The efficacy of Herceptin therapies is influenced by the expression of other erbB receptors, their ligands and the activation of downstream signalling proteins

ErbB2 and EGFR are attractive oncology therapeutic targets as their overexpression in tumors predicts a poorer clinical outcome in a variety of epithelial malignancies. However, clinical results with therapeutic compounds targeting these receptors have been mixed. Therefore, there is a need for improved predictive biomarkers for these targeted therapeutics. In this study we analysed tissue microarrays of patients treated with combination chemotherapy and Herceptin for expression or phosphorylation of signalling proteins associated with erbB receptors to identify protein biomarkers that are predictive of breast cancer patient response. A comparison of expression or phosphorylation of these markers with patient outcome revealed that response to Herceptin depended not only on expression levels of erbB2 but also on expression of EGFR, expression of erbB ligands, expression of other receptors and phosphorylation of downstream proteins. Elucidating the biological effects of EGFR/erbB2 targeted therapeutics will enable patient tumor profiling to identify likely responders and the determination of biologically effective doses that allows chronic administration of these agents in order to maximise efficacy

Structural and Functional Insights into the Malaria Parasite Moving Junction Complex

Members of the phylum Apicomplexa, which include the malaria parasite Plasmodium, share many features in their invasion mechanism in spite of their diverse host cell specificities and life cycle characteristics. The formation of a moving junction (MJ) between the membranes of the invading apicomplexan parasite and the host cell is common to these intracellular pathogens. The MJ contains two key parasite components: the surface protein Apical Membrane Antigen 1 (AMA1) and its receptor, the Rhoptry Neck Protein (RON) complex, which is targeted to the host cell membrane during invasion. In particular, RON2, a transmembrane component of the RON complex, interacts directly with AMA1. Here, we report the crystal structure of AMA1 from Plasmodium falciparum in complex with a peptide derived from the extracellular region of PfRON2, highlighting clear specificities of the P. falciparum RON2-AMA1 interaction. The receptor-binding site of PfAMA1 comprises the hydrophobic groove and a region that becomes exposed by displacement of the flexible Domain II loop. Mutations of key contact residues of PfRON2 and PfAMA1 abrogate binding between the recombinant proteins. Although PfRON2 contacts some polymorphic residues, binding studies with PfAMA1 from different strains show that these have little effect on affinity. Moreover, we demonstrate that the PfRON2 peptide inhibits erythrocyte invasion by P. falciparum merozoites and that this strong inhibitory potency is not affected by AMA1 polymorphisms. In parallel, we have determined the crystal structure of PfAMA1 in complex with the invasion-inhibitory peptide R1 derived by phage display, revealing an unexpected structural mimicry of the PfRON2 peptide. These results identify the key residues governing the interactions between AMA1 and RON2 in P. falciparum and suggest novel approaches to antimalarial therapeutics

Dendritic cell vaccination and immune monitoring

We exploited dendritic cells (DC) to vaccinate melanoma patients. We recently demonstrated a statistical significant correlation between favorable clinical outcome and the presence of vaccine-related tumor antigen-specific T cells in delayed type hypersensitivity (DTH) skin biopsies. However, favorable clinical outcome is only observed in a minority of the treated patients. Therefore, it is obvious that current DC-based protocols need to be improved. For this reason, we study in small proof of principle trials the fate, interactions and effectiveness of the injected DC

Seasonal and inter-annual patterns in primary production, respiration, and net ecosystem metabolism in three estuaries in the Northeast Gulf of Mexico

Measurements of primary production and respiration provide fundamental information about the trophic status of aquatic ecosystems, yet such measurements are logistically difficult and expensive to sustain as part of long-term monitoring programs. However, ecosystem metabolism parameters can be inferred from high frequency water quality data collections using autonomous logging instruments. For this study, we analyzed such time series datasets from three Gulf of Mexico estuaries: Grand Bay, MS; Weeks Bay, AL; and Apalachicola Bay, FL. Data were acquired from NOAA's National Estuarine Research Reserve System Wide Monitoring Program and used to calculate gross primary production (GPP), ecosystem respiration (ER), and net ecosystem metabolism (NEM) using Odum's open water method. The three systems represent a diversity of estuaries typical of the Gulf of Mexico region, varying by as much as two orders of magnitude in key physical characteristics, such as estuarine area, watershed area, freshwater flow, and nutrient loading. In all three systems, GPP and ER displayed strong seasonality, peaking in summer and being lowest during winter. Peak rates of GPP and ER exceeded 200 mmol O₂ m⁻² day⁻¹ in all three estuaries. To our knowledge, this is the first study examining long-term trends in rates of GPP, ER, and NEM in estuaries. Variability in metabolism tended to be small among sites within each estuary. Nitrogen loading was highest in Weeks Bay, almost two times greater than that in Apalachicola Bay and 35 times greater than to Grand Bay. These differences in nitrogen loading were reflected in average annual GPP rates, which ranged from 825 g C m⁻² year⁻¹ in Weeks Bay to 401 g C m⁻² year⁻¹ for Apalachicola Bay and 377 g C m⁻² year⁻¹ in Grand Bay. Despite the strong inter-annual patterns in freshwater flow and salinity, variability in metabolic rates was low, perhaps reflecting shifts in the relative importance of benthic and phytoplankton productivity, during different flow regimes. The advantage of the open water method is that it uses readily available and cost-effective sonde monitoring technology to estimate these fundamental estuarine processes, thus providing a potential means for examining long-term trends in net carbon balance. It also provides a historical benchmark for comparison to ongoing and future monitoring focused on documenting the effect of human activities on the coastal zone.final article publishedJournal Articl