The goal of this project is to evaluate the effect of a moderate- level brain injury on risk for opioid abuse using preclinical models of abuse- related behaviors in rats. Thus far we have assessed the effect of brain injury on the rewarding effects of oxycodone in two rat self-administration procedures. We have found that there are significant differences in the acquisition and maintenance of oxycodone intravenous self-administration behavior between brain-injured and control rats. Data collected to date suggest brain injured rats have a greater sensitivity to the rewarding effects of oxycodone and a greater tolerance for the use-limiting effects of oxycodone (eg. sedation, motor impairment, dysphoria). Conversely, it appears that there is no difference between brain-injured and sham controls in a model of relapse to oxycodone self-administration. Preliminary testing of oxycodone for analgesic strength and development of tolerance also has shown no difference between sham controls and brain injured subjects. Additional self- administration studies to determine the rewarding strength of oxycodone as well as development of physical dependence are ongoing.

关键词：药物滥用；创伤性脑损伤；镇痛药

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Adoptive cell therapy (ACT) of cancer with ex vivo activated/expanded T-cells is one of the promising treatments currently being tested in patients. One challenge of the approach is that the transferred T cells become functionally anergic in the tumor environment, limiting their anti- tumor effect. We have investigated whether tumor-mediated immune suppression can be overcome by arming tumor-specific T cells with cytokine/immunostimulator- loaded nanoparticles carried by each cell. Specially, we have defined the role of CD70 and CD80/CD86 in dendritic cell-mediated activation of tumor tolerized CD8 T cells, discovered the effect of CD8 T cell responses in selecting for antigen-negative tumor cells, and develop a better prostate model for monitoring T cell responses to prostate cancer in mice. Findings from our studies identify molecular interactions that are important for maintaining T cell function in the tumor environment, suggesting possible interventions to enhance T cell functionality during ACT. Our results that CD8 T cells are effective in eliminating antigen-bearing prostate tumor cells but they also can select for the outgrowth of antigen-negative tumor cells provide insights into the requirements for an effective cancer immunotherapy: not only inducing potent immune responses but also avoiding selection and outgrowth of antigen- negative tumor cells.

关键词：免疫治疗；前列腺癌；T淋巴细胞

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The purpose of this project is to characterize the breast cancer in women affected with Neurofibromatosis type 1 (NF1) in a multi-institutional setting. The first aim is to assess the incidence of breast cancer in this cohort and the clinical features of NF1 associated with breast cancer. The second aim is to investigate any characteristic NF1 gene germline mutations in women with breast cancer and to carry out whole exome sequencing to search for other germline mutations. For the third aim, at tumor tissue level, we plan to characterize the selected signaling pathway on archived breast cancer tissue from women with NF1 utilizing immunohistochemistry (IHC) methods. At the somatic level, we plan to study loss of heterozygosity (LOH) of the NF1 gene using ion semiconductor sequencing and MLPA copy number analysis. We also plan to sequence 30 other breast cancer genes on the breast cancer specimens. By the end of May, 2013, a total of 242 cases of NF1 women have been reviewed. Eleven women have had a diagnosis of breast cancer. Statistical analysis was conducted. The presence of plexiform neurofibroma has a trend to be inversely related to the occurrence of breast cancer, however, it has not reached statistical significance (p=0.083). A family history of cancer is associated with a personal history of breast cancer (p=0.000119). No other clinical features or family histories were found to be associated with the occurrence of breast cancer. To date, germline NF1 mutations has been investigated in 10 women with breast cancer. No significant pattern of mutation has been discovered. Nine breast cancer specimens have been collected and are ready to be analyzed. We are expected to review up to 450 cases of women with NF1 and collect another 5 archived breast cancer specimens. The analyses listed in the above aims are to be completed once the final cases have been reviewed and the specimens have been collected.

关键词：乳腺癌；临床医学；基因

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Numbers dendritic cells (DCs) increased 100-fold in pancreata of mice with acute pancreatitis to account for nearly 15% of intrapancreatic leukocytes. Intrapancreatic DCs acquired a distinct immune phenotype in mice with acute pancreatitis; they expressed higher levels of major histocompatibility complex II and CD86 and increased production of interleukin- 6, MCP 1, and TNFa. DC over-expansion exacerbated disease. However, DCs were also required for pancreatic viability; the exocrine pancreas died in mice that were depleted of DCs and challenged with caerulein or L-arginine. All mice with pancreatitis that were depleted of DCs died from acinar cell death within 4 days. These data suggest that DC have simultaneous paradoxical pro-inflammatory and protective effects in pancreatitis. Further investigations are warranted.

关键词：树突状结构；胰腺炎；细胞（生物学）

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To realize three-dimensional optical imaging using transillumination images, we developed a novel technique to suppress the scattering effect in a transillumination image and reconstruct a three-dimensional image of the internal light-absorbing structure of animal body from two-dimensional images. The problem of the depth-dependence of PSF was solved by the calculation of the projection image in the filtered back-projection method. The feasibility and the effectiveness of the proposed technique were verified in the experiments with tissue-equivalent phantom and a living mouse. Using the proposed technique, the high-absorption organs such as kidneys and the bottom parts of the liver were reconstructed in a 3D image of mouse abdomen.

关键词：透照成像；散射抑制；反褶积

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Prostate cancer is the second leading cause of cancer death among American men. Although radiotherapy (RT) is one of the two curative treatments for prostate cancer patients, approximately 10% of low-risk cancer patients and 30-60% of high-risk prostate cancer patients experience biochemical recurrence within five years, among them 20% die in 10 years. The proposed research is based on the hypothesis that targeting protein arginine methyltransferase 5 (PRMT5) can sensitize primary and recurrent prostate cancer cells to RT. During the first grant period, we have successfully demonstrated that knockdown of PRMT5 or inhibition of PRMT5 by a specific inhibitor can sensitize prostate cancer cells (LNCaP, DU-145 and PC-3) to radiation in vitro. This radiosensitization is likely due to the involvement of PRMT5 in the regulation of the DNA damage response. These results collectively suggest that targeting PRMT5 can sensitize prostate cancer cells to radiation. We are currently isolating stably integrated clones to inducibly knock down PRMT5 for proposed in vivo experiments. We have also successfully isolated 3 radiation-resistant sublines from DU-145 after 40 Gy of fractionated ionizing radiation. These resistant cell sublines along with previously isolated LNCaP radiation- resistant sublines will be used for PRMT5 targeting experiments. In addition, we found that PRMT5 regulates prostate cancer cell growth in an AR-dependent manner, and this effect is likely medicated by epigenetic regulation of AR transcription. We will continue to pursue this novel and exciting finding.

关键词：电离辐射；前列腺癌；放射疗法

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This study will recruit wounded warriors with severe extremity trauma, which places them at high risk for heterotopic ossification (HO); bone formation at abnormal sites, which causes pain, limits motion and/or limits the use of a prosthetic device. There are three goals: (1) to understand the mechanisms involved in HO; (2) to define accurate and practical methods to predict where HO will develop; and (3) to define potential therapies for prevention or mitigation of HO.

关键词：骨；创伤和损伤；精度；诊断

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The purpose of the report is to determine in a mouse model of high and low fear whether a protein expressed in the lateral amygdala is more abundance in the high fear mice following Pavlovian fear conditioning. These data provide important insights into the micro network mechanisms in the barin that may lead to different levels of fear response in individuals. Understanding the circuit level cellular and molecular mechanisms that underlie individual variability in far learning is critical for the development of effective treatment of fear-related illnesses such as PTSD. These data suggest that inhibiting a specific protein in a precise brain region in mice may reduce high fear memory to the level of low fear memory. This information may begin to provide foundations for the understanding and eventual treatment of pathological fear. Using a mouse model of very high and very low fear memory to understand cellular mechanisms in specific brain regions involved in forming fear memories may help identify novel ways to predict individuals at risk for fear related illness. This can potentially lead to targeted treatments for fear-related disorders such as PTSD.

关键词：恐惧；内存；老鼠；脑细胞（生物学）

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The aim of our research is to find novel targeted therapy for NF2 patients to provide a new therapeutic option for meningiomas that includes the standard of care radiotherapy. We have successfully selected four small molecule compounds that preferentially inhibit NF2 mutant meningioma cells in vitro. These are FDA approved drugs that are safe for human use. We validated these drugs by using human cell lines and tested their ability to synergize with radiation. We have optimized the use of a CT-guided conformal radiation that can be tested in a meningioma mouse model. The findings from this research project will be valuable for future clinical trials, for testing combinatorial approaches to treat meningioma patients.

关键词：脑膜；治疗；细胞（生物学）；临床试验

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Recombinant human lubricin (rhPRG4) was expressed by transfected CHO- S cells at a level of secretion that may be scalable and thus enable production of a GMP protein for clinical use. A purification bioprocess was created that achieved a purity level of approximately 95%. A total of 5 bands visualized on SDS-PAGE are all product related following excision and analysis by LC-MS. Full- length and truncated rhPRG4 constructs are being tested for chondroprotective ability. The full-length construct significantly lowers friction between pressurized discs of articular cartilage. The coefficient of friction in the presence of rhPRG4 was 0.03 whereas for a saline control 0.07. Study of the 5 other constructs and confirmation of reduction in levels of chondrocyte apoptosis attributable to lower friction is ongoing. The posttranslational glycosylations on rhPRG4 appear to be (1-3)GalNac-Gal (2,3) NeuAc judging by sequential enzymatic deglycosylation and molecular weight shift on SDS-PAGE. Western blotting of rhPRG4 with mAb 9G3, which reacts with the glycosylations in the lubricin mucin domain, also confirms that the glycosylations are consistent with lubricin secreted by human synovial fibroblasts. Analytical ultracentrifugation shows that there are 5 distinct species in rhPRG4, 3 of which are major forms: monomer, dimer and a higher order tetramer. This biophysical form of analysis may also prove useful in a quality assurance program in GMP protein production as the relative amounts of each major form can be monitored.

关键词：关节炎；软骨；细胞凋亡

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