Mitogen-Activated Protein Kinase Kinase 5 Regulates Proliferation and Biosynthetic Processes in Procyclic Forms of Trypanosoma brucei.

The pathogenic protozoan T. brucei alternates into distinct developmental stages in the mammalian and insect hosts. The mitogen-activated protein kinase (MAPK) signaling pathways transduce extracellular stimuli into a range of cellular responses, which ultimately lead to the adaptation to the external environment. Here, we combined a loss of function approach with stable isotope labeling with amino acids in cell culture (SILAC)-based mass spectrometry (MS) to investigate the role of the mitogen-activated protein kinase kinase 5 (MKK5) in T. brucei. The silencing of MKK5 significantly decreased the proliferation of procyclic forms of T. brucei. To shed light on the molecular alterations associated with this phenotype, we measured the total proteome and phosphoproteome of cells silenced for MKK5. In the total proteome, we observed a general decrease in proteins related to ribosome and translation as well as down-regulation of several components of the fatty acids biosynthesis pathway. In addition, we observed alterations in the protein levels and phosphorylation of key metabolic enzymes, which point toward a suppression of the oxidative metabolism. Taken together, our findings show that the silencing of MKK5 alters cell growth, energy metabolism, protein and fatty acids biosynthesis in procyclic T. brucei.

Modified native electrophoresis protocol for the solubilization and separation of mitochondrial protein complexes.

To isolate mitochondrial complexes, we have combined elements from the classic Laemmli protocol and blue native polyacrylamide gel electrophoresis (BN-PAGE) methods to develop a straightforward modified native electrophoresis protocol. This modified protocol presented good resolution for native electrophoresis of inner mitochondrial membrane proteins, where bands were easily visualized with no leftover stain or gel lanes overlap. Enzymatic tests revealed that complexes I and V remain active in the gel. This protocol, designed to overcome specific limitations of the standard protocols, provides a potential methodology to study membrane proteins in their functional form.

[Practical application of the nursing process in an adolescent suffering from a chronic disease]. / Aplicaçã o prática do processo de enfermagem a uma adolescente portadora de doenca crônica.

The purpose of this case study was to apply nursing diagnoses and interventions based on the North American Nursing Diagnosis Association (NANDA) Taxonomy of Nursing Diagnoses and the Nursing Interventions Classification (NIC) on an adolescent with secondary mellitus to cystic fibrosis and on her caretaker in order to provide them full care and guidelines. The nursing interventions were based on the following diagnoses: ineffective confrontation and stress syndrome due to alterations. These references helped us develop effective nursing interventions that provided a framework for individualized care, in addition to promoting a standardized nursing language.