Cadherin-dependent mechanotransduction depends on ligand identity but not affinity.

This study investigates the relationship between classical cadherin binding affinities and mechanotransduction through cadherin-mediated adhesions. The mechanical properties of cadherin-dependent intercellular junctions are generally attributed to differences in the binding affinities of classical cadherin subtypes that contribute to cohesive energies between cells. However, cell mechanics and mechanotransduction may also regulate intercellular contacts. We used micropipette measurements to quantify the two-dimensional affinities of cadherins at the cell surface, and two complementary mechanical measurements to assess ligand-dependent mechanotransduction through cadherin adhesions. At the cell surface, the classical cadherins investigated in this study form both homophilic and heterophilic bonds with two-dimensional affinities that differ by less than threefold. In contrast, mechanotransduction through cadherin adhesions is strongly ligand dependent such that homophilic, but not heterophilic ligation mediates mechanotransduction, independent of the cadherin binding affinity. These findings suggest that ligand-selective mechanotransduction may supersede differences in cadherin binding affinities in regulating intercellular contacts.

N-glycosylation alters cadherin-mediated intercellular binding kinetics.

We present direct evidence that the N-glycosylation state of neural cadherin impacts the intrinsic kinetics of cadherin-mediated intercellular binding. Micropipette manipulation measurements quantified the effect of N-glycosylation mutations on intercellular binding dynamics. The wild-type protein exhibits a two-stage binding process in which a fast, initial binding step is followed by a short lag and second, slower transition to the final binding stage. Mutations that ablate N-glycosylation at three sites on the extracellular domains 2 and 3 of neural cadherin alter this kinetic fingerprint. Glycosylation does not affect the affinities between the adhesive N-terminal domains, but instead modulates additional cadherin interactions, which govern the dynamics of intercellular binding. These results, together with previous findings that these hypo-glycosylation mutations increase the prevalence of cis dimers on cell membranes, suggest a binding mechanism in which initial adhesion is followed by additional cadherin interactions, which enhance binding but are modulated by N-glycosylation. Given that oncogene expression drives specific changes in N-glycosylation, these results provide insight into possible mechanisms altering cadherin function during tumor progression.

Cadherin point mutations alter cell sorting and modulate GTPase signaling.

This study investigated the impact of cadherin binding differences on both cell sorting and GTPase activation. The use of N-terminal domain point mutants of Xenopus C-cadherin enabled us to quantify binding differences and determine their effects on cadherin-dependent functions without any potential complications arising as a result of differences in cytodomain interactions. Dynamic cell-cell binding measurements carried out with the micropipette manipulation technique quantified the impact of these mutations on the two-dimensional binding affinities and dissociation rates of cadherins in the native context of the cell membrane. Pairwise binding affinities were compared with in vitro cell-sorting specificity and ligation-dependent GTPase signaling. Two-dimensional affinity differences greater than five-fold correlated with cadherin-dependent in vitro cell segregation, but smaller differences failed to induce cell sorting. Comparison of the binding affinities with GTPase signaling amplitudes further demonstrated that differential binding also proportionally modulates intracellular signaling. These results show that differential cadherin affinities have broader functional consequences than merely controlling cell-cell cohesion.

Skin Temperature Increase Mediated By Wearable, Long Duration, Low-Intensity Therapeutic Ultrasound.

One of the safety concerns with the delivery of therapeutic ultrasound is overheating of the transducer-skin interface due to poor or improper coupling. The objective of this research was to define a model that could be used to calculate the heating in the skin as a result of a novel, wearable long-duration ultrasound device. This model was used to determine that the maximum heating in the skin remained below the minimum threshold necessary to cause thermal injury over multiple hours of use. In addition to this model data, a human clinical study used wire thermocouples on the skin surface to measure heating characteristics during treatment with the sustained ultrasound system. Parametric analysis of the model determined that the maximum temperature increase is at the surface of the skin ranged from 40-41.8° C when perfusion was taken into account. The clinical data agreed well with the model predictions. The average steady state temperature observed across all 44 subjects was 40°C. The maximum temperature observed was less than 44° C, which is clinically safe for over 5 hours of human skin contact. The resultant clinical temperature data paired well with the model data suggesting the model can be used for future transducer and ultrasound system design simulation. As a result, the device was validated for thermal safety for typical users and use conditions.

Sustained Acoustic Medicine: A Novel Long Duration Approach to Biomodulation Utilizing Low Intensity Therapeutic Ultrasound.

Therapeutic ultrasound is an established technique for biomodulation used by physical therapists. Typically it is used to deliver energy locally for the purpose of altering tissue plasticity and increasing local circulation. Access to ultrasound therapy has been limited by equipment and logistic requirements, which has reduced the overall efficacy of the therapy. Ultrasound miniaturization allows for development of portable, wearable, self-applied ultrasound devices that sidestep these limitations. Additionally, research has shown that the timescale of acoustic stimulation matters, and directly affects the quality of result. This paper describes a novel, long duration approach to therapeutic ultrasound and reviews the current data available for a variety of musculoskeletal conditions.

Human Factors Engineering and testing for a wearable, long duration ultrasound system self-applied by an end user.

One of the major challenges in the design of a new class of medical device is ensuring that the device will have a safe and effective user interface for the intended users. Human Factors Engineering addresses these concerns through direct study of how a user interacts with newly designed devices with unique features. In this study, a novel long duration, low intensity therapeutic ultrasound device is tested by 20 end users representative of the intended user population. Over 90% of users were able to operate the device successfully. The therapeutic ultrasound device was found to be reasonably safe and effective for the intended users, uses, and use environments.

Pilot Clinical Studies of Long Duration, Low Intensity Therapeutic Ultrasound for Osteoarthritis.

Osteoarthritis is one of the leading causes of disability in the aging population. Long duration, low intensity therapeutic ultrasound has had promising impact in animal models to slow the progression of the disease and provide joint relief. Two pilot studies were conducted using a novel, wearable platform for delivering ultrasound to evaluate the potential clinical benefits of ultrasound therapy on knee osteoarthritis. There was a pain reduction effect from using ultrasound, as high as fifty two percent in one study. As well, initial data demonstrates that mobility may be increased for patients experiencing mild to moderate arthritis of the knee.

Design and evaluation of a wearable self-applied therapeutic ultrasound device for chronic myofascial pain.

Ultrasound therapy for pain and healing is a versatile treatment modality for musculoskeletal conditions that is used daily in rehabilitation clinics around the world. Our group designed and constructed a wearable, battery-operated, low-intensity therapeutic ultrasound (LITUS) device that patients could self-apply and operate during daily activity for up to 6 h. Thirty patients with chronic trapezius myofascial pain evaluated the LITUS system in a double-blind, placebo-controlled, 10-d study under institutional review board approval. While continuing their prescribed medication regimen, patients with the active device reported on average 1.94× reduction in pain and 1.58× improvement in health relative to placebo devices after 1 h of treatment. Both of these results were statistically significant (p < 0.05) for the first 2 d of the study. Male patients reported the majority of benefit, and there is a sex-treatment confound in the sample. The study indicates that wearable, long-duration LITUS technology improves mobile access to drug-free pain relief.