PDF file - 346K, Analysis of CTC03 of patient #22: The array-CGH ratio profile demonstrates losses on chromosomes 3, 4, 5, 8p, and 18 and gains on chromosomes 7p, 17q, and 20.

PDF file - 92K, Identification of mutations in cancer candidate (CAN) genes in patient samples using next-generation sequencing. Mutations that are shared between primary tumor and/or metastasis and/or CTCs are highlighted in bold.

PDF file - 868K, Ratio bin distributions for CTC12 (a) and CTC03 (b) from patient #6. The large bin 13 peak indicates the frequency of oligonucleotides with balanced ratio values. The bins to the right of 13 represent the bins for oligonucleotides with deleted, the bins to the left oligonucleotides with increased values. (a) In CTC12 only one of the bins for deleted values was increased (i.e. bin 15), suggesting that this cell is diploid. In a diploid cell the log2 ratio for balanced values ranges from 0.295 to -0.293 and therefore extends to bins adjacent to 13, which is reflected by the small increases in the bins 12 and 14. The peaks at bins 11 and 12 reflect trisomies (i.e. a 3:2 ratio or an integer copy number of 3), the peak at 9 indicates tetrasomies (i.e. 4:2) or 4 copies). (b) CTC03 had clearly more discrete deleted copy number states visible at bins 15 to 18, suggesting that this cell is tetraploid. In a tetraploid cell the log2 ratio range for balanced values is narrower (i.e. 0.161 to -0.208) as in a diploid cell, which is reflected in the almost empty bins 12 and 14. Bin 15 represents oligonucleotides with ratio values translating to 3 copies, bins 16 and 17 to two copies and bin 18 to one copy. Bin 10 represents 5 copies, bins 8 and 9 six copies, bin 7 seven copies, and bin 4 eight copies.

PDF file - 188K, Sequencing of UCR41 and flanking regions: UCR41 is a 217-bp-long ultra-conserved region depicted in dark green. The gray bar indicates the extremely conserved sequence. The region is flanked by two SNPs that are frequent in the European population (blue arrows). The black curve indicates the mutability, which is higher in non-conserved regions compared with conserved segments. The black bars illustrate the 5 overlapping amplicons used for amplification. Mutations identified in our samples are indicated with red arrows. The genomic coordinates refer to the hg18 assembly of the human genome. (The graph is reproduced with permission from De Grassi et al.)

PDF file - 940K, Evaluation of available material from patient #6. (a-b) Array CGH profiles of the primary tumor (a) and metastasis (b). The multicolor bar codes at the top or bottom of the ratio profiles illustrate the results obtained during the iterative calculations with various window sizes, the single green and red bars summarize the regions which were gained or lost based on all calculations (Supplementary Methods). Black parts in the profile represent balanced regions, lost regions appear in red and gained regions in green.

PDF file - 1303K, Evaluation of available material from patient #9. (a-b) Array CGH profiles of the primary tumor (a) and metastasis (peritoneal carcinomatosis) (b). Regarding the color bar codes please see legend to Supp. Fig. 2.

PDF file - 95K, List of 68 genes, which, according to the COSMIC database, are frequently mutated in colorectal carcinoma (CRC) and analyzed in our study.

PDF file - 3383K, Calculation of different integer copy number profiles for CTC03 from patient #6 assuming diploidy (a), triploidy (b), or tetraploidy (c) and the respective array CGH profile (d).

PDF file - 1362K, Evaluation of available material from patient #38. (a-b) Array CGH profiles of the primary tumor (a) and liver metastasis (b).

PDF file - 1233K, Additional CTCs from patient #26. Individual CTC-integer number profiles from CTCs 05 (a), 24 (b), 25 (c), 28 (d).

PDF file - 146K, Additional clinical data and detailed Material & Methods section

PDF file - 98K, Summary of clinical characteristics of the 6 colorectal carcinoma (CRC) patients.

PDF file - 231K, Evaluation of available material from patient #6. (c) Integer copy number profiles of CTC14.

Summary The purpose of this observational study was to determine the prevalence of comorbid conditions in cancer patients with solid tumours selected for specific treatment at 12 divisions of medical oncology in Austria. Data from 1137 patients were collected using a standardized questionnaire; of these, 1036 datasets were evaluable for further analysis. Data were prospectively collected from patients during an in- or outpatient hospital visit over a 4-month period in 2011. Of these patients 42% had gastrointestinal cancer, 31% had breast cancer, 9% lung cancer and the remaining had urogenital cancer, sarcoma or other types of rare cancers. Around two-thirds of patients had metastatic disease (59%), confined to a single organ site in 55% of patients. A high proportion of patients had a good performance status (Eastern Cooperative Oncology Group [ECOG] 0, 1: 82%). Comorbid conditions were classified according to the Charlson scheme score and were present in 86% of patients with a median age of 64 years. The predominant conditions were cardiovascular diseases (57%), metabolic diseases (44%), endocrinological diseases (30%), gastrointestinal diseases (26%), neurological (23%) and respiratory diseases (23%). As has been reported by others we found a clear association between number of comorbid conditions and age. While 60% of the whole population had at least 2 comorbidities, most patients of the elderly population (89%) had more than three comorbidities. The high proportion of patients with comorbidities and accompanying medication represents a substantial challenge for medical oncologists in selecting the optimal cancer-specific treatment especially in the era of novel targeted and immunotherapies. Comorbid conditions and accompanying comedications require special precautions concerning potential interactions and unexpected adverse reactions from prescribed tumour-specific treatment.

Objective The aim of this study was to determine a potential benefit of the specific psychoeducational intervention Learning to Live with Cancer (LTLWC) for patients with operated nonmetastatic breast cancer, with respect to psychological variables and endocrine and immune parameters. Methods Fifty-two postmenopausal women with operated stage I to III breast cancer were randomized to either a breast cancer intervention group (BCIG, n = 30) who immediately began participating in the LTLWC intervention program or to a breast cancer control group (BCCG, n = 22). Matched healthy women were asked to participate as a noncancer comparison group (n = 26). All participants were evaluated at three different time points (t1-t3) using a set of standardized questionnaires and blood samples were taken to analyze immune cell subsets and stress hormone levels. Results A significant reduction in trait anxiety/State Trait Anxiety Inventory score was observed in the BCIG (t1: median = 35.0 [interquartile range = 28.0-38.0] versus t3: median = 26.0 [interquartile range = 18.5-37.0], p = .0001) compared with the BCCG (t1: median = 41.0 [interquartile range =32.75-49.0]; t3: median = 38.5 [interquartile range = 30.75-46.5], p = .01524; p interaction = .001). In parallel, a significant rise of serotonin levels (t1: median = 66.5 ng/ml [interquartile range = 11.50-106.00] versus t3: median = 80.5 ng/ml [interquartile range =59.00-118.00], p = .00008) as well as a significant reduction of the elevated number of Treg cells at baseline (t1: median = 4.45% [interquartile range = 4.00-5.33] versus t3: median = 2.80% [interquartile range = 2.68-3.13], p < .00001) were observed in the BCIG versus no change in the BCCG. A significant statistical association between reduced trait anxiety and decreased Treg cell number could be demonstrated in the BCIG (r = .62, p < .01). Conclusions The observed results of this study provide preliminary support for the efficacy of the LTLWC program in significantly improving psychoneuroimmunological parameters in patients with nonmetastatic breast cancer.

AIM:To evaluate feasibility and safety of neoadjuvant chemotherapy with capecitabine, oxaliplatin and bevacizumab followed by concomitant standard chemoradiation and surgical resection in patients with high-risk locally advanced rectal cancer.PATIENTS AND METHODS:Magnetic resonance imaging (MRI)-defined high-risk cT3/4 rectal cancer patients were treated with 3 cycles of neoadjuvant chemotherapy with capecitabine (1,000 mg/m2 twice daily days 1-14, 22-35, 43-56), oxaliplatin (130 mg/sqm on days 1, 22, 43) and bevacizumab (7.5 mg/kg on days 1, 22, 43) followed by capecitabine (825 mg/m2 twice daily on radiotherapy days week 1-4) concomitantly with radiotherapy (1.8 Gy daily up to 45 Gy in 5 weeks) and surgical resection by total mesorectal excision. Feasibility, safety, response rate and postoperative morbidity were evaluated.RESULTS:Twenty-five patients were recruited. Median age was 62 years (range=24-78 years) and all patients had Eastern Cooperation Oncology Group (ECOG) performance status 0. From all patients, 79.2% finished neoadjuvant chemotherapy. Twenty patients underwent surgery. Pathologic complete remission rate, R0 resection and T-downstaging were achieved in 25%, 95% and 54.2% of the "intention to treat" (ITT) patients. The most common grade 3 adverse events (AEs) during neoadjuvant chemotherapy were diarrhea (16.6%) and mucositis (12.5%). In one patient, a grade 4 acute renal failure occurred (4.2%). During chemoradiation, skin reactions (5.3%) were the most common grade 3 AEs. Two major perioperative complications required re-intervention.CONCLUSION:Neoadjuvant chemotherapy with bevacizumab, capecitabine and oxaliplatin followed by concomitant standard chemoradiation is feasible in patients with high-risk locally advanced rectal cancer (LARC) and resulted in complete pathologic remission (pCR) rate of 25% and neoadjuvant chemotherapy completion rate of 80%.

Background Patients with testicular germ cell tumors (TGCT) have an increased risk for venous thromboembolism (VTE). We identified risk factors for VTE in this patient cohort and developed a clinical risk model. Methods In this retrospective cohort study at the Medical University of Graz we included 657 consecutive TGCT patients across all clinical stages. A predictive model for VTE was developed and externally validated in 349 TGCT patients treated at the University Hospital Zurich. Results Venous thromboembolic events occurred in 34 (5.2%) patients in the Graz cohort. In univariable competing risk analysis, higher clinical stage (cS) and a retroperitoneal lymphadenopathy (RPLN) were the strongest predictors of VTE (p<0.0001). As the presence of a RPLN with more than 5cm in greatest dimension without coexisting visceral metastases is classified as cS IIC, we constructed an empirical VTE risk model with the following four categories (12-month-cumulative incidence): cS IA-B 8/463 patients (1.7%), cS IS-IIB 5/86 patients (5.9%), cS IIC 3/21 patients (14.3%) and cS IIIA-C 15/70 patients (21.4%). This risk model was externally validated in the Zurich cohort (12-month-cumulative incidence): cS IA-B (0.5%), cS IS-IIB (6.0%), cS IIC (11.1%) and cS IIIA-C (19.1%). Our model had a significantly higher discriminatory performance than a previously published classifier (RPLN-VTE-risk-classifier) which is based on the size of RPLN alone (AUC-ROC: 0.75 vs. 0.63, p = 0.007). Conclusions According to our risk stratification, TGCT patients with cS IIC and cS III disease have a very high risk of VTE and may benefit from primary thromboprophylaxis for the duration of chemotherapy.

We investigated the hypothesis that the varying treatment efficacy of adjuvant 5-fluorouracil (5FU) in stage III colon cancer is linked to the TP53 mutational status.ABCSG-90 was a prospective randomized trial in which effect of adjuvant 5FU was studied in stage III colon cancer patients. Tumor material of 70% of these patients (389/572) was available for analysis of the biomarker TP53 using a TP53-gene-specific Sanger sequencing protocol.Median follow-up was 88 months. TP53 mutation frequency was 33%. A significant interaction between TP53 status, outcomes and nodal category was found (P = 0.0095). In the N1 category, TP53 wildtype patients had significantly better overall survival than TP53 mutated (81.0% vs. 62.0% overall survival at 5 years; HR = 2.131; 95% CI: 1.344-3.378; P = 0.0010). In the N2 category, the TP53 status did not affect survival (P = 0.4992). In TP53 wildtype patients, the prognostic significance of N category was significantly enhanced (P = 0.0002). In TP53 mutated patients, survival curves of N1 and N2 patients overlapped and nodal category was no longer prognostic. The biomarker TP53 independently predicted effect of adjuvant 5FU in N1 colon cancer patients. TP53 was not predictive in N2 patients, in whom 5FU is known to have no effect. (C) 2015 The Authors. Published by Elsevier B.V.