BACKGROUND: Helicobacter pylori-associated peptic ulcer is a frequent complication in cirrhotic patients and its morbidity rate is high. In spite of this, diagnostic methods for H. pylori infection have not been fully evaluated in these patients. AIM: To evaluate H. pylori diagnostic methods in patients with liver cirrhosis. METHODS: One hundred and one cirrhotic patients were included in the study. Three antral and two corpus biopsies were obtained for rapid urease test of the antral mucosa, and Giemsa stain and immunohistochemistry were performed for both the corpus and antrum. Serology, 13C-urea breath test and faecal H. pylori antigen determination were also carried out. RESULTS: Sixty-two patients were positive and 35 were negative for H. pylori infection; four were indeterminate. The sensitivity and specificity were 90.4% and 100%, respectively, for antral histology, 100% and 100% for gastric body histology, 90.4% and 100% for antral immunohistochemistry, 96.2% and 96.7% for body immunochemistry, 85.7% and 97% for rapid urease test, 83.6% and 55.9% for serology, 96.4% and 97.1% for 13C-urea breath test and 75.4% and 94.1% for faecal antigen. CONCLUSION: The most reliable tests for H. pylori infection in cirrhotic patients were the 13C-urea breath test and gastric body histology.

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

BACKGROUND AND AIM: The majority of mismatch repair (MMR) gene mutations causing Lynch syndrome (LS) occur either in MLH1 or MSH2. However, the relative contribution of PMS2 is less well defined. The aim of this study was to evaluate the role of PMS2 in LS by assessing the pathogenicity of variants of unknown significance (VUS) detected in the mutational analysis of PMS2 in a series of Spanish patients. METHODS: From a cohort of 202 LS suspected patients, 13 patients showing loss of PMS2 expression in tumours were screened for germline mutations in PMS2, using a long range PCR based strategy and multiplex ligation dependent probe amplification (MLPA). Pathogenicity assessment of PMS2 VUS was performed evaluating clinicopathological data, frequency in control population and in silico and in vitro analyses at the RNA and protein level. RESULTS: Overall 25 different PMS2 DNA variants were detected. Fourteen were classified as polymorphisms. Nine variants were classified as pathogenic: seven alterations based on their molecular nature and two after demonstrating a functional defect (c.538-3C>G affected mRNA processing and c.137G>T impaired MMR activity). The c.1569C>G variant was classified as likely neutral while the c.384G>A remained as a VUS. We have also shown that the polymorphic variant c.59G>A is MMR proficient. CONCLUSIONS: Pathogenic PMS2 mutations were detected in 69% of patients harbouring LS associated tumours with loss of PMS2 expression. In all, PMS2 mutations account for 6% of the LS cases identified. The comprehensive functional analysis shown here has been useful in the classification of PMS2 VUS and contributes to refining the role of PMS2 in LS.

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.

Chemoresistance is the main cause of treatment failure in advanced colorectal cancer (CRC). However, molecular mechanisms underlying this phenomenon remain to be elucidated. In a previous work we identified low levels of PKM2 as a putative oxaliplatin-resistance marker in HT29 CRC cell lines and also in patients. In order to assess how PKM2 influences oxaliplatin response in CRC cells, we silenced PKM2 using specific siRNAs in HT29, SW480 and HCT116 cells. MTT test demonstrated that PKM2 silencing induced resistance in HT29 and SW480 cells and sensitivity in HCT116 cells. Same experiments in isogenic HCT116 p53 null cells and double silencing of p53 and PKM2 in HT29 cells failed to show an influence of p53. By using trypan blue stain and FITC-Annexin V/PI tests we detected that PKM2 knockdown was associated with an increase in cell viability but not with a decrease in apoptosis activation in HT29 cells. Fluorescence microscopy revealed PKM2 nuclear translocation in response to oxaliplatin in HCT116 and HT29 cells but not in OXA-resistant HTOXAR3 cells. Finally, by using a qPCR Array we demonstrated that oxaliplatin and PKM2 silencing altered cell death gene expression patterns including those of BMF, which was significantly increased in HT29 cells in response to oxaliplatin, in a dose and time-dependent manner, but not in siPKM2-HT29 and HTOXAR3 cells. BMF gene silencing in HT29 cells lead to a decrease in oxaliplatin-induced cell death. In conclusion, our data report new non-glycolytic roles of PKM2 in response to genotoxic damage and proposes BMF as a possible target gene of PKM2 to be involved in oxaliplatin response and resistance in CRC cells.

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.

We report a case of a 68-year-old man who developed an advanced Triple-Negative Breast Cancer (TNBC) presented as a Carcinoma of Unknown Primary (CUP) at diagnoses. After progressing to first line therapy, the molecular platform EPICUP® was performed and the results suggested breast cancer as origin, although non breast primary tumor was found by imaging techniques; and the patient was treated according to this origin. During the evolution of the disease a nodule appeared in the left breast, and a biopsy confirmed the diagnosis of an infiltrating ductal carcinoma. Male breast cancer (MBC) represents an infrequent and heterogeneous disease, but TNBC in men represents less than 0.02% of total breast cancer cases. Unfortunately, there is no standard of care for MBC, and due to the development of new treatment strategies are focused on women BC, there are less opportunities to participate in clinical trials for MBC. Through the review of a clinical case, we show how the molecular platform could help in finding the origin of the CUP. Moreover, further coordinated multicenter international collaboration is necessary to conduct larger and successful therapeutic clinical trials in MBC, and specifically, male TNBC trials to better understand this rare entity and guide future management strategies.

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.

Abstract Background/purpose: Despite remaining one of the most active drugs in the treatment of advanced colorectal cancer (aCRC), oxaliplatin (OXA) resistance still is a multifactorial and complex process to decipher. In a previous work we identified low levels of PKM2 (mRNA and protein) as a putative OXA-resistance marker in CRC cell lines and patients. The purpose of this work was to identify molecular mechanisms underlying PKM2 role in resistance to OXA. Material and methods: PKM2 transient gene silencing was achieved by transfecting specific siRNAs (Ambion) in HT29 and HCT116 CRC cell lines. Gene silencing was validated by qPCR and Western Blot with specific Taqman primers and probes (assay no. Hs00762869_s1) and antibody (Cell signaling), respectively. Cell proliferation was measured by the MTT assay (Roche Diagnostics). Inhibitory concentrations were determined in each cell line by the median-effect line method. Cell viability was studied with the use of trypan blue stain and apoptosis was detected with FITC Annexin V Apoptosis Detection Kit I (BD Pharmingen). PKM2 subcellular localization was studied by fluorescence microscopy in an Axiovision Z1 by using Apotome system at 40x immersion oil lens (Carl Zeiss, Heidelberg, Germany). Expression patterns of cell death genes were analyzed through Human Cell Death Pathway Finder PCR Array 384 HT (PAHS-212Z, SA Biosciences). Results: In order to assess how PKM2 expression influences OXA response in CRC cells, we silenced PKM2 by using specific siRNAs in HT29 and HCT116 cells. Cells were then treated with OXA at different concentrations and we observed that PKM2 silencing induced resistance in HT29 (p53 mutated) cells and sensitivity in HCT116 (p53 wild type) cells. Moreover, PKM2 knockdown was associated with an increase in cell viability but not with a decrease in apoptosis activation in HT29 cells. Fluorescence microscopy revealed PKM2 nuclear translocation in response to OXA in HCT116 and HT29 cells but not in OXA-resistant HTOXAR3 cells. Finally, by using a qPCR Array we demonstrated that OXA and PKM2 silencing altered cell death gene expression patterns including BMF, which was significantly increased in HT29 cells in response to OXA in a dose and time-dependent manner but not in siPKM2-HT29 and HTOXAR3 cells. Conclusion: Our data report new non-glycolytic roles of PKM2 in response to genotoxic damage, which depend on p53 mutational status and proposes BMF as a possible target gene of PKM2 to be involved in OXA response and resistance in CRC cells. Citation Format: Alba Ginés, Anna Martínez-Cardús, Vicenç Ruiz de Porras, Eva Musulén, José Luis Manzano, Laura Layos, Cristina Bugés, Albert Abad, Eva Martinez-Balibrea. PKM2 subcellular localization is involved in oxaliplatin resistance acquisition in human colorectal cancer cell lines. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3752. doi:10.1158/1538-7445.AM2014-3752

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.

PURPOSE: To evaluate proton fat-water chemical shift fast low-angle shot magnetic resonance (MR) imaging for differentiation of fat-containing hyperechoic liver nodules from hyperechoic liver nodules without a fatty component. MATERIALS AND METHODS: T1-weighted fast low-angle shot fat-water chemical shift gradient-echo MR imaging was performed in 96 patients without cirrhosis with 138 hyperechoic liver nodules. In-phase and opposed-phase breath-hold images were acquired. The percentage of signal intensity variation between in-phase and opposed-phase images and the spleen-to-lesion contrast ratio were used to differentiate liver nodules. RESULTS: Chemical shift MR images showed fat in 15 (11%) hyperechoic nodules (two angiomyolipomas and 13 nodular fatty infiltrations of the liver). The mean percentage of signal intensity variation between in-phase and opposed-phase images was 156% (standard error, 43.5%) in nodules with fat and -0.16% (standard error, 0.96%) in nodules without fat (P = .003). Spleen-to-lesion contrast was similar on in- and opposed-phase images in lesions without fat (mean difference, -0.0107; standard error, 0.012), whereas the mean difference in fat-containing nodules was 0.805 (standard error, 0.225; P = .003). The area under the receiver operating characteristic curve was 0.97 for signal intensity variation. CONCLUSION: Hyperechogenicity in certain liver nodules is caused by fat. Chemical shift MR imaging allows accurate differentiation between these and other hyperechoic lesions with no fat component.