Objective. CD248 is a transmembrane glycoprotein expressed on the surface of activated perivascular and fibroblast-like cells. This study was undertaken to explore the function of CD248 and its cytoplasmic domain in arthritis.Methods. Synovial tissue biopsy samples from healthy controls, from patients with psoriatic arthritis (PsA), and from patients with rheumatoid arthritis (RA) were stained for CD248. Transgenic mice that were CD248-deficient (CD248-knockout [CD248 KO/KO ]) or mice with CD248 lacking the cytoplasmic domain (CD248 CyD/CyD ) were generated. Collagen antibodyinduced arthritis (CAIA) was induced in these mice and in corresponding wild-type (WT) mice as controls. Clinical signs and histologic features of arthritis were evaluated. Cytokine levels were determined by enzymelinked immunosorbent assay, and the number of infiltrating inflammatory cells was quantified by immunohistochemistry. In vitro studies were performed with fibroblasts from CD248-transgenic mouse embryos to explain the observed effects on inflammation.Results. Immunostaining of synovium from patients with PsA and patients with RA and that from mice after the induction of CAIA revealed strong CD248 expression in perivascular and fibroblast-like stromal cells. CD248 KO/KO and CD248 CyD/CyD mice had less severe arthritis, with lower plasma levels of proinflammatory cytokines, as compared with WT controls. Moreover, the joints of these mice had less synovial hyperplasia, reduced accumulation of inflammatory cells, and less articular cartilage and bone damage. Tumor necrosis factor ␣-induced monocyte adhesion to CD248 CyD/CyD fibroblasts was impaired. CD248 CyD/CyD fibroblasts exhibited reduced expression of hypoxia-inducible factor 1␣, placental growth factor, vascular endothelial growth factor, and matrix metalloproteinase 9 activity in response to transforming growth factor ␤.Conclusion. CD248 contributes to synovial hyperplasia and leukocyte accumulation in inflammatory arthritis, the effects of which are mediated partly via its cytoplasmic domain. CD248 is therefore a potential new target in the treatment of arthritis.

Genetic analysis of BRCA1 by sequencing is often preceded by a scanning method like denaturing gradient gel electrophoresis (DGGE), protein truncation test (PTT) or DHPLC. High-resolution melting curve (HRM) analysis is a promising and economical method for high-throughput mutation scanning. The EuroGentest network (www.eurogentest.org) aims to assist with the introduction of novel technologies in the diagnostic setting. Therefore, we have performed a thorough and highstandard interlaboratory evaluation and validation of HRM, in collaboration with Idaho Technology, the manufacturer of the LightScanner TM (LS). Through this detailed study of 170 variants, we have generated guidelines for easy setup and implementation of HRM as a scanning technique for new genes, which are adaptable to the quality system of an individual diagnostic laboratory. This validation study includes the description of a BRCA1-specific mutation screening test using the 96-well LS. This assay comprises 40 amplicons and was evaluated using a statistically significant elaborate panel of variants and control DNA samples. All heterozygous variants were detected. Moreover, genotype analysis for nine common polymorphisms created a fast screening and detection method for these frequently occurring nonpathogenic variants. A blind study using a total of 28 patient-derived DNA samples resulted also in 100% detection and showed an average specificity of 98%, indicating a low incidence of false positives (FPs).

We have investigated diffusion and activation of boron implanted with 6 keV energy to a maximum concentration of 8.0×1020atoms∕cm3 in crystalline germanium (c-germanium) and preamorphized germanium, employing rapid thermal annealing in the range of 400–600 °C. As-implanted boron profiles in preamorphized germanium are shallower than the ones in c-germanium due to channeling suppression. While boron diffusion is not observed either in c-germanium or during the germanium regrowth from amorphous state, the boron activation level achieved from the two starting phases is significantly different. A boron activation level of 2.4×1020atoms∕cm3 has been found in regrown germanium, while a level of only 1.2×1019atoms∕cm3 is observed in c-germanium. Remarkably, there is no evidence of any residual extended defectivity at the original crystalline/amorphous interface, when preamorphization is performed.

BackgroundStromal fibroblasts participate in the development of a permissive environment for tumor growth, yet molecular pathways to therapeutically target fibroblasts are poorly defined. CD248, also known as endosialin or tumor endothelial marker 1 (TEM1), is a transmembrane glycoprotein expressed on activated fibroblasts. We recently showed that the cytoplasmic domain of CD248 is important in facilitating an inflammatory response in a mouse model of arthritis. Others have reported that CD248 gene inactivation in mice results in dampened tumor growth. We hypothesized that the conserved cytoplasmic domain of CD248 is important in regulating tumor growth.MethodsMice lacking the cytoplasmic domain of CD248 (CD248CyD/CyD) were generated and evaluated in tumor models, comparing the findings with wild-type mice (CD248WT/WT).ResultsAs compared to the response in CD248WT/WT mice, growth of T241 fibrosarcomas and Lewis lung carcinomas was significantly reduced in CD248CyD/CyD mice. Tumor size was similar to that seen with CD248-deficient mice. Conditioned media from CD248CyD/CyD fibroblasts were less effective at supporting T241 fibrosarcoma cell survival. In addition to our previous observation of reduced release of activated matrix metalloproteinase (MMP)-9, CD248CyD/CyD fibroblasts also had impaired PDGF-BB-induced migration and expressed higher transcripts of tumor suppressor factors, transgelin (SM22α), Hes and Hey1.ConclusionsThe multiple pathways regulated by the cytoplasmic domain of CD248 highlight its potential as a therapeutic target to treat cancer.

We have studied implant-induced damage, defect annealing, and recrystallization of B, Ga, P, As, and Sb introduced in Ge by ion implantation at high doses, such that dopant chemical concentrations are above the corresponding solubility in Ge, with energies that target about 100-nm junction depths. It is shown that the amount of damage induced in the Ge lattice increases with the mass of the implanted ion, as expected. Implanted B produces local amorphous regions, although crystalline Ge zones are present in the implanted layer. P is a self-amorphizing ion, creating a continuous amorphous layer during implantation. However, a low thermal budget is sufficient to fully regrow the amorphous layer, without evidence of residual extended defects, as evaluated by crosssectional transmission electron microscopy. Conversely, high concentrations of As cause a significant decrease of the regrowth rate of the damaged layer during rapid thermal annealing in the 400-600°C range studied. Finally, high-dose implantation of heavy ions such as Sb induces dramatic morphologic changes in Ge that are not recovered by post-implant rapid thermal annealing.