Using the BLT Humanized Mouse as a Stem Cell based Gene Therapy Tumor Model

Small animal models such as mice have been extensively used to study human disease and to develop new therapeutic interventions. Despite the wealth of information gained from these studies, the unique characteristics of mouse immunity as well as the species specificity of viral diseases such as human immunodeficiency virus (HIV) infection led to the development of humanized mouse models. The earlier models involved the use of C. B 17 scid/scid mice and the transplantation of human fetal thymus and fetal liver termed thy/liv (SCID-hu) ^{1, 2} or the adoptive transfer of human peripheral blood leukocytes (SCID-huPBL) ³. Both models were mainly utilized for the study of HIV infection.One of the main limitations of both of these models was the lack of stable reconstitution of human immune cells in the periphery to make them a more physiologically relevant model to study HIV disease. To this end, the BLT humanized mouse model was developed. BLT stands for bone marrow/liver/thymus. In this model, 6 to 8 week old NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) immunocompromised mice receive the thy/liv implant as in the SCID-hu mouse model only to be followed by a second human hematopoietic stem cell transplant ⁴. The advantage of this system is the full reconstitution of the human immune system in the periphery. This model has been used to study HIV infection and latency ^5-8.We have generated a modified version of this model in which we use genetically modified human hematopoietic stem cells (hHSC) to construct the thy/liv implant followed by injection of transduced autologous hHSC ^{7, 9}. This approach results in the generation of genetically modified lineages. More importantly, we adapted this system to examine the potential of generating functional cytotoxic T cells (CTL) expressing a melanoma specific T cell receptor. Using this model we were able to assess the functionality of our transgenic CTL utilizing live positron emission tomography (PET) imaging to determine tumor regression (9).The goal of this protocol is to describe the process of generating these transgenic mice and assessing in vivo efficacy using live PET imaging. As a note, since we use human tissues and lentiviral vectors, our facilities conform to CDC NIH guidelines for Biosafety Level 2 (BSL2) with special precautions (BSL2+). In addition, the NSG mice are severely immunocompromised thus, their housing and maintenance must conform to the highest health standards (http://jaxmice.jax.org/research/immunology/005557-housing.html).     

Absolute quantification of somatic DNA alterations in human cancer

We describe a computational method that infers tumor purity and malignant cell ploidy directly from analysis of somatic DNA alterations. The method, named ABSOLUTE, can detect subclonal heterogeneity and somatic homozygosity, and it can calculate statistical sensitivity for detection of specific aberrations. We used ABSOLUTE to analyze exome sequencing data from 214 ovarian carcinoma tumor-normal pairs. This analysis identified both pervasive subclonal somatic point-mutations and a small subset of predominantly clonal and homozygous mutations, which were overrepresented in the tumor suppressor genes TP53 and NF1 and in a candidate tumor suppressor gene CDK12. We also used ABSOLUTE to infer absolute allelic copy-number profiles from 3,155 diverse cancer specimens, revealing that genome-doubling events are common in human cancer, likely occur in cells that are already aneuploid, and influence pathways of tumor progression (for example, with recessive inactivation of NF1 being less common after genome doubling). ABSOLUTE will facilitate the design of clinical sequencing studies and studies of cancer genome evolution and intra-tumor heterogeneity.     

棉叶中棉酚的快速提取与测定方法研究

棉酚(G)及其相关物甲氧基半棉酚(DHG)、半棉酚酮(HGQ)、半棉酚(HG)、杀实夜蛾素(H1-4)等是棉花中重要的抗虫性萜烯类次生物质.利用高效液相色谱法(HPLC)对棉酚及其相关物进行了分离,测定了棉叶中的棉酚含量,讨论了不同的提取方法和测定条件对结果的影响,给出了一套简便、快速的分析测试方法,同时与紫外-可见分光光度法(苯胺法)的结果进行了对比,认为对于棉花的抗虫性的研究来说,HPLC是比较适宜的方法.     

Self‐Reported Body Mass Index and Health‐Related Quality of Life: Findings from the Behavioral Risk Factor Surveillance System

Objective: To examine the relationship between self‐reported body mass index (BMI) and health‐related quality of life in the general adult population in the United States. Research Methods and Procedures: Using data from 109,076 respondents in the 1996 Behavioral Risk Factor Surveillance System, we examined how self‐reported BMI is associated with five health‐related quality of life measures developed by the Centers for Disease Control and Prevention for population health surveillance. Results: After adjusting for age, gender, race or ethnicity, educational attainment, employment status, smoking status, and physical activity status, participants with a self‐reported BMI of <18.5 kg/m² and participants with a self‐reported BMI of ≥30 kg/m² reported impaired quality of life. Compared with persons with a self‐reported BMI of 18.5 to <25 kg/m², odds ratios (ORs) of poor or fair self‐rated health increased among persons with self‐reported BMIs of <18.5 (1.57, 95% confidence interval [CI]: 1.31 to 1.89), 25 to <30 kg/m² (1.12, 95% CI: 1.04 to 1.20), 30 to <35 kg/m² (1.65, 95% CI: 1.50 to 1.81), 35 to <40 kg/m² (2.58, 95% CI: 2.21 to 3.00), and ≥40 kg/m² (3.23, 95% CI: 2.63 to 3.95); ORs for reporting ≥14 days of poor physical health during the previous 30 days were 1.44 (95% CI: 1.21 to 1.72), 1.04 (95% CI: 0.96 to 1.14), 1.32 (95% CI: 1.19 to 1.47), 1.80 (95% CI: 1.52 to 2.13), and 2.37 (95% CI: 1.90 to 2.94), respectively; ORs for having ≥14 days of poor mental health during the previous 30 days were 1.18 (95% CI: 0.97 to 1.42), 1.02 (95% CI: 0.95 to 1.11), 1.22 (95% CI: 1.10 to 1.36), 1.68 (95% CI: 1.42 to 1.98), and 1.66 (95% CI: 1.32 to 2.09), respectively. Discussion: In the largest study to date, low and increased self‐reported BMI significantly impaired health‐related quality of life. Particularly, deviations from normal BMI affected physical functioning more strongly than mental functioning.     

Effect of latent human immunodeficiency virus infection on cell surface phenotype.

Highly active antiretroviral therapy has succeeded in many cases in suppressing virus production in patients infected with human immunodeficiency virus (HIV); however, once treatment is discontinued, virus replication is rekindled. One reservoir capable of harboring HIV in a latent state and igniting renewed infection once therapy is terminated is a resting T cell. Due to the sparsity of T cells latently infected with HIV in vivo, it has been difficult to study viral and cellular interactions during latency. The SCID-hu (Thy/Liv) mouse model of HIV latency, however, provides high percentages of latently infected cells, allowing a detailed analysis of phenotype. Herein we show that latently infected cells appear phenotypically normal. Following cellular stimulation, the virus completes its life cycle and induces phenotypic changes, such as CD4 and major histocompatibility complex class I down-regulation, in the infected cell. In addition, HIV expression following activation did not correlate with expression of the cellular activation marker CD25. The apparently normal phenotype and lack of HIV expression in latently infected cells could prevent recognition by the immune response and contribute to the long-lived nature of this reservoir.     

Spatial and Temporal Considerations in Restoring Habitat for Wildlife

An accumulated body of theory and empirical evidence suggests that habitat selection by animals is a scale‐dependent, hierarchical process. Hierarchy theory predicts that habitat suitability is influenced by the interaction of factors at multiple spatial scales from the microsite to the landscape and that higher‐order factors impose constraints at lower levels. For instance, large‐scale factors such as landscape context may make a site unsuitable for a species even if the vegetation structure and composition are appropriate. In addition, the spatial arrangement of habitat elements at all scales must be considered when planning restoration efforts. For example, the presence of snags does not ensure that the site will be suitable for snag‐dependent species. The size, age, and spacing of snags and their juxtaposition to other habitat elements must also be considered. Finally, all habitats are dynamic, and therefore the ecological processes that contribute to those dynamics must be maintained or suitable substitutes included in the recovery plan. When considering restoring habitat for wildlife, we recommend that managers: (1) identify the wildlife species they want to target for restoration efforts, (2) consider the size and landscape context of the restoration site and whether it is appropriate for the target species, (3) identify the habitat elements that are necessary for the target species, (4) develop a strategy for restoring those elements and the ecological processes that maintain them, and (5) implement a long‐term monitoring program to gauge the success of the restoration efforts.     

A Geospatial Comparison of Distributed Solar Heat and Power in Europe and the US

The global trends for the rapid growth of distributed solar heat and power in the last decade will likely continue as the levelized cost of production for these technologies continues to decline. To be able to compare the economic potential of solar technologies one must first quantify the types and amount of solar resource that each technology can utilize; second, estimate the technological performance potential based on that resource; and third, compare the costs of each technology across regions. In this analysis, we have performed the first two steps in this process. We use physical and empirically validated models of a total of 8 representative solar system types: non-tracking photovoltaics, 2d-tracking photovoltaics, high concentration photovoltaics, flat-plate thermal, evacuated tube thermal, concentrating trough thermal, concentrating solar combined heat and power, and hybrid concentrating photovoltaic/thermal. These models are integrated into a simulation that uses typical meteorological year weather data to create a yearly time series of heat and electricity production for each system over 12,846 locations in Europe and 1,020 locations in the United States. Through this simulation, systems composed of various permutations of collector-types and technologies can be compared geospatially and temporally in terms of their typical production in each location. For example, we see that silicon solar cells show a significant advantage in yearly electricity production over thin-film cells in the colder climatic regions, but that advantage is lessened in regions that have high average irradiance. In general, the results lead to the conclusion that comparing solar technologies across technology classes simply on cost per peak watt, as is usually done, misses these often significant regional differences in annual performance. These results have implications for both solar power development and energy systems modeling of future pathways of the electricity system.     

Costimulation of Naive CD8+ Lymphocytes Induces CD4 Expression and Allows Human Immunodeficiency Virus Type 1 Infection

Human immunodeficiency virus type 1 (HIV-1) infection requires cell surface expression of CD4. Costimulation of CD8⁺/CD4⁻ T lymphocytes by anti-CD3 and anti-CD28 antibodies or by allogeneic dendritic cells induced expression of CD4 and rendered these CD8 cells susceptible to HIV-1 infection. Naive CD45RA⁺ cells responded with greater expression of CD4 than did CD45RO⁺ cells. CD8⁺ lymphocytes derived from fetal or newborn sources exhibited a greater tendency to express CD4, consistent with their naive states. This mechanism of infection suggests HIV-induced perturbation of the CD8 arm of the immune response and could explain the generally rapid disease progression seen in HIV-infected children.     

A biosystematic study of the Interiores species group of the genus Vernonia (Compositae)

The taxa of the Interiores species group are treated as two species, one of which is divided into two subspecies.Vernonia missurica, a widespread taxon, has large flower heads (32-58 flowers per head), obtuse involucral bract tips, and a characteristic flavonoid Chromatographic profile.Vernonia baldwinii ssp.baldwinii, occurring throughout the Ozarks and Ouachitas, andV. baldwinii ssp.interior of the Central Plains possess smaller flower heads (17-34 flowers per head), acute or acuminate involucral bracts, and a different Chromatographic profile.Vernonia baldwinii ssp.baldwinii is distinguished from ssp.interior by the recurved involucral bract tips and by the possession of more flowers per head. Morphological data reveal that an intergradation zone exists between the population systems of the two subspecies along the edges of the Ozarks and Ouachitas. Although much gene exchange occurs between these two taxa, they maintain a level of genetic integrity that permits their ready recognition throughout wide ranges. All three taxa have a gametic chromosome number ofn = 17 and hybridize extensively with sympatric species.