Significant effect of the posterior tibial slope on the weight-bearing, midflexion in vivo kinematics after cruciate-retaining total knee arthroplasty.

The purpose of the present study was to compare weight bearing (WB) and non-WB conditions, and to evaluate the effect of the posterior tibial slope (PTS) on the in vivo kinematics of 21 knees after posterior cruciate ligament-retaining total knee arthroplasty during midflexion using 2-dimensional/3-dimensional registration. During WB, medial pivot and bicondylar rollback were observed. During non-WB, both the medial and lateral condyles moved significantly more anteriorly as compared to the WB state. These patients were divided into 2 groups according to their PTS. The large PTS group showed a significant posterior displacement of the medial femoral condyle as compared with the small PTS group, but no significant difference was observed at the lateral femoral condyle during both WB and non-WB. The PTS influenced knee kinematics through gravity (124/125).

In vivo kinematics of three-component mobile-bearing total ankle replacement in rheumatoid ankle with talocalcaneal arthrodesis and spontaneous talocalcaneal fusion.

OBJECTIVE: It is often that patients with rheumatoid arthritis (RA) who require ankle surgeries already have the degeneration of talocalcaneal joints. When talocalcaneal joint was fused, whether operatively or spontaneously, ankle kinematics would be affected. The purpose of this paper was to study in vivo kinematics of mobile-bearing total ankle replacement (TAR) in rheumatoid ankle with concomitant talocalcaneal arthrodesis or with preexisting spontaneous talocalcaneal fusion. METHODS: Thirteen TARs in ten patients with RA, in whom talocalcaneal joints had already been fused spontaneously or surgically, were studied. Fluoroscopic images were obtained while each patient was walking with full weightbearing on the implanted ankle. Thereafter tibio-talar motion was analyzed by 2D/3D registration technique. RESULTS: Average tibio-talar motion was 4.0 ± 5.3° for plantarflexion and 6.6 ± 0.3° for dorsiflexion. Average range of internal/external rotation, inversion/eversion and AP translation was 3.8 ± 1.3°, 2.7 ± 1.0° and 1.6 ± 0.6 mm, respectively. CONCLUSIONS: Mobility of mobile-bearing TAR with talocalcaneal fusion was small during the stance phase of gait, but clinically measured ROM was mostly preserved. The movements of internal/external rotation and AP translation were allowed to a certain degree, but not of inversion/eversion. Even though the movement of inversion/eversion is limited, talocalcaneal arthrodesis could be accompanied with mobile-bearing TAR in rheumatoid ankles.

Structural analysis of the mechanism of inhibition and allosteric activation of the kinase domain of HER2 protein.

Aberrant signaling of ErbB family members human epidermal growth factor 2 (HER2) and epidermal growth factor receptor (EGFR) is implicated in many human cancers, and HER2 expression is predictive of human disease recurrence and prognosis. Small molecule kinase inhibitors of EGFR and of both HER2 and EGFR have received approval for the treatment of cancer. We present the first high resolution crystal structure of the kinase domain of HER2 in complex with a selective inhibitor to understand protein activation, inhibition, and function at the molecular level. HER2 kinase domain crystallizes as a dimer and suggests evidence for an allosteric mechanism of activation comparable with previously reported activation mechanisms for EGFR and HER4. A unique Gly-rich region in HER2 following the α-helix C is responsible for increased conformational flexibility within the active site and could explain the low intrinsic catalytic activity previously reported for HER2. In addition, we solved the crystal structure of the kinase domain of EGFR in complex with a HER2/EGFR dual inhibitor (TAK-285). Comparison with previously reported inactive and active EGFR kinase domain structures gave insight into the mechanism of HER2 and EGFR inhibition and may help guide the design and development of new cancer drugs with improved potency and selectivity.

Trimerization of murine TNF ligand family member LIGHT increases the cytotoxic activity against the FM3A mammary carcinoma cell line.

LIGHT is a member of the tumor necrosis factor ligand superfamily, which plays important roles in inflammatory and immune responses. LIGHT forms a membrane-anchored homotrimeric complex on the cell surface and is often processed as a soluble protein. Recombinant soluble human LIGHT produced by mammalian cells or Escherichia coli is functional at nanomolar concentrations. However, there is little information about the biological activity of mouse LIGHT (mLIGHT) because of the difficulty in producing bioactive soluble mLIGHT. In this study, recombinant trimeric soluble mLIGHT, or Foldon-mLIGHT, was produced by fusing mLIGHT with the trimerization domain foldon from bacteriophage T4 fibritin. Foldon-mLIGHT was secreted from 293F cells as a 68-kDa trimeric protein. The recombinant protein potently inhibited the growth of the FM3A mouse mammary carcinoma cell line with an IC(50) of 77 pM; however, the monomer or dimer forms of mLIGHT produced by E. coli or mammalian cell systems showed weak or no inhibitory activity. These data clearly indicated that trimerization of soluble mLIGHT is essential for its biological activity.

Comparison of in vivo kinematics of total knee arthroplasty between cruciate retaining and cruciate substituting insert.

BACKGROUND: The decision to choose cruciate retaining (CR) insert or cruciate substituting (CS) insert during total knee arthroplasty (TKA) remains a controversial issue. We hypothesized that there are different knee kinematics between CR and CS inserts and that a raised anterior lip design would offer a potential minimization of the paradoxical movement and provide joint stability. The objective of this study was to evaluate and compare kinematics of a CR and CS TKA of the same single-radius design. METHODS: We investigated the in vivo knee kinematics of 20 knees with a CR TKA (10 knees in the CR insert and 10 knees in the CS insert). Patients were examined during deep knee flexion using fluoroscopy and femorotibial motion was determined using a 2- to 3-dimensional registration technique, which used computer-assisted design models to reproduce the spatial positions of the femoral and tibial components. We evaluated the knee range of motion (ROM), femoral axial rotation relative to the tibial component, anteroposterior translation, and kinematic pathway of the nearest point of the medial and lateral femoral condyles on the tibial tray. RESULTS: The average ROM was 121.0 ± 17.3° in CR and 110.8 ± 12.4° in CS. The amount of femoral axial rotation was 7.2 ± 3.9° in CR, and 7.4 ± 2.7° in CS. No significant difference was observed in the amount of anterior translation between CR and CS. The CR and CS inserts had a similar kinematic pattern up to 100° flexion that was central pivot up to 70° flexion and then paradoxical anterior femoral movement until 100° flexion. CONCLUSION: The present study demonstrated that there was no significant difference between the inserts in knee kinematics. These kinematic results suggested that the increased anterior lip could not control anterior movement in the CS insert.

Patellar resurfacing has minimal impact on in vitro tibiofemoral kinematics during deep knee flexion in total knee arthroplasty.

BACKGROUND: While patellar resurfacing can affect patellofemoral kinematics, the effect on tibiofemoral kinematics is unknown. We hypothesized that patellar resurfacing would affect tibiofemoral kinematics during deep knee flexion due to biomechanical alteration of the extensor mechanism. METHODS: We performed cruciate-retaining TKA in fresh-frozen human cadaveric knees (N = 5) and recorded fluoroscopic kinematics during deep knee flexion before and after the patellar resurfacing. To simulate deep knee flexion, cadaver knees were tested on a dynamic, quadriceps-driven, closed-kinetic chain simulator based on the Oxford knee rig design under loads equivalent to stair climbing. To measure knee kinematics, a 2-dimensional to 3-dimensional fluoroscopic registration technique was used. Component rotation, varus-valgus angle, and anteroposterior translation of medial and lateral contact points of the femoral component relative to the tibial component were calculated over the range of flexion. RESULTS: There were no significant differences in femoral component external rotation (before patellar resurfacing: 6.6 ± 2.3°, after patellar resurfacing: 7.2 ± 1.8°, p = 0.36), and less than 1° difference in femorotibial varus-valgus angle between patellar resurfacing and non-resurfacing (p = 0.01). For both conditions, the medial and lateral femorotibial contact points moved posteriorly from 0° to 30° of flexion, but not beyond 30° of flexion. At 10° of flexion, after patellar resurfacing, the medial contact point was more anteriorly located than before patellar resurfacing. CONCLUSION: Despite the potential for alteration of the knee extensor biomechanics, patellar resurfacing had minimal effect on tibiofemoral kinematics. Patellar resurfacing, if performed adequately, is unlikely to affect postoperative knee function.

Effective expression and purification of bioactive recombinant soluble LIGHT.

LIGHT (TNFSF14, CD258) is a member of the tumor necrosis factor (TNF) ligand superfamily, which is involved in innate and adaptive immune responses as well as in regulation of cell survival and proliferation. LIGHT forms a membrane-anchored homotrimeric complex on the cell surface and is often processed as a soluble protein. In this study, we established an effective strategy for producing bioactive soluble forms of human LIGHT (s-hLIGHT), which is an extracellular region (Ile(84)-Val(240)) of human LIGHT (hLIGHT), and soluble forms of mouse LIGHT (s-mLIGHT), which is an extracellular region (Asp(72)-Val(239)) of mouse LIGHT (mLIGHT). To enhance the refolding of s-hLIGHT from inclusion bodies in Escherichia coli, we added L-cysteine to the denaturation buffer, which significantly improved the refolding efficiency of s-hLIGHT. However, there was little information available about the biological activity of mLIGHT in the literature because of the difficulty in producing bioactive s-mLIGHT. We produced trimeric s-mLIGHT by fusing s-mLIGHT with a trimerization domain termed "foldon" from bacteriophage T4 fibritin (Foldon-mLIGHT). We further demonstrated that Foldon-mLIGHT inhibited the growth of mouse carcinoma cells at the picomolar range, indicating that trimerization of s-mLIGHT is essential for its biological activity.

In vivo kinematic analysis of posterior-stabilized total knee arthroplasty for the valgus knee operated by the gap-balancing technique.

BACKGROUND: Most in vivo kinematic studies of total knee arthroplasty (TKA) report on the varus knee. The objective of the present study was to evaluate in vivo kinematics of a posterior-stabilized fixed-bearing TKA operated on a valgus knee during knee bending in weight-bearing (WB) and non-weight-bearing (NWB). METHODS: A total of sixteen valgus knees in 12 cases that underwent TKA with Scorpio NRG PS knee prosthesis and that were operated on using the gap balancing technique were evaluated. We evaluated the in vivo kinematics of the knee using fluoroscopy and femorotibial translation relative to the tibial tray using a 2-dimensional to 3-dimensional registration technique. RESULTS: The average flexion angle was 111.3°±7.5° in WB and 114.9° ± 8.4° in NWB. The femoral component demonstrated a mean external rotation of 5.9° ± 5.8° in WB and 7.4° ± 5.2° in NWB. In WB and NWB, the femoral component showed a medial pivot pattern from 0° to midflexion and a bicondylar rollback pattern from midflexion to full flexion. The medial condyle moved similarly in the WB condition and in the NWB condition. The lateral condyle moved posteriorly at a slightly earlier angle during the WB condition than during the NWB condition. CONCLUSIONS: We conclude that similar kinematics after TKA can be obtained with the gap balancing technique for the preoperative valgus deformity when compared to the kinematics of a normal knee, even though the magnitude of external rotation was small. LEVEL OF EVIDENCE: IV.

Role of the liver in glucose homeostasis in PI 3-kinase p85alpha-deficient mice.

Phosphoinositide 3-kinase (PI3K) p85alpha-deficient mice exhibit hypoglycemia as a result of increased insulin sensitivity and glucose uptake in peripheral tissues. Although PI3K is central to the metabolic actions of insulin, its mechanism of action in liver is not well understood. In the present study, we investigated hepatic insulin signaling and glucose homeostasis in p85alpha-deficient and wild-type mice. In the livers of p85alpha-deficient mice, p50alpha played a compensatory role in insulin-stimulated PI3K activation by binding to insulin receptor substrate (IRS)-1/2. In p85alpha-deficient mice, the ratio of p50alpha over p110 catalytic subunit of PI3K in the liver was higher than in the muscles. PI3K activity associated with IRS-1/2 was not affected by the lack of p85alpha in the liver. Insulin-stimulated Akt and phosphatase and tensin homologue deleted on chromosome 10 (PTEN) activities in the liver were similar in p85alpha-deficient and wild-type mice. A hyperinsulinemic-euglycemic clamp study revealed that the glucose infusion rate and the rate of disappearance were higher in p85alpha-deficient mice than in wild-type mice but that endogenous glucose production tended to be higher in p85alpha-deficient mice than in wild-type mice. Consistent with this finding, the expression of glucose-6-phosphatase and phosphoenolpyruvate carboxykinase in livers after fasting was higher in p85alpha-deficient mice than in wild-type mice. After mice were fasted, the intrahepatic glucose-6-phosphate level was almost completely depleted in p85alpha-deficient mice. The glycogen content fell to nearly zero as a result of glycogenolysis shortly after the initiation of fasting in p85alpha-deficient mice. The absence of an increase in insulin-stimulated PI3K activation in the liver of p85alpha-deficient mice, unlike the muscles, may be associated with the molecular balance between the regulatory subunit and the catalytic subunit of PI3K. Gluconeogenesis was rather elevated in p85alpha-deficient mice, compared with in wild-type mice, and the liver seemed to partially compensate for the increase in glucose uptake in peripheral tissues.