Intratumoral heterogeneity of microRNA expression in rectal cancer

Introduction: An increasing number of studies have investigated microRNAs (miRNAs) as potential markers of diagnosis, treatment and prognosis. So far, agreement between studies has been minimal, which may in part be explained by intratumoral heterogeneity of miRNA expression. The aim of the present study was to assess the heterogeneity of a panel of selected miRNAs in rectal cancer, using two different technical approaches. Materials and Methods: The expression of the investigated miRNAs was analysed by real-time quantitative polymerase chain reaction (RT-qPCR) and in situ hybridization (ISH) in tumour specimens from 27 patients with T3-4 rectal cancer. From each tumour, tissue from three different luminal localisations was examined. Inter- and intra-patient variability was assessed by calculating intraclass correlation coefficients (ICCs). Correlations between RT-qPCR and ISH were evaluated using Spearman's correlation. Results: ICCsingle (one sample from each patient) was higher than 50% for miRNA-21 and miRNA-31. For miRNA-125b, miRNA-145, and miRNA-630, ICCsingle was lower than 50%. The ICCmean (mean of three samples from each patient) was higher than 50% for miRNA-21(RT-qPCR and ISH), miRNA-125b (RT-qPCR and ISH), miRNA-145 (ISH), miRNA-630 (RT-qPCR), and miRNA-31 (RT-qPCR). For miRNA-145 (RT-qPCR) and miRNA-630 (ISH), ICCmean was lower than 50%. Spearman correlation coefficients, comparing results obtained by RT-qPCR and ISH, respectively, ranged from 0.084 to 0.325 for the mean value from each patient, and from -0.085 to 0.515 in the section including the deepest part of the tumour. Conclusion: Intratumoral heterogeneity may influence the measurement of miRNA expression and consequently the number of samples needed for representative estimates. Our findings with two different methods suggest that one sample is sufficient for adequate assessment of miRNA-21 and miRNA-31, whereas more samples would improve the assessment of miRNA-125b, miRNA-145, and miRNA-630. Interestingly, we found a poor correlation between the expression estimates obtained by RT-qPCR and ISH, respectively

Expression Profiling of Rectal Tumors Defines Response to Neoadjuvant Treatment Related Genes

To date, no effective method exists that predicts the response to preoperative chemoradiation (CRT) in locally advanced rectal cancer (LARC). Nevertheless, identification of patients who have a higher likelihood of responding to preoperative CRT could be crucial in decreasing treatment morbidity and avoiding expensive and time-consuming treatments. The aim of this study was to identify signatures or molecular markers related to response to pre-operative CRT in LARC. We analyzed the gene expression profiles of 26 pre-treatment biopsies of LARC (10 responders and 16 non-responders) without metastasis using Human WG CodeLink microarray platform. Two hundred and fifty seven genes were differentially over-expressed in the responder patient subgroup. Ingenuity Pathway Analysis revealed a significant ratio of differentially expressed genes related to cancer, cellular growth and proliferation pathways, and c-Myc network. We demonstrated that high Gng4, c-Myc, Pola1, and Rrm1 mRNA expression levels was a significant prognostic factor for response to treatment in LARC patients (p<0.05). Using this gene set, we were able to establish a new model for predicting the response to CRT in rectal cancer with a sensitivity of 60% and 100% specificity. Our results reflect the value of gene expression profiling to gain insight about the molecular pathways involved in the response to treatment of LARC patients. These findings could be clinically relevant and support the use of mRNA levels when aiming to identify patients who respond to CRT therapy.C, CC and AB were supported by projects CTS2200 and PI-0710-2013 of Junta de Andalucía, TIN2013-41990-R of Programa Estatal I+D+i MINECO, and GREIB PYR_2010-02 and 2010_05 of University of Granada

Isolated limb perfusion for unresectable extremity cutaneous squamous cell carcinoma; an effective limb saving strategy

Background: A small minority of patients present with locally advanced cutaneous Squamous Cell Carcinoma (cSCC). The aim of this study was to evaluate the effectiveness of Tumour necrosis factor α (TNF) and melphalan based isolated limb perfusion (TM-ILP) as a limb saving strategy for locally advanced extremity cSCC. Methods: A retrospective search from prospectively maintained databases, at two tertiary referral centers, was performed to identify patients treated with TM-ILP for locally advanced cSSC of an extremity between 2000 and 2015. Results: A total of 30 patients treated with TM-ILP for cSCC were identified, with a median age of 71 years (36–92) and 50% female. Response could not be evaluated in 3 patients. After a median follow up of 25 months, the overall response rate was 81% (n = 22), with 16 patients having a complete response (CR, 59%). A total of 7 patients developed local recurrence, with a median time to recurrence of 9 months (Interquartile Range 7–10). Progressive disease was observed in 5 patients (19%). Limb salvage rate was 80%. The overall 2-year survival was 67%. Conclusions: TM-ILP should be considered as an option in patients with locally advanced cSCC in specialised centers, resulting in a high limb salvage rate

The 1944 eruption of Vesuvius, Italy: combining contemporary accounts and field studies for a new volcanological reconstruction

We integrate the different contemporary sources together with new field data on the pyroclastic deposits to make a new volcanological reconstruction of the explosive phases of the 1944 Vesuvius eruption. We adopt the four successive phases of the eruption first defined by Imb`o (1945), who made the most detailed contemporary description of the eruption: Phase 1 – effusive, Phase 2 – lava fountains, Phase 3 – mixed explosions and Phase 4 – seismic-explosive. Phase 1 consisted of four days of effusive activity. Phase 2 generated eight successive lava fountains which formed agglutinated spatter in a restricted area around the crater. At distances of > 1 km from the crater, reverse graded, well-sorted, scoria lapilli with up to 94 wt % juvenile material and calculations indicate a volume of 8.2 × 106 m3 DRE (Dense Rock Equivalent) for Phase 2. A short pause in scoria fallout was observed that coincides with the transition between Phases 2 and 3 of the eruption. On the crater rim there is clear evidence for the different phases, owing to the stratification of the deposits; however, away from the crater, stratigraphic breaks suggesting any discontinuity in the eruptive activity are absent. The beginning of Phase 3 is marked by the appearance of abundant dense scoria fragments, coincident with the coarsest part of the lapilli. High-density scoria forms 10 wt % of juvenile material in Phase 2, increasing to 45 % in the upper part of Phase 3. Isopach maps derived from field measurements indicate a mean volume of 40.2 × 106 m3 DRE for Phase 3. Distal ash, mainly formed during Phase 3, was dispersed to the SE as far as Albania, and calculations yield a volume of 102 × 106 m3 DRE. Intermittent activity associated with Phase 4 generated ash-rich plumes dispersed towards the SWand contemporary thickness descriptions yield a bulk volume of 4.2 × 106 m3 (2.5 × 106 m3 DRE). Small pyroclastic density currents (PDCs) were observed during Phases 3 and 4. The deposits (200 m from the crater rim) of these currents have been identified on the flanks of the cone. Thin, massive and poorly sorted ash layers, that occur up to 2.5 km from the crater rim, are interpreted to represent the distal facies of these PDCs. Mass discharge rate (MDR) estimates for the paroxysmal phase (end of Phase 2 and start of Phase 3) of this event are around 3.5 × 106 kg/s, however, this increases to> 107 kg/s if the mass of distal ash is taken into account. Column height estimates from fallout isopleths associated with the eruption’s paroxysmal phase are> 10 km. Based on the contemporaneous chronicles, we were able to define the type and extent of damage associated with the different styles (or temporal phases) of the eruption. Our calculations demonstrate that the present-day population at risk has doubled compared to 1944. The contemporaneous (and also subsequent) scientific literature underestimated the magnitude and intensity of this eruption and very little attention has been dedicated to the damage that occurred. We suggest that this is at least partly related to the extensive destruction of Neapolitan area and the deaths of tens of thousands of civilians related to the Second World War