Novel and recurrent TRPV4 mutations and their association with distinct phenotypes within the TRPV4 dysplasia family.

BACKGROUND: Mutations in TRPV4, a gene that encodes a Ca(2+) permeable non-selective cation channel, have recently been found in a spectrum of skeletal dysplasias that includes brachyolmia, spondylometaphyseal dysplasia, Kozlowski type (SMDK) and metatropic dysplasia (MD). Only a total of seven missense mutations were detected, however. The full spectrum of TRPV4 mutations and their phenotypes remained unclear. OBJECTIVES AND METHODS: To examine TRPV4 mutation spectrum and phenotype-genotype association, we searched for TRPV4 mutations by PCR-direct sequencing from genomic DNA in 22 MD and 20 SMDK probands. RESULTS: TRPV4 mutations were found in all but one MD subject. In total, 19 different heterozygous mutations were identified in 41 subjects; two were recurrent and 17 were novel. In MD, a recurrent P799L mutation was identified in nine subjects, as well as 10 novel mutations including F471del, the first deletion mutation of TRPV4. In SMDK, a recurrent R594H mutation was identified in 12 subjects and seven novel mutations. An association between the position of mutations and the disease phenotype was also observed. Thus, P799 in exon 15 is a hot codon for MD mutations, as four different amino acid substitutions have been observed at this codon; while R594 in exon 11 is a hotspot for SMDK mutations. CONCLUSION: The TRPV4 mutation spectrum in MD and SMDK, which showed genotype-phenotype correlation and potential functional significance of mutations that are non-randomly distributed over the gene, was presented in this study. The results would help diagnostic laboratories establish efficient screening strategies for genetic diagnosis of the TRPV4 dysplasia family diseases.

Morphometric analysis of acetabular dysplasia in cerebral palsy.

We carried out a morphometric analysis of acetabular dysplasia in patients with cerebral palsy requiring hip surgery using three-dimensional CT. We evaluated 54 hips in 27 patients. The contralateral normal hips of ten patients with unilateral Perthes' disease were used as a control group. The acetabular defects were assessed qualitatively as anterior, posterior or global. Quantitative assessments were made using three-directional acetabular indices (anterosuperior, superolateral and posterosuperior) and measured by multiplanar reformation, from which we calculated the acetabular volume. In the qualitative study, posterior defects were most common in the subluxation group whereas global defects predominated in the dislocation group. In the quantitative study, all acetabular indices in both the subluxation and dislocation groups were higher than those in the control group and the superolateral indices showed a tendency to elevation in the dislocation group. The acetabular volume was largest in the control group, smallest in the dislocation group, and intermediate between the two in the subluxation group.

The bicompartmental acetabulum in Perthes' disease: 3D-CT and MRI study.

The bicompartmental acetabulum is one of the morphological changes which may be seen in children with Legg-Calvé-Perthes' disease. Three-dimensional CT and MRI were used to analyse the detailed morphology of the acetabulum with special reference to its inner surface, in 16 patients with Perthes' disease and a bicompartmental acetabulum.The bicompartmental appearance was seen on the coronal plane image through the acetabular fossa. The lunate surface was seen to grow laterally resulting in an increased mediolateral thickness of the triradiate cartilage. On the horizontal plane images, the acetabular fossa had deepened and had a distinct prominence at its posterior border. The combination of these morphological changes resulted in a bicompartmental appearance on plain radiography. Acetabular bicompartmentalisation appears to be the result of an imbalance of growth between the cartilage-covered lunate surface and the cartilage-devoid acetabular fossa.

Edge contacts of graphene formed by using a controlled plasma treatment.

Despite the fact that the outstanding properties of graphene are well known, the electrical performance of the material is limited by the contact resistance at the metal-graphene interface. In this study, we demonstrate the formation of "edge-contacted" graphene through the use of a controlled plasma processing technique that generates a bond between the graphene edge and the contact metal. This technique controls the edge structure of the bond and significantly reduces the contact resistance. This simple approach requires no additional post-processing and has been proven to be very effective. In addition, controlled pre-plasma processing was applied in order to produce CVD-graphene field effect transistors with an enhanced adhesion and improved carrier mobility. The contact resistance attained by using pre-plasma processing was 270 Ω µm, which is a decrease of 77%.

Evaluation of In-Stent Restenosis After Stent Implantation in the Vertebral Artery Ostium by Multislice Computed Tomography Angiography: Factors Affecting Accurate Diagnosis.

PURPOSE: Few articles have evaluated vertebral artery ostium stents using multislice computed tomography (CT). The purpose of our study was to evaluate the diagnostic performance of 64- and 16-slice CT for detecting significant in-stent restenosis after vertebral artery ostium stenting, and to identify factors affecting the accurate diagnosis by CT. METHODS: We reviewed 57 stents scanned using 64-slice CT and 34 stents using 16-slice CT. The accuracy of CT for diagnosing significant in-stent restenosis (≥ 50% diameter narrowing) was calculated using conventional angiography as a reference standard. Possible factors influencing the diagnostic performance of CT were analyzed, such as CT scanner, image quality, and stent characteristics. RESULTS: With 64-slice CT, 46 (80.7%) of 57 stents were classified as evaluable, while with 16-slice CT, 28 (82.3%) of 34 stents were classified as evaluable. No stents with diameters ≤ 2.75 mm were evaluable. The respective results for 64- versus 16-slice CT were sensitivity 87.5% (95% confidence interval [CI] 47.3-99.7%) versus 100% (95% CI 15.8-100.0%), specificity 94.7% (95% CI 82.3%-99.4%) versus 96.2% (95% CI 80.4-99.9%). Factors reducing the accurate diagnosis were those associated with poor image quality, a diameter ≤ 2.75 mm, and drug-eluting stent type (p < 0.05). CONCLUSIONS: 64-slice and 16-slice CT scans are adequate in stents with diameters > 2.75 mm for the evaluation of in-stent restenosis after stent implantation in the vertebral artery ostium.

Valgus femoral osteotomy for hinge abduction in Perthes' disease. Decision-making and outcomes.

We studied, clinically and radiologically, the growth and remodelling of 21 hips after valgus femoral osteotomy with both rotational and sagittal correction for hinge abduction in 21 patients (mean age, 9.7 years) with Perthes' disease. The exact type of osteotomy performed was based on the pre-operative clinical and radiological assessment and the results of intra-operative dynamic arthrography. The mean IOWA hip score was 66 (34 to 76) before surgery and 92 (80 to 100) at a mean follow-up of 7.1 years (3.0 to 15.0). Radiological measurements revealed favourable remodelling of the femoral head and improved hip joint mechanics. Valgus osteotomy, with both rotational and sagittal correction, can improve symptoms, function and remodelling of the hip in patients with Perthes' disease.

Transfer of extensor carpi radialis longus or brevis for opponensplasty.

For the restoration of thumb opposition many types of tendon transfer techniques have been described. The flexor digitorum superficialis (FDS) of the ring finger is commonly selected as a motor. On occasion, however, the quality of the flexor muscles of the fingers or wrist is not good enough for tendon transfer and another available muscle must be selected. In this situation, we have preferred to use an extensor carpi radialis longus (ECRL) or brevis (ECRB) transfer to restore opposition of the thumb. Follow-up examination, at an average 5 years and 10 months after operation, showed that the results of ten of 11 transfers were excellent and the other was good.

Development of a fibre-optic dosemeter to measure the skin dose and percentage depth dose in the build-up region of therapeutic photon beams.

In this study, a fibre-optic dosemeter (FOD) using an organic scintillator with a diameter of 0.5 mm for photon-beam therapy dosimetry was fabricated. The fabricated dosemeter has many advantages, including water equivalence, high spatial resolution, remote sensing and real-time measurement. The scintillating light generated from an organic-dosemeter probe embedded in a solid-water stack phantom is guided to a photomultiplier tube and an electrometer via 20 m of plastic optical fibre. Using this FOD, the skin dose and the percentage depth dose in the build-up region according to the depths of a solid-water stack phantom are measured with 6- and 15-MV photon-beam energies with field sizes of 10 × 10 and 20 × 20 cm(2), respectively. The results are compared with those measured using conventional dosimetry films. It is expected that the proposed FOD can be effectively used in radiotherapy dosimetry for accurate measurement of the skin dose and the depth dose distribution in the build-up region due to its high spatial resolution.

Feasibility study on development of Cerenkov fiber-optic dosimeter for radiotherapy application.

To obtain real-time dose information in photon-beam therapy using a clinical linear accelerator, we fabricated a novel Cerenkov fiber-optic dosimeter using two plastic optical fibers without employing a scintillator. In this study, the light intensity and spectrum of Cerenkov radiation induced by a high-energy photon beam were measured as functions of the irradiation angle and the length difference between the two plastic optical fibers in the dosimeter probe. Also, we obtained a percentage depth dose curve for a 6 MV photon beam with a field size of 10 × 10 cm(2) according to the depth of the solid water phantom. Based on the results of this study, it is anticipated that the proposed Cerenkov fiber-optic dosimeter can be developed as a useful dosimeter to accurately obtain dose information prior to conducting radiotherapy.

Fracture in long bones stabilised by telescopic intramedullary rods in patients with osteogenesis imperfecta.

We investigated the fracture-free survival of long bones stabilised by a telescopic intramedullary rod (TIMR) in patients with osteogenesis imperfecta with respect to the remodelling status of fracture or osteotomy sites and TIMR regions, in order to identify risk factors for fracture. A total of 44 femora and 28 tibiae in 25 patients with a mean age of 5.0 years (1.9 to 10.5) at presentation were studied. There were six patients with Sillence type I, five with type III, 13 with type IV and one with type V osteogenesis imperfecta. All received bisphosphonate treatment at the same stage during the mean follow-up of 7.3 years (0.5 to 18.1). The fracture-free survival was estimated at 6.2 years (95% confidence interval 5.1 to 7.3) by Kaplan-Meier analysis. More than half the fracture or osteotomy sites remained in a less-remodelled state at the latest follow-up or time of fracture. Of the 33 fractures, 29 (87.9%) occurred in long bones containing a less-remodelled site, and these fractures were located at this site. The relative fracture risk at the rod tip was significantly greater than in any other TIMR region (p < 0.001), and this was higher in bone segments having a less-remodelled site. This study shows a persistent fracture risk in TIMR-stabilised long bones, especially at less-remodelled fracture or osteotomy sites and at the rod tip.

Shelf acetabuloplasty for children with Perthes' disease and reducible subluxation of the hip: prognostic factors related to hip remodelling.

We evaluated radiological hip remodelling after shelf acetabuloplasty and sought to identify prognostic factors in 25 patients with a mean age of 8.9 years (7.0 to 12.3) who had unilateral Perthes' disease with reducible subluxation of the hip in the fragmentation stage. At a mean follow-up of 6.7 years (3.2 to 9.0), satisfactory remodelling was observed in 18 hips (72%). The type of labrum in hip abduction, as determined by intra-operative dynamic arthrography, was found to be a statistically significant prognostic factor (p = 0.012). Shelf acetabuloplasty as containment surgery seems to be best indicated for hips in which there is not marked collapse of the epiphysis and in which the extruded epiphyseal segment slips easily underneath the labrum on abduction, without imposing undue pressure on the lateral edge of the acetabulum.

Residual pelvic rotation after single-event multilevel surgery in spastic hemiplegia.

Our aim in this retrospective study of 52 children with spastic hemiplegia was to determine the factors which affected the amount of residual pelvic rotation after single-event multilevel surgery. The patients were divided into two groups, those who had undergone femoral derotation osteotomy and those who had not. Pelvic rotation improved significantly after surgery in the femoral osteotomy group (p < 0.001) but not in the non-femoral osteotomy group. Multiple regressions identified the following three independent variables, which significantly affected residual pelvic rotation: the performance of femoral derotation osteotomy (p = 0.049), the pre-operative pelvic rotation (p = 0.003) and the post-operative internal rotation of the hip (p = 0.001). We concluded that there is a decrease in the amount of pelvic rotation after single-event multilevel surgery with femoral derotation osteotomy. However, some residual rotation may persist when patients have severe rotation before surgery.

Morphometric changes in the acetabulum after Dega osteotomy in patients with cerebral palsy.

We carried out a morphometric analysis of the acetabulum following Dega osteotomy in patients with cerebral palsy using three-dimensional CT. We assessed 17 acetabula in 12 patients with instability of the hip. A Dega osteotomy and varus derotation femoral osteotomy were performed in all 17 hips. Three-dimensional CT scans were taken before and approximately one year after operation. Acetabular cover was evaluated using anterosuperior, superolateral and posterosuperior acetabular indices, and the change in the acetabular volume was calculated. Inter- and intra-observer reliability was assessed using the intraclass correlation coefficient. After the osteotomy, the anterosuperior, superolateral and posterosuperior cover had improved significantly towards the value seen in a control group. The mean acetabular volume increased by 68%.

Refracture after Ilizarov osteosynthesis in atrophic-type congenital pseudarthrosis of the tibia.

We investigated patterns of refracture and their risk factors in patients with congenital pseudarthrosis of the tibia after Ilizarov osteosynthesis. We studied 43 cases in 23 patients. Temporal and spatial patterns of refracture and refracture-free survival were analysed in each case. The refracture-free rate of cumulative survival was 47% at five years and did not change thereafter. Refracture occurred at the previous pseudarthrosis in 16 of 19 cases of refracture. The risk of refracture was significantly higher when osteosynthesis was performed below the age of four years, when the tibial cross-sectional area was narrow, and when associated with persistent fibular pseudarthrosis. Refracture occurs frequently after successful osteosynthesis in these patients. Delaying osteosynthesis, maximising the tibial cross-sectional area and stabilising the fibula may reduce the risk of refracture.

Expression and role of interleukin-6 in distraction osteogenesis.

Distraction osteogenesis is a special form of bone healing in which well-controlled distraction stresses and consequent tensile strains within callus tissue induce very efficient new bone formation. Proinflammatory cytokines are involved during the early phase of fracture healing and callus remodeling. Temporal expression patterns of proinflammatory cytokines were assessed in Sprague-Dawley rat tibial models of distraction osteogenesis and acute lengthening, and only interleukin-6 (IL-6) was found to be specifically induced during the distraction phase. IL-6 immunoreactivity was detected not only in hemopoietic cells and osteoblasts but also in the spindle-shaped cells of the fibrous interzone, where most of the tensile strains are concentrated. In vitro study revealed that IL-6 did not affect the proliferation of C3H10T1/2 cells, mouse bone marrow stromal cells (MSCs), or MC3T3-E1 cells; but its blocking antibody reduced the proliferation of C3H10T1/2 cells and MSCs. The mRNA expression of COL1A1 and osteopontin were not changed by IL-6 or its blocking antibody, but the alkaline phosphatase activities of MC3T3-E1 cells were increased by IL-6 and decreased by its blocking antibody. These findings indicate that IL-6 is a proinflammatory cytokine that responds to tensile strain during distraction osteogenesis. IL-6 negatively affects the proliferation of primitive mesenchymal cells, whereas the differentiation of more mature osteoblastic lineage cells is enhanced by IL-6 in vitro. IL-6 appears to be one of the cytokines involved in the complex network of signal cascades evoked during distraction osteogenesis and may differentially affect immature and mature osteoblastic lineage cells.

Distraction osteogenesis for segmental bone defect. Physeal change after acute bone shortening followed by gradual lengthening in a rabbit tibia model.

We investigated physeal change after acute bone shortening followed by distraction osteogenesis in a rabbit tibia model. Distraction osteogenesis was performed on acutely shortened right rabbit tibia after removing a diaphyseal segment until distraction reached the length of the excised segment. Sixty animals were divided into three groups (n=20) according to the amount of shortening (20, 30, and 40% of original length) and radiographic, immunohistochemical, and histomorphometric analyses were performed to evaluate the physeal activity. The results showed no differences between groups and all animals could recover the original length without growth retardation. There was no notable venous congestion or lymphatic kinking associated with acute shortening. These results suggest that distraction osteogenesis at the primary site of shortening may have little adverse effects on the growth plate, when the shortening can provide sufficient soft tissue without vascular compromise and the amount of lengthening does not exceed that of shortening.

Matrix and modifier effects in the supercritical fluid extraction of cocaine and benzoylecgonine from human hair.

The supercritical fluid extraction (SFE) behavior of cocaine and its major metabolite benzoylecgonine (BZE) was investigated and found to be highly dependent upon the chemical nature of the matrix and the manner in which the target drug analytes are incorporated into or on the matrix. The recovery of cocaine from Teflon wool, filter paper, drug-fortified hair, and drug user hair was studied using a variety of CO2/modifier mixtures. Incorporation of a triethylamine (TEA)/water modifier mixture provided dramatic improvements in the recovery of cocaine from interactive matrixes. The results suggest that the SF extractability of cocaine is not limited by analyte solubility; rather, desorption of cocaine from hair binding sites is a rate-limiting step in the SFE process. A displacement SFE mechanism is hypothesized in which TEA (as the triethylammonium cation) competes with cocaine for negatively charged hair binding sites. The dependence of extractability on hair/drug binding interactions allows the differentiation of cocaine present at different discrete sites in hair based on differences in SFE behavior. These findings suggest the potential for distinguishing exogenous (i.e., environmental) from endogenous (i.e., physiological) sources of drugs in hair. In contrast to the results observed for cocaine, SFE recoveries of BZE were poor from all matrixes and under all conditions studied. Its increased polarity, the presence of an additional binding site, and the possibility of multiple charged states suggest that poor BZE recoveries may be due to both poor analyte solubility and failure to desorb the analyte from hair binding sites under the conditions employed.