No description available.

Les myopathies sont une maladie rare qui entraine une déficience musculaire. Il apparait que la plupart des myopathies présentent une modification dans la biodistribution et la concentration des collagènes. Le diagnostic actuel de ces maladies repose sur l’étude de protéines déficientes par des outils de biologie moléculaire dont les résultats sont incertains. Nous proposons de développer la spectro-imagerie IRTF pour différencier les cinq principaux types de collagène dans le tissu conjonctif musculaire. Pour cela, nous avons appliqué deux méthodes : la déconvolution spectrale de l’amide I et la détermination des coefficients d’extinction anisotropes des fibres. Nous avons ensuite mis au point une matrice synthétique mimant la composition moléculaire d’un tissu musculaire squelettique d’un muscle de bœuf. Cette matrice a pour but de calibrer la spectro-imagerie IRTF et de quantifier à moyen terme les collagènes dans les tissus. / Myopathies are a rare disease which leads a muscular deficiency. It seems that most of the myopathies present a modification in the biodistribution and in the concentration of collagens. The current diagnosis of these diseases is based on the study of deficient proteins by molecular biology techniques whose the results are uncertain. We suggest developing the spectro-imaging IRTF to differentiate five main types of collagen in the muscular connective tissue. For that, we applied two methods: the spectral déconvolution of the amide I and the determination of the anisotropic extinction coefficient of the fibers. Then, we have set up a synthetic matrix miming the molecular composition of a squeletic muscular tissue of beef muscle. This matrix aims at calibrating the spectro-imaging IRTTF and at quantifying in the medium term the collagen in tissues.

The chicken eggshell possesses physical and chemical barriers to protect the embryo from pathogens. The avian eggshell cuticle is the outmost layer of the eggshell whose protein constituents remain largely unknown. Since eggs with incomplete or absent cuticle are more susceptible to bacterial contamination, we hypothesize that cuticle protein components play an important role in microbial resistance. In our study, at least 47 proteins were identified by LC/MS/MS in the non-calcified cuticle layer. Similar to Kunitz-like protease inhibitor (also annotated as ovocalyxin-25, OCX-25) and ovocalyxin-32 (OCX-32) were two of most abundant proteins of the cuticle proteins. Some proteins that have antimicrobial activity were also detected in the proteomic results, such as lysozyme C, ovotransferrin, ovocalyxin-32, cystatin, ovoinhibitor. This study represents the first comprehensive report of the cuticle proteome. Since the sequence similarity of the kunitz motif in OCX-25 is similar to that of BPTI, it is predicted that it will have the same trypsin inhibitory and antimicrobial activity against Gram-positive and/or Gram-negative bacteria. In order to test the antimicrobial property and trypsin inhibitor activity of OCX-25, cuticle proteins were extracted by 1N HCl. Antimicrobial activity was monitored using the Bioscreen C instrument; and antimicrobial activity was identified primarily against Staphylococcus aureus. Trypsin inhibitor activity was studied by using a specific trypsin assay, and the assay indicated that the cuticle proteins could inhibit the reaction of trypsin and substrate. Therefore, the current research has provided some insight into the antimicrobial and enzymatic aspects of the cuticle proteins, and its function for egg protection. Eggshell membranes are another important component of the chicken eggshell.Due to its insoluble and stable properties, there are still many questions regarding formation and constituents of the eggshell membranes. The purpose of our study was to identify eggshell membrane proteins, particularly these responsible for its structural features, by examining the transcriptome of the white isthmus during its formation. Bioinformatics tools were applied to analyze the differentially expressed genes as well as their encoded proteins. Some interesting proteins were encoded by the over-expressed genes in the white isthmus during the formation of eggshell membranes, such as Collagen X, and similar to spore coat protein SP75. These proteins may have potential applications. Our study provides a detailed description of the chicken white isthmus transcriptome during formation of the eggshell membranes; it could lead to develop the strategies to improve food safety of the table egg.

The chicken eggshell possesses physical and chemical barriers to protect the embryo from pathogens. The avian eggshell cuticle is the outmost layer of the eggshell whose protein constituents remain largely unknown. Since eggs with incomplete or absent cuticle are more susceptible to bacterial contamination, we hypothesize that cuticle protein components play an important role in microbial resistance. In our study, at least 47 proteins were identified by LC/MS/MS in the non-calcified cuticle layer. Similar to Kunitz-like protease inhibitor (also annotated as ovocalyxin-25, OCX-25) and ovocalyxin-32 (OCX-32) were two of most abundant proteins of the cuticle proteins. Some proteins that have antimicrobial activity were also detected in the proteomic results, such as lysozyme C, ovotransferrin, ovocalyxin-32, cystatin, ovoinhibitor. This study represents the first comprehensive report of the cuticle proteome. Since the sequence similarity of the kunitz motif in OCX-25 is similar to that of BPTI, it is predicted that it will have the same trypsin inhibitory and antimicrobial activity against Gram-positive and/or Gram-negative bacteria. In order to test the antimicrobial property and trypsin inhibitor activity of OCX-25, cuticle proteins were extracted by 1N HCl. Antimicrobial activity was monitored using the Bioscreen C instrument; and antimicrobial activity was identified primarily against Staphylococcus aureus. Trypsin inhibitor activity was studied by using a specific trypsin assay, and the assay indicated that the cuticle proteins could inhibit the reaction of trypsin and substrate. Therefore, the current research has provided some insight into the antimicrobial and enzymatic aspects of the cuticle proteins, and its function for egg protection. Eggshell membranes are another important component of the chicken eggshell.Due to its insoluble and stable properties, there are still many questions regarding formation and constituents of the eggshell membranes. The purpose of our study was to identify eggshell membrane proteins, particularly these responsible for its structural features, by examining the transcriptome of the white isthmus during its formation. Bioinformatics tools were applied to analyze the differentially expressed genes as well as their encoded proteins. Some interesting proteins were encoded by the over-expressed genes in the white isthmus during the formation of eggshell membranes, such as Collagen X, and similar to spore coat protein SP75. These proteins may have potential applications. Our study provides a detailed description of the chicken white isthmus transcriptome during formation of the eggshell membranes; it could lead to develop the strategies to improve food safety of the table egg.

The chicken eggshell possesses physical and chemical barriers to protect the embryo from pathogens. The avian eggshell cuticle is the outmost layer of the eggshell whose protein constituents remain largely unknown. Since eggs with incomplete or absent cuticle are more susceptible to bacterial contamination, we hypothesize that cuticle protein components play an important role in microbial resistance. In our study, at least 47 proteins were identified by LC/MS/MS in the non-calcified cuticle layer. Similar to Kunitz-like protease inhibitor (also annotated as ovocalyxin-25, OCX-25) and ovocalyxin-32 (OCX-32) were two of most abundant proteins of the cuticle proteins. Some proteins that have antimicrobial activity were also detected in the proteomic results, such as lysozyme C, ovotransferrin, ovocalyxin-32, cystatin, ovoinhibitor. This study represents the first comprehensive report of the cuticle proteome. Since the sequence similarity of the kunitz motif in OCX-25 is similar to that of BPTI, it is predicted that it will have the same trypsin inhibitory and antimicrobial activity against Gram-positive and/or Gram-negative bacteria. In order to test the antimicrobial property and trypsin inhibitor activity of OCX-25, cuticle proteins were extracted by 1N HCl. Antimicrobial activity was monitored using the Bioscreen C instrument; and antimicrobial activity was identified primarily against Staphylococcus aureus. Trypsin inhibitor activity was studied by using a specific trypsin assay, and the assay indicated that the cuticle proteins could inhibit the reaction of trypsin and substrate. Therefore, the current research has provided some insight into the antimicrobial and enzymatic aspects of the cuticle proteins, and its function for egg protection. Eggshell membranes are another important component of the chicken eggshell.Due to its insoluble and stable properties, there are still many questions regarding formation and constituents of the eggshell membranes. The purpose of our study was to identify eggshell membrane proteins, particularly these responsible for its structural features, by examining the transcriptome of the white isthmus during its formation. Bioinformatics tools were applied to analyze the differentially expressed genes as well as their encoded proteins. Some interesting proteins were encoded by the over-expressed genes in the white isthmus during the formation of eggshell membranes, such as Collagen X, and similar to spore coat protein SP75. These proteins may have potential applications. Our study provides a detailed description of the chicken white isthmus transcriptome during formation of the eggshell membranes; it could lead to develop the strategies to improve food safety of the table egg.

Le gliome est le type le plus agressif et mortel de tumeur cérébrale. Ces tumeurs se caractérisent par la présence conjointe de phénotypes solides (de bas grade, moins invasif, hautement vascularisé) et diffus (haut grade, très envahissant et diffus) des glioblastomes multiformes. Les collagènes sont des composants majeurs de la MEC des cellules tumorales des gliomes, et sont également présents dans la membrane basale des vaisseaux sanguins, mais avec une composition différente entre vasculatures saine et tumorale. L'abondance et la typologie des collagènes dans la MEC des cellules tumorales et la vasculature représentent donc un marqueur potentiel de diagnostic pour la gradation des tumeurs gliales. Nous avons développé la spectro-imagerie infrarouge à transformée de Fourier pour déterminer les modifications morphologiques et moléculaires apparaissant dans les formes solides et diffuses de gliomes, ainsi que dans les vasculatures saine et tumorale. Nous avons d'abord mis en évidence les vasculatures saine et tumorale en utilisant des nanoparticules injectées dans le système sanguin. Ensuite, nous avons appliqué des méthodes de reconstruction spectrale pour distinguer les tissus sains vs. ceux des formes solide et diffuse de tumeurs sur la base de leurs contenus en collagène de la MEC. Enfin, nous avons déterminé les changements de types du collagène au cours de la progression tumorale, validant ainsi la notion que l’analyse de ces contenus est potentiellement un marqueur diagnostic pour la gradation des gliomes. / The glioma is the most aggressive and lethal type of brain tumor. Such tumor is characterized both by solid (low grade, less invasive, highly vascularized) and diffuse (high grade, very invasive and diffuse) phenotypes in high-grades. Collagens are major components of ECM in glioma tumor cells, and are also present in basement membrane of blood vessels in vasculature, but with different composition between healthy and tumor capillaries. The abundance and typology of collagens in tumor cell ECM and vasculature is thus a potential diagnostic marker for grading glioma tumors. We developed Fourier transform infrared (FTIR) spectro-imaging as a functional technique to determine the morphological and molecular changes occurring in solid and diffuse form of tumor tissues as well as in healthy and tumor vasculatures. We first highlighted healthy and tumor vasculatures using nanoparticles injected in blood system. Then, we applied curve-fitting methods to distinguish between healthy tissue vs. solid and diffuse tumor tissues on the basis of the collagen contents found in ECM. Finally, we determined collagen typology changes during tumor progression, thus validating that collagen contents analysis is potentially a diagnostic marker for glioma grading.

No description available.

A doença pulmonar obstrutiva crônica (DPOC) é caracterizada por inflamação crônica e alterações estruturais que levam a obstrução das pequenas vias aéreas e destruição do parênquima alveolar. A composição da matriz extracelular (MEC) nos pulmões tem um importante papel em prover e sustentar a arquitetura pulmonar. No entanto, não há uma descrição abrangente da composição da matriz extracelular no trato respiratório de indivíduos portadores de DPOC. No presente estudo investigou-se a composição da MEC das vias aéreas grandes (VAG), pequenas (VAP) e do parênquima pulmonar de pacientes com DPOC. Utilizando imunohistoquímica e análise de imagem analisou-se a área fracionada de fibras elásticas, colágenos I, III e IV, versicam, decorina, biglicano, lumicam, fibronectina e tenascina nas VAG, VAP e no parênquima peribrônquico e distal de 26 indivíduos com DPOC e comparou-se à área fracionada nos pulmões de 26 fumantes sem DPOC e 16 indivíduos não fumantes. A área fracionada de fibras elásticas foi significante maior no grupo de fumantes não obstruídos em comparação com os demais grupos, em todos os compartimentos analisados. Houve menor expressão de colágeno I na camada interna das VAG e nas camadas interna, muscular e externa das VAP dos indivíduos com DPOC e na camada externa das VAP dos fumantes não obstruídos quando comparados ao grupo controle. A área fracionada de versicam mostrou-se menor apenas no parênquima distal do grupo DPOC comparado ao grupo controle. O estudo da matriz de glicoproteínas mostrou maior área fracionada de fibronectina nas camadas interna, muscular e externa das VAP dos indivíduos com DPOC comparados aos demais grupos, assim como maior área fracionada de tenascina foi observado na membrana basal das VAG e na camada interna das VAP do grupo DPOC comparados aos controles. Além disso, a composição da MEC correlacionou-se com valores funcionais, como o VEF1 (% predito). A partir desses resultados, concluímos que a DPOC é caracterizada por complexas alterações nas principais proteínas estruturais nas pequenas e grandes vias aéreas. Tais alterações podem contribuir para a lesão tecidual persistente e com a obstrução ao fluxo aéreo observado na DPOC / COPD is characterized by chronic inflammation and structural alterations leading to small airway obstruction and to destruction of the lung parenchyma. The extracellular matrix (ECM) composition of the lungs has an important role in determining airway structure. However, there are no comprehensive descriptions of the ECM composition along the respiratory tract in COPD patients. We postulated that the ECM composition in large and small airways and in lung parenchyma of COPD patients differs from that observed in smoking and non-smoking controls. Using immunohistochemistry and image analysis, fractional areas of elastic fibers, type-I, -III and IV collagen, the proteoglycans versican, decorin, biglycan and lumican; fibronectin and tenascin were quantified in the large (LA) and small airways (SA), in peribronchiolar (PP) and distal parenchyma (DP) of 26 COPD patients and compared to 26 smokers without COPD and 16 non-smoking controls. The fractional area of elastic fibers was higher in non-obstructed smokers than in COPD and non-smoking controls subjects, in all lung compartments. Type-I collagen fractional area was lower in the inner layer of LA and in the inner, muscle and outer layer (OL) of SA of COPD patients and in the OL of SA of non-obstructed smokers when compared to non-smoking controls. The versican fractional area was lower in DP of COPD patients than non-smokers. Fibronectin fractional área was higher in the inner, muscle and outer layer of SA of COPD patients compared to non-smokers. Tenascin fractional area was higher in the subepithelial area of LA and inner layer of SA of COPD when compared to non-smoking controls. Furthermore, ECM composition correlated with FEV1% predicted. Architectural alterations due to an altered ECM composition in COPD are likely to contribute to the persistent tissue injury and to the airflow obstruction characteristic of this disease

A doença pulmonar obstrutiva crônica (DPOC) é caracterizada por inflamação crônica e alterações estruturais que levam a obstrução das pequenas vias aéreas e destruição do parênquima alveolar. A composição da matriz extracelular (MEC) nos pulmões tem um importante papel em prover e sustentar a arquitetura pulmonar. No entanto, não há uma descrição abrangente da composição da matriz extracelular no trato respiratório de indivíduos portadores de DPOC. No presente estudo investigou-se a composição da MEC das vias aéreas grandes (VAG), pequenas (VAP) e do parênquima pulmonar de pacientes com DPOC. Utilizando imunohistoquímica e análise de imagem analisou-se a área fracionada de fibras elásticas, colágenos I, III e IV, versicam, decorina, biglicano, lumicam, fibronectina e tenascina nas VAG, VAP e no parênquima peribrônquico e distal de 26 indivíduos com DPOC e comparou-se à área fracionada nos pulmões de 26 fumantes sem DPOC e 16 indivíduos não fumantes. A área fracionada de fibras elásticas foi significante maior no grupo de fumantes não obstruídos em comparação com os demais grupos, em todos os compartimentos analisados. Houve menor expressão de colágeno I na camada interna das VAG e nas camadas interna, muscular e externa das VAP dos indivíduos com DPOC e na camada externa das VAP dos fumantes não obstruídos quando comparados ao grupo controle. A área fracionada de versicam mostrou-se menor apenas no parênquima distal do grupo DPOC comparado ao grupo controle. O estudo da matriz de glicoproteínas mostrou maior área fracionada de fibronectina nas camadas interna, muscular e externa das VAP dos indivíduos com DPOC comparados aos demais grupos, assim como maior área fracionada de tenascina foi observado na membrana basal das VAG e na camada interna das VAP do grupo DPOC comparados aos controles. Além disso, a composição da MEC correlacionou-se com valores funcionais, como o VEF1 (% predito). A partir desses resultados, concluímos que a DPOC é caracterizada por complexas alterações nas principais proteínas estruturais nas pequenas e grandes vias aéreas. Tais alterações podem contribuir para a lesão tecidual persistente e com a obstrução ao fluxo aéreo observado na DPOC / COPD is characterized by chronic inflammation and structural alterations leading to small airway obstruction and to destruction of the lung parenchyma. The extracellular matrix (ECM) composition of the lungs has an important role in determining airway structure. However, there are no comprehensive descriptions of the ECM composition along the respiratory tract in COPD patients. We postulated that the ECM composition in large and small airways and in lung parenchyma of COPD patients differs from that observed in smoking and non-smoking controls. Using immunohistochemistry and image analysis, fractional areas of elastic fibers, type-I, -III and IV collagen, the proteoglycans versican, decorin, biglycan and lumican; fibronectin and tenascin were quantified in the large (LA) and small airways (SA), in peribronchiolar (PP) and distal parenchyma (DP) of 26 COPD patients and compared to 26 smokers without COPD and 16 non-smoking controls. The fractional area of elastic fibers was higher in non-obstructed smokers than in COPD and non-smoking controls subjects, in all lung compartments. Type-I collagen fractional area was lower in the inner layer of LA and in the inner, muscle and outer layer (OL) of SA of COPD patients and in the OL of SA of non-obstructed smokers when compared to non-smoking controls. The versican fractional area was lower in DP of COPD patients than non-smokers. Fibronectin fractional área was higher in the inner, muscle and outer layer of SA of COPD patients compared to non-smokers. Tenascin fractional area was higher in the subepithelial area of LA and inner layer of SA of COPD when compared to non-smoking controls. Furthermore, ECM composition correlated with FEV1% predicted. Architectural alterations due to an altered ECM composition in COPD are likely to contribute to the persistent tissue injury and to the airflow obstruction characteristic of this disease

Streptococcus mutans é considerado um agente etiológico primário da cárie dentária e um importante agente etiológico da endocardite infecciosa. O microrganismo pode ser classificado em quatro sorotipos (c, e, f e k), sendo o sorotipo c o mais comumente prevalente na população. S. mutans também pode acessar a corrente sanguínea e colonizar células endoteliais das artérias coronárias, através de sua ligação e posterior invasão. O gene cnm, codificador de uma proteína de ligação ao colágeno encontrado em algumas cepas de S. mutans pode ser relacionado com sua virulência. Objetivos: (1) formação de biofilmes em superfícies tratadas com colagénio e a expressão de genes a formação de biofilme chave spaP e brpA, (2) aderência (30 min e 2 h) , invasão (5h) e persistência (24h) em células endoteliais e (3) desmineralização da dentina. Tanto os experimentos de formação de biofilme dependente de colágeno quanto os de aderência, invasão e persistência em células primárias Human Coronary Artery Endothelial Cells foram realizadas utilizando cepas de S. mutans UA159 (sorotipo c), B14 (sorotipo e), OM50E (sorotipo e), LM7 (sorotipo e), OMZ175 (sorotipo f), NCTC 11060 (sorotipo f) e os isolados clínicos deste estudo 7,1 (sorotipo c), C2A4 (sorotipo e) e 61 (sorotipo k). Mutantes deletérios do gene cnm foram construídas em todas as cepas cnm positivas, exceto para C2A4. Cepas OMZ175 e 61 e seus respectivos mutantes também foram empregados para avaliar a expressão de genes spaP e brpA por PCR em Tempo Real e a desmineralização da dentina através de Tomografia de Coerência Óptica (OCT). Os dados mostraram que o gene cnm é significativo para a formação de biofilme em superfícies tratadas com colágeno e invasão de células endoteliais, mas não apresenta influência na adesão celular. A maior parte das cepas cnm positivas foram capazes de persistir intra-celular 24h. O modelo de cárie experimental não foi capaz de demonstrar a importância do gene cnm na desmineralização da dentina. O gene cnm foi necessário para o desenvolvimento de biofilme de e invasão do microrganismo à células endoteliais, mas não para a adesão às HCAEC. / Streptococcus mutans is considered a primary etiological agent of dental caries and an important etiological agent of infectious endocarditis. It can be classified into four serotypes (c, e, f and k), which serotype c is the most commonly prevalent in the population. The recent breakthrough that the cnm gene, which encodes a collagen binding protein found in some S. mutans strains, has set off new studies on this species, such as biofilm formation and bacteria-cell interactions. Aims: (1) biofilm formation on collagen-treated surfaces and the expression of biofilm formation key genes spaP and brpA; (2) adhesion (30 min and 2h), invasion (5h) and persistence (24h) in endothelial cells and (3) dentin demineralization. Experiments on collagen-dependent biofilm formation, and adhesion, invasion and persistence in HCAEC cells (primary Human Coronary Artery Endothelial Cells) were performed using S. mutans strains UA159 (serotype c), B14 (serotype e), OM50E (serotype e), LM7 (serotype e), OMZ175 (serotype f), NCTC 11060 (serotype f) and the clinical isolates of this study 7.1 (serotype c), C2A4 (serotype e) and 61 (serotype k). Mutants cnm were constructed in all strains presenting the gene, except for C2A4. Strains OMZ175 and 61 and their respective mutants were also employed to evaluate both the expression of spaP and brpA genes by PCR Real Time and the dentin demineralization through Optical Coherence Tomography (OCT). One hundred forty four clinical isolates of S. mutans were collected from 47 patients. Our data have shown that the cnm gene is significant for both biofilm formation on collagen-treated surfaces and invasion of endothelial cells, but it has no influence in cell adhesion. In addition, most of the cnm positive strains were able to persist up to 24h intracellularly. Likewise, the model of experimental caries was not able to demonstrate the importance of cnm in dentin demineralization. Thus, gene cnm is required for S. mutans on collagen-dependent biofilm formation and invasion of endothelial cells, but it was not required for adhesion to HCAEC.