The effect of lymphadenectomy and radiotherapy on recurrence and survival in endometrial carcinoma. Experience in a population reference centre

Aim

To evaluate the effect of lymphadenectomy and/or radiotherapy on recurrence and survival patterns in endometrial carcinoma (EC) in a radiotherapy reference centre population.

Material and Methods

A retrospective population-based review was conducted on 261 patients with stages I–III EC. Univariate and multivariate analyses were carried out. Both recurrence and survival were analysed according to patient age, FIGO stage, tumour size, myometrial invasion, tumour grade, lymphadenectomy, external beam irradiation (EBI), and brachytherapy (BT).

Results

Median age: 64.8 years. Median follow-up: 151 months. The following treatments were administered: surgery, 97.32%; lymph-node dissection, 54.4%; radiotherapy, 162 patients (62%) (EBI and BT: 64.1%, BT alone: 30.2%, EBI alone: 5.6%).Twenty-six patients (9.96%) suffered loco-regional recurrence, whilst 27 (10.34%) suffered distant failure. The 5-year overall survival (OS) for all stages was 80.1%. The 5-year disease free survival (DFS) was 92.1% for all patients. The 10-year DFS was 89.9%.The independent significant prognostic factors for a good outcome identified through the multivariate analysis were: age <75 years (p = 0.001); tumour size ≤2 cm (p = 0.003); myometrial invasion ≤50% (p = 0.011); lymphadenectomy (p = 0.02); EBI (p = 0.001); and BT (p = 0.031).Toxicity occurred in 114 of the 162 patients who received radiotherapy (70.5%). The toxicity was mainly acute, and late in only 28.3% (n = 45) of cases. The majority experienced G1-2 toxicity, and only 3% of patients experienced G3 late toxicity (5/162).

Conclusions

Our results suggest that age <75 years, tumour size ≤2 cm, myometrial invasion ≤50%, lymphadenectomy, EBI, and BT, are predictors of a good outcome in EC.     

Development of biosensor for phenolic compounds containing PPO in β-cyclodextrin modified support and iridium nanoparticles

A biosensor based on the iridium nanoparticles dispersed in ionic liquid (IL) 1-butyl-3-methylimidazolium hexafluorophosphate (Ir-BMI·PF₆) and a celery (Apium graveolens) extract as a source of polyphenol oxidase (PPO) was constructed. A modified support based on β-cyclodextrin (β-CDEP) was used for enzyme immobilization. The behavior of phenolic compounds was investigated by square-wave voltammetry and rutin was selected by presenting the greatest signal. The best performance was obtained with a composition of 70:10:10:10% (w/w/w/w) of the graphite powder:β-CDEP:Nujol:Ir-BMI·PF₆ composition, a PPO concentration of 500 units mL^?1, in 0.1 M phosphate buffer solution (pH 6.0) with frequency, pulse amplitude and scan increment at 100 Hz, 60 mV, and 3.0 mV, respectively. Under optimized conditions, the cathodic currents increased linearly for the rutin concentration range of 1.3 × 10^?7–2.0 × 10^?6 M with a detection limit of 7.9 × 10^?8 M. This sensor demonstrated acceptable repeatability and reproducibility and the results for the rutin recovery ranged from 92.8 to 103.4%. A relative error of 0.7% was obtained in the rutin determination in simulated samples.     

BRAF mutation and RASSF1A expression in thyroid carcinoma of southern Italy

Aim of this work is to provide a detailed comparison of clinical‐pathologic features between well‐differentiated and poorly differentiated tumors according to their BRAF and RASSF1A status. We analyzed RASSF1A methylation by MSP and BRAF mutation by LCRT‐PCR with LightMix® kit BRAF V600E in neoplastic thyroid tissues. Immunohistochemical evaluation of RASSF1A expression was also performed by standard automated LSAB‐HRP technique. An overall higher degree of RASSF1A over‐expression than normal thyroid parenchyma surrounding tumors (P < 0.05) has been found in all malignant well‐differentiated lesions. Moreover, statistically significant higher levels of RASSF1A expression were observed in differentiated cancers associated to an inflammatory autoimmune background (P = 0.01). Amplifiable DNA for LC PCR with LightMix® kit BRAF V600E was obtained in nine PTCs, four FVPTCs, five ATCs, and one control. The V600E mutation was found in 13 of 18 (72%) tumors. BRAF was mutated in 6 of 9 (66%) classical PTC, in 2 of 4 (50%) follicular variant PTC and in all ACs (100%). The overall frequency of RASSF1A promoter methylation observed was 20.5% (9 cases out 44). Hypermethylation of RASSF1A in primary tumors was variable according to histotypes ranging from100% (5/5) in ACs to only 12.5% (4/32) in PTCs. We show a correlation between RASSF1A methylation status and RASSF1A protein expression. Finally, we conclude that BRAF V600E mutation and RASSF1A methylation were pathogenetic event restricted to a subgroup of PTC/FVPTCs in early stage and to clinically aggressive ATCs. J. Cell. Biochem. 114: 1174–1182, 2013. © 2012 Wiley Periodicals, Inc.     

Amazonian biogenic volatile organic compounds under global change

Biogenic volatile organic compounds (BVOCs) play important roles at cellular, foliar, ecosystem and atmospheric levels. The Amazonian rainforest represents one of the major global sources of BVOCs, so its study is essential for understanding BVOC dynamics. It also provides insights into the role of such large and biodiverse forest ecosystem in regional and global atmospheric chemistry and climate. We review the current information on Amazonian BVOCs and identify future research priorities exploring biogenic emissions and drivers, ecological interactions, atmospheric impacts, depositional processes and modifications to BVOC dynamics due to changes in climate and land cover. A feedback loop between Amazonian BVOCs and the trends of climate and land‐use changes in Amazonia is then constructed. Satellite observations and model simulation time series demonstrate the validity of the proposed loop showing a combined effect of climate change and deforestation on BVOC emission in Amazonia. A decreasing trend of isoprene during the wet season, most likely due to forest biomass loss, and an increasing trend of the sesquiterpene to isoprene ratio during the dry season suggest increasing temperature stress‐induced emissions due to climate change.     

Evidence of mesenchymal stromal cell adaptation to local microenvironment following subcutaneous transplantation

Subcutaneous transplantation of mesenchymal stromal cells (MSC) emerged as an alternative to intravenous administration because it avoids the pulmonary embolism and prolongs post‐transplantation lifetime. The goal of this study was to investigate the mechanisms by which these cells could affect remote organs. To this aim, murine bone marrow–derived MSC were subcutaneously transplanted in different anatomical regions and the survival and behaviour have been followed. The results showed that upon subcutaneous transplantation in mice, MSC formed multicellular aggregates and did not migrate significantly from the site of injection. Our data suggest an important role of hypoxia‐inducible signalling pathways in stimulating local angiogenesis and the ensuing modulation of the kinetics of circulating cytokines with putative protective effects at distant sites. These data expand the current understanding of cell behaviour after subcutaneous transplantation and contribute to the development of a non‐invasive cell‐based therapy for distant organ protection.     

Red porgy (Pagrus pagrus) larval feeding performance and behavior at the onset of exogenous feeding

The feeding performance and behavior at the onset of exogenous feeding, 3 to 4 days after hatching (DAH), were studied in red porgy Pagrus pagrus larvae. Similar feeding efficiency and intensity were achieved for two feeding treatments (live or freeze-dried rotifers) suggesting that prey movement is not decisive for their detection and capture and demonstrating that at first feeding red porgy larvae can ingest inert food. Larvae feeding performance was not affected by a diet shift between treatments. Based on maximum rotifers consumption and gut evacuation time at 18 °C, the daily ration was estimated as 14.035 μg, considering 14 h of feeding and a 25% egg:female rotifer ratio. Larval swimming activity measured by video recording showed a close association with gut fullness and similar swimming patterns for 3 and 4 DAH larvae. However, 20.3% larger mouth gape and 54.6% higher swimming speed of the older larvae should provide a better feeding performance and more energy needed for growth.     

Volatile organic compounds emissions in Norway spruce (Picea abies) in response to temperature changes

Volatile organic compound (VOC) emissions from Norway spruce ( Picea abies ) saplings were monitored in response to a temperature ramp. Online measurements were made with a proton transfer reaction – mass spectrometer under controlled conditions, together with plant physiological variables. Masses corresponding to acetic acid and acetone were the most emitted VOCs. The emission rates of m137 (monoterpenes), m59 (acetone), m33 (methanol), m83 (hexanal, hexenals), m85 (hexanol) and m153 (methyl salicylate, MeSa) increased exponentially with temperature. The emission of m61 (acetic acid) and m45 (acetaldehyde), however, increased with temperature only until saturation around 30°C, closely following the pattern of transpiration rates. These results indicate that algorithms that use only incident irradiance and leaf temperature as drivers to predict VOC emission rates may be inadequate for VOCs with lower H, and consequently higher sensitivity to stomatal conductance.     

Cross-species Proteomics Reveals Specific Modulation of Signaling in Cancer and Stromal Cells by Phosphoinositide 3-kinase (PI3K) Inhibitors

The tumor microenvironment plays key roles in cancer biology, but its impact on the regulation of signaling pathway activity in cancer cells has not been systemically investigated. We designed an analytical strategy that allows differential analysis of signaling between cancer and stromal cells present in tumor xenografts. We used this approach to investigate how in vivo growth conditions and PI3K inhibitors regulate pathway activities in both cancer and stromal cell populations. We found that, despite inducing more modest changes in protein expression, in vivo growing conditions extensively rewired protein kinase networks in cancer cells. As a result, different sets of phosphorylation sites were modulated by PI3K inhibitors in cancer cells growing in tumors relative to when these cells were in culture. The p110δ PI3K-selective compound CAL-101 (Idelalisib) did not inhibit markers of PI3K activity in cancer or stromal cells; however, unexpectedly, it induced phosphorylation on SQ motifs in both subpopulations of tumor cells in vivo but not in vitro. Thus, the interaction between cancer cells and the stroma modulated the ability of PI3K inhibitors to induce the activation of apoptosis in solid tumors. Our study provides proof-of-principle of a proteomics workflow for measuring signaling specifically in cancer and stromal cells and for investigating how cancer biochemistry is modulated in vivo.Solid tumors contain a heterogeneous population of cells. Transformed epithelial cells recruit different types of somatic cells to the tumor microenvironment where they influence varying aspects of cancer biology. The role of heterotypic communication between normal stromal cells and transformed cancer cells is well established (1, 2). Different somatic cell types, including fibroblasts, epithelial cells, and cells of the immune system—all of which are found in tumors—promote cancer cell development by means of gap-junction intercellular communication, direct cell-to-cell contacts, and by the release of growth factors, enzymes, and cytokines that act on neighboring malignant cells (3–6).The tumor microenvironment determines the ability of cancer cells to survive in specific organs and their ability to proliferate and metastasize (7–9). Growth factors released from tumor-associated stromal cells also influence how cancer cells respond to drug administration (10). Therefore, the advancement of targeted cancer therapies requires an understanding of how the tumor microenvironment modulates the biochemistry of transformed cancer cells. In addition, targeting the tumor stroma is emerging as an intriguing concept for the development of anti-cancer therapies (11). It is therefore important to investigate specific effects of compounds in clinical development on stromal cells in addition to those exerted toward malignant cancer cells (12).Here we investigated the effects that changes in growing conditions from a two-dimensional cell culture to an in vivo three-dimensional tumor environment had in modulating protein and phosphoprotein expression in human cancer cells. For this, we used mass spectrometry (MS) to specifically measure cancer and stromal proteomes and phosphoproteomes within mouse tumor xenografts.We also investigated the effects that the pharmacological inhibitors of PI3K, namely GDC-0941 or CAL-101, would have on the phosphoproteomes of stromal cells relative to cancer cells in solid tumors. GDC-0941 is an inhibitor with specificity for class I PI3Ks, whereas CAL-101 specificity is restricted to the p110δ isoform of PI3K (13, 14), which in untransformed tissues is mainly found in leukocytes (15). The PI3K signaling pathway is often deregulated in different cancer types (16), including colorectal cancer (17), and both compounds used in this study are in different stages of clinical development (18–20). PI3K signaling has also been implicated in mediating the effects that the microenvironment has on cancer cells (21).We found that in vivo growth conditions had profound effects on phosphoprotein expression, which was reflected on the phosphorylation sites modulated by PI3K inhibitors in vivo relative to in vitro and in their ability to induce apoptotic markers across these two cell culture conditions.     

2,9-disubstituted-N6-(arylcarbamoyl)-8-azaadenines as new selective A3 adenosine receptor antagonists: synthesis, biochemical and molecular modelling studies

A number of N6-(N-arylcarbamoyl)-2-substituted-9-benzyl-8-azaadenines, obtained by a modification of the synthetic scheme used to prepare selective A1 ligands, by only three or two steps, are described. At first we prepared a series of 2-phenyl-9-benzyl-8-azaadenines having as N6 substituent a variously substituted N-phenylcarbamoyl group. Some of these derivatives demonstrated good affinity towards the A3 subtype but low selectivity. Compounds having p-CF3, p-F and p-OCH3, as substituents on the phenylcarbamoyl group were selected as lead compounds for the second part of this study. Without modifying the N6 substituent, which would assure A3 affinity, we varied the 9 and 2 positions on these molecules to enhance selectivity. Some compounds having a p-methyl group on the 2-phenyl substituent showed a very good affinity and selectivity for the A3 subtype, revealing the first class of A3 adenosine receptor selective antagonists with a bicyclic structure strictly correlated to the adenine nucleus. The molecular modelling work, carried out using the DOCK program, supplied two models which may be useful for a better understanding of the binding modes. Both models highlighted the preferred interacting tautomeric forms of the antagonists for human A1 and A3 receptors.