HK-2 human renal proximal tubule cells (RPTC) are commonly used in the in vitro study of “normal” RPTCs. We recently discovered that HK-2 cells are uncoupled from dopamine-1 receptor (D1R) adenylyl cyclase (AC) stimulation. We hypothesized that G protein coupled receptor kinase type 4 (GRK4) single nucleotide polymorphisms (SNPs) may be responsible for the D1R/AC uncoupling in HK-2. This hypothesis was tested by genotyping GRK4 SNPs, measuring D1-like receptor agonist (fenoldopam)stimulated cAMP accumulation, quantifying D1R inhibition of sodium transport, and testing the ability of GRK4 siRNA to reverse the D1R/AC uncoupling. We compared HK-2 to 2 normally coupled human RPTC cell lines (nRPTC) and 2 uncoupled RPTC cell lines (uRPTC). The HK-2 cell line was found to have 4 out of 6 potential GRK4 SNPs known to uncouple the D1R from AC (namely R65L, A142V, and A486V). AC response to fenoldopam stimulation was increased in the two nRPTC cell lines (FEN 2.02±0.05-fold and 2.33±0.19-fold over control, P<0.001, N=4), but not in the two uncoupled or HK-2 cell lines. GRK4 siRNA rescued the fenoldopam-mediated AC stimulation in the uncoupled cells, including HK-2. The expected fenoldopam -mediated inhibition of sodium hydrogen exchanger type 3 was absent in HK-2 (N=6) and uRPTCs (N=6), but was observed in the two nRPTCs (−25.41±4.7% and −27.36±2.70% (P<0.001, N=6)), which express wild-type GRK4. Despite the fact that HK-2 cells retain many functional characteristics of RPTCs, they are not normal from the perspective of dopaminergic function.

Chronic lateral ankle instability often causes adults to require a surgical intervention with subsequent physical therapy to assist with returning to their prior level of function. This systematic review is hoping to provide an up to date understanding of surgical procedures performed to correct chronic lateral ankle instability and establish a protocol for others to follow when treating adults who are status-post chronic lateral ankle instability surgery. This review looked at level I to III research studies that included surgical interventions to correct chronic lateral ankle instability as well as a rehabilitation protocol. This study found implementation of a rehabilitation protocol after surgical intervention could improve balance and subjective functional outcomes. It also determined that early weight-bearing may allow for early strengthening as range of motion returns faster. Further research is required utilizing larger randomized studies to better evaluate the outcomes of specific rehabilitation protocols in this patient population.

Category: Basic Sciences/Biologics; Ankle; Hindfoot Introduction/Purpose: The tall Controlled Ankle Motion (CAM) boot and the short CAM boot are commonly used devices to immobilize the foot and ankle. However, the effect of these devices on joint contact pressures is unknown. The objective of this study is to assess the effect of the tall CAM boot and short CAM boot on contact pressures of the ankle, subtalar, talonavicular, and calcaneocuboid joints. We hypothesize that both the tall CAM boot and short CAM boot will reduce contact pressures of the ankle and hindfoot joints, with the tall CAM boot having the greatest effect. Methods: Eight lower extremity cadaver specimens were mounted on a servohydraulic test frame. The specimens were loaded to 700 N at a cyclical frequency of 1 Hz with the posterior tibial, peroneus longus, peroneus brevis, flexor hallucis longus, flexor digitorum longus, and Achilles tendon physiologically tensioned. TekScan (TekScan, Boston, MA) pressure sensors were placed in the ankle, subtalar, talonavicular, and calcaneocuboid joints. In the sagittal plane, the specimens were loaded on a neutral surface, followed by 20o of dorsiflexion. Each specimen served as its own control, with contact pressures measured with no immobilization (control), followed by placement in a short CAM boot and tall CAM boot. In addition, contact pressures in the immobilized limbs were measured at muscle loads both equal to and half of the load applied to the control in order to account for decreased muscle activation during immobilization. Results: There was no difference in the average and peak contact pressures of the ankle, subtalar, talonavicular and calcaneocuboid joints when comparing the short CAM boot to no immobilization at equal tendon loads. The tall CAM boot significantly decreased average and peak contact pressures of the ankle, subtalar, and talonavicular joints when compared to no immobilization. The tall CAM decreased the contact pressures of the talonavicular and subtalar joint to a greater degree than the ankle joint. The reduction in contact pressures was accentuated when the load applied to the tendons was decreased in accordance with diminished muscle activation during immobilization. Neither immobilization device decreased the contact pressures of the calcaneocuboid joint at equal tendon loads. Neither CAM boot changed the center of pressure of any joint. Conclusion: Immobilization in a tall CAM boot decreases contact pressures of the ankle and hindfoot in both a neutral position and in dorsiflexion. A tall CAM boot should be used clinically if the goal of its use is to maximally reduce contact pressures of the ankle and hindfoot. The tall CAM boot is better at reducing the contact pressures of the subtalar and talonavicular joint than the ankle joint.