Plos One
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Phenotypic assays using human primary cells are highly valuable tools for target discovery and validation in drug discovery. Expression knockdown (KD) of such targets in these assays allows the investigation of their role in models of disease processes. Therefore, efficient and fast modes of protein KD in phenotypic assays are required. ⋯ A direct comparison between co-TD of separate Cas9 and gRNA AdV, versus TD with a single "all-in-one" Cas9/gRNA AdV, revealed that both methods achieve similar levels of indel formation. These data demonstrate that AdV CRISPR/Cas9 is a useful and efficient tool for protein KD in human primary cell phenotypic assays. The use of AdV CRISPR/Cas9 may offer significant advantages over the current existing tools and should enhance target discovery and validation opportunities.
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Basic knowledge about the thoracic spinal flexibility is limited and to the authors' knowledge, no in vitro studies have examined the flexibility of every thoracic spinal segment under standardized experimental conditions using pure moments. In our in vitro study, 68 human thoracic functional spinal units including the costovertebral joints (at least n = 6 functional spinal units per segment from T1-T2 to T11-T12) were loaded with pure moments of ±7.5 Nm in flexion/extension, lateral bending, and axial rotation in a custom-built spine tester to analyze range of motion (ROM) and neutral zone (NZ). ROM and NZ showed symmetric motion behavior in all loading planes. ⋯ Comparing these ROM values with those in literature, they agree that ROM is lowest in flexion/extension and highest in axial rotation, as well as decreasing in the lower segments in axial rotation. Differences were found in flexion/extension and lateral bending in the lower segments, where, in contrast to the literature, no increase of the ROM from superior to inferior segments was found. The data of this in vitro study could be used for the validation of numerical models and the design of further in vitro studies of the thoracic spine without the rib cage, the verification of animal models, as well as the interpretation of already published human in vitro data.
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OWL2 semantics are becoming increasingly popular for the real domain applications like Gene engineering and health MIS. The present work identifies the research gap that negligible attention has been paid to the performance evaluation of Knowledge Base Systems (KBS) using OWL2 semantics. To fulfil this identified research gap, an OWL2 benchmark for the evaluation of KBS is proposed. ⋯ The proposed benchmark is tested on memory based, file based, relational database and graph based KBS for performance and scalability measures. The results show that the proposed benchmark is able to evaluate the behaviour of different state of the art KBS on OWL2 semantics. On the basis of the results, the end users (i.e. domain expert) would be able to select a suitable KBS appropriate for his domain.
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The anaphylatoxin receptor C5aR1 plays an important role not only in innate immune responses, but also in bone metabolism and fracture healing, being highly expressed on immune and bone cells, including osteoblasts and osteoclasts. C5aR1 induces osteoblast migration, cytokine generation and osteoclastogenesis, however, the exact role of C5aR1-mediated signaling in osteoblasts is not entirely known. Therefore, we hypothesized that osteoblasts are essential target cells for C5a and that fracture healing should be disturbed in mice with an osteoblast-specific C5aR1 overexpression (Col1a1-C5aR1). ⋯ C5aR1 signaling in osteoblasts might possibly affect RANKL/OPG balance, leading to increased bone resorption. Additional trauma significantly impaired fracture healing, particularly in Col1a1-C5aR1 mice. In conclusion, the data indicate that C5aR1 signaling in osteoblasts plays a detrimental role in bone regeneration after fracture.
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Standard open and percutaneous minimally invasive surgical procedures co-exist in the treatment of fractures of the thoracolumbar spine. Shorter skin incisions just above the pedicles are used in minimally invasive procedures. Full-length skin incisions and invasive preparations are applied in the standard open approach. ⋯ Pair-wise comparison of the types of surgical procedure for each measurement point revealed a significantly higher flow value in the minimally invasive group at one of the measurement points located between the incisions (P = .041). The point-wise analyses of SO2 and Hb did not show significant differences between the approaches. In conclusion, significantly albeit moderately higher flow values could be found in minimally invasive procedures compared to open operations of thoracolumbar fractures in the area of skin that is spared by the incisions.