A novel tumor suppressor gene, PTEN/MMAC1, has been recently shown to be mutated in gliomas, breast, prostate, kidney cancers and melanomas. Loss-ofheterozygosity studies in melanoma have suggested the presence of at least one chromosome 10q locus lost early in tumor progression. In this study, we screened 45 melanoma cell lines and 17 paired uncultured metastatic melanoma and peripheral blood specimens for PTEN/ MMAC1 alterations using PCR-SSCP and direct sequencing. We found nine melanoma cell lines with homozygous deletions (®ve with intragenic loss) and four cell lines with mutations (one nonsense and one frameshift; two intronic); from among our uncultured melanoma specimens, we found one tumor with a somatic 17 bp duplication in exon 7 leading to a premature stop codon and one tumor with a possible homozygous deletion. Furthermore, we have identi®ed a novel intragenic polymorphism within intron 4 of PTEN/ MMAC1. Taken together, these data suggest that PTEN/MMAC1 may be a chromosome 10q tumor suppressor important in melanoma tumor formation or progression.

Summary 1.Widely observed macro-ecological patterns in log abundance vs. log body mass of organisms can be explained by simple scaling theory based on food (energy) availability across a spectrum of body sizes. The theory predicts that when food availability falls with body size (as in most aquatic food webs where larger predators eat smaller prey), the scaling between log N vs. log m is steeper than when organisms of different sizes compete for a shared unstructured resource (e.g. autotrophs, herbivores and detritivores; hereafter dubbed 'detritivores'). 2. In real communities, the mix of feeding characteristics gives rise to complex food webs. Such complexities make empirical tests of scaling predictions prone to error if: (i) the data are not disaggregated in accordance with the assumptions of the theory being tested, or (ii) the theory does not account for all of the trophic interactions within and across the communities sampled. 3. We disaggregated whole community data collected in the North Sea into predator and detritivore components and report slopes of log abundance vs. log body mass relationships. Observed slopes for fish and epifaunal predator communities (-1·2 to -2·25) were significantly steeper than those for infaunal detritivore communities (-0·56 to -0·87). 4. We present a model describing the dynamics of coupled size spectra, to explain how coupling of predator and detritivore communities affects the scaling of log N vs. log m . The model captures the trophic interactions and recycling of material that occur in many aquatic ecosystems. 5. Our simulations demonstrate that the biological processes underlying growth and mortality in the two distinct size spectra lead to patterns consistent with data. Slopes of log N vs. log m were steeper and growth rates faster for predators compared to detritivores. Size spectra were truncated when primary production was too low for predators and when detritivores experienced predation pressure. 6. The approach also allows us to assess the effects of external sources of mortality (e.g. harvesting).Removal of large predators resulted in steeper predator spectra and increases in their prey (small fish and detritivores). The model predictions are remarkably consistent with observed patterns of exploited ecosystems.

A new time-dependent continuous model of biomass size spectra is developed. In this model, predation is the single process governing the energy flow in the ecosystem, as it causes both growth and mortality. The ratio of predator to prey is assumed to be distributed: predators may feed on a range of prey sizes. Under these assumptions, it is shown that linear size spectra are stationary solutions of the model. Exploited fish communities are simulated by adding fishing mortality to the model: it is found that realistic fishing should affect the curvature and stability of the size spectrum rather than its slope.

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Accumulating evidence suggests an important role for cyclooxygenase-2 (COX-2) in the pathogenesis of a wide range of malignancies. Here we tested the hypothesis that the COX-2 product prostaglandin E 2 (PGE 2 ) increases cellular invasive potential by inducing matrix metalloproteinase-2 (MMP-2) expression and activity through an extracellular signalregulated kinase (ERK)/Ets-1-dependent mechanism in pancreatic cancer. PANC-1 and MIAPaCa-2 pancreatic cancer cells were treated with PGE 2 or rofecoxib, a selective COX-2 inhibitor. MMP-2 expression and activity were assayed using Western blot analysis and zymography, respectively. MMP-2 promoter activity was analyzed with a luciferase-based assay. Ets-1 activity was analyzed using gel shift assay. Ets-1 expression was specifically silenced using RNA interference. Cellular invasive and migratory potentials were determined using a Boyden chamber assay with or without Matrigel, respectively. Exogenous PGE 2 induced MMP-2 expression and activity and increased ERK1/2 phosphorylation, Ets-1 binding activity, and MMP-2 promoter activity. PGE 2 also increased cellular migratory and invasive potentials. The mitogen-activated protein kinase kinase inhibitor PD98059 and Ets-1 silencing each abolished PGE 2 -induced increases in MMP-2 expression. PD98059 and Ets-1 silencing each abrogated the effect of PGE 2 on cellular invasive potential but not on cellular migratory potential. Rofecoxib suppressed MMP-2 expression and activity, Ets-1 binding activity, MMP-2 promoter activity, and cellular migratory and invasive potentials. These results suggest that PGE 2 mediates pancreatic cancer cellular invasiveness through an ERK/Ets-1-dependent induction of MMP-2 expression and activity. They also suggest that COX-2 inhibition may represent a strategy to inhibit invasive potential in pancreatic cancer.

Most patients with pancreatic adenocarcinoma present with surgically incurable disease. Gemcitabine, the principal agent used to treat such patients, has little impact on outcome. Overexpression of carcinoembryonic antigen-related cell adhesion molecule (CEACAM) 6, a feature of this malignancy, is associated with resistance to anoikis and increased metastasis. The purpose of this study was to determine the role of CEACAM6 in cellular chemoresistance to gemcitabine. CEACAM6 was stably overexpressed in Capan2 cells, which inherently express very low levels of the protein. Suppression of CEACAM6 expression was achieved in BxPC3 cells, which inherently overexpress CEACAM6, by stable transfection of a CEACAM6 small interfering RNA-generating vector. The effects of modulating CEACAM6 expression on gemcitabine-induced cytotoxicity were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide cytotoxicity assay, flow cytometric apoptosis quantification, caspase profiling, and Western analysis of cytoplasmic cytochrome c release. The roles of Akt and c-Src kinases as downstream targets of CEACAM6 signaling were examined. Stable overexpression of CEACAM6 in Capan2 increased gemcitabine chemoresistance, whereas CEACAM6 gene silencing in BxPC3 markedly increased the sensitivity of these cells to gemcitabine. Differential expression of CEACAM6 modulates Akt activity in a c-Src-dependent manner, and CEACAM6 overexpression appears to protect cells from cytochrome c-induced caspase 3 activation and apoptosis.

BackgroundAutologous bone marrow-derived stem cells have been ascribed an important therapeutic role in No-Option Critical limb Ischemia (NO-CLI). One primary endpoint for evaluating NO-CLI therapy is major amputation (AMP), which is usually combined with mortality for AMP-free survival (AFS). Only a trial which is double blinded can eliminate physician and patient bias as to the timing and reason for AMP. We examined factors influencing AMP in a prospective double-blinded pilot RCT (2:1 therapy to control) of 48 patients treated with site of service obtained bone marrow cells (BMAC) as well as a systematic review of the literature.MethodsCells were injected intramuscularly in the CLI limbs as either BMAC or placebo (peripheral blood). Six month AMP rates were compared between the two arms. Both patient and treating team were blinded of the assignment in follow-up examinations. A search of the literature identified 9 NO-CLI trials, the control arms of which were used to determine 6 month AMP rates and the influence of tissue loss.ResultsFifteen amputations occurred during the 6 month period, 86.7% of these during the first 4 months. One amputation occurred in a Rutherford 4 patient. The difference in amputation rate between patients with rest pain (5.6%) and those with tissue loss (46.7%), irrespective of treatment group, was significant (p = 0.0029). In patients with tissue loss, treatment with BMAC demonstrated a lower amputation rate than placebo (39.1% vs. 71.4%, p = 0.1337). The Kaplan-Meier time to amputation was longer in the BMAC group than in the placebo group (p = 0.067). Projecting these results to a pivotal trial, a bootstrap simulation model showed significant difference in AFS between BMAC and placebo with a power of 95% for a sample size of 210 patients. Meta-analysis of the literature confirmed a difference in amputation rate between patients with tissue loss and rest pain.ConclusionsBMAC shows promise in improving AMP-free survival if the trends in this pilot study are validated in a larger pivotal trial. The difference in amp rate between Rutherford 4 & 5 patients suggests that these patients should be stratified in future RCTs.