A total of 5 full-scale Reinforced Concrete (RC) frames were tested to develop new models for predicting the structural behavior of corroded RC frames and to verify the accuracy of the models mostly developed for RC columns. Four corroded and one non-corroded RC frame were tested under cyclic loading. Corroded RC frames were tested under displacement-controlled cyclic loading at four corrosion levels and for a constant applied axial load ratio of 0.20, with the same concrete compression strength level and reinforcement bar layout. Actual corrosion ratios were obtained following the loading tests, breaking all RC columns and beams and extracting all reinforcement bars from concrete. The test results demonstrated that the corrosion level played an active role in the structural capacity of the RC frames. Initial corrosion crack widths had more significant effects on structural performance. Some models proposed for corroded RC columns can also be used for corroded RC frames in terms of yield and ultimate load capacities according to the corrosion levels of the columns and beam composing the RC frame. It was also concluded from the test results that the displacement-based ductility ratio for corroded RC frames could still be misleading if they were not computed according to energy-based or bilateral failure criteria. Structural properties such as initial/secant stiffness and energy absorption capacity calculated based on experimental measurements for RC frames had further disclosed corrosion effects

In this update of the consensus of the Spanish Society of Medical Oncology (Sociedad Española de Oncología Médica-SEOM) and the Spanish Society of Pathology (Sociedad Española de Anatomía Patológica-SEAP), advances in the analysis of biomarkers in advanced colorectal cancer (CRC) as well as susceptibility markers of hereditary CRC and molecular biomarkers of localized CRC are reviewed. Recently published information on the essential determination of KRAS, NRAS and BRAF mutations and the convenience of determining the amplification of human epidermal growth factor receptor 2 (HER2), the expression of proteins in the DNA repair pathway and the study of NTRK fusions are also evaluated. From the pathological point of view, the importance of analysing the tumour budding and poorly differentiated clusters, and its prognostic value in CRC is reviewed, as well as the impact of molecular lymph node analysis on lymph node staging in CRC. The incorporation of pan-genomic technologies, such as next-generation sequencing (NGS) and liquid biopsy in the clinical management of patients with CRC is also outlined. All these aspects are developed in this guide, which, like the previous one, will remain open to any necessary revision in the future.

9718 Background: Epidermal growth factor receptor (EGFR) is a protoncogen that is found overexpressed in colorectal carcinomas (CRC) and it correlates with a worse prognosis. The aim was to describe EGFR overexpression patterns in non-metastatic CRC and to correlate these data with follow-up. Methods: We analyzed a series of 56 CRC. Inclusion criteria were: a) resected primary adenocarcinoma; b) curative surgery; c) pT3 N0–2 M0 without progression during the first 6 months post surgery; d) minimum follow-up over 3 years. EGFR overexpression was analyzed by immunohistochemistry (IHC) using the Dako PharmaDx kit (Glostrup, Denmark). As positive control the Dako slides and a bloc cell of A431-AAM cells were used. Presence of cytoplasmic and membrane patterns (intensity 1(+), 2(+) and 3(+)) were evaluated as well as the percentage of positive cells. Statistical analysis: association between qualitative variables was analyzed by Fisher's exact test. Disease-free and overall survival distributions were estimated by the Kaplan-Meier method and were analyzed with the log rank test. All P values are from two-sided statistical tests. Results: Characteristics of the patients (pts) were as follows: mean age: 68,4 years (range: 28–91); sex: 24 male and 32 female; location: 19 right colon and 37 left colon; follow-up: 44 pts alive without disease (78,5%), 9 died of disease (16,5%) and 3 died of other causes (5%). 10 pts had metastasis in the follow-up. IHC: Forty-two cases were positive (75%) and were classified as follows: 17 with >50% of positive cells (30%), 7 with 25–50% of positive cells (13%) and 18 cases with < 25% of positive cells. Patterns of staining were cytoplasmic predominance in 27 cases (48%) and 15 with membrane predominance (27%): 4 with intensity 1(+), 9 with 2(+) and 2 with 3(+). Only those patients with tumors harboring membrane positivity 2(+) and 3(+) had more probability to present metastasis (p= 0.067). Conclusions: EGFR overexpression evaluated as a membrane pattern with 2(+) and 3(+) intensity is related with metastatic development in colon carcinoma. Suported by a grant of C.I.R. Hospital de Sabadell. Consorci Sanitari Parc Taulí. Sabadell. Spain No significant financial relationships to disclose.

4089 Background: Overexpression of HER2 is an important biomarker in gastroesophageal junction (GEJ) and gastric cancer (GC) and a predictive factor for trastuzumab (T, Herceptin). The ToGA phase III trial showed the benefit in overall survival of adding trastuzumab to chemotherapy inHER2-positive advanced GEJ/GC patients, following validated scoring criteria by a central laboratory. This study examines the incidence of HER2 positivity (local laboratories) in GEJ/GC according to EMA Herceptin label. Methods: HER-EAGLE was an epidemiological, non–interventional, international study assessing HER2 status by IHC/ISH in tumor samples from any stage GEJ/GC patients. Neutral buffered 10% formalin embedded tissues (surgical excision specimens or minimum 6-8 biopsies) analyzed via validated Ventana and Dako methods and scoring criteria used in ToGA: HER2-positive if IHC 3+ or IHC 2+ (FISH/SISH confirmed; HER2/CEP17 ratio ≥ 2.0); HER2-negative if IHC 0 or IHC 1+. Overall and subgroup estimates calculated with 95%CI. Data from the Spanish cohort are presented. Results: The Spanish cohort of HER-EAGLE included 1,954 participants (63.7% males) from 33 hospitals (Dec2010 to Aug2011), with 469 biopsies (24.0%) and 1,479 excisions (75.7%). Samples were 13.7% GEJ and 86.3% GC, and the adenocarcinomas were 1,137 intestinal (58.2%), 510 diffused (26.1%), 163 mixed (8.3%; Laureen classification), and 144 not available (7.3%). Median time from sampling to HER2 analysis of 5.1 years (range from days to 12 years). Total of 210 cases tested IHC 3+ (10.7%), 215 IHC 2+ (11.0%), 308 IHC 1+ (15.8%), and 1,221 IHC 0 (62.5%). Overall, HER2 positivity (IHC 3+ or IHC2+/ISH+) was 14.1% (95%CI 12.61 – 15.75). HER2 positivity by histological type was higher in intestinal adenocarcinomas (19.5%) compared to diffused (4.5%) or in mixed (9.2%). Conclusions: HER-EAGLE confirmed the feasibility of community based HER2 testing in GC as the first study with a high number of samples analyzed by local laboratories. The incidence of HER2-positivity (EMA label definition) was similar to the ToGA screening population, and it was again higher in adenocarcinomas of intestinal type (19.5%).

AIMS: To compare the immunohistochemical expression of prognostic markers p27(Kip1), p45(Skp2) and Ki67 in Merkel cell carcinoma (primary neuroendocrine carcinoma of the skin, MCC), small cell neuroendocrine carcinoma of lung and urinary bladder (SNC), and cutaneous squamous cell carcinoma (SCC). METHODS AND RESULTS: Immunohistochemistry was performed using antibodies directed against p27(Kip1), p45(Skp2) and Ki67 on 72 tumour cases: 24 MCC, 25 SCC, and 23 SNC (15 from the lung and eight from the urinary bladder). Percentages of positive cells were determined for each marker and statistically analysed. Expression profiles on MCC and SCC were significantly different for all three markers. MCC and SNC exhibited significant similarities in their p27(Kip1) and p45(Skp2) expression profiles. In contrast, MCC and SNC differed significantly in their Ki67 proliferation indices, which were much higher in SNC. Additionally, MCC cases showed an association between increased proliferation indices and the appearance of local recurrence(s) and/or metastases. CONCLUSION: The immunohistochemical profile of MCC differs from that of SCC, in spite of their common oncogenesis and the supposed metaplastic origin of MCC, and resembles that of SNC, except for Ki67 levels, which were higher in the latter (characterized by greater biological aggressiveness). High levels of Ki67 also appear to be a prognostic factor in MCC.

Fibrous benign proliferations of the testis and paratesticular tissues are an uncommon and heterogeneous group of lesions that can mimic true neoplasms. Among them, those considered to be reactive proliferations have been included in the category of fibrous pseudotumors and are often associated with trauma, hydrocele, or infection. We report 2 cases of nodular and diffuse fibrous proliferation. This condition is part of the spectrum of reactive benign fibrous lesions and has previously been described in the testicular tunics and paratesticular region only. One of our 2 cases had the peculiarity of being located in the penile shaft, whereas the other involved the tunica vaginalis. To the best of our knowledge, this is the first report of a diffuse and nodular fibrous tumor involving the penis.

Idiopathic pylephlebitis and primary sclerosing peritonitis are two highly unusual entities. To our knowledge, the association of the two diseases has not been described previously. We report a 42-year-old patient with a protein S deficiency who presented with fever and chills, in whom idiopathic pylephlebitis was diagnosed. A year later, the patient was readmitted because of recurrent vomiting and weight loss. An exploratory laparotomy yielded diagnosis of sclerosing peritonitis, which resolved after surgery. The short time interval between the processes suggests that they were related to each other, and also to the protein S deficiency.

Squamous cell carcinomas have a range of histopathological manifestations. The parameters that determine this clinically observed heterogeneity are not fully understood. Here, we report the generation of a cell culture model that reflects part of this heterogeneity. We have used the catalytic subunit of human telomerase hTERT and large T to immortalize primary UV-unexposed keratinocytes. Then, mutant HRAS G12V has been introduced to transform these immortal keratinocytes. When injected into immunosuppressed mice, transformed cells grew as xenografts with distinct histopathological characteristics. We observed three major tissue architectures: solid, sarcomatoid and cystic growth types, which were primarily composed of pleomorphic and basaloid cells but in some cases displayed focal apocrine differentiation. We demonstrate that the cells generated represent different stages of skin cancerogenesis and as such can be used to identify novel tumor-promoting alterations such as the overexpression of the PADI2 oncogene in solid-type SCC. Importantly, the cultured cells maintain the characteristics from the xenograft they were derived from while being amenable to manipulation and analysis. The availability of cell lines representing different clinical manifestations opens a new tool to study the stochastic and deterministic factors that cause case-to-case heterogeneity despite departing from the same set of oncogenes and the same genetic background.

Recently, the copper efflux transporters ATP7B and ATP7A have been implicated in the transport of and resistance to platinum drugs in breast and ovarian cancers. Because of the extensive use of oxaliplatin in colorectal cancer (CRC), we examined the expression of both transporters in tumors from CRC patients treated with oxaliplatin/5FU and sought to determine whether their expression can predict clinical outcome in these patients. ATP7B and ATP7A levels were determined by quantitative real-time PCR in 50 primary tumors of previously untreated patients with advanced colorectal adenocarcinoma who were subsequently treated with oxaliplatin/5FU. Additionally, ATP7B protein expression was assessed by immunohistochemical staining using a tissue microarray. Patients with the lowest mRNA expression levels of ATP7B had a significantly longer time to progression (TTP) (p = 0.0009) than patients with the highest levels (12.14 months vs. 6.43 months) who also had an increased risk of progression (HR = 3.56; 95% CI, 1.6-7.9; p = 0.002). Furthermore, patients with low levels of both protein and mRNA of ATP7B derived the maximum benefit from oxaliplatin/5FU with the longest TTP as compared with patients with high levels of ATP7B protein and mRNA (14.64 months vs. 4.63 months, respectively, p = 0.01) and showed a nonsignificant trend toward a lower response rate (37.5% and 75%, respectively). In conclusion, ATP7B mRNA and protein expression in colorectal tumors is associated with clinical outcome to oxaliplatin/5FU. Prospective studies are required to evaluate the role of this marker in tailoring chemotherapy.

Introduction Cdk5 has been related to cancer progression and metastasis and to response to DNA damage in cancer. In colorectal cancer (CRC), our previous results showed that Cdk5 was overexpressed in CRC tumours as compared to normal tissues. CDK5 gene knockdown was associated with decreased migration and invasion of colon cancer cells but has no effect on cell proliferation; this effect was not not observed in cell lines with BRAF or KRAS mutations. In cell lines with acquired resistance to oxaliplatin (OXA) CDK5 knockdown overcame this resistance. The aim of this work was to study the involvement of Cdk5 on migration and invasion according to KRAS status and to investigate the prognostic and predictive value of Cdk5 in representative patients cohorts. Material and methods SW48 (KRAS WT) and the isogenic SW48 KI G12V (KRAS mutant) cell lines were used (Horizon Discovery Ltd). CDK5 gene was silenced with specific siRNA (Ambion, 5 nM) to study its role in migration and invasion using Boyden chamber assays (HTS transwell, Corning). Comparisons between different experimental conditions were carried out using the T test. For clinical studies, we used 342 tumour samples from CRC patients (Non-treated stage II-III frozen n=135; stage IV FFPE=207). Stage IV tumours corresponded to patients treated with schedules containing irinotecan (n=139) or OXA (n=68) without antitarget drugs. Cdk5 expression was analysed by qPCR (Taqman assay) or by immunohistochemistry. Log rank (LR) and cox regression (CR) tests were used to study differences in disease-free (DFS) overall survival (OS) or time to progression (TTP); Chi-square or Fisher’s tests were used to study differences in response rates (ORR). Results and discussions CDK5 gene silencing was associated with decreased cell migration only in SW48 G12V cells (p=0.035). High Cdk5 mRNA levels was a bad prognostic factor in stage II (DFS CR HR=4.15; p=0.022; OS CR HR=8.25, p=0.048) and III (DFS LR 24 months vs. not reached; p=0.048). In stage II, Cdk5 prognostic value was associated with KRAS mutant status (CR p=0.045). In stage IV CRC, Cdk5 positive IHC staining was associated with worse TTP (LR 8 vs. 18 months, p=0.043; CR HR=2.17) and ORR (53% vs. 87%, p=0.029) only in OXA-treated patients. Conclusion These data confirm previous results establishing Cdk5 as a tumor-promoting factor in CRC, especially in a KRAS-mutated environment. Cdk5 immunostaining predicts outcome of stage IV CRC patients treated with OXA. Extensive data will be presented during the EACR meeting.

BACKGROUND: Non-hereditary colorectal cancer (CRC) is a complex disorder resulting from the combination of genetic and non-genetic factors. Genome-wide association studies (GWAS) are useful for identifying such genetic susceptibility factors. However, the single loci so far associated with CRC only represent a fraction of the genetic risk for CRC development in the general population. Therefore, many other genetic risk variants alone and in combination must still remain to be discovered. The aim of this work was to search for genetic risk factors for CRC, by performing single-locus and two-locus GWAS in the Spanish population. RESULTS: A total of 801 controls and 500 CRC cases were included in the discovery GWAS dataset. 77 single nucleotide polymorphisms (SNP)s from single-locus and 243 SNPs from two-locus association analyses were selected for replication in 423 additional CRC cases and 1382 controls. In the meta-analysis, one SNP, rs3987 at 4q26, reached GWAS significant p-value (p = 4.02×10(-8)), and one SNP pair, rs1100508 CG and rs8111948 AA, showed a trend for two-locus association (p = 4.35×10(-11)). Additionally, our GWAS confirmed the previously reported association with CRC of five SNPs located at 3q36.2 (rs10936599), 8q24 (rs10505477), 8q24.21(rs6983267), 11q13.4 (rs3824999) and 14q22.2 (rs4444235). CONCLUSIONS: Our GWAS for CRC patients from Spain confirmed some previously reported associations for CRC and yielded a novel candidate risk SNP, located at 4q26. Epistasis analyses also yielded several novel candidate susceptibility pairs that need to be validated in independent analyses.

BACKGROUND To assess the putative prognostic value of K-ras mutations in nonmucinous ovarian tumors, the authors looked for K-ras point mutations at codons 12 and 13 in 144 nonmucinous ovarian tumors. METHODS A series of 144 consecutive, unselected, archival, nonmucinous ovarian tumors (35 benign, 12 borderline, and 97 malignant) were studied. K-ras mutations at codons 12 and 13 were determined by polymerase chain reaction using the restriction fragment length polymorphism method with mismatched nested primers. Extensive clinicopathologic and follow-up data on all patients were evaluated. RESULTS The overall prevalence of K-ras mutations at codons 12 and 13 was 30.5% (44/144). In benign tumors, it was 20% (7/35); in borderline tumors, 25% (3/12); and in carcinomas, 35% (34/97). The presence of K-ras point mutations did not correlate with survival. Among the benign tumors, K-ras mutations were detected in three Brenner tumors with a mucinous component. CONCLUSIONS These results indicate that K-ras mutations are not initial events in the pathogenesis of nonmucinous ovarian tumors and do not appear to be related to survival. Cancer 1998;82:1088-95. © 1998 American Cancer Society.

3750 Background: Colon cancer is the second neoplasm in men and women. Its prognostic depends on TNM staging at diagnosis. The treatment seems to improve the survival. The aim of this study is to analyze our series of surgical colon cancer from 1996 to 2001. Methods: We analyzed 422 surgical colon cancer. 70 patients were metastatic at the time of diagnosis or behind surgery (5 months). The remainder 352 patients without metastases were analyzed. The prognosis factors were: age at diagnosis, gender, locations in the large bowel, kind of surgery, pT, pN, stage, use of chemotherapy, disease free survival (DFS) and overall survival (OS). Results: The 352 non metastatic surgical colon cancer presented a mean age at diagnosis of 70,22 years (24 to 101). The ratio male/female was 197/155. The location in the bowel was: right 149 cases and left 203 cases. pT: pTis 6, pT114, pT2 43, pT3 244, pT4 41. pN: pN0 + pNX 229, pN1 70, pN2 52. 145 patients received adjuvant chemotherapy. Stage: I:49; II:171; III:122.pT: DFS was pT1=100%, pT2=93%, pT3=73%, pT4=51,2%. OS was pT1=71,4%, pT2=83,7%, pT3=66%, pT4=48,8%.pN: DFS was pN0=83,4%, pN1=64,3%,pN2=50%. OS was pN0=70,3%, pN1=61,4%, pN2=55,8%. Stage: OS was I=80%, II=68%, III=59%. Conclusions: The main prognostic factor for survival in colon cancer was the staging at diagnosis. Patients with pT4 or pN2 should received aggressive treatments for the high risk of metastasis. No significant financial relationships to disclose.

We aimed to gain further understanding of the molecular mechanisms involved in oxaliplatin resistance in colorectal cancer by using a proteomic approach. A 5-fold oxaliplatin-resistant cell line, HTOXAR3, was compared with its parental cell line, HT29, using two-dimensional PAGE. Mass spectrometry, Western blot, and real-time quantitative PCR confirmed the down-regulation of pyruvate kinase M2 (PK-M2) in HTOXAR3 cells. In a panel of eight colorectal cancer cell lines, we found a negative correlation between oxaliplatin resistance and PK-M2 mRNA levels (Spearman r=-0.846, P=0.008). Oxaliplatin exposure in both HT29 and HTOXAR3 led to PK-M2 mRNA up-regulation. PK-M2 mRNA levels were measured by real-time quantitative PCR in 41 tumors treated with oxaliplatin/5-fluorouracil. Tumors with the lowest PK-M2 levels attained the lowest response rates (20% versus 64.5%, P=0.026). High PK-M2 levels were associated with high p53 levels (P=0.032). In conclusion, the data provided clearly link PK-M2 expression and oxaliplatin resistance mechanisms and further implicate PK-M2 as a predictive marker of response in patients with oxaliplatin-treated colorectal cancer.

BACKGROUND: Progression from actinic keratosis (AK) to invasive squamous cell carcinoma (iSCC) of the skin is thought to occur after the development of full thickness epidermal neoplasia, as in the classic pathway of cervical cancer. Nevertheless, cutaneous iSCC may also directly arise from a proliferation of atypical basaloid cells limited mostly to the epidermal basal layer (AK I), akin to what happens in the 'differentiated pathway' of iSCC of the vulva, oral cavity and other locations. OBJECTIVE: To evaluate the prevalence of classic and differentiated pathways in the development of cutaneous iSCC. METHODS: The epidermis adjacent to and overlying iSCC, assumed to be representative of pre-existing lesions, was histologically studied in 196 skin biopsy specimens showing iSCC. RESULTS: AK I, AK II and AK III lesions overlying iSCC were present in 63.8%, 17.9% and 18.4% of cases respectively. The corresponding percentages in the epidermis adjacent to iSCC were 77.9%, 6.6% and 8.3% respectively (stage could not be assessed in 8.1% of cases). Focal epidermal ulceration overlying iSCC was seen in 32% of AK I, 28.6% of AK II and 33.3% of AK III instances. Adnexal involvement by atypical keratinocytes (proliferative AK) was present more frequently in cases with overlying AK I (39/125, 31.2%) than with AK II (8/35, 22.9%) and AKII I (5/36, 13.9%). CONCLUSION: Direct invasion from proliferating basaloid atypical keratinocytes limited to the epidermal basal layer (AK I), known as the differentiated pathway, was the most common form of progression to cutaneous iSCC in our series. On the other hand, stepwise progression from AK I to AK II and AK III (classic pathway) was seen to be operative in a substantial proportion of iSCC cases. All AK lesions, irrespective of intraepidermal neoplasia thickness, are therefore potentially invasive and tumour advance along adnexal structures might facilitate iSCC development from AK I lesions.