Mutation of the RB-1 and p53 tumor suppressors is associated with the development of human osteosarcoma. With the goal of generating a mouse model of this disease, we used conditional and transgenic mouse strains to inactivate Rb and/or p53 specifically in osteoblast precursors. The resulting Rb;p53 double mutant (DKO) animals are viable but develop early onset osteosarcomas with complete penetrance. These tumors display many of the characteristics of human osteosarcomas, including being highly metastatic. We established cell lines from the DKO osteosarcomas to further investigate their properties. These immortalized cell lines are highly proliferative and they retain their tumorigenic potential, as judged by their ability to form metastatic tumors in immunocompromised mice. Moreover, they can be induced to differentiate and, depending on the inductive signal, will adopt either the osteogenic or adipogenic fate. Consistent with this multipotency, a significant portion of these tumor cells express Sca-1, a marker that is typically associated with stem cells/uncommitted progenitors. By assaying sorted cells in transplant assays, we demonstrate that the tumorigenicity of the osteosarcoma cell lines correlates with the presence of the Sca-1 marker. Finally, we show that loss of Rb and p53 in Sca-1-positive mesenchymal stem/progenitor cells is sufficient to yield transformed cells that can initiate osteosarcoma formation in vivo.osx-cre ͉ Sca-1 ͉ hibernoma mouse model O steosarcomas account for Ϸ30% of malignant bone tumors and 3-4% of all childhood malignancies (1, 2). They arise primarily around the knee joint, lower femur and upper tibia, which are all regions of active bone growth and repair. These tumors are predominantly osteoblastic in nature, although there is a correlation between loss of differentiation and poor prognosis. The generation of new therapeutic treatments for osteosarcoma has improved the 5-year survival rate of affected individuals. However, like other mesenchymal neoplasms, osteosarcomas are predisposed to metastasize via the hematogenous route, and thus, pulmonary metastasis is a major cause of death. Analyses of both sporadic and hereditary tumors show that inactivation of the p53 and RB-1 tumor suppressors plays a key role in the development of this tumor type (1, 2). Li-Fraumeni patients, who often carry germ-line mutations in p53, are predisposed to a variety of tumors, 12% of which are bone sarcomas (3, 4). p53 mutations are also observed in 20-60% of sporadic osteosarcomas (5-7). Similarly, patients carrying germline mutations in RB-1 have an Ϸ500-fold higher incidence of osteosarcoma than the general population (8). Moreover, RB-1 mutations are detected in 70% of all adolescent osteosarcomas (9). Finally, human osteosarcomas can carry mutations in both p53 and RB-1 (10).Mouse models have provided considerable insight into the role of p53 in bone development and tumorigenesis. Experiments from three different settings suggest that p53 plays an important role in bone development by modul...

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The results and interpretation of the modest nower ( 90 kW) lower hybrid heating experiment on Alcator A are presented.The expected results from linear waveguide-plasma coupling theory are outlined, and the possible effects of parametric instabilities, scattering from density fluctuations, and imperfect energetic ion confinement are addressed.It is found experimentally that good coupling and the absence of rf breakdown are achieved with a double wavecuide array 2 at available rf power densities Prf < 2.5 kW/cm , the waveguide vacuum windows being outside the toroidal field magnets; a waveguide array having vacuum windows near the waveguide mouth so that the w = wc. layer can be pressurizedshows no breakdown at P > 8 kW/cm2 when a single waveguide is energized. Energetic ion wroduction and a factor of 50 increase in the :usion neutron rate are ozserved to take place at well defined values of central plasma density; below these densities electron heating occurs. The ion tail production is found to be independent of the relative phase of the double waveguide array employed. This ion heating occurs at a lower density than theoretically expected; together with the electron heating this indicates waves having n 1 5 being absorbed near the plasma center. Probes at the plasma edge observe a frequency downshifted and broadened rf pump signal that .is strongly attenuated as the plasma density increases through the neutron production band. These anomalous heating results and probe signals can be explained by a parametric decay of the pump wave into higher n lower hybrid waves near the plasma edge. An alternate qualitative explanation would be the poloidal scattering of the lower hybrid waves at the plasma periphery due to density fluctuations; the n,, of the scattered lower hybrid waves would then increase as they propagated inward due to magnetic shear. The neutron rate decay times imply that the rf creates ion tails having a substantial fraction of their energy above 50 keV.The neutron decay times and rates strongly depend on plasma current and indicate the expected influence of ion confinement on rf heating efficiencies. Finally the rf heating efficiencies are assessed.2

The retinoblastoma tumor-suppressor protein, pRb, is a member of the pocket protein family that includes p107 and p130. These proteins have well-defined roles in regulating entry into and exit from the cell cycle and also have cell cycle-independent roles in facilitating differentiation. Here we investigate the overlap between pocket protein's function during embryonic development by using conditional mutant alleles to generate Rb;p107 doublemutant embryos (DKOs) that develop in the absence of placental defects. These DKOs die between e13.5 and e14.5, much earlier than either the conditional Rb or the germline p107 single mutants, which survive to birth or are largely viable, respectively. Analyses of the e13.5 DKOs shows that p107 mutation exacerbates the phenotypes resulting from pRb loss in the central nervous system and lens, but not in the peripheral nervous system. In addition, these embryos exhibit novel phenotypes, including increased proliferation of blood vessel endothelial cells, and heart defects, including double-outlet right ventricle (DORV). The DORV is caused, at least in part, by a defect in blood vessel endothelial cells and/or heart mesenchymal cells. These findings demonstrate novel, overlapping functions for pRb and p107 in numerous murine tissues.cell cycle ͉ heart ͉ p107 ͉ retinoblastoma