RESUMO
Streptococcus gordonii is a commensal oral organism. Harmless in the oral cavity, S. gordonii is an opportunistic pathogen. S. gordonii adheres to body surfaces using surface adhesive proteins (adhesins), which are critical to subsequent formation of biofilm communities. As in most Gram-positive bacteria, S. gordonii surface proteins containing the C-terminal LPXTG motif cleavage sequence are processed by sortase A (SrtA) to become covalently attached to the cell wall. To characterize the functional diversity and redundancy in the family of SrtA-processed proteins, an S. gordonii DL1 markerless deletion mutant library was constructed of each of the 26 putative SrtA-processed proteins. Each library member was evaluated for growth in rich medium, biofilm formation on plastic, saliva and salivary fractions, cell surface hydrophobicity (CSH), hemagglutination, and integration into an ex vivo plaque biofilm community. Library members were compared to the non-SrtA-processed adhesins AbpA and AbpB. While no major growth differences in rich medium were observed, many S. gordonii LPXTG/A proteins impacted biofilm formation on one or more of the substrates. Several mutants showed significant differences in hemagglutination, hydrophobicity, or fitness in the ex vivo plaque model. From the identification of redundant and unique functions in these in vitro and ex vivo systems, functional stratification among the LPXTG/A proteins is apparent.IMPORTANCES. gordonii interactions with its environment depend on the complement of cell wall proteins. A subset of these cell wall proteins requires processing by the enzyme sortase A (SrtA). The identification of SrtA-processed proteins and their functional characterization will help the community to better understand how S. gordonii engages with its surroundings, including other microbes, integrates into the plaque community, adheres to the tooth surface, and hematogenously disseminates to cause blood-borne infections. This study identified 26 putative SrtA-processed proteins through creation of a markerless deletion mutant library. The library was subject to functional screens that were chosen to better understand key aspects of S. gordonii physiology and pathogenesis.
Assuntos
Aminoaciltransferases/metabolismo , Proteínas de Bactérias/fisiologia , Biofilmes/crescimento & desenvolvimento , Cisteína Endopeptidases/metabolismo , Streptococcus gordonii/fisiologia , Aminoaciltransferases/química , Animais , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Cisteína Endopeptidases/química , Placa Dentária/microbiologia , Deleção de Genes , Hemaglutinação , Humanos , Interações Hidrofóbicas e Hidrofílicas , Boca/microbiologia , Saliva/microbiologia , Ovinos/sangue , Streptococcus gordonii/genética , Streptococcus gordonii/crescimento & desenvolvimentoRESUMO
Neutrophils contribute to lung injury in acute pneumococcal pneumonia. The interleukin 17 receptor E (IL-17RE) is the functional receptor for the epithelial-derived cytokine IL-17C, which is known to mediate innate immune functions. The aim of this study was to investigate the contribution of IL-17RE/IL-17C to pulmonary inflammation in a mouse model of acute Streptococcus pneumoniae pneumonia. Numbers of neutrophils and the expression levels of the cytokine granulocyte colony-stimulating factor (G-CSF) and tumor necrosis factor alpha (TNF-α) were decreased in lungs of IL-17RE-deï¬cient (Il-17re-/- ) mice infected with S. pneumoniae Numbers of alveolar macrophages rapidly declined in both wild-type (WT) and Il-17re-/- mice and recovered 72 h after infection. There were no clear differences in the elimination of bacteria and numbers of blood granulocytes between infected WT and Il-17re-/- mice. The fractions of granulocyte-monocyte progenitors (GMPs) were significantly reduced in infected Il-17re-/- mice. Numbers of neutrophils were significantly reduced in lungs of mice deficient for IL-17C 24 h after infection with S. pneumoniae These data indicate that the IL-17C/IL-17RE axis promotes the recruitment of neutrophils without affecting the recovery of alveolar macrophages in the acute phase of S. pneumoniae lung infection.
Assuntos
Interleucina-17/metabolismo , Neutrófilos/fisiologia , Pneumonia Pneumocócica/metabolismo , Receptores de Interleucina-17/metabolismo , Animais , Diferenciação Celular , Feminino , Granulócitos , Interleucina-17/genética , Camundongos , Camundongos Knockout , Pneumonia Pneumocócica/microbiologia , Receptores de Interleucina-17/genética , Streptococcus pneumoniaeRESUMO
Patients who suffer from foot drop have impaired gait pattern functions and a higher risk of stumbling and falling. Therefore, they are usually treated with an assistive device, a so-called ankle-foot orthosis. The support of the orthosis should be in accordance with the motor requirements of the patient and should only be provided when needed, which is referred to as assistance-as-needed. Thus, in this publication, an approach is presented to determine the assistance-as-needed support using musculoskeletal human models. Based on motion capture recordings of multiple subjects performing gaits at different speeds, a parameter study varying the optimal force of a reserve actuator representing the ankle-foot orthosis added in the musculoskeletal simulation is conducted. The results show the dependency of the simulation results on the selected optimal force of the reserve actuator but with a possible identification of the assistance-as-needed support required from the ankle-foot orthosis. The required increase in support due to the increasing severity of foot drop is especially demonstrated with the approach. With this approach, information for the required support of individual subjects can be gathered, which can further be used to derive the design of an ankle-foot orthosis that optimally assists the subjects.
Assuntos
Órtoses do Pé , Neuropatias Fibulares , Humanos , Tornozelo , Braquetes , PacientesRESUMO
The individualization of patient-specific ankle joint orthoses is becoming increasingly important and can be ideally realized by means of additive manufacturing. However, currently, there are no functional additively manufactured fiber-reinforced products that are used in the field of orthopedic treatment. In this paper, an approach as to how additively manufactured orthopedic products can be designed and produced quickly and flexibly in the future is presented. This is demonstrated using the example of a solid ankle-foot orthosis. For this purpose, test results on PETG-CF15, which were determined in a previous work, were integrated into a material map for an FEA simulation. Therewith, the question can be answered as to whether production parameters that were determined at the test specimen level can also be adapted to real, usable components. Furthermore, gait recordings were used as loading conditions to obtain exact results for the final product. In order to perfectly adapt the design of the splint to the user, a 3D scan of a foot was performed to obtain a perfect design space for topology optimization. This resulted in a patient-specific and stiffness-optimized product. Subsequently, it was demonstrated that the orthosis could be manufactured using fused layer modelling. Finally, a comparison between the conventional design and the consideration of AM-specific properties was made. On this basis, it can be stated that the wearing comfort of the patient-specific design is very good, but the tightening of the splint still needs to be improved.