Can Dynamic Compression in the Absence of Growth Factors Induce Chondrogenic Differentiation of Bone Marrow Derived MSCs Encapsulated in Agarose Hydrogels?

The objectives of this study were twofold; to determine if cartilage specific matrix synthesis by mesenchymal stem cells (MSCs) is regulated by the magnitude and/or duration of dynamic compression in the absence of growth factors, and to investigate if expanding MSCs in the presence of both fibroblast growth factor-2 (FGF-2) and transforming growth factor β-3 (TGF-β3) would influence their subsequent response to dynamic compression following encapsulation in agarose hydrogels. Porcine bone marrow derived MSCs were suspended in agarose and cast to produce cylinders (Ø5×3mm). Constructs were maintained in a chemically defined medium. Dynamic compression was applied at 1 Hz at strain amplitudes of 5%, 10% and 5% superimposed upon a 5% pre-strain for durations of 1, 3 and 12 hours. MSCs were also expanded in the presence of FGF-2 and TGF-β3. The biochemical constituents of constructs were analyzed. Under strain magnitudes of 5% and 10% and durations of 1 and 3 hours small increases in sGAG accumulation relative to unloaded controls were observed. However this was orders of magnitude lower than that induced by TGF-β3 stimulation. Expansion in FGF-2 and TGF-β3 did not positively modulate chondrogenesis of MSCs in either unloaded or loaded culture

Cx30 exhibits unique characteristics including a long half-life when assembled into gap junctions

In the present study we investigated the life cycle, trafficking, assembly and cell surface dynamics of a poorly characterized connexin family member, connexin 30 (Cx30; also known as GJB6), which plays a critical role in skin health and hearing. Unexpectedly, Cx30 localization at the cell surface and gap junctional intercellular communication was not affected by prolonged treatments with the endoplasmic reticulum (ER)-Golgi transport inhibitor brefeldin A or the protein synthesis inhibitor cycloheximide, whereas Cx43 (also known as GJA1) was rapidly cleared. Fluorescent recovery after photobleaching revealed that Cx30 plaques were rebuilt from the outer edges in keeping with older channels residing in the inner core of the plaque. Expression of a dominant-negative form of Sar1 GTPase led to the accumulation of Cx30 within the ER, in contrast to a report that Cx30 traffics via a Golgi-independent pathway. Co-expression of Cx30 with Cx43 revealed that these connexins segregate into distinct domains within common gap junction plaques, suggesting that their assembly is governed by different mechanisms. In summary, Cx30 was found to be an unusually stable, long-lived connexin (half-life >12 h), which may underlie its specific role in the epidermis and cochlea

The Influence of Foot-Strike Technique on the Neuromechanical Function of the Foot

This is the author accepted manuscript. The final version is available from Lippincott, Williams & Wilkins via the DOI in this recordPURPOSE: The aim of this study was to investigate the influence of foot-strike technique on longitudinal arch mechanics and intrinsic foot muscle function during running. METHODS: Thirteen healthy participants ran barefoot on a force-instrumented treadmill at 2.8 ms with a forefoot (FFS) and rearfoot (RFS; habitual) running technique, whereas kinetic, kinematic, and electromyographic data from the intrinsic foot muscles were collected simultaneously. The longitudinal arch was modeled as a single "midfoot" joint representing motion of the rearfoot (calcaneus) relative to the forefoot (metatarsals). An inverse dynamic analysis was performed to estimate joint moments generated about the midfoot, as well as mechanical work and power. RESULTS: The midfoot was more plantar flexed (higher arch) at foot contact when running with a forefoot running technique (RFS 0.2 ± 1.8 vs FFS 6.9 ± 3.0°, effect size (ES) = 2.7); however, there was no difference in peak midfoot dorsiflexion in stance (RFS -11.6 ± 3.0 vs FFS -11.4 ± 3.4°, ES = 0.63). When running with a forefoot technique, participants generated greater moments about the midfoot (27% increase, ES = 1.1) and performed more negative work (240% increase, ES = 2.2) and positive work (42% increase, ES = 1.1) about the midfoot. Average stance-phase muscle activation was greater for flexor digitorum brevis (20% increase, ES = 0.56) and abductor hallucis (17% increase, ES = 0.63) when running with a forefoot technique. CONCLUSIONS: Forefoot running increases loading about the longitudinal arch and also increases the mechanical work performed by the intrinsic foot muscles. These findings have substantial implications in terms of injury prevention and management for runners who transition from a rearfoot to a forefoot running technique.Funding for this study was provided via an industry research grant from Asics Oceania (grant identification number 2014000885

Biofabrication of vasculature in microphysiological models of bone

Bone contains a dense network of blood vessels that are essential to its homoeostasis, endocrine function, mineral metabolism and regenerative functions. In addition, bone vasculature is implicated in a number of prominent skeletal diseases, and bone has high affinity for metastatic cancers. Despite vasculature being an integral part of bone physiology and pathophysiology, it is often ignored or oversimplified in in vitro bone models. However, 3D physiologically relevant vasculature can now be engineered in vitro, with microphysiological systems (MPS) increasingly being used as platforms for engineering this physiologically relevant vasculature. In recent years, vascularised models of bone in MPSs systems have been reported in the literature, representing the beginning of a possible technological step change in how bone is modelled in vitro. Vascularised bone MPSs is a subfield of bone research in its nascency, however given the impact of MPSs has had in in vitro organ modelling, and the crucial role of vasculature to bone physiology, these systems stand to have a substantial impact on bone research. However, engineering vasculature within the specific design restraints of the bone niche is significantly challenging given the different requirements for engineering bone and vasculature. With this in mind, this paper aims to serve as technical guidance for the biofabrication of vascularised bone tissue within MPS devices. We first discuss the key engineering and biological considerations for engineering more physiologically relevant vasculature in vitro within the specific design constraints of the bone niche. We next explore emerging applications of vascularised bone MPSs, and conclude with a discussion on the current status of vascularised bone MPS biofabrication and suggest directions for development of next generation vascularised bone MPSs

Intrinsic foot muscles contribute to elastic energy storage and return in the human foot

 This is the author accepted manuscript. The final version is available from American Physiological Society via the DOI in this recordData Availability: Data from this study is available at https://www.dropbox.com/sh/okbsab120jsc2az/AAD05Q1dtaY6MLdi_dkaqUjsa?dl=0The human foot is uniquely stiff to enable forward propulsion, yet also possesses sufficient elasticity to act as an energy store, recycling mechanical energy during locomotion. Historically this dichotomous function has been attributed to the passive contribution of the plantar aponeurosis. However, recent evidence highlights the potential for muscles to actively modulate the energetic function of the foot. Here we test the hypothesis that the central nervous system can actively control the foot's energetic function, via activation of the muscles within the foot's longitudinal arch. We used a custom-built loading apparatus to deliver cyclical loads to human feet in-vivo, in order to deform the arch in a manner similar to that observed in locomotion. We recorded foot motion and forces, alongside muscle activation and ultrasound images from flexor digitorum brevis (FDB), an intrinsic foot muscle that spans the arch. When active, the FDB muscle fascicles contracted in an isometric manner, facilitating elastic energy storage in the tendon, in addition to the energy stored within the plantar aponeurosis. We propose that the human foot is akin to an active suspension system for the human body, with mechanical and energetic properties that can be actively controlled by the central nervous system.Australian Research CouncilNational Health & Medical Research Council (NHMRC

Shoes alter the spring-like function of the human foot during running

This is the author accepted manuscript. The final version is available from the Royal Society via the DOI in this recordThe capacity to store and return energy in legs and feet that behave like springs is crucial to human running economy. Recent comparisons of shod and barefoot running have led to suggestions that modern running shoes may actually impede leg and foot-spring function by reducing the contributions from the leg and foot musculature. Here we examined the effect of running shoes on foot longitudinal arch (LA) motion and activation of the intrinsic foot muscles. Participants ran on a force-instrumented treadmill with and without running shoes. We recorded foot kinematics and muscle activation of the intrinsic foot muscles using intramuscular electromyography. In contrast to previous assertions, we observed an increase in both the peak (flexor digitorum brevis +60%) and total stance muscle activation (flexor digitorum brevis +70% and abductor hallucis +53%) of the intrinsic foot muscles when running with shoes. Increased intrinsic muscle activation corresponded with a reduction in LA compression (-25%). We confirm that running shoes do indeed influence the mechanical function of the foot. However, our findings suggest that these mechanical adjustments are likely to have occurred as a result of increased neuromuscular output, rather than impaired control as previously speculated. We propose a theoretical model for foot-shoe interaction to explain these novel findings.Funding for this study was provided by Asics Oceania Pty Ltd. Grant ID number 2014000885

The functional importance of human foot muscles for bipedal locomotion (article)

This is the final version. Available from the national Academy of Sciences via the DOI in this record.The dataset associated with this article is located in ORE at: https://doi.org/10.24378/exe.1084Human feet have evolved to facilitate bipedal locomotion, losing an opposable digit that grasped branches in favor of a longitudinal arch (LA) that stiffens the foot and aids bipedal gait. Passive elastic structures are credited with supporting the LA, but recent evidence suggests that plantar intrinsic muscles (PIMs) within the foot actively contribute to foot stiffness. To test the functional significance of the PIMs, we compared foot and lower limb mechanics with and without a tibial nerve block that prevented contraction of these muscles. Comparisons were made during controlled limb loading, walking, and running in healthy humans. An inability to activate the PIMs caused slightly greater compres- sion of the LA when controlled loads were applied to the lower limb by a linear actuator. However, when greater loads were experienced during ground contact in walking and running, the stiffness of the LA was not altered by the block, indicating that the PIMs ’ contribution to LA stiffness is minimal, probably because of their small size. With the PIMs blocked, the distal joints of the foot could not be stiffened sufficiently to provide normal push-off against the ground during late stance. This led to an increase in stride rate and compensatory power generated by the hip muscu- lature, but no increase in the metabolic cost of transport. The results reveal that the PIMs have a minimal effect on the stiffness of the LA when absorbing high loads, but help stiffen the distal foot to aid push-off against the ground when walking or running bipedally.Australian Research Council Discovery GrantNational Health and Medical Research Council Early Career Researcher Fellowshi

Second trimester hepatic rupture in a 35 year old nulliparous woman with HELLP syndrome: a case report

The HELLP syndrome (haemolysis, elevated liver blood tests and low platelets) is a serious complication in pregnancy characterized by haemolysis, elevated liver enzymes and low platelet count occurring in 0.5 to 0.9% of all pregnancies and in 10–20% of cases with severe preeclampsia. Hepatic capsular rupture is a rare yet dramatic complication of HELLP syndrome. The majority of cases occur in multiparous women over the age of 30. Classically it presents with acute onset right upper quadrant pain in the presence of constitutional symptoms such as vomiting and pyrexia. However, symptoms and signs are usually non specific. Spontaneous hepatic rupture can be preceded by signs of hypovolaemic shock; yet the diagnosis is infrequently made prior to emergent laparotomy. We present the case of a 35 year old nulliparous woman with a second trimester gestational hepatic rupture associated with HELLP syndrome. We briefly discuss the aetiology, diagnostic difficulties and treatment options associated with this rare presentation

A multicopper oxidase (Cj1516) and a CopA homologue (Cj1161) are major components of the copper homeostasis system of Campylobacter jejuni

© American Society for Microbiology, 2008. Post-print version of article deposited in accordance with SHERPA RoMEO guidelines.Metal ion homeostasis mechanisms in the food-borne human pathogen Campylobacter jejuni are poorly understood. The Cj1516 gene product is homologous to the multicopper oxidase CueO, which is known to contribute to copper tolerance in Escherichia coli. Here we show, by optical absorbance and electron paramagnetic resonance spectroscopy, that purified recombinant Cj1516 contains both T1 and trinuclear copper centers, which are characteristic of multicopper oxidases. Inductively coupled plasma mass spectrometry revealed that the protein contained approximately six copper atoms per polypeptide. The presence of an N-terminal "twin arginine" signal sequence suggested a periplasmic location for Cj1516, which was confirmed by the presence of p-phenylenediamine (p-PD) oxidase activity in periplasmic fractions of wild-type but not Cj1516 mutant cells. Kinetic studies showed that the pure protein exhibited p-PD, ferroxidase, and cuprous oxidase activities and was able to oxidize an analogue of the bacterial siderophore anthrachelin (3,4-dihydroxybenzoate), although no iron uptake impairment was observed in a Cj1516 mutant. However, this mutant was very sensitive to increased copper levels in minimal media, suggesting a role in copper tolerance. This was supported by increased expression of the Cj1516 gene in copper-rich media. A mutation in a second gene, the Cj1161c gene, encoding a putative CopA homologue, was also found to result in copper hypersensitivity, and a Cj1516 Cj1161c double mutant was found to be more copper sensitive than either single mutant. These observations and the apparent lack of alternative copper tolerance systems suggest that Cj1516 (CueO) and Cj1161 (CopA) are major proteins involved in copper homeostasis in C. jejuni

An Electrocorticographic Brain Interface in an Individual with Tetraplegia

Brain-computer interface (BCI) technology aims to help individuals with disability to control assistive devices and reanimate paralyzed limbs. Our study investigated the feasibility of an electrocorticography (ECoG)-based BCI system in an individual with tetraplegia caused by C4 level spinal cord injury. ECoG signals were recorded with a high-density 32-electrode grid over the hand and arm area of the left sensorimotor cortex. The participant was able to voluntarily activate his sensorimotor cortex using attempted movements, with distinct cortical activity patterns for different segments of the upper limb. Using only brain activity, the participant achieved robust control of 3D cursor movement. The ECoG grid was explanted 28 days post-implantation with no adverse effect. This study demonstrates that ECoG signals recorded from the sensorimotor cortex can be used for real-time device control in paralyzed individuals