Glucose restriction in Saccharomyces cerevisiae modulates the phosphorylation pattern of the 20S proteasome and increases its activity

Caloric restriction is known to extend the lifespan and/or improve diverse physiological parameters in a vast array of organisms. In the yeast Saccharomyces cerevisiae, caloric restriction is performed by reducing the glucose concentration in the culture medium, a condition previously associated with increased chronological lifespan and 20S proteasome activity in cell extracts, which was not due to increased proteasome amounts in restricted cells. Herein, we sought to investigate the mechanisms through which glucose restriction improved proteasome activity and whether these activity changes were associated with modifications in the particle conformation. We show that glucose restriction increases the ability of 20S proteasomes, isolated from Saccharomyces cerevisiae cells, to degrade model substrates and whole proteins. In addition, threonine 55 and/or serine 56 of the α5-subunit, were/was consistently found to be phosphorylated in proteasomes isolated from glucose restricted cells, which may be involved in the increased proteolysis capacity of proteasomes from restricted cells. We were not able to observe changes in the gate opening nor in the spatial conformation in 20S proteasome particles isolated from glucose restricted cells, suggesting that the changes in activity were not accompanied by large conformational alterations in the 20S proteasome but involved allosteric activation of proteasome catalytic site.

The optical analysis of the abdominal wall using the biospeckle after implants of polypropylene mesh in rats.

PURPOSE: To evaluate an experimental animal model to study the abdominal tissue activity considering its interaction with a polypropylene mesh, through the use of one of the optical phenomena of light Laser, the biospeckle. METHODS: Fifty Wistar male rats were divided into four groups: Group 1: ten animals not submitted to surgery; Group 2: ten animals submitted to surgery without polypropylene mesh; Group 3: 20 animals submitted to surgery followed by the mesh placement; Group 4: (sham) with ten animals. None of the animals presented post surgical complications being submitted to the optical tests at the 20th postoperative day. RESULTS: The analysis from the biospeckle tests, comparing the medians and standard deviations with T Student test, indicated that no significative difference was observed on the abdominal wall tissue activity in the four groups considered, with and without polypropylene mesh prosthesis implantation. CONCLUSION: The animal model is viable and the biospeckle open ways for a great number of experiments to be developed in evaluating tissue activity.

The optical analysis of the abdominal wall using the biospeckle after implants of polypropylene mesh in rats / Análise ótica da parede abdominal pelo biospeckle após implante de prótese de polipropileno em ratos

PURPOSE: To evaluate an experimental animal model to study the abdominal tissue activity considering its interaction with a polypropylene mesh, through the use of one of the optical phenomena of light Laser, the biospeckle. METHODS: Fifty Wistar male rats were divided into four groups: Group 1: ten animals not submitted to surgery; Group 2: ten animals submitted to surgery without polypropylene mesh; Group 3: 20 animals submitted to surgery followed by the mesh placement; Group 4: (sham) with ten animals. None of the animals presented post surgical complications being submitted to the optical tests at the 20th postoperative day. RESULTS: The analysis from the biospeckle tests, comparing the medians and standard deviations with T Student test, indicated that no significative difference was observed on the abdominal wall tissue activity in the four groups considered, with and without polypropylene mesh prosthesis implantation. CONCLUSION: The animal model is viable and the biospeckle open ways for a great number of experiments to be developed in evaluating tissue activity.

OBJETIVO: Avaliar o modelo experimental animal para o estudo da atividade tecidual da parede abdominal em sua interação com a prótese de polipropileno, através da utilização de um dos fenômenos ópticos da luz Laser, o biospeckle. MÉTODOS: Foram utilizados 50 ratos Wistar machos, divididos em quatro grupos: Grupo 1 com 10 animais não operados e controles; Grupo 2 com 10 animais submetidos a cirurgia sem a colocação de prótese; Grupo 3 com 20 animais submetidos a cirurgia com colocação de prótese de polipropileno pré-peritonial; Grupo 4 (Sham) com 10 animais. Nenhum animal apresentou complicações pós-operatórias, sendo que no 20º PO foram submetidos ao ensaio óptico. RESULTADOS: A comparação das médias e o desvio padrão das medidas da atividade tecidual da parede abdominal, com e sem implantação da prótese de polipropileno obtidas dos grupos de animais, pelo teste T de Student, não evidenciou diferença estatística significativa (p>0.05). CONCLUSÃO: A análise dos dados obtidos permitiu concluir que o modelo animal é viável e que o biospeckle abre caminhos para toda uma linha de experimentos a ser desenvolvida em avaliar atividade tecidual.