Hasil untuk "nucl-ex"

M. Grossman, Daniel J. Rader, David W.M. Muller et al.

V. Shirvani, Rodica Ouatu-Lascar, Baljeet S. Kaur et al.

R. Dahlstrom, A. Nygaard

E. Shpall, R. Quinones, R. Giller et al.

Umbilical cord blood (CB) from unrelated donors is increasingly used to restore hematopoiesis after myeloablative therapy. CB transplants are associated with higher rates of delayed and failed engraftment than are bone marrow transplants, particularly for adult patients. We studied the ex vivo expansion of CB in an attempt to improve time to engraftment and reduce the graft failure rate in the recipients. In this feasibility study, 37 patients (25 adults, 12 children) with hematologic malignancies (n = 34) or breast cancer (n = 3) received high-dose therapy followed by unrelated allogeneic CB transplantation. A fraction of each patient's CB allograft was CD34-selected and cultured ex vivo for 10 days prior to transplantation in defined media with stem cell factor, granulocyte colony-stimulating factor, and megakaryocyte growth and differentiation factor. The remainder of the CB graft was infused without further manipulation. Two sequential cohorts of patients were accrued to the study. The first cohort had 40% and the second cohort had 60% of their CB graft expanded. Patients received a median of 0.99 x 10(7) total nucleated cells (expanded plus unexpanded) per kilogram. The median time to engraftment of neutrophils was 28 days (range, 15-49 days) and of platelets was 106 days (range, 38-345 days). All evaluable patients who were followed for 28 days or longer achieved engraftment of neutrophils. Grade III/IV acute GVHD was documented in 40% and extensive chronic GVHD in 63% of patients. At a median follow-up of 30 months, 13 (35%) of 37 of patients survived. This study demonstrates that the CD34 selection and ex vivo expansion of CB prior to transplantation of CB is feasible. Additional accrual will be required to assess the clinical efficacy of expanded CB progenitors.

V. Govindarajan, P. Kopalle

Keren Levanon, V. Ng, H. Piao et al.

Recent studies suggest that some serous ovarian carcinomas (SOCs) arise from the fallopian tube (FT) epithelium rather than the ovarian surface epithelium. This hypothesis places emphasis on the FT secretory epithelial cell as a cell-of-origin. Herein, we report the development of a novel ex vivo primary human FT epithelium culture system that faithfully recapitulates the in vivo epithelium, as shown by morphological, ultrastructural and immunophenotypic analyses. Mass spectrometry-based proteomics reveal that these cultures secrete proteins previously identified as biomarkers for ovarian cancer. We also use this culture system to study the response of the FT epithelium to genotoxic stress and find that the secretory cells exhibit a distinct response to DNA damage when compared with neighboring ciliated cells. The secretory cells show a limited ability to resolve the damage over time, potentially leaving them more susceptible to accumulation of additional mutagenic injury. This divergent response is confirmed with in situ studies using tissue samples, further supporting the use of this ex vivo culture system to investigate FT epithelial pathobiology. We anticipate that this novel culture system will facilitate the study of SOC pathogenesis, and propose that similar culture systems could be developed for other organ site-specific epithelia.

A. Veerapathran, J. Pidala, F. Beato et al.

O. N. Chuang-Smith, C. Wells, M. Henry-Stanley et al.

Infectious endocarditis involves formation of a microbial biofilm in vivo. Enterococcus faecalis Aggregation Substance (Asc10) protein enhances the severity of experimental endocarditis, where it has been implicated in formation of large vegetations and in microbial persistence during infection. In the current study, we developed an ex vivo porcine heart valve adherence model to study the initial interactions between Asc10+ and Asc10− E. faecalis and valve tissue, and to examine formation of E. faecalis biofilms on a relevant tissue surface. Scanning electron microscopy of the infected valve tissue provided evidence for biofilm formation, including growing masses of bacterial cells and the increasing presence of exopolymeric matrix over time; accumulation of adherent biofilm populations on the cardiac valve surfaces during the first 2–4 h of incubation was over 10-fold higher than was observed on abiotic membranes incubated in the same culture medium. Asc10 expression accelerated biofilm formation via aggregation between E. faecalis cells; the results also suggested that in vivo adherence to host tissue and biofilm development by E. faecalis can proceed by Asc10-dependent or Asc10-independent pathways. Mutations in either of two Asc10 subdomains previously implicated in endocarditis virulence reduced levels of adherent bacterial populations in the ex vivo system. Interference with the molecular interactions involved in adherence and initiation of biofilm development in vivo with specific inhibitory compounds could lead to more effective treatment of infectious endocarditis.

Maxine Burkett

N. Katabi, D. Gomez, D. Klimstra et al.

F. Filippi, A. Nuzzolo, A. Comi et al.

Many city public authorities have implemented measures to alleviate the negative effects of freight transport in urban areas, but these have often proved ineffective. The literature contains studies related to ex-post assessment of urban freight transport policies. This paper proposes a methodology for ex-ante assessment of their effects. The focus is the assessment of pollutant emissions. The application of the methodology to the inner urban area of Rome shows that an urban distribution centre can be more effective in reducing environmental externalities than policies based on vehicle fleet renewal.

M. E. Dulloo, D. Hunter, T. Borelli

The effective conservation and use of agricultural biodiversity is vital for creating and maintaining sustainable increases in the productivity of healthy food for mankind, as well as contributing to the increased resilience of agricultural systems. Major advances in the two main complementary strategies for agricultural biodiversity conservation, namely ex situ and in situ, over the last decade are presented to reflect on their current global status and trends. The FAO Second State of the World Report on Plant Genetic Resources for Food and Agriculture reports that the total number of accessions conserved in ex situ collections is about 7.4 million, in over 1750 genebanks around the world. There has also been increasing awareness of the importance and value of conserving crop wild relatives (CWR) in situ and a greater understanding of the scientific issues surrounding on farm management of genetic diversity. Recent research outputs produced by Bioversity International to ensure the effective and efficient conservation and use of genetic diversity are cited. These have involved development of best practices for genebank management and the development of enhanced technologies and methodologies for conserving and promoting the use of the genetic diversity. Bioversity International has led the development of methodologies for on farm conservation, and promoted the drafting of policies and strategies for the in situ conservation of crop wild relatives and their management inside and outside protected areas. Also an outlook of the research priorities and needs for conservation and use of agricultural biodiversity is described.

H. Yalcin, Akshay Shekhar, Ajinkya A. Rane et al.

Understanding the relationships between genetic and microenvironmental factors that drive normal and malformed embryonic development is fundamental for discovering new therapeutic strategies. Advancements in imaging technology have enabled quantitative investigation of the organization and maturing of the body plan, but later stage embryonic morphogenesis is less clear. Chicken embryos are an attractive vertebrate animal model system for this application because of its ease of culture and surgical manipulation. Early embryos can be cultured for a short time on filter paper rings, which enables complete optical access for cell patterning and fate studies. Studying advanced developmental processes such as cardiac morphogenesis are traditionally performed through a window of the eggshell, but this technique limits optical access due to window size. We previously developed a simple method to culture whole embryos ex-ovo on hexagonal weigh boats for up to 10 days, which enabled high resolution imaging via ultrasonography. These cultures were difficult to transport, limiting the types of imaging tools available for live experiments. We here present an improved shell-less culture system with a cost-effective, portable environmental chamber. Eggs were cracked onto a hammock created by a polyurethane membrane (cling wrap) affixed circumferentially to a plastic cup partially filled with sterile water. The dimensions of the circumference and depth of the hammock were both critical to maintain surface tension, while the mechanics of the hammock and water beneath helped dampen vibrations induced by transportation. A small footprint circulating water bath was also developed to enable continuous temperature control during experimentation. We demonstrate the ability to culture embryos in this way for at least 14 days without morphogenic defect or delay and employ this system in several microsurgical and imaging applications.

C. Gollier, Alexander Muermann

K. Liechty

M. Bernoux, G. Branca, A. Carro et al.

EX-ACT (EX-Ante Carbon-balance Tool) is a tool developed by the Food and Agriculture Organization of the United Nations (FAO). It provides ex-ante measurements of the mitigation impact of agriculture and forestry development projects, estimating net C balance from GHG emissions and Carbon (C) sequestration. EX-ACT is a land-based accounting system, measuring C stocks, stock changes per unit of land, and CH 4 and N 2 O emissions expressed in t CO 2 -eq per hectare and year. The main output of the tool is an estimation of the C-balance associated with the adoption of improved land management options, as compared with a "business as usual" scenario. EX-ACT has been developed using primarily the IPCC 2006 Guidelines for National Greenhouse Gas Inventories, complemented by other existing methodologies and reviews of default coefficients. Default values for mitigation options in the agriculture sector are mostly from the 4 th Assessment Report of IPCC (2007). Thus, EX-ACT allows for the carbon-balance appraisal of new investment programmes by ensuring an appropriate method available for donors and planning officers, project designers, and decision makers within agriculture and forestry sectors in developing countries. The tool can also help to identify the mitigation impacts of various investment project options, and thus provide an additional criterion for consideration in project selection.

A. Majewicz, T. Wedlick, K. B. Reed et al.

Yolande Hiriart, D. Martimort, J. Pouyet

S. Tung, S. Parmar, S. Robinson et al.

M. Sperandio