In this work, we report evidences that the association of phosphofructokinase and F-actin can be affected by insulin stimulation in rabbit skeletal muscle homogenates and that this association can be a mechanism of phosphofructokinase regulation. Through co-sedimentation techniques, we observed that on insulin-stimulated tissues, approximately 70% of phosphofructokinase activity is co-located in an actin-enriched fraction, against 28% in control. This phenomenon is accompanied by a 100% increase in specific phosphofructokinase activity in stimulated homogenates. Purified F-actin causes an increase of 230% in phosphofructokinase activity and alters its kinetic parameters. The presence of F-actin increases the affinity of phosphofructokinase for fructose 6-phosphate nevertheless, with no changes in maximum velocity (V max ). Here we propose that the modulation of cellular distribution of phosphofructokinase may be one of the mechanisms of control of glycolytic flux in mammalian muscle by insulin.

Salmonella enterica serovar Typhimurium is an animal welfare and public health concern due to its ability to parasite livestock and potentially contaminate pork products. To reduce Salmonella shedding and the risk of pork contamination, antibiotic therapy is used and can contribute to antimicrobial resistance. Here we hypothesized that immune system education by the microbiota can play a role in intestinal resilience to infection. We used amoxicillin (15mg/Kg) to modulate the intestinal microbiome of 10 piglets, paired with same age pigs that received a placebo (n = 10) from 0 to 14 days of age. Animals were euthanized at 4weeks old. Each pig donated colon sections for ex vivo culture (n = 20 explants/pig). Explants were inoculated with S. Typhimurium, PBS or LPS (n = 6 explants/pig/group, plus technical controls). The gut bacteriome was characterized by sequencing of the 16S rRNA at 7, 21 days of age, and upon in vitro culture. Explants response to infection was profiled through high-throughput mRNA sequencing. In vivo antibiotic treatment led to β-diversity differences between groups at all times (P<0.05), while α-diversity did not differ between amoxicillin and placebo groups on day 21 and at euthanasia (P<0.03 on day 7). Explant microbiomes were not different from in vivo. In vitro challenge with S. Typhimurium led to lower necrosis scores in explants from amoxicillin-treated pigs, when compared to explants placebo-treated pigs (P<0.05). This was coupled with the activation of immune-related pathways in explants from amoxicillin-treated pigs (IL-2 production, NO production, BCR activation), when compared to placebo-treated pigs. In addition, several DNA repair and intestinal wound healing pathways were also only activated in explants from amoxicillin-treated pigs. Taken together, these findings suggest that immune education by the amoxicillin-disturbed microbiota may have contributed to mitigate intestinal lesions following pathogen exposure.

Introdução: Limitações físicas e psicológicas decorrentes do repouso prolongado no leito são complicações comumente encontradas no paciente crítico e impactam diretamente na capacidade funcional. Objetivos: Avaliar a evolução da capacidade funcional, da força muscular periférica e respiratória de pacientes ventilados mecanicamente internados no CTI Geral do Hospital Universitário Pedro Ernesto (HUPE). Metodologia: No período de dezembro de 2016 a fevereiro de 2017 observamos os pacientes admitidos no CTI. Pacientes que não evoluíram para ventilação mecânica (VM) foram excluídos. Os pacientes foram avaliados em quatro momentos: admissão no CTI geral (A1), primeiro dia de VM (A2), desmame da VM (A3) e alta do CTI (A4). Avaliamos a força muscular respiratória periférica, respiratória e a capacidade funcional através da escala PERME. Avaliamos a funcionalidade pré-internação hospitalar através do Índice de Katz no momento da admissão. Resultados: Foram incluídos 26 pacientes e 9 completaram as quatro avaliações. A mediana de idade foi de 63 (58 – 70) anos, SAPS III de 64 (44,75 – 68,50) e índice de Katz de 1 (0 - 1). Os valores do MRC, PImax e PEmax aumentaramprogressivamente. Os valores na escala Perme aumentaram de 2 (1,5-7,25) em A1 para 10 (6,75–22) em A4. Conclusão: A capacidade funcional, força muscular periférica e respiratória dos pacientes ventilados mecanicamente melhorou ao longo da internação no CTI Geral do HUPE. A escala Perme não foi aplicada em todos os momentos estipulados.Descritores: Fisioterapia; Unidade de terapia intensiva; Cuidados críticos; Polineuropatias; Limitação da mobilidade; Força muscular.