We hypothesized that an exogenous bone growth factor could augment healing of a tendon graft in a bone tunnel in a rabbit anterior cruciate ligament-reconstruction model. Seventy rabbits underwent bilateral anterior cruciate ligament reconstructions with a semitendinosus tendon graft. One limb received a collagen sponge carrier vehicle containing a mixture of bone-derived proteins while the contralateral limb was treated with either no sponge or a sponge without bone-derived proteins. The reconstruction was evaluated at 2, 4, or 8 weeks with histologic, biomechanical, and magnetic resonance imaging analysis. Histologic analysis demonstrated that specimens treated with bone-derived proteins had a more consistent, dense interface tissue and closer apposition of new bone to the graft, with occasional formation of a fibrocartilaginous interface, when compared with control specimens. The treated specimens had significantly higher load-to-failure rates than did control specimens. Treatment with bone-derived proteins resulted in an average increase in tensile strength of 65%. The treated specimens were stronger than control specimens at each time point, but the difference was greatest at 8 weeks. On the basis of signal characteristics and new bone formation, magnetic resonance imaging was useful for predicting which limb was treated, the site of failure, and the limbs with higher load-to-failure values. This study demonstrates the potential for augmenting tendon healing in an intraarticular bone tunnel using an osteoinductive growth factor.

The successful establishment of human embryonic stem cell (hESC) lines has inaugurated a new era in regenerative medicine by facilitating the transplantation of differentiated ESCs to specific organs. However, problems with the safety and efficacy of hESC therapy in vivo remain to be resolved. Preclinical studies using animal model systems, including nonhuman primates, are essential to evaluate the safety and efficacy of hESC therapies. Previously, we demonstrated that common marmosets are suitable laboratory animal models for preclinical studies of hematopoietic stem cell therapies. As this animal model is also applicable to preclinical trials of ESC therapies, we have established novel common marmoset ESC (CMESC) lines. To obtain marmoset embryos, we developed a new embryo collection system, in which blastocysts can be obtained every 3 weeks from each marmoset pair. The inner cell mass was isolated by immunosurgery and plated on a mouse embryonic feeder layer. Some of the CMESC lines were cultured continuously for more than 1 year. These CMESC lines showed alkaline phosphatase activity and expressed stage-specific embryonic antigen (SSEA)-3, SSEA-4, TRA-1-60, and TRA-1-81. On the other hand, SSEA-1 was not detected. Furthermore, our novel CMESCs are pluripotent, as evidenced by in vivo teratoma formation in immunodeficient mice and in vitro differentiation experiments. Our established CMESC lines and the common marmoset provide an excellent experimental model system for understanding differentiation mechanisms, as well as the development of regenerative therapies using hESCs. Stem Cells 2005;23:1304-1313 This material is protected by U.S.

We investigated the effect of diet on abdominal autofluorescence detected by in vivo fluorescence imaging (FLI) of living mice.Groups of mice were fed a regular, alfalfa-free, or purified diet, and whole-body FLI was performed without the administration of fluorescent probes. In addition, quantum dots were injected intravenously into mice fed one of the three diets, and FLI was performed 3 and 24 hours later. Intense autofluorescence originating from the animals' intestinal contents was observed in mice fed the regular diet. Intestinal autofluorescence decreased substantially after feeding with the alfalfa-free diet and further after feeding with the purified diet. The decline was rapid and took only 1 to 2 days; however, it may have been affected by an intake of feces. The reticuloendothelial system was clearly delineated using a low dose of quantum dots in mice fed the purified diet. On the other hand, intestinal autofluorescence was visible 24 hours postinjection in mice given the alfalfa-free diet and definitely impaired the image quality in mice fed the regular diet. The use of a low-fluorescence diet, especially a purified diet, rapidly reduces intestinal autofluorescence and is expected to enhance the potential of in vivo FLI.

In addition to rare injection failure, the IP injection of D: -luciferin led to the overestimation of signals from IP tissues. For BLI, SC injection was shown to be a convenient alternative to IP injection.

No abstract

Patients with Philadelphia chromosomepositive acute lymphoblastic leukemia (Ph ؉ ALL) have poor prognosis despite intensive therapeutic intervention. Recently, imatinib, a BCR-ABL tyrosine kinase inhibitor, has been proven to be an effective treatment for Ph ؉ ALL, but nearly all patients rapidly acquire resistance. High-dose imatinib administration might overcome this resistance; however, systemic toxicities would likely limit this approach. Therefore, a new delivery system allowing for the specific targeting of imatinib is urgently needed. Because almost all Ph ؉ ALL cells express CD19 on their surface, we have developed an immunoliposome carrying anti-CD19 antibody (CD19-liposomes). The internalization efficiency of the CD19-liposomes approached 100% in all Ph ؉ ALL cells but was very low in CD19 ؊ cells. The cytocidal effect of imatinib-encapsulated CD19-liposomes (imatinib-CD19-liposomes) on Ph ؉ ALL cell lines and primary leukemia cells from patients with Ph ؉ ALL was much greater than that of imatinib with or without control liposomes. Importantly, the imatinib-CD19-liposomes did not affect the colony formation of CD34 ؉ hematopoietic cells, even at inhibitory concentration of free imatinib. Taken together, these data clearly demonstrate that the imatinib-CD19-liposomes induced specific and efficient death of Ph ؉ ALL cells. This new therapeutic approach might be a useful treatment for Ph ؉ ALL with fewer side effects than free imatinib. IntroductionDespite the successes of high-dose chemotherapy followed by hematopoietic stem cell transplantation for the treatment of leukemia, Philadelphia chromosome-positive (Ph ϩ ) acute lymphoblastic leukemia (ALL) remains refractory to most therapeutic modalities currently available. The Philadelphia chromosome includes one of several forms of fusion genes between bcr and c-abl, and these fusions play a substantial role in the pathogenesis of chronic myelogenous leukemia (CML) and ALL. 1 The p190 bcr-abl fusion gene is detected in 20% to 35% of patients with ALL, and the prognosis of these patients is particularly poor. [2][3][4] Allogenic stem cell transplantation is the only curative therapeutic option today. Long-term survival rates are between 35% and 65% in cases of first complete remissions, but survival rates quickly decline in cases of second and third remissions. [4][5][6][7][8][9] Additionally, the relapse rate for Ph ϩ ALL remains quite high even with transplantation. [4][5][6][7][8][9] Recently, targeted molecular therapy has been introduced to treat such refractory cases, and a BCR-ABL tyrosine kinase inhibitor, imatinib (Glivec, STI571), proved to be a useful agent for Ph ϩ ALL as well as CML. 10 However, in contrast to patients with CML, most of the patients with Ph ϩ ALL quickly become resistant to imatinib. 10,11 In a phase 2 clinical trial examining cases of relapsed or refractory Ph ϩ ALL, only 6% of patients maintained a complete response for more than 4 weeks, in contrast to an overall response rate of 29%. 12 Moreover, the median estimated time...

Vaecination with crude lipopolysaccharide (LPS) induced better proteetion against infection with Aeromonas hydrophila in carp than vaccination with formalin killed vaccine. Dipping fish in vaccine for 2h at 25X was more effective than intraperitoneal injection of the vaccine in procedural simphcity, lower stress loading and the degree of protection acquired. In carp immunized with crude LPS by the dip method, antibodies were not detected by bacterial agglutination, passive haemaggiutination and the agar diffusion tests. The results indicate that the proteetion against A. hydrophila infeetion in carp is not dependent on humoral immunity.

OBJECTIVE AND IMPORTANCE:A solitary fibrous tumor (SFT) is a rare neoplasm of probable mesenchymal origin that was first reported in the pleura but can occur in different sites. Only six cases of SFT arising from the spinal cord have been reported. CLINICAL PRESENTATION: We report a case of primary SFT occurring in the thoracic spinal cord in a 64-year-old man with Brown-Séquard syndrome. Magnetic resonance imaging revealed an intradural mass at the level of T2-T3. INTERVENTION: Total T2-T3 laminectomies were performed. The tumor appeared to be adherent to the right lateral aspect of the cord but not attached to the meninges. On histological examination, the tumor exhibited spindle cell proliferation with abundant dense collagen but without a hemangiopericytomatous pattern. Immunohistochemically, the tumor cells were reactive with CD34 and vimentin only. CONCLUSION: We report a rare case of SFT occurring in the thoracic spinal cord. Histologically and immunohistochemically, we confirmed the diagnosis of SFT. Low signal intensity on T1-and T2-weighted images corresponded to the histological findings. When a spinal cord tumor exhibits a signal pattern similar to this, SFT should be included in the differential diagnosis. Because of the rarity of reports on this condition, the clinical manifestations and course of SFT of the spinal cord are unknown, and careful long-term follow-up is recommended.