire aspetti evolutivamente conservati. L’analisi strutturale e funzionale dei fattori di traduzione (con elevata omologia tra Archea ed Eucarioti) è fondamentale per determinare non solo il loro ruolo nella sintesi proteica, ma anche per evidenziare un possibile coinvolgimento in processi cellulari fisiologici e patologici. Questo progetto di dottorato è finalizzato allo studio del fattore di traduzione aIF5A, il cui coinvolgimento nella sintesi proteica risulta ancora oggi elusivo, utilizzando come organismo modello un archeo ipertermofilo Sulfolobus solfataricus. Tale fattore è altamente conservato fra Eucarioti e Archea, inoltre anche l’ortologo batterico (EF-P) è stato caratterizzato. Il fattore eIF5A è l’unica proteina soggetta ad una peculiare modifica post-traduzionale, l’ipusinazione, catalizzata da due diversi enzimi, la deossiipusina sintetasi (DHS) e la deossiipusina idrossilasi (DOHH); la proteina batterica EF-P subisce invece una diversa modifica post-traduzionale chiamata β-lisilazione. Attraverso un modello sperimentale che permette di studiare la formazione del primo legame peptidico, eIF5A / EF-P sono stati identificati come fattori di inizio, ma studi successivi hanno evidenziato un loro coinvolgimento anche nella fase di allungamento, terminazione della traduzione, l’associazione a molecole di RNA e infine l’implicazione in processi di proliferazione cellulare. In questo studio abbiamo identificato la modifica post-traduzionale di aIF5A in Sulfolobus solfataricus, dimostrando che la proteina è completamente ipusinata e, in questa forma, associata alle subunità ribosomiali durante la sintesi proteica. Inoltre, l'analisi strutturale e funzionale della deossiipusina sintasi ha evidenziato come tale processo di modificazione post-traduzionale risulti evolutivamente conservato fra Archea ed Eucarioti. Dall'individuazione dei partner di interazione proteici, in particolare la scoperta di un'interessante associazione con il fattore di recupero della traduzione (Trf) e dalla caratterizzazione dell'attività ribonucleolitica di aIF5A, sono emerse caratteristiche del tutto inedite di questo fattore, in Sulfolobus solfataricus. Le nostre evidenze sperimentali rivelano per la prima volta un probabile coinvolgimento di aIF5A nel processo di stabilità / turnover dell'RNA, aprendo la strada ad ulteriori indagini.The characterization of the process of protein synthesis in the three primary domains of life is an interesting challenge which entails, among other things, a full understanding of the function of translation factors. The archaeal translational apparatus, shares several characteristics with the eukaryal counterparts, therefore the study of the process of translation in Archaea could help to clarify several aspects which have been conserved during the evolution. The structural and functional study of translation factors (with high homology between Archaea and Eukarya) is important to determine not only their role in protein synthesis but also to highlight their possible involvement in physiological and pathological cellular processes. This PhD project is aimed at the elucidation of the functions of S. solfataricus translation factor aIF5A, whose involvement in protein synthesis has remained elusive. This factor is highly conserved in Eukarya and Archaea while an orthologue has been found in bacteria (EF-P). The eukaryotic factor contains the unique modified amino acid, hypusine, formed by the post-translational modification of a lysine, a reaction carried out by two different enzymes, the deoxyhypusine synthase (DHS) and the deoxyhypusine hydroxylase (DOHH); the bacterial protein EF-P undergoes a different post-translational modification called β-lysylation. In a model assay for the first peptide bond formation eIF5A/EF-P were identified as initiation factors, but subsequent studies have highlighted their implication in translation elongation, translation termination, binding to RNA molecules and cell proliferation. We identified the post-translational modification of Sulfolobus solfataricus aIF5A, showing that the protein is fully hypusinated and bound to translating ribosomes. Moreover the structural and functional analysis of the deoxyhypusine synthase allowed us to conclude that this unique post-translational modification pathway is evolutionarily conserved in archaea and eukaryotes. From the identification of protein interacting partners, in particular the discovery of an interesting association with the translation recovery factor (Trf) and the characterization of aIF5A ribonucleolytic activity arise unreleased features of this factor in Sulfolobus solfataricus. For the first time we provide new hints of a probable involvement of aIF5A in the RNA stability/turnover process, a finding which paves the way for further investigation

O encontro dos desencontros : a população em situação de rua e os edifícios abandonados no centro de Curitiba

Orientadora: Profa. Dra. Daniele Regina PontesDissertação (mestrado) - Universidade Federal do Paraná, Setor de Tecnologia, Programa de Pós-Graduação em Planejamento Urbano. Defesa : Curitiba, 20/10/2021Inclui referências: p. 108-113Resumo: Num cenário de crise econômica, observa-se a ampliação do déficit habitacional e o aumento de pessoas em situação de rua nas capitais do Brasil. Em Curitiba, não é diferente. A cidade possui histórico excludente e de estigmatização dessa população, que em março de 2021 já contava com 2832 pessoas e que aumenta diante do contexto instável. A crítica à cidade modelo está assentada no silenciar e na insuficiência das políticas públicas adotadas, tanto no campo assistencial, com a não adequação dos usuários aos equipamentos disponibilizados, habitacional com a oferta ínfima de unidades habitacionais, e urbanísticas com inações administrativas para o cumprimento das funções sociais da cidade e da propriedade. Considerando a contradição que existe observando que a área de maior localização de pessoas em situação de rua coincide com o bairro de maior concentração de unidades habitacionais ociosas, esta pesquisa busca verificar as relações existentes entre estes dois temas. Para tanto, foram realizados dois levantamentos para a verificação de concentrações de pessoas em situação de rua, sendo um in loco e o segundo com dados da Central 156. Também foi realizado levantamento quanto as edificações ociosas no Centro de Curitiba, com espacialização e tabulação das metragens estimadas. Concluiu-se não ser possível afirmar que a ocorrência de concentrações de edifícios não utilizados ou subutilizados e pessoas em situação de rua são causa-efeito, no entanto, verificou-se que o Centro é dividido em duas partes, onde a porção com maior caráter público, com praças e equipamentos, também possui menor valor venal, maior concentração de edifícios ociosos e maior concentração de pessoas em situação de rua. Além disso, verificou-se permissividade por parte do município de Curitiba para o descumprimento da função social da propriedade, não apenas ao não aplicar instrumentos que oneram os proprietários de imóveis ociosos, mas ao solicitar que estes edifícios sejam lacrados para evitar o uso por pessoas em situação de rua. Portanto, verifica-se que a questão da população em situação de rua não é inerente apenas ao setor de assistência social, como é realizado, mas muito permeia aspectos do Planejamento Urbano.Abstract: In a financial crisis situation, the housing deficit and the homelessness increase seemingly over Brazilian large cities. Curitiba is not an exception. There were 2832 homeless people in March, 2021, number that keeps escalating, coming from a history of prejudice and stigmatization toward this population. The status of an International Model City is often questioned, considering the history of social silencing and lack of effective public policies that address people fighting through life without a roof. Furthermore, there is a noticeable inertia of the government to apply regulations that guarantee the social purpose of the city and the property, as well as a difficulty to officially discuss numbers and formal features of the provided public equipment. By examining this subject, there is a drastic contradiction that affects Curitiba's downtown: the biggest concentration of homeless people overlaps the area with the largest amount of vacant residential properties, thus, the following Research is aimed at verifying the association of these two subjects. Two extensive surveys were conducted to check on the groups of homeless people, the first one being a field research based on technical observation and the second conducted through the municipality's given data. On top of that, a different field survey was lead to estimate how many vacant unities currently exist in Curitiba's downtown, providing the results in square metres. After a comprehensive analysis of the gathered material, it is not possible to state that the concentration of vacant residential units and the large number of homeless people in the same area are to be considered "cause-effect", nonetheless it was found that the city's downtown has two distinct typologies, of which the one with the largest area of public spaces, also with the lesser economic value, is the one with both the largest amount of vacant buildings and homeless people. Additionally, the municipal government is far too lenient with the non observance of social purpose of the property, not only by ignoring the regulations that were created to guarantee that landlords consistently give use to units, but also by requiring vacant buildings to be kept safely closed in order to keep them from being used by the homeless population. As a final observation, the homelessness subject is not only inherently connected to social assistance programs, but also deeply rooted to aspects of Urban Planning

FOOD CONSUMPTION OF CIRRHOTIC PATIENTS, COMPARISON WITH THE NUTRITIONAL STATUS AND DISEASE STAGING

ABSTRACT Background Patients with liver disease often present protein-energy malnutrition. The assessment of food intake is very important in the investigation regarding the "health-disease" process. Objective To assess dietary intake of cirrhotic patients through food record during 3 days, correlating it with the nutritional status of the patient and the stage of the disease. Methods Cirrhotic outpatients from the Santa Casa de Misericórdia Hospital, RS, Brazil, were assessed. Nutritional assessment was performed by anthropometry; non-dominating handgrip strength; adductor pollicis muscle thickness; phase angle by bioelectrical impedance analysis; and Subjective Global Assessment. For analysis of food consumption we used the food records of 3 days using scales for weighing of all foods. Results We evaluated 25 (68%) patients, in which there was a prevalence of cirrhosis by hepatitis C virus. The upper arm circumference, handgrip strength and phase angle by bioelectrical impedance analysis diagnosed 56% of malnourished. Phase angle by bioelectrical impedance analysis and upper arm circumference were associated with Child-Pugh score (P<0.05). The average consumption of calories, carbohydrates, proteins and lipids were within the recommended. However, sodium was above the recommendations, 106±57.2 mEq, and was inversely associated with Child-Pugh score (rs=-0.410; P=0.042). Conclusion Food intake did not have a significant difference between the Child-Pugh scores and nutritional status. In addition, food intake did not vary neither according to the stage of the disease, nor to the nutritional assessment, by the phase angle by bioelectrical impedance analysis

Catabolite repression control protein antagonist, a novel player in Pseudomonas aeruginosa carbon catabolite repression control.

Peer reviewed: TrueAcknowledgements: The authors would like to thank Ralph Baldrian for technical assistance.In the opportunistic human pathogen Pseudomonas aeruginosa (Pae), carbon catabolite repression (CCR) orchestrates the hierarchical utilization of N and C sources, and impacts virulence, antibiotic resistance and biofilm development. During CCR, the RNA chaperone Hfq and the catabolite repression control protein Crc form assemblies on target mRNAs that impede translation of proteins involved in uptake and catabolism of less preferred C sources. After exhaustion of the preferred C-source, translational repression of target genes is relieved by the regulatory RNA CrcZ, which binds to and acts as a decoy for Hfq. Here, we asked whether Crc action can be modulated to relieve CCR after exhaustion of a preferred carbon source. As Crc does not bind to RNA per se, we endeavored to identify an interacting protein. In vivo co-purification studies, co-immunoprecipitation and biophysical assays revealed that Crc binds to Pae strain O1 protein PA1677. Our structural studies support bioinformatics analyzes showing that PA1677 belongs to the isochorismatase-like superfamily. Ectopic expression of PA1677 resulted in de-repression of Hfq/Crc controlled target genes, while in the absence of the protein, an extended lag phase is observed during diauxic growth on a preferred and a non-preferred carbon source. This observations indicate that PA1677 acts as an antagonist of Crc that favors synthesis of proteins required to metabolize non-preferred carbon sources. We present a working model wherein PA1677 diminishes the formation of productive Hfq/Crc repressive complexes on target mRNAs by titrating Crc. Accordingly, we propose the name CrcA (catabolite repression control protein antagonist) for PA1677

Indications for a moonlighting function of translation factor aIF5A in the crenarchaeum Sulfolobus solfataricus

Translation factor a/eIF5A is highly conserved in Eukarya and Archaea. The eukaryal eIF5A protein is required for transit of ribosomes across consecutive proline codons, whereas the function of the archaeal orthologue remains unknown. Here, we provide a first hint for an involvement of Sulfolobus solfataricus (Sso) aIF5A in translation. CRISPR-mediated knock down of the aif5A gene resulted in strong growth retardation, underlining a pivotal function. Moreover, in vitro studies revealed that Sso aIF5A is endowed with endoribonucleolytic activity. Thus, aIF5A appears to be a moonlighting protein that might be involved in protein synthesis as well as in RNA metabolism

{\textquotedblleft}Dual Anta-Inhibitors{\textquotedblright} of the A2A Adenosine Receptor and Casein Kinase {CK}1delta: Synthesis, Biological Evaluation, and Molecular Modeling Studies

Based on a screening of a chemical library of A2A adenosine receptor (AR) antagonists, a series of di- and tri-substituted adenine derivatives were synthesized and tested for their ability to inhibit the activity of the enzyme casein kinase 1 delta (CK1δ) and to bind adenosine receptors (ARs). Some derivatives, here called “dual anta-inhibitors”, demonstrated good CK1δ inhibitory activity combined with a high binding affinity, especially for the A2AAR. The N6-methyl-(2-benzimidazolyl)-2-dimethyamino-9-cyclopentyladenine (17, IC50 = 0.59 μM and KiA2A = 0.076 μM) showed the best balance of A2AAR affinity and CK1δ inhibitory activity. Computational studies were performed to simulate, at the molecular level, the protein–ligand interactions involving the compounds of our series. Hence, the dual anta-inhibitor 17 could be considered the lead compound of new therapeutic agents endowed with synergistic effects for the treatment of chronic neurodegenerative and cancer diseases