A Role for the Retinoblastoma Protein As a Regulator of Mouse Osteoblast Cell Adhesion: Implications for Osteogenesis and Osteosarcoma Formation

The retinoblastoma protein (pRb) is a cell cycle regulator inactivated in most human cancers. Loss of pRb function results from mutations in the gene coding for pRb or for any of its upstream regulators. Although pRb is predominantly known as a cell cycle repressor, our data point to additional pRb functions in cell adhesion. Our data show that pRb regulates the expression of a wide repertoire of cell adhesion genes and regulates the assembly of the adherens junctions required for cell adhesion. We conducted our studies in osteoblasts, which depend on both pRb and on cell-to-cell contacts for their differentiation and function. We generated knockout mice in which the RB gene was excised specifically in osteoblasts using the cre-lox P system and found that osteoblasts from pRb knockout mice did not assemble adherens junction at their membranes. pRb depletion in wild type osteoblasts using RNAi also disrupted adherens junctions. Microarrays comparing pRb-expressing and pRb-deficient osteoblasts showed that pRb controls the expression of a number of cell adhesion genes, including cadherins. Furthermore, pRb knockout mice showed bone abnormalities consistent with osteoblast adhesion defects. We also found that pRb controls the function of merlin, a well-known regulator of adherens junction assembly, by repressing Rac1 and its effector Pak1. Using qRT-PCR, immunoblots, co-immunoprecipitation assays, and immunofluorescent labeling, we observed that pRb loss resulted in Rac1 and Pak1 overexpression concomitant with merlin inactivation by Pak1, merlin detachment from the membrane, and adherens junction loss. Our data support a pRb function in cell adhesion while elucidating the mechanism for this function. Our work suggests that in some tumor types pRb inactivation results in both a loss of cell cycle control that promotes initial tumor growth as well as in a loss of cell-to-cell contacts, which contributes to later stages of metastasis

A922 Sequential measurement of 1 hour creatinine clearance (1-CRCL) in critically ill patients at risk of acute kidney injury (AKI)

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Identification of Potential Sources of Airborne Olea Pollen in the Southwest Iberian Peninsula

This study aims to determine the potential origin of Olea pollen recorded in Badajoz in the Southwest of the Iberian Peninsula during 2009–2011. This was achieved using a combination of daily average and diurnal (hourly) airborne Olea pollen counts recorded at Badajoz (south-western Spain) and Évora (south-eastern Portugal), an inventory of olive groves in the studied area and air mass trajectory calculations computed using the HYSPLIT model. Examining olive pollen episodes at Badajoz that had distinctly different diurnal cycles in olive pollen in relation to the mean, allowed us to identify three different scenarios where olive pollen can be transported to the city from either distant or nearby sources during conditions with slow air mass movements. Back trajectory analysis showed that olive pollen can be transported to Badajoz from the West on prevailing winds, either directly or on slow moving air masses, and from high densities of olive groves situated to the Southeast (e.g. Andalucía). Regional scale transport of olive pollen can result in increased nighttime concentrations of this important aeroallergen. This could be particularly important in Mediterranean countries where people can be outdoors during this time due to climate and lifestyle. Such studies that examine sources and the atmospheric transport of pollen are valuable for allergy sufferers and health care professionals because the information can be incorporated into forecasts, the outputs of which are used for avoiding exposure to aeroallergens and planning medication. The results of studies of this nature can also be used for examining gene flow in this important agricultural crop