AVALIAÇÃO DA QUALIDADE PAISAGÍSTICA PARA O USO TURÍSTICO DO PARQUE DO INGÁ, MARINGÁ (PR)

Objetivando avaliar a qualidade visual da paisagem, para a melhora do gerenciamento do uso turístico no Parque do Ingá, localizado na região central da cidade de Maringá (PR), a presente pesquisa utilizou o método de análise da qualidade visual da paisagem, que foi dividido em duas etapas. A primeira consistiu na avaliação técnica da paisagem, por meio de fotografias, pelo método direto, por intermédio dos componentes de cada fotografia. A segunda etapa, pelas fotografias selecionadas na etapa anterior, consistiu na realização de entrevistas com os turistas que frequentam o Parque. As entrevistas permitiram determinar a preferência paisagística destes. Observou-se que as duas paisagens mais valoradas foram dois atrativos turísticos: o Jardim Japonês, como primeira colocada; e a Gruta Nossa Senhora da Glória, como segunda. Foi possível identificar a preferência por paisagens organizadas e com harmonia entre os elementos, e as que apresentaram poucos elementos e desarmonia foram avaliadas como menos qualitativa. Algumas fotografias apresentaram elementos depreciadores da qualidade visual do Parque. Também foi possível apontar o fator subjetivo, individual de cada pessoa, como determinante para a ordem de preferências. A presente metodologia permitiu conhecer os pontos de maior qualidade visual e os que depreciam a qualidade visual do Parque do Ingá

Turismo e comunidades tradicionais: uma reflexão acerca da Ilha do Mel, PR

O turismo faz parte do consumo global que responde às dinâmicas econômicas e sociais do capitalismo mundial potencializando aspectos culturais, econômicos, geográficos e sociais de destinos turísticos. Dessa forma, a atividade turística pode ser considerada um importante fator para o desenvolvimento socioeconômico das comunidades tradicionais, cuja compreensão está cada vez mais na busca pela qualidade de vida de determinada comunidade, tendo os indivíduos que a compõem uma efetiva participação em todo o processo. Nesse sentido, este estudo teve como objetivo compreender como o turismo impacta no desenvolvimento sociocultural das comunidades caiçaras da Ilha do Mel, Paraná (PR). Para isso, foi discutido o conceito de comunidades tradicionais caiçaras enfatizando as relações entre áreas naturais protegidas e o fomento do turismo. O procedimento metodológico pautou-se na pesquisa bibliográfica e entrevistas individuais. Como resultados, foram identificados que ao mesmo tempo em que o turismo contribuiu para o estabelecimento de novas relações sociais e posteriores transformações culturais, também têm estimulado as comunidades a ressignificarem traços culturais que definem seus membros como caiçaras, como nativos da Ilha do Mel. Por fim, constata-se que esse estímulo por meio do turismo materializa-se com propostas de oficinas culturais, cultura da pesca e a prática de festividades religiosas

A novel in-frame deletion in MYOT causes an early adult onset distal myopathy

Missense mutations in MYOT encoding the sarcomeric Z-disk protein myotilin cause three main myopathic phenotypes including proximal limb-girdle muscular dystrophy, spheroid body myopathy, and late-onset distal myopathy. We describe a family carrying a heterozygous MYOT deletion (Tyr4_His9del) that clinically was characterized by an early-adult onset distal muscle weakness and pathologically by a myofibrillar myopathy (MFM). Molecular modeling of the full-length myotilin protein revealed that the 4-YERPKH-9 amino acids are involved in local interactions within the N-terminal portion of myotilin. Injection of in vitro synthetized mutated human MYOT RNA or of plasmid carrying its cDNA sequence in zebrafish embryos led to muscle defects characterized by sarcomeric disorganization of muscle fibers and widening of the I-band, and severe motor impairments. We identify MYOT novel Tyr4_His9 deletion as the cause of an early-onset MFM with a distal myopathy phenotype and provide data supporting the importance of the amino acid sequence for the structural role of myotilin in the sarcomeric organization of myofibers

Effects of alirocumab on types of myocardial infarction: insights from the ODYSSEY OUTCOMES trial

Aims  The third Universal Definition of Myocardial Infarction (MI) Task Force classified MIs into five types: Type 1, spontaneous; Type 2, related to oxygen supply/demand imbalance; Type 3, fatal without ascertainment of cardiac biomarkers; Type 4, related to percutaneous coronary intervention; and Type 5, related to coronary artery bypass surgery. Low-density lipoprotein cholesterol (LDL-C) reduction with statins and proprotein convertase subtilisin–kexin Type 9 (PCSK9) inhibitors reduces risk of MI, but less is known about effects on types of MI. ODYSSEY OUTCOMES compared the PCSK9 inhibitor alirocumab with placebo in 18 924 patients with recent acute coronary syndrome (ACS) and elevated LDL-C (≥1.8 mmol/L) despite intensive statin therapy. In a pre-specified analysis, we assessed the effects of alirocumab on types of MI. Methods and results  Median follow-up was 2.8 years. Myocardial infarction types were prospectively adjudicated and classified. Of 1860 total MIs, 1223 (65.8%) were adjudicated as Type 1, 386 (20.8%) as Type 2, and 244 (13.1%) as Type 4. Few events were Type 3 (n = 2) or Type 5 (n = 5). Alirocumab reduced first MIs [hazard ratio (HR) 0.85, 95% confidence interval (CI) 0.77–0.95; P = 0.003], with reductions in both Type 1 (HR 0.87, 95% CI 0.77–0.99; P = 0.032) and Type 2 (0.77, 0.61–0.97; P = 0.025), but not Type 4 MI. Conclusion  After ACS, alirocumab added to intensive statin therapy favourably impacted on Type 1 and 2 MIs. The data indicate for the first time that a lipid-lowering therapy can attenuate the risk of Type 2 MI. Low-density lipoprotein cholesterol reduction below levels achievable with statins is an effective preventive strategy for both MI types.For complete list of authors see http://dx.doi.org/10.1093/eurheartj/ehz299</p

A Solve-RD ClinVar-based reanalysis of 1522 index cases from ERN-ITHACA reveals common pitfalls and misinterpretations in exome sequencing

Purpose Within the Solve-RD project (https://solve-rd.eu/), the European Reference Network for Intellectual disability, TeleHealth, Autism and Congenital Anomalies aimed to investigate whether a reanalysis of exomes from unsolved cases based on ClinVar annotations could establish additional diagnoses. We present the results of the “ClinVar low-hanging fruit” reanalysis, reasons for the failure of previous analyses, and lessons learned. Methods Data from the first 3576 exomes (1522 probands and 2054 relatives) collected from European Reference Network for Intellectual disability, TeleHealth, Autism and Congenital Anomalies was reanalyzed by the Solve-RD consortium by evaluating for the presence of single-nucleotide variant, and small insertions and deletions already reported as (likely) pathogenic in ClinVar. Variants were filtered according to frequency, genotype, and mode of inheritance and reinterpreted. Results We identified causal variants in 59 cases (3.9%), 50 of them also raised by other approaches and 9 leading to new diagnoses, highlighting interpretation challenges: variants in genes not known to be involved in human disease at the time of the first analysis, misleading genotypes, or variants undetected by local pipelines (variants in off-target regions, low quality filters, low allelic balance, or high frequency). Conclusion The “ClinVar low-hanging fruit” analysis represents an effective, fast, and easy approach to recover causal variants from exome sequencing data, herewith contributing to the reduction of the diagnostic deadlock

Effect of alirocumab on mortality after acute coronary syndromes. An analysis of the ODYSSEY OUTCOMES randomized clinical trial

Background: Previous trials of PCSK9 (proprotein convertase subtilisin-kexin type 9) inhibitors demonstrated reductions in major adverse cardiovascular events, but not death. We assessed the effects of alirocumab on death after index acute coronary syndrome. Methods: ODYSSEY OUTCOMES (Evaluation of Cardiovascular Outcomes After an Acute Coronary Syndrome During Treatment With Alirocumab) was a double-blind, randomized comparison of alirocumab or placebo in 18 924 patients who had an ACS 1 to 12 months previously and elevated atherogenic lipoproteins despite intensive statin therapy. Alirocumab dose was blindly titrated to target achieved low-density lipoprotein cholesterol (LDL-C) between 25 and 50 mg/dL. We examined the effects of treatment on all-cause death and its components, cardiovascular and noncardiovascular death, with log-rank testing. Joint semiparametric models tested associations between nonfatal cardiovascular events and cardiovascular or noncardiovascular death. Results: Median follow-up was 2.8 years. Death occurred in 334 (3.5%) and 392 (4.1%) patients, respectively, in the alirocumab and placebo groups (hazard ratio [HR], 0.85; 95% CI, 0.73 to 0.98; P=0.03, nominal P value). This resulted from nonsignificantly fewer cardiovascular (240 [2.5%] vs 271 [2.9%]; HR, 0.88; 95% CI, 0.74 to 1.05; P=0.15) and noncardiovascular (94 [1.0%] vs 121 [1.3%]; HR, 0.77; 95% CI, 0.59 to 1.01; P=0.06) deaths with alirocumab. In a prespecified analysis of 8242 patients eligible for ≥3 years follow-up, alirocumab reduced death (HR, 0.78; 95% CI, 0.65 to 0.94; P=0.01). Patients with nonfatal cardiovascular events were at increased risk for cardiovascular and noncardiovascular deaths (P<0.0001 for the associations). Alirocumab reduced total nonfatal cardiovascular events (P<0.001) and thereby may have attenuated the number of cardiovascular and noncardiovascular deaths. A post hoc analysis found that, compared to patients with lower LDL-C, patients with baseline LDL-C ≥100 mg/dL (2.59 mmol/L) had a greater absolute risk of death and a larger mortality benefit from alirocumab (HR, 0.71; 95% CI, 0.56 to 0.90; Pinteraction=0.007). In the alirocumab group, all-cause death declined wit h achieved LDL-C at 4 months of treatment, to a level of approximately 30 mg/dL (adjusted P=0.017 for linear trend). Conclusions: Alirocumab added to intensive statin therapy has the potential to reduce death after acute coronary syndrome, particularly if treatment is maintained for ≥3 years, if baseline LDL-C is ≥100 mg/dL, or if achieved LDL-C is low. Clinical Trial Registration: URL: https://www.clinicaltrials.gov. Unique identifier: NCT01663402

Incorporation of beta alanine in polypeptides

Les cellules vivantes utilisent 20 acides α aminés canoniques lors de la synthèse ribosomale des protéines, même si dans de rares occasions, d’autres acides aminés (selenocysteine ou pyrrolysine) peuvent être utilisés. Le répertoire des acides aminés utilisables est donc assez restreint. Cela limite la possibilité de construire des protéines ayant de nouvelles propriétés. Une tendance forte dans le domaine de l’ingénierie des protéines est la construction de systèmes permettant d’incorporer des acides aminés non canoniques pendant la biosynthèse in vivo. Ces travaux ont connu d’importants succès, mais tous les acides aminés incorporés jusqu'aux années 2010 étaient des acides aminés α. L’incorporation d’acides aminés β à des sites spécifiques créerait une flexibilité supérieure à celle des acides aminés α dans la chaîne principale, ce qui augmenterait les possibilités de repliement de la protéine. Il a été montré récemment en utilisant un système de traduction in vitro de synthèse protéique qu’il était possible d’incorporer des acides aminés β à des positions spécifiques (Katoh and Suga, 2018). Pour parvenir à transposer ce système in vivo, la principale limitation est l’aminoacylation des ARNt avec les acides aminés β. Il a été récemment proposé que certaines aminoacyl-ARNt synthetases étaient capables d’utiliser les acides aminées β, mais cette capacité reste limitée.Le but de cette thèse est d’incorporer la β-méthionine dans des polypeptides in vivo. Pour cela nous avons caractérisé la reconnaissance et l’utilisation de la L-β-homométhionine par la méthionyl-ARNt synthetase (MetRS) d’E. coli. Nous avons en particulier déterminé une structure cristallographique à haute résolution du complexe MetRS: β -Met. En utilisant la spectroscopie de fluorescence ainsi que la spectroscopie de masse, nous avons pu mettre en évidence l’activation de la -Met en adénylate ainsi que son estérification à un ARNtMet. Toutefois, les efficacités mesurées sont très petites par rapport à ce qui avait été publié. Une contamination de la -Met commerciale par de la méthionine pourrait expliquer ces différences. Une étude in silico basée sur la structure du complexe MetRS:β-Met a été menée en collaboration avec l’équipe de Bio-informatique du laboratoire afin de rechercher des enzymes mutantes plus efficaces vis à vis des acides aminés β. Enfin, nous avons amorcé la mise en place d’une méthode d’évolution dirigée destinée à améliorer l’efficacité d’incorporation des acides aminés β in vivo.Living cells use 20 canonical α amino acids during ribosomal protein synthesis, although in rare cases, other amino acids such as selenocysteine or pyrrolysine can be used. The repertoire of usable amino acids is therefore quite limited. This limits the ability to build proteins having new properties. A dominant trend in the field of protein engineering is the construction of systems capable of incorporating non-canonical amino acids during in vivo protein synthesis. These studies have been very successful, but all amino acids incorporated until the 2010s were α-amino acids. Incorporation of β-amino acids at discrete sites would create unprecedented flexibility in the main chain, which would increase the potential for new protein folds. It has recently been shown using an in vitro protein synthesis system that it is possible to incorporate β-amino acids at specific positions (Katoh and Suga, 2018). In order to transpose this system in vivo, the main limitation is the aminoacylation of tRNAs with β -amino acids. It has recently been proposed that some aminoacyl-tRNA synthetases can use β-amino acids, but this capacity remains limited. The aim of this thesis is to incorporate β-methionine in polypeptides in vivo. For this purpose, we have characterized the recognition and use of L-β-homomethionine by methionyl-tRNA synthetase (MetRS) from E. coli. In particular, we have determined a high-resolution crystallographic structure of the MetRS:β-Met complex. Using fluorescence spectroscopy and mass spectroscopy, we were able to demonstrate the activation of -Met into adenylate as well as its esterification onto tRNAMet. However, the measured efficiencies are very small compared to what was published. Contamination of commercial β-Met with methionine may explain these differences. An in silico study based on the structure of the MetRS:β-Met complex was conducted in collaboration with the laboratory's bioinformatics team in order to search for mutant enzymes more efficient in the use of β-amino acids. Finally, we initiated the implementation of a directed evolution method to improve the efficiency of incorporation of amino acids in vivo

Incorporation de la beta alanine dans des polypeptides

Living cells use 20 canonical α amino acids during ribosomal protein synthesis, although in rare cases, other amino acids such as selenocysteine or pyrrolysine can be used. The repertoire of usable amino acids is therefore quite limited. This limits the ability to build proteins having new properties. A dominant trend in the field of protein engineering is the construction of systems capable of incorporating non-canonical amino acids during in vivo protein synthesis. These studies have been very successful, but all amino acids incorporated until the 2010s were α-amino acids. Incorporation of β-amino acids at discrete sites would create unprecedented flexibility in the main chain, which would increase the potential for new protein folds. It has recently been shown using an in vitro protein synthesis system that it is possible to incorporate β-amino acids at specific positions (Katoh and Suga, 2018). In order to transpose this system in vivo, the main limitation is the aminoacylation of tRNAs with β -amino acids. It has recently been proposed that some aminoacyl-tRNA synthetases can use β-amino acids, but this capacity remains limited. The aim of this thesis is to incorporate β-methionine in polypeptides in vivo. For this purpose, we have characterized the recognition and use of L-β-homomethionine by methionyl-tRNA synthetase (MetRS) from E. coli. In particular, we have determined a high-resolution crystallographic structure of the MetRS:β-Met complex. Using fluorescence spectroscopy and mass spectroscopy, we were able to demonstrate the activation of -Met into adenylate as well as its esterification onto tRNAMet. However, the measured efficiencies are very small compared to what was published. Contamination of commercial β-Met with methionine may explain these differences. An in silico study based on the structure of the MetRS:β-Met complex was conducted in collaboration with the laboratory's bioinformatics team in order to search for mutant enzymes more efficient in the use of β-amino acids. Finally, we initiated the implementation of a directed evolution method to improve the efficiency of incorporation of amino acids in vivo.Les cellules vivantes utilisent 20 acides α aminés canoniques lors de la synthèse ribosomale des protéines, même si dans de rares occasions, d’autres acides aminés (selenocysteine ou pyrrolysine) peuvent être utilisés. Le répertoire des acides aminés utilisables est donc assez restreint. Cela limite la possibilité de construire des protéines ayant de nouvelles propriétés. Une tendance forte dans le domaine de l’ingénierie des protéines est la construction de systèmes permettant d’incorporer des acides aminés non canoniques pendant la biosynthèse in vivo. Ces travaux ont connu d’importants succès, mais tous les acides aminés incorporés jusqu'aux années 2010 étaient des acides aminés α. L’incorporation d’acides aminés β à des sites spécifiques créerait une flexibilité supérieure à celle des acides aminés α dans la chaîne principale, ce qui augmenterait les possibilités de repliement de la protéine. Il a été montré récemment en utilisant un système de traduction in vitro de synthèse protéique qu’il était possible d’incorporer des acides aminés β à des positions spécifiques (Katoh and Suga, 2018). Pour parvenir à transposer ce système in vivo, la principale limitation est l’aminoacylation des ARNt avec les acides aminés β. Il a été récemment proposé que certaines aminoacyl-ARNt synthetases étaient capables d’utiliser les acides aminées β, mais cette capacité reste limitée.Le but de cette thèse est d’incorporer la β-méthionine dans des polypeptides in vivo. Pour cela nous avons caractérisé la reconnaissance et l’utilisation de la L-β-homométhionine par la méthionyl-ARNt synthetase (MetRS) d’E. coli. Nous avons en particulier déterminé une structure cristallographique à haute résolution du complexe MetRS: β -Met. En utilisant la spectroscopie de fluorescence ainsi que la spectroscopie de masse, nous avons pu mettre en évidence l’activation de la -Met en adénylate ainsi que son estérification à un ARNtMet. Toutefois, les efficacités mesurées sont très petites par rapport à ce qui avait été publié. Une contamination de la -Met commerciale par de la méthionine pourrait expliquer ces différences. Une étude in silico basée sur la structure du complexe MetRS:β-Met a été menée en collaboration avec l’équipe de Bio-informatique du laboratoire afin de rechercher des enzymes mutantes plus efficaces vis à vis des acides aminés β. Enfin, nous avons amorcé la mise en place d’une méthode d’évolution dirigée destinée à améliorer l’efficacité d’incorporation des acides aminés β in vivo

Recognition of l-β-homomethionine by methionyl-trna synthetase

International audienc