Evaluation of a Smart Knee Brace for Range of Motion and Velocity Monitoring during Rehabilitation Exercises and an Exergame.

Anterior cruciate ligament (ACL) injuries often require a lengthy duration of rehabilitation for patients to return to their prior level of function. Adherence to rehabilitation during this prolonged period can be subpar due to the treatment duration and poor adherence to home exercises. This work evaluates whether a smart instrumented knee brace system is capable of monitoring knee range of motion and velocity during a series of common knee rehabilitation exercises and an exergame. A total of 15 healthy participants completed a series of common knee rehabilitation exercises and played an exergame while wearing a smart instrumented knee brace. The range of motion (ROM) and velocity of the knee recorded by the knee brace was compared to a reference optoelectronic system. The results show good agreement between the knee brace system and the reference system for all exercises performed. Participants were able to quickly learn how to play the exergame and scored well within the game. The system investigated in this study has the potential to allow rehabilitation to occur outside of the clinic with the use of remote monitoring, and improve adherence and outcomes through the use of an exergame.

Adaptation of a clinical fixation device for biomechanical testing of the lumbar spine.

In-vitro biomechanical testing is widely performed for characterizing the load-displacement characteristics of intact, injured, degenerated, and surgically repaired osteoligamentous spine specimens. Traditional specimen fixture devices offer an unspecified rigidity of fixation, while varying in the associated amounts and reversibility of damage to and "coverage" of a specimen - factors that can limit surgical access to structures of interest during testing as well as preclude the possibility of testing certain segments of a specimen. Therefore, the objective of this study was to develop a specimen fixture system for spine biomechanical testing that uses components of clinically available spinal fixation hardware and determine whether the new system provides sufficient rigidity for spine biomechanical testing. Custom testing blocks were mounted into a robotic testing system and the angular deflection of the upper fixture was measured indirectly using linear variable differential transformers. The fixture system had an overall stiffness 37.0, 16.7 and 13.3 times greater than a typical human functional spine unit for the flexion/extension, axial rotation and lateral bending directions respectively - sufficient rigidity for biomechanical testing. Fixture motion when mounted to a lumbar spine specimen revealed average motion of 0.6, 0.6, and 1.5° in each direction. This specimen fixture method causes only minimal damage to a specimen, permits testing of all levels of a specimen, and provides for surgical access during testing.

Unique model evokes the supination/pronation deficits found after Mason II fractures.

A rapid prototyping model of Mason II fracture was used to investigate baseline recommendations for surgical intervention founded on kinematic forearm rotational blockage. Exact replicas of the radial heads in nine cadaveric specimens were produced and specimens were tested in a physiologic elbow simulator. After testing supination/pronation, the rotations were repeated with native replicas and with replicas modeling 3 mm depressed Mason II fractures with and without a gap of 1 mm between the body and fragment. The fragments were located circumferentially around the radial head at 10, 2 and 6 o'clock positions. There was no statistical difference between the range of motion of the native case and the native replica without fracture. After inclusion of the fracture, seven of the nine specimens showed rotational blockages. A two-way ANOVA found no statistical difference due to type of Mason II fracture (p > 0.87) or fracture location (p > 0.27). A χ-square analysis showed that presence of a kinematic deficit with a fractured radial head was significant (p < 0.03). The results support continued surgical intervention for a 3 mm depressed fracture and also establish the use of the rapid prototype as a model for kinematic investigation of fractures in a cadaveric model when ligamentous attachments are preserved.

Femoral bone strains during antegrade nailing: a comparison of two entry points with identical nails using finite element analysis.

BACKGROUND: Antegrade femoral nailing has become the standard treatment for diaphyseal femoral shaft fractures. Concerns linger that improper location of the nail entry point may lead to iatrogenic fracture and further complications. This study used finite element analysis to compare the strain magnitude and distribution resulting from each of two entry points in the proximal femur during antegrade nailing. METHODS: A finite element model was created from a CT scan of a 37 year old male femur and of a standard antegrade nail. Using implicit time-stepping, the nail was inserted through piriformis and trochanteric entry points and strain was computed at 9 anatomic locations. FINDINGS: The strain levels were higher overall when inserting a nail through the trochanteric starting point. The highest strain occurred immediately medial and lateral to the trochanteric entry point. The posterior greater trochanter also showed very high strain levels during nail insertion. All strain values for nail insertion through the piriformis entry point were less than 2000 µm/m. INTERPRETATION: The trochanteric entry will have a much greater potential of iatrogenic fracture of the proximal femur during insertion of a nail. Strains with this entry point exceed the yield level of bone and the repeated loading with the progression of the nail could cause fissures or fractures. Caution should be taken during insertion of an antegrade nail when utilizing a lateral trochanteric starting point secondary to an increased risk of trochanteric fracture and lateral cortex fracture.

The history of medicine and the scientific revolution.

The "new philosophy" of the seventeenth century has continued to be explained mainly on its own terms: as a major philosophical turn. Twentieth-century modernism gave pride of place to big ideas and reinforced the tendency to explain the rise of science in light of new ideas. Such orientations subordinated medicine (and technology) to sciences that appeared to be more theoretical. In attempts to persuade historians of science of the importance of medicine, then, many authors took an approach arguing that the major changes in the history of medicine during the so-called scientific revolution arose from philosophical commitments. Yet because medicine is also intimately connected to other aspects of life, its histories proved to be recalcitrant to such reductions and so continue to offer many possibilities for those who seek fresh means to address histories of body and mind united rather than divided.

Expression of MARCKS effector domain mutants alters phospholipase D activity and cytoskeletal morphology of SK-N-MC neuroblastoma cells.

Stable overexpression of myristoylated alanine-rich C-kinase substrate (MARCKS) is known to enhance phorbol ester stimulation of phospholipase D (PLD) activity and protein kinase Calpha (PKCalpha) levels in SK-N-MC neuroblastoma cells. In contrast, expression of MARCKS mutants (S152A or S156A) lacking key PKC phosphorylation sites within the central basic effector domain (ED) had no significant effect on PLD activity or PKCalpha levels relative to vector control cells. Like control cells, those expressing wild type MARCKS were elongated and possessed longitudinally oriented stress fibers, although these cells were more prone to detach from the substratum and undergo cell death upon phorbol ester treatment. However, cells expressing MARCKS ED mutants were irregularly shaped and stress fibers were either shorter or less abundant, and cell adhesion and viability were not affected. These results suggest that intact phosphorylation sites within the MARCKS ED are required for PLD activation and influence both membrane-cytoskeletal organization and cell viability.

The phospholipid scramblase PLSCR1 increases UV induced apoptosis primarily through the augmentation of the intrinsic apoptotic pathway and independent of direct phosphorylation by protein kinase C delta.

Cell death by apoptosis can be caused by the DNA mutagen UV light whose exposure causes the direct activation of both the caspase 9 regulated cell damage intrinsic pathway and the caspase 8 regulated plasma membrane extrinsic pathway. We determined that increased activity of the plasma membrane phospholipid scramblase, PLSCR1, amplified UV mediated apoptosis primarily through the activation of the intrinsic apoptotic pathway. The caspase 8 inhibitor z-IETD-fmk was not as effective an inhibitor of PLSCR1 augmented UV induced apoptosis compared to treatment with caspase 3, caspase 9, or pan-caspase inhibitors. The inability of the caspase 8 inhibitor to decrease UV induced apoptosis was dependent on PLSCR1, as UV induced apoptosis was decreased by a similar amount in the control cells in the presence of inhibitors of caspase 8, caspase 9, caspase 3, or the pan-caspase inhibitor. PKC-delta directly phosphorylates human PLSCR1 resulting in increased PLSCR1 scramblase activity. PKC-delta can also be activated by caspase mediated cleavage resulting in the release of a constitutively active kinase domain. We observed that replacing the PKC-delta phosphorylation site of PLSCR1 with an alanine did not affect the ability of PLSCR1 to enhance UV induced apoptosis implying that PKC-delta does not directly phosphorylate PLSCR1 to increase plasma membrane scramblase activity during apoptosis. Cells transfected with a PLSCR1 mutant that contained an alanine substitution at its known PKC-delta phosphorylation site underwent UV induced apoptosis at a level similar to those transfected with wild type PLSCR1. The combined results indicate that UV exposure in cells possessing PLSCR1 increases apoptosis primarily by enhancement of the intrinsic apoptotic pathway, and also imply that the increased apoptosis observed upon exposure to UV light is not through direct phosphorylation of PLSCR1 by PKC-delta.

Resistance to UV-induced apoptosis in Chinese-hamster ovary cells overexpressing phosphatidylserine synthases.

Externalization of PtdSer (phosphatidylserine) is an important event in signalling removal of apoptotic cells. In contrast with previous work [Yu, Byers, Ridgway, McMaster and Cook (2000) Biochim. Biophys. Acta 1487, 296-308] with U937 cells showing that specific stimulation of PtdSer biosynthesis during apoptosis was caspase dependent, PtdSer biosynthesis in CHO (Chinese-hamster ovary)-K1 increased 2.5-fold during UV-induced apoptosis but was not reversed by a caspase inhibitor, Z-VAD-FMK (benzyloxycarbonyl-Val-Ala-DL-Asp-fluoromethylketone). Also, in CHO-K1 cells, stimulation of synthesis was less specific for PtdSer as similar levels of stimulation were observed for sphingomyelin biosynthesis. Involvement of PtdSer synthase isoforms was tested in CHO-K1 cells overexpressing PSS I (PtdSer synthase I) and PSS II. Both types of transformed cells showed resistance to UV-induced apoptosis based on the decreased levels of caspase 3 activation and morphology changes; externalization of PtdSer was reduced with UV treatment even though expression of endogenous scramblase increased slightly. Serine-labelling experiments showed that PSS I- or PSS II-expressing cells had higher basal levels of PtdSer biosynthesis compared with vector control cells. When cells were exposed to UV light to induce apoptosis, PtdSer biosynthesis was further stimulated 1.5- and 2-fold in PSS I- and PSS II-expressing cells respectively compared with UV-treated vector cells. Caspase activation was not required, as Z-VAD-FMK did not change PtdSer synthesis. Although enhanced PtdSer synthesis was supposed to facilitate apoptosis, cells overexpressing PSS I and II were actually resistant to UV-induced apoptosis. Whereas enhanced PtdSer synthesis was associated with apoptosis, potential anti-apoptotic effects were observed when excess activity of these synthetic enzymes was present. This suggests a tightly regulated role for PtdSer synthesis and/or an important dependence on compartmentation of PSS enzymes in association with scramblase facilitated enrichment of this phospholipid at the cell surface.

The MARCKS family of phospholipid binding proteins: regulation of phospholipase D and other cellular components.

Myristoylated alanine-rich C kinase substrate (MARCKS) and MARCKS-related protein (MRP) are essential proteins that are implicated in coordination of membrane-cytoskeletal signalling events, such as cell adhesion, migration, secretion, and phagocytosis in a variety of cell types. The most prominent structural feature of MARCKS and MRP is a central basic effector domain (ED) that binds F-actin, Ca2+-calmodulin, and acidic phospholipids; phosphorylation of key serine residues within the ED by protein kinase C (PKC) prevents the above interactions. While the precise roles of MARCKS and MRP have not been established, recent attention has focussed on the high affinity of the MARCKS ED for phosphatidylinositol 4,5-bisphosphate (PIP2), and a model has emerged in which calmodulin- or PKC-mediated regulation of these proteins at specific membrane sites could in turn control spatial availability of PIP2. The present review summarizes recent progress in this area and discusses how the above model might explain a role for MARCKS and MRP in activation of phospholipase D and other PIP2-dependent cellular processes.

Induction of protein kinase C substrates, Myristoylated alanine-rich C kinase substrate (MARCKS) and MARCKS-related protein (MRP), by amyloid beta-protein in mouse BV-2 microglial cells.

Microglial activation by amyloid beta-protein in senile plaques contributes to neurodegeneration in Alzheimer disease. In BV-2 microglial cells, amyloid beta-protein 1-40 (Abeta 1-40) elicited a dose-dependent increase (3-4 fold) of Myristoylated alanine-rich C kinase substrate (MARCKS) and MARCKS-related protein (MRP), two protein kinase C substrates implicated in membrane-cytoskeletal alterations underlying microglial adhesion, migration, secretion, and phagocytosis. Neither MARCKS nor MRP was induced by the amyloid fragment Abeta 25-35, although both Abeta 1-40 and Abeta 25-35 caused extensive aggregation of BV-2 cells. Interferon-gamma synergistically enhanced the induction by Abeta 1-40 of inducible nitric oxide synthase, but not MARCKS or MRP. Our results suggest that MARCKS and MRP may play important roles in microglia activated by amyloid peptides.

Stimulation of phosphatidylserine biosynthesis and facilitation of UV-induced apoptosis in Chinese hamster ovary cells overexpressing phospholipid scramblase 1.

Members of the phospholipid scramblase (PLSCR) family play active roles in altering lipid asymmetry at the plasma membrane including phosphatidylserine (PtdSer) exposure on the cell surface. To determine whether PtdSer biosynthesis and externalization are altered by PLSCR activities during apoptosis, Chinese hamster ovary K1 cell lines stably overexpressing PLSCR1 and PLSCR2 were established. PLSCR1 was localized on the plasma membrane, whereas PLSCR2 was predominantly in the nucleus. Cells overexpressing PLSCR1 showed suppressed growth, altered cell morphology, and higher basal levels of cell death. Following UV irradiation, these cells showed earlier and enhanced PtdSer exposure, increased caspase-3 activation, apoptotic nuclear changes, and PARP cleavage indicative of apoptosis. UV irradiation in cells overexpressing PLSCR1 led to a 4-fold stimulation of PtdSer synthesis (accompanied by increased movement of newly made PtdSer into microvesicles) relative to untreated PLSCR1 cells, whereas PtdSer formation in UV-irradiated vector control cells increased only by 2-fold. No differences in these responses were observed between PLSCR2-expressing cells and vector controls. PtdSer synthesis and its transbilayer movement stimulated by PLSCR1 overexpression were blocked by a caspase inhibitor along with progression of apoptosis. Thus, our studies showed that overexpression of PLSCR1 in Chinese hamster ovary K1 cells stimulated caspase-dependent PtdSer externalization and synthesis, implying an up-regulation of PtdSer formation in response to enhanced outward movement of this phospholipid to the cell surface during apoptosis. PLSCR1 also appears to influence progression of UV-induced apoptosis and could be a point of regulation or intervention during programmed cell death.