The morphology of the glenoid cavity is highly variable, and no consensus exists regarding how to classify the different forms. We examined 98 dry scapulae to identify a common morphological entity and to define reproducible bony references of the glenoid cavity. The glenoid cavities were photographed perpendicularly in a standardized fashion. The bony peripheral rim was studied on these two-dimensional images, defined by randomly chosen points in order to define one or more circles. This study showed that only the peripheral rim of the inferior quadrants of the articular surface was found to be located on a circle ( P=0.926) with a radius of 12.8 mm (SD 1.3 mm). Defining the center of this circle appeared to be more reliable (ICC 0.98) than determining the middle point of the longitudinal axis (0,0) between the most cranial and most caudal points, defined as Saller's line (ICC 0.89). The distance of the center of this projected circle to the middle point of Saller's line had a unimodal distribution, suggesting the existence of only one glenoid cavity morphotype. We then investigated the relationship between the center of the circle and the area of subchondral bone thickening under the bare spot, the so-called tubercle of Assaki. Ten phenolized cadaveric glenoid cavities were examined with computed tomography. A circle was projected on the first image showing the bony peripheral rim, and this circle was copied on the consecutive slices until the tubercle of Assaki came across. The center of the circle was located within the area of the tubercle of Assaki, in all but one specimen. To investigate the clinical implications of this finding, the cadaver specimens were used to compare the position of the center of the circle with the postulated center of implantation according to the literature, and to the reference guide for a commonly used total shoulder prosthesis. The center of the circle was consistently situated more distal than the postulated center of the guide (mean 5.5 mm, range 4-8 mm) and the middle point of the glenoid cavity (mean 2 mm, range 1-3 mm). These findings could offer a reproducible point of reference for the glenoid cavity in osseous anthropometry and a valuable reference in shoulder replacement surgery, and might help in the definition of osseous glenohumeral instability.

• DCE-MRI plays an important role in differentiating benign from malignant cartilage tumours. • Retrospective study defined a threshold for 100 % detection of chondrosarcoma with DCE-MRI. • The threshold values were relative enhancement = 2 and slope = 4.5. • One hundred per cent chondrosarcoma detection corresponds with 36.7 % false-positive diagnosis of enchondroma. • Standard MRI is complementary to DCE-MRI in differentiating cartilaginous tumours.

We retrospectively reviewed 107 patients with 108 malignant or locally aggressive bone tumours treated between 1978 and 2009 by extracorporeal irradiation with 300 Gy to eradicate the tumour, and reimplantation of the bone as an orthotopic autograft. Patient subgroups were defined according to resection type. We describe the local recurrence rate, the graft infection rate and the factors affecting graft healing and graft survival. No local recurrences were detected in the irradiated grafts. At fiveyear follow-up, graft healing had occurred in 64% of patients, providing a stable and lasting reconstruction. For various reasons, 11% of grafts were removed, although no single factor was predictive of failure. All patient subgroups had comparable results. Early infection predicted the development of pseudarthrosis. Pelvic reconstructions had a worse graft survival. Rigid fixation and bridging of the graft appeared to be important technical points.

The secretory Rab27B small GTPase promotes invasive growth and metastasis in estrogen receptor (ER) a-positive breast cancer cells by orchestrating the peripheral targeting of vesicles secreting proinvasive growth regulators. Increased Rab27B expression is associated with poor prognosis in breast cancer patients. The molecular mechanisms of peripheral Rab27B secretory vesicle distribution are poorly understood. Mass spectrometry analysis on green fluorescent protein (GFP)-Rab27B vesicles prepared from GFP-Rab27B transfected MCF-7 human breast cancer cells detected eight subunits of the vacuolar H(1)-ATPase (V-ATPase) and the presence of V 0 a1 and V 0 d1 subunits was confirmed by Western blot analysis. Reversible inhibition of V-ATPase activity by bafilomycin A1 or transient silencing of V 0 a1 or V 0 d1 subunits demonstrated that V-ATPase controls peripheral localization and size of Rab27B vesicles. V-ATPase expression and activity further controls Rab27B-induced collagen type I invasion, cell-cycle progression and invasive growth in the chorioallantoic membrane assay. In agreement, Rab27B-dependent extracellular heat shock protein90a release and matrix metalloprotease-2 activation is markedly reduced by bafilomycin A1 and transient silencing of V 0 a1 and V 0 d1 subunits. Poor prognosis ERa-positive primary breast tumors expressing high levels of Rab27B also expressed multiple V-ATPase subunits and showed a strong cytoplasmic and peripheral V-ATPase V 1 E expression. In conclusion, inhibiting V-ATPase activity by interfering agents and drugs might be an effective strategy for blocking Rab27B-dependent proinvasive secretory vesicle trafficking in ERa-positive breast cancer patients.

Citation: Sys, G.M.L., Lapeire, L., Stevens, N., Favoreel, H., Forsyth, R., Bracke, M., De Wever, O. The In ovo CAM-assay as a Xenograft Model for Sarcoma. J. Vis. Exp. (77), e50522, doi:10.3791/50522 (2013). AbstractSarcoma is a very rare disease that is heterogeneous in nature, all hampering the development of new therapies. Sarcoma patients are ideal candidates for personalized medicine after stratification, explaining the current interest in developing a reproducible and low-cost xenotransplant model for this disease. The chick chorioallantoic membrane is a natural immunodeficient host capable of sustaining grafted tissues and cells without species-specific restrictions. In addition, it is easily accessed, manipulated and imaged using optical and fluorescence stereomicroscopy. Histology further allows detailed analysis of heterotypic cellular interactions.This protocol describes in detail the in ovo grafting of the chorioallantoic membrane with fresh sarcoma-derived tumor tissues, their single cell suspensions, and permanent and transient fluorescently labeled established sarcoma cell lines (Saos-2 and SW1353). The chick survival rates are up to 75%. The model is used to study graft-(viability, Ki67 proliferation index, necrosis, infiltration) and host (fibroblast infiltration, vascular ingrowth) behavior. For localized grafting of single cell suspensions, ECM gel provides significant advantages over inert containment materials. The Ki67 proliferation index is related to the distance of the cells from the surface of the CAM and the duration of application on the CAM, the latter determining a time frame for the addition of therapeutic products. Video LinkThe video component of this article can be found at