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Mutations of the tumor suppressor E-cadherin and overexpression of the receptor tyrosine kinase epidermal growth factor receptor (EGFR) are among the most frequent genetic alterations associated with diffuse-type gastric carcinoma. Accumulating evidence suggests a functional relationship between E-cadherin and EGFR that regulates both proteins. We report that somatic mutation of E-cadherin is associated with increased activation of EGFR followed by enhanced recruitment of the downstream acting signaling components growth factor receptor binding protein 2 and Shc, and activation of Ras. Reduced complex formation of mutant E-cadherin — with an in frame deletion of exon 8 in the extracellular domain resulting in reduced adhesion and increased motility — with EGFR was observed compared with wild-type E-cadherin. We conclude that reduced binding of mutant E-cadherin to EGFR in a multicomponent complex or reduced stability of the complex may enhance EGFR surface motility, thereby facilitating EGFR dimerization and activation. Furthermore, reduced surface localization due to enhanced internalization of mutant E-cadherin compared with the wild-type protein was observed. The internalization of EGFR was decreased in response to epidermal growth factor stimulation in cells expressing mutant E-cadherin, suggesting that mutation of E-cadherin also influences the endocytosis of EGFR. Moreover, we show increased activation of EGFR in gastric carcinoma samples with mutant E-cadherin lacking exons 8 or 9. In summary, we describe activation of EGFR by mutant E-cadherin as a novel mechanism in tumor cells that explains the enhanced motility of tumor cells in the presence of an extracellular mutation of E-cadherin. [Cancer Res 2008;68(3):707–14]

Supplementary Tables S1-S3 from Enhanced Activation of Epidermal Growth Factor Receptor Caused by Tumor-Derived E-Cadherin Mutations

Supplementary Results, Figure 1, Methods and Materials from Enhanced Activation of Epidermal Growth Factor Receptor Caused by Tumor-Derived E-Cadherin Mutations

Doxorubicin represents the standard first-line treatment for metastatic soft-tissue sarcoma. We assessed the efficacy and safety of trofosfamide in elderly patients. In this controlled phase II trial, we randomly (1:2) assigned 120 previously untreated patients with soft-tissue sarcoma, older than 60 years, with an Eastern Cooperative Oncology Group score of 0–2, to receive either doxorubicin for 6 cycles (arm A) or oral trofosfamide (arm B). The primary end-point was a 6-month progression-free rate (PFR) in the experimental arm (clinical trial information: NCT00204568). Between August 2004 and October 2012, forty and 80 patients were randomly assigned to arm A and arm B, respectively, in 16 centres. The median age was 70 years (range, 60–89). The primary study end-point (6-month PFR) was exceeded, with 27.6% in arm B (95% confidence interval [CI], 18.0–39.1) and 35.9% in arm A: (95% CI, 21.2–52.8). Survival data in terms of progression-free survival were 4.3 months (95% CI, 2.2–6.3) and 2.8 months (95% CI, 1.7–3.6) and in terms of overall survival were 9.8 months (95% CI, 6.7–11.6) and 12.3 months (95% CI, 9.6–16.2), respectively. The number of serious adverse event (SAE) was 59% in arm A and 30.3% in arm B (p = 0.005). Trofosfamide caused more often dyspnoea and low-grade fatigue, whereas with doxorubicin, more often leukocytopenia, neutropenia and mucositis were seen. Discontinuation rates for reasons other than disease progression were 15.4% (arm A) vs. 7.9% (arm B). In an elderly population of patients, oral trofosfamide achieved the estimated primary end-point 6-month PFR and was associated with a favourable toxicity profile compared with doxorubicin.

Emerging evidence shows that severe coronavirus disease 2019 (COVID-19) can be complicated with coagulopathy, namely disseminated intravascular coagulation, which has a rather prothrombotic character with high risk of venous thromboembolism. The incidence of venous thromboembolism among COVID-19 patients in intensive care units appears to be somewhat higher compared to that reported in other studies including such patients with other disease conditions. D-dimer might help in early recognition of these high-risk patients and also predict outcome. Preliminary data show that in patients with severe COVID-19, anticoagulant therapy appears to be associated with lower mortality in the subpopulation meeting sepsis-induced coagulopathy criteria or with markedly elevated d-dimer. Recent recommendations suggest that all hospitalized COVID-19 patients should receive thromboprophylaxis, or full therapeutic-intensity anticoagulation if such an indication is present.

Longitudinal data on the E6/E7 mRNA-based Aptima human papillomavirus (AHPV) assay exceeding three years in comparison to the gold standard Digene Hybrid Capture 2 (HC2) test are not available. We previously reported the cross-sectional data of the German AHPV Screening Trial (GAST) in which 10,040 women were recruited and tested by liquid-based cytology, the HC2 assay, and the AHPV assay., Longitudinal data on the E6/E7 mRNA-based Aptima human papillomavirus (AHPV) assay exceeding three years in comparison to the gold standard Digene Hybrid Capture 2 (HC2) test are not available. We previously reported the cross-sectional data of the German AHPV Screening Trial (GAST) in which 10,040 women were recruited and tested by liquid-based cytology, the HC2 assay, and the AHPV assay. Four hundred eleven test-positive women were followed for up to six years. In addition, 3,295 triple-negative women were screened after a median time of six years. Overall, 28 high-grade cervical intraepithelial neoplasia (CIN3) cases were detected. The absolute risk of developing high-risk HPV-positive CIN3+ over six years among those women that tested negative at baseline was 2.2 (95% confidence interval [95% CI], 1.0 to 4.9) and 3.1 (95% CI, 1.7 to 5.7) per 1,000 women screened by the HC2 and the AHPV tests; the additional risk for those with AHPV-negative compared with HC2-negative results was 0.9 (95% CI, −0.2 to 2.1) per 1,000. In comparison, the absolute risk following a negative LBC test was 9.3 (95% CI, 2.9 to 30.2). The relative sensitivity of AHPV compared to HC2 was 91.5% for CIN3+, and the negative predictive values were 99.8% (95% CI, 99.5 to 99.9%) for HC2 and 99.7% (95% CI, 99.4 to 99.8%) for AHPV. Our data show that the longitudinal performance of the AHPV test over six years is comparable to the performance of the HC2 test and that the absolute risk of CIN3+ over six years following a negative AHPV result in a screening population is low. (This study is registered at ClinicalTrials.gov under registration number NCT02634190.)

Testing for E6/E7 mRNA in cells infected with high-risk (HR) human papillomavirus (HPV) might improve the specificity of HPV testing for the identification of cervical precancerous lesions. Here we compared the RNA-based Aptima HPV (AHPV) assay (Hologic) and the DNA-based Hybrid Capture 2 (HC2) HPV test (Qiagen) to liquid-based cytology (LBC) for women undergoing routine cervical screening. A total of 10,040 women, 30 to 60 years of age, were invited to participate in the study, 9,451 of whom were included in the analysis. Specimens were tested centrally by LBC, the AHPV test, and the HC2 test, and women who tested positive on any test were referred for colposcopy. Genotyping was performed on all HR-HPV-positive samples. Test characteristics were calculated based on histological review. As a result, we identified 90 women with cervical intraepithelial neoplasia grade 2+ (CIN2+), including 43 women with CIN3+. Sensitivity differences between the AHPV test and the HC2 test in detecting CIN2+ ( P = 0.180) or CIN3+ ( P = 0.0625) lesions were statistically nonsignificant. Of three CIN3 cases that were missed with the AHPV test, two cases presented lesion-free cones and one had a non-HR HPV67 infection. The specificity (P < 0.001) than those of the HC2 test. The overall agreement between the tests was substantial (κ = 0.77). Finally, we present results for several possible triage strategies, based on the primary screening test being either the AHPV test or the HC2 test. In summary, the AHPV assay is both specific and sensitive for the detection of high-grade precancerous lesions and may be used in primary cervical cancer screening for women ≥30 years of age.

11507Background: DOX is still the standard in metastatic STS. We assessed the efficacy and safety of oral TRO. Methods: This is a randomized phase 2 trial at 15 german and 1 french centers. We incl...

Objective To measure perfusion in different lung cancer subtypes and compare results with histopathological/immunohistochemical results. Methods Seventy-two consecutive untreated patients with lung cancer (40 adenocarcinomas, 20 squamous cell, and 12 small cell lung cancers) were enrolled. A 40-second volume perfusion computed tomography of the tumor bulk was obtained. Blood flow (BF), blood volume (BV), and transit constant were determined. Tumor volume and tumor necrosis were determined on contrast-enhanced computed tomography. Pathologic specimens were assessed for microvessel density (MVD), hypoxia-induced transcription (hif-1/-2), and proliferation (Ki-67). Results Higher MVD is associated with higher BF and BV. Higher tumor grade leads to lower BF but increased necrosis and tumor volume. Markers of hypoxia were independent from perfusion parameters, extent of necrosis or MVD. Blood flow, BV, and MVD were not significantly different among lung cancer subtypes. Transit constant was significantly reduced in small cell lung cancer versus adenocarcinoma. Conclusions Perfusion values are related to MVD and tumor grade but vary considerably among lung cancer subtypes.

Background: Cryoextraction is a procedure used for the recanalization of obstructed airways caused by visible exophytic endobronchial tumor. Biopsy samples obtained by this technique have been shown to be useful for histological assessment. Objectives: The aim of the present animal study was to systematically evaluate biopsy size, histological quality and bleeding risk after cryobiopsy with new, flexible cryoprobes in comparison with forceps biopsy, serving as the gold standard. Methods: Biopsies were obtained from anesthetized pigs with the flexible bronchoscopy technique, and evaluated histologically with respect to their size and quality. Bleeding frequency, bleeding duration and histological changes in the biopsy bed were also recorded. Results: Cryobiopsies were significantly larger than forceps biopsies. The size of cryobiopsies was dependent on the freezing time. The histological quality of the cryobiopsy specimenswas not impaired by the freezing process, whereas forceps biopsies showed typical crush artifacts. Despite the larger defects left in the tracheobronchial system after cryobiopsy, bleeding frequency and duration were not higher compared to forceps biopsy. Conclusions: Since cryobiopsy sampling is not associated with a higher bleeding risk compared with forceps biopsy, this new biopsy technique offers – in addition to a good specimen quality – a safe and valuable tool with the potential of improving the outcome of diagnostic endoscopy.

Background For endoscopic submucosal dissection (ESD), adequate exposure and visualization of the submucosa for controlled dissection is of eminent importance. Objective To determine the feasibility and safety of percutaneously assisted endoscopic surgery (PA-ES) with a new prototype PEG-minitrocar (PMT) for advanced ESD in a porcine model. Interventions Placement of the PMT was done in all pigs by the use of a modified pull-through technique. After endoscopic incision of the mucosa, traction was provided for ESD by grasping the incisional margins of the mucosa with a rigid forceps introduced through the PMT, enabling stepwise dissection of the exposed submucosa under direct vision. Main Outcome Measurements Feasibility and safety of the new PMT for PA-ES and en bloc resection of prespecified mucosal areas. Results The study started with acute experiments in 8 animals, followed by a 10-day survival study in another 8 pigs. A total of 20 mucosal pieces were resected. The sizes of the resected pieces varied up to 7.5 × 4.0 cm ex vivo. All but one could be resected en bloc. Percutaneous assistance resulted in an excellent exposure of the submucosal space and enabled stepwise dissection of the submucosal connective tissue. Neither the PMT nor advanced ESD led to relevant complications. Conclusions We demonstrated the feasibility and safety of a new PMT for advanced ESD. With the use of PA-ES, mucosal pieces of various sizes can be resected en bloc in gastric locations that are difficult to access by flexible endoscopy alone.

BACKGROUND: Accumulation of advanced glycation end products, that is, N(epsilon)-carboxymethyllysine (CML), induces oxidative stress and inflammation, and is present in chronic renal failure. Proximal tubular cells (PTCs) take up advanced glycation end products-bound proteins by apical megalin-receptors and degrade them. We hypothesized that renal transplant dysfunction affects renal CML homeostasis. Therefore, tubular and glomerular deposition of CML was investigated in a rat transplantation model, and in human allograft biopsies. METHODS: Fisher 344 kidneys were orthotopically transplanted into Lewis recipients. Recipients were treated with placebo, angiotensin II type 1 receptor blocker (candsartan 5 mg/kg/day), or calcium channel blocker (lacidipine 1 mg/kg/day) more than 28 weeks posttransplantation. Grafts were harvested at 12, 20, and 28 weeks posttransplantation. Sixty-two renal transplant patients underwent graft biopsy because of creatinine increase. Biopsies were graded according to interstitial fibrosis and tubular atrophy. N(epsilon)-carboxymethyllysine and megalin were semiquantitatively investigated in rats and humans using immunohistochemistry. RESULTS: In Fisher grafts, the development of transplant dysfunction was associated with a longitudinal increase in CML deposition in PTCs (week 12: 1.0+/-0.0, week 20: 1.5+/-0.3, week 28: 2.1+/-0.2, P

Rho GTPases are a family of major regulators of E-cadherin-mediated cell adhesion that are implicated in the carcinogenic process by deregulated expression of the family members itself or of upstream modulators or downstream effectors. Combined investigation of the Rho GTPase Rac1, the effector protein IQGAP1 and the activator Tiam1 in relation to expression or mutation of E-cadherin in gastric adenocarcinomas has not been reported. The aim of the study was to determine the expression and prognostic significance of Rac1, IQGAP1, Tiam1 and E-cadherin in gastric adenocarcinomas. Gastric carcinomas of 76 patients were investigated immunohistochemically in a tissue microarray study for expression of Rac1, IQGAP1, Tiam1 and E-cadherin. Correlations with clinical and follow-up data were examined. Moderate or strong reactivity for Rac1 was observed in 46% and for Tiam1 in 56% of tumors. Expression of IQGAP1 was present in 59% and of E-cadherin in 87% of tumors. While Rac1 and E-cadherin expression were not related to prognosis, a trend was observed between a lack of IQGAP1 expression (log-rank 0.088) as well as presence of Tiam1 (log-rank 0.097) and favorable prognosis in Kaplan-Meier survival analysis. Expression of Rac1 was positively linked to IQGAP1 expression (P=0.007, r=0.343) and tended to be inversely associated with expression of E-cadherin (P=0.055, r=-0.245). In conclusion, we observed deregulated expression of Rac1, IQGAP1, Tiam1 and E-cadherin in gastric cancer. We present evidence that either upregulation (for Rac1 and IQGAP1) or downregulation (for Tiam1 and E-cadherin) occurs. Rac1 and E-cadherin expression were not related to prognosis, while trends pointing to favorable prognosis of patients with Tiam1 expression and a lack of IQGAP1 expression were observed. These results indicate that the investigated regulators of E-cadherin-mediated cell adhesion play a role in gastric carcinogenesis.

BACKGROUND Matrix metalloproteinases (MMP) are involved in the development of interstitial fibrosis and tubular atrophy (IF/TA) in renal disease. The synthesis of MMP is activated by the extracellular matrix metalloproteinases inducer protein (EMMPRIN). To analyze the role of EMMPRIN in IF/TA, we retrospectively detected EMMPRIN expression in specimens of human renal allografts with various levels of IF/TA. METHODS Immunohistochemistry was performed to detect EMMPRIN expression. In a retrospective analysis, a total cohort of 50 specimens were divided according to BANFF-classification into four subgroups (0-3): no, mild (≤ 25%), moderate (26-50%), or severe (>50%) IF/TA. Among other parameters, renal function was analyzed and compared to EMMPRIN expression. RESULTS In 24 of 38 biopsies, we detected positive EMMPRIN staining. All nephrectomy (n = 12) samples were negative for EMMPRIN. Positive staining in the biopsy samples was detectable on the basolateral side of tubular epithelial cells. EMMPRIN staining was negatively correlated with IF/TA (p < 0.001). We found significant differences between the mean EMMPRIN expression in IF/TA groups 0 and 3 (p = 0.021) and groups 1 and 3 (p = 0.004). Furthermore, we found significant correlations between EMMPRIN staining and renal function. CONCLUSION Our data suggest that EMMPRIN is involved in the pathophysiology of IF/TA.

Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) is a standard procedure for intrathoracic lymph node biopsies. The newly developed cryo-needle operates in a similar way to the EBUS-TBNA but is able to obtain specimens for histological evaluation. The purpose of this animal study was to evaluate the feasibility, effect, and safety of the cryo-needle biopsies. Four EBUS-guided cryo-needle biopsies were obtained from a mediastinal lymph node of a healthy pig. In an open surgery approach, cryo-needle biopsies using activation times of 1, 2, and 3 s (A1/A2/A3) and needle biopsies using a 21-gauge EBUS-TBNA needle were obtained from mesenteric lymph nodes. Cryo-needle biopsies A2 were performed with (A2+) and without (A2−) an oversheath. The size, weight, percentage of lymphatic tissue and artefact-free area of each cryobiopsy were evaluated. Smears were made with the TBNA-needle aspirates to determine the number of lymphocytes per high-power field (HPF). The bleeding duration was measured. We successfully obtained EBUS-guided cryo-needle biopsies. The area and weight of the biopsies A3 and A2+ were significantly larger compared with A1 (1.7 ± 0.8 and 1.4 ± 0.3 vs. 0.9 ± 0.4 mm2; 5.2 ± 2.4 and 3.4 ± 1.8 vs. 1.5 ± 0.7 mg). The percentage of lymphatic tissue of the cryobiopsies was 90 ± 25 and 98 % of samples were artefact-free. The number of lymphocytes/HPF of TBNA-needle smears was 128 ± 54.3. There was no difference in bleeding duration between the techniques. The cryo-needle yields large histological specimens of high quality.

Advanced adult soft-tissue sarcomas (STSs) are rare tumors with a dismal prognosis and limited systemic treatment options. STSs may originate from mesenchymal stem cells (MSCs); the latter have mainly been isolated from adult bone marrow as plastic-adherent cells with differentiation capacity into mesenchymal tissues. Recently, a panel of antibodies has been established that allows for the prospective isolation of primary MSCs with high selectivity. Similar to cancer stem cells in other malignancies, sarcoma stem cells may bear immunophenotypic similarity with the corresponding precursor, that is, MSCs. We therefore set out to establish the expression pattern of MSC markers in sarcoma cell lines and primary tumor samples by flow cytometry. In addition, fibroblasts from different sources were examined. The results document a significant amount of MSC markers shared by sarcoma cells. The expression pattern includes uniformly expressed markers, as well as MSC markers that only stained subpopulations of sarcoma cells. Expression of W5C5, W8B2 (tissue nonspecific alkaline phosphatase [TNAP]), CD344 (frizzled-4), and CD271 marked subpopulations displaying increased proliferation potential. Moreover, CD271+ cells displayed in vitro doxorubicin resistance and an increased capacity to form spheres under serum-free conditions. Interestingly, another set of antigens, including the bona fide progenitor cell markers CD117 and CD133, were not expressed. Comparative expression patterns of novel MSC markers in sarcoma cells, as well as fibroblasts and MSCs, are presented. Our data suggest a hierarchical cytoarchitecture of the most common adult type sarcomas and introduce W5C5, TNAP, CD344, and CD271 as potential sarcoma progenitor cell markers.

To evaluate the influence of regional hyperthermia on rates of complete pathological response (pCR) and sphincter-sparing surgery in the context of an up-to-date radiochemotherapy protocol for locally advanced rectal cancer.Between 2007 and 2010, 106 patients with locally advanced cancer of the middle and lower rectum were admitted to neoadjuvant radiochemotherapy either with (n = 61) or without (n = 45) regional hyperthermia. A retrospective comparison was performed between two groups: 45 patients received standard treatment consisting of 5040 cGy in 28 fractions to the pelvis and 5-fluorouracil (RCT group) and 61 patients received the same treatment in combination with regional hyperthermia (HRCT group). Target temperature was 40.5°C for at least 60 min. Total mesorectal excision was performed routinely.pCR was seen in 6.7% of patients in the RCT group and 16.4% in the HRCT group. Patients who received at least four hyperthermia treatments (n = 40) achieved a significantly higher pCR rate (22.5%) than the remaining 66 patients (p = 0.043). Rates of sphincter-sparing surgery were similar in both groups with 64% in the RCT group and 66% in HRCT. When considering only low-lying tumours located within 8 cm of the anal verge prior to treatment, the rate of sphincter-sparing surgery was 57% in the HRCT group compared with 35% in the RCT group (p = 0.077).The combination of regional hyperthermia and neoadjuvant radiochemotherapy may lead to an increased pCR rate in locally advanced rectal cancer. Patients with low-lying tumours especially may benefit when additional downsizing allows sphincter-preserving surgery.

Major causes of morbidity in intravenous drug users are infections. In infective endocarditis, the tricuspid valve is mainly involved. Masses can cause septic embolisms and, in rare cases, they are associated with mycotic aneurysms of pulmonary arteries that lead to severe haemorrhage. We report the case of a young woman with a history of intravenous drug abuse and prolonged infective tricuspid valve endocarditis. Initially, echocardiography showed large masses on the anterior leaflet of the tricuspid valve and severe tricuspid regurgitation; blood cultures revealed staphylococcus and streptococcus species. Eight months after initial diagnosis, she presented with severe haemoptysis and fever. CT revealed a ruptured mycotic aneurysm of the right pulmonary artery. Lobectomy was performed immediately. Postoperatively, the patient fully recovered. After continued antibiotic treatment, follow-up examinations showed negative echocardiographic findings and blood cultures results.

The expression of Heat Shock Proteins (HSPs) is increased in various cancers and has been shown to correlate with biological tumor behaviour. This study aimed to investigate the impact of HSP70, HSP60 and HSP27 expression in colon cancer. HSP expression was determined by immunohistochemistry on a tissue microarray with 355 primary resected colon carcinomas of all stages. Expression patterns were correlated with pathologic features (UICC pTNM category, tumor grading) and survival. Expression of HSP27, HSP60 and HSP70 ranged from negative to high. There was no correlation between HSP27, HSP60 and HSP70 expression among each other and with UICC pT category, presence of lymph node or distant metastases or tumor grading. High HSP70 expression was associated with worse overall survival (p

Background Renal cell carcinoma can cause various paraneoplastic syndromes including metabolic and hematologic disturbances. Paraneoplastic hypereosinophilia has been reported in a variety of hematologic and solid tumors. We present the first case in the literature of severe paraneoplastic hypereosinophilia in a patient with renal cell carcinoma. Case presentation A 46 year-old patient patient with a history of significant weight loss, reduced general state of health and coughing underwent radical nephrectomy for metastasized renal cell carcinoma. Three weeks after surgery, the patient presented with excessive peripheral hypereosinophilia leading to profound neurological symptoms due to cerebral microinfarction. Systemic treatment with prednisolone, hydroxyurea, vincristine, cytarabine, temsirolimus and sunitinib led to reduction of peripheral eosinophils but could not prevent rapid disease progression of the patient. At time of severe leukocytosis, a considerable increase of cytokines associated with hypereosinophilia was measurable. Conclusions Paraneoplastic hypereosinophilia in patients with renal cell carcinoma might indicate poor prognosis and rapid disease progression. Myelosuppressive therapy is required in symptomatic patients.

The aim of this study was to investigate correlations between glucose metabolism registered by 18F-FDG PET/CT and tumor perfusion quantified by volume perfusion CT and immunohistochemical markers Ki67 and microvessel density (MVD) in patients with non–small cell lung cancer (NSCLC). Methods: Between February 2010 and April 2011, 24 consecutive patients (21 women, 3 men; mean age ± SD, 67.6 ± 6.8 y; age range, 55.6–81.3 y) with histologically proven NSCLC (14 adenocarcinoma, 9 squamous cell lung carcinoma [SCC], and 1 mixed adenocarcinoma and SCC) underwent 18F-FDG PET/CT and additional volume perfusion CT. Maximum standardized uptake value (SUVmax), mean SUV, and the metabolic tumor volume were used for 18F-FDG uptake quantification. Blood flow (BF), blood volume (BV), flow extraction product (Ktrans), and standardized perfusion value (SPV) were determined as CT perfusion parameters. Both perfusion parameters and 18F-FDG uptake values were subsequently related to the histologic subtypes, proliferation marker Ki67, MVD according to CD34 staining, and total tumor volume. Results: Mean SUV, SUVmax, and the metabolic tumor volume (mL) were 5.8, 8.7, and 32.3, respectively, in adenocarcinoma and 8.5, 12.9, and 16.8, respectively, in SCC. Mean BF (mL/100 mL/min), mean BV (mL/100 mL), and Ktrans (mL/100 mL/min) were 35.4, 7.3, and 27.8, respectively, in adenocarcinoma and 35.5, 10.0, and 27.8, respectively, in SCC. Moderate correlations were found between the 18F-FDG PET/CT parameters and Ki67 as well as between CT perfusion parameters and MVD but not vice versa. For all tumors, the following correlations were found: between SUVmax and Ki67, r = 0.762 (P = 0.017); between SUVmax and MVD, r = −0.237 (P = 0.359); between mean BF and Ki67, r = −0.127 (P = 0.626); and between mean BF and MVD, r = 0.467 (P = 0.059). Interestingly, correlations between the BF–metabolic relationship and total tumor volume were higher in SCC (r = 0.762, P = 0.017) than in adenocarcinoma (r = −0.0791, P = 0.788). Conclusion:18F-FDG uptake correlates with Ki67, whereas BF, BV, and Ktrans correlate with MVD. Therefore, 18F-FDG uptake and perfusion parameters provide complementary functional information. An improved tumor profiling will be beneficial for both prognosis and therapy response evaluation in these tumors.

Tissues are complicated three-dimensional structures, composed of different types of interacting cells. Since the cell population of interest might constitute only a minor fraction of the total tissue volume, the problem of tissue heterogeneity has been a major barrier to the molecular analysis of normal versus diseased tissue. Thus, tissue microdissection represents one of the most promising techniques in molecular pathology offering the link between morphology and genetic analysis since it was established in the early 1970s. These first applications and further developments in the techniques enable preparation of morphologically well described and circumscribed cell populations of either tumor cells or surrounding tissue or even cytology specimens without contamination of unwanted cells. Laser capture microdissection is suitable for the dissection of both paraffin embedded and fresh frozen material. Further applications of the dissected genomic material are isolation of DNA and RNA as described later on followed by PCR or RT-PCR and sequencing.

Tissues are complicated three-dimensional structures, composed of different types of interacting cells. Since the cell population of interest might constitute only a minor fraction of the total tissue volume, the problem of tissue heterogeneity has been a major barrier to the molecular analysis of normal versus diseased tissue. Thus, tissue microdissection represents one of the most promising techniques in molecular pathology offering the link between morphology and genetic analysis since it was established in the early 1970s. These first applications and further developments in the techniques enable preparation of morphologically well described and circumscribed cell populations of either tumor cells or surrounding tissue or even cytology specimens without contamination of unwanted cells. Laser capture microdissection is suitable for the dissection of both paraffin embedded and fresh frozen material. Further applications of the dissected genomic material are isolation of DNA and RNA as described later on followed by PCR or RT-PCR and sequencing.

Paratesticular fibrous pseudotumor (nodular periorchitis, inflammatory pseudotumor of the spermatic cord) is a rare, benign condition of unknown etiology characterized by solitary or multiple intrascrotal nodules composed of dense fibrous tissue with a variable, sometimes sparse inflammatory infiltrate. Based on certain similarities to other fibroinflammatory disorders characterized by infiltrates of IgG4-expressing plasma cells and recently subsumed under the heading of IgG4-mediated diseases, we investigated the plasma cell distribution and immunoglobulin isotypes in three cases of paratesticular fibrous and inflammatory pseudotumor. All three cases showed a high number of IgG4-positive plasma cells with an IgG4 to IgG ratio of 44–48%. This finding indicates that paratesticular fibrous pseudotumor might belong to the growing list of IgG4-related diseases, which by now includes such diverse entities as retroperitoneal fibrosis, sclerosing pancreatitis and cholangitis, Riedel’s thyroiditis, or sclerosing sialadenitis.

A 33-year-old obese woman presented with a retroperitonealmassasanincidentalfindingduringherniasurgery.Aphysicalexam-inationrevealednopathologicfinding.Obesitypreventedpalpationofany abdominal mass. At the time of presentation to our hospital atEberhard-Karls-University(Tu¨bingen,Germany),laboratoryparam-eters were at normal levels. A computed tomography scan showedbilateral giant adrenal tumors with predominantly fatty regions (Fig1A;straightwhitearrows)interspersedwithsolid,septa-liketissue(Fig1A; curved white arrow) demonstrating mild enhancement after in-travenouscontrastadministration.Themasseswereencapsulatedandmeasured19.1 10.5 22.6cmattheleftand12.3 8.7 19.3cmat the right side, compressing retroperitoneal vessels. The tumor waswell circumscribed and extended to the surrounding tissue withoutinvasion. The suspected diagnosis of bilateral adrenal myelolipomaswas histologically confirmed after a computed tomography–guidedbiopsy. The microscopic analysis of biopsy samples revealed adipo-cytes with clear cytoplasm, interspersed with cells of hematopoiesis.One half of the cells were erythroid nests with dense blue nuclei (Fig1B,whitestar).Theothercellswereprimarilyofmyeloidlineagewithmaturation to segmented granulocytes (Fig 1B; white arrow). Onemultilobulatedmegakaryocytewasalsopresentinthebonemarrow–likespecimen.Giventheabsenceofclinicalsymptoms,thetreatmentstrategychosenwasawatchandwaitregimen.First described in 1905, adrenal myelolipomas are rare, hor-monally inactive, benign tumors that are composed of mature fattissue and hematopoietic elements.

Because (18)F-FDG PET alone has only limited value in metastatic germ cell tumors (GCTs), we investigated the addition of 3'-deoxy-3'-(18)F-fluorothymidine (FLT) to (18)F-FDG for early response monitoring and prediction of the histology of residual tumor masses in patients with metastatic GCT.Eleven patients with metastatic GCT were examined with both (18)F-FDG PET/CT and (18)F-FLT PET/CT before chemotherapy, after the first cycle of chemotherapy (early response), and 3 wk after completion of chemotherapy. In 1 patient with negative (18)F-FLT PET/CT results before chemotherapy, no further (18)F-FLT scanning was performed. PET images were analyzed visually and, using standardized uptake values (SUVs), semiquantitatively. The results were compared with the findings of CT and tumor marker levels and validated by histopathologic examination of resected residual masses, including Ki-67 immunostaining (7 patients), or by clinicoradiologic follow-up for at least 6 mo (4 patients). A responder was defined as a patient showing the presence of necrosis, a complete remission, or a marker-negative partial remission within a minimum progression-free interval of 6 mo. Early treatment response was judged according to the criteria of the European Organization for Research and Treatment of Cancer.Before chemotherapy, reference lesions showed increased (18)F-FDG uptake (mean SUV, 8.8; range, 2.9-15.0) in all patients and moderate (18)F-FLT uptake (mean SUV, 3.7; range, 1.7-9.7) in 10 of 11 patients. After 1 cycle of chemotherapy, mean SUV decreased in responders and nonresponders by 64% and 60%, respectively, for (18)F-FDG (P = 0.8) and by 58% and 48%, respectively, for (18)F-FLT (P = 0.5). After the end of chemotherapy, mean SUV decreased in responders and nonresponders by 85% and 73%, respectively, for (18)F-FDG (P = 0.1) and by 68% and 65%, respectively, for (18)F-FLT (P = 0.8). The results of early and final PET were inconsistent in 6 of 11 patients for (18)F-FDG and in 4 of 10 patients for (18)F-FLT. Both patients with teratoma had false-negative results on both (18)F-FDG and (18)F-FLT. The sensitivity, specificity, positive predictive value, and negative predictive value for detection of viable tumor after 1 cycle of chemotherapy were 60%, 33%, 43%, and 50%, respectively, for (18)F-FDG and 60%, 80%, 75%, and 67%, respectively, for (18)F-FLT PET/CT. The respective values after the end of chemotherapy were 20%, 100%, 100%, and 60% for (18)F-FDG and 0%, 100%, 0%, and 50% for (18)F-FLT PET/CT.PET-negative residual masses after chemotherapy of metastatic GCT still require resection, since the low negative predictive value of (18)F-FDG PET for viable tumor cannot be improved by application of (18)F-FLT.

Precision-cut liver tissue slices (PCLS) have been used for decades to study pharmacological metabolism as well as toxicology and efficacy of novel substances on primary material under standardized conditions. Slicing of primary liver tissue has been done using different slicing machines. Since there has been great variability in the results, we sought to compare the reproducibility of tissue slices generated using the newly developed Leica VT1200 S vibrating blade microtome with Vibrocheck (LV) and the Krumdieck tissue slicer (KD) which has been the standard apparatus for this application so far. Liver samples from five different species (human, pig, cattle, rat, mouse) were cut and the reproducibility of slice thickness was analyzed by cross sectioning the PCLS. The quality of the sliced tissue was determined via measurement of the ATP content. As a result, we found an improved accuracy and reproducibility of rat, mouse and human tissue slices using the new Leica vibrating blade microtome.

Cytoreductive surgery followed by hyperthermic intraperitoneal chemotherapy (HIPEC) is associated with significantly longer survival in patients with peritoneal carcinomatosis (PC). So far, no morphological imaging method has proven to accurately assess the intra-abdominal tumor spread. This study was designed to predict tumor load in patients with PC using dual-modality (18)FDG-PET/CT and to compare the results with those of PET and CT alone by correlating imaging findings with intraoperative staging.Twenty-two patients with PC from gastrointestinal (n = 13), ovarian cancer (n = 8), and mesothelioma (n = 1) underwent contrast-enhanced (18)FDG-PET/CT before surgery and HIPEC. In a retrospective analysis PET, CT, and fused PET/CT were separately and blindly reviewed for the extent of peritoneal involvement using the Peritoneal Cancer Index (PCI). Imaging results were correlated with the intraoperative PCI using Pearson's correlation coefficient and linear regression analysis.There was a strong correlation between the PCI obtained with PET/CT and the surgical PCI with respect to the total score (r = 0.951) as well as in the regional analysis (small bowel, r = 0.838; other, r = 0.703). The correlation was slightly lower for CT alone (total score, r = 0.919; small bowel, r = 0.754; other, r = 0.666) and significantly lower (p = 0.002) for PET alone (total score, r = 0.793; small bowel, r = 0.553, other, 0.507).Contrast-enhanced CT is superior compared with PET alone to predict the extent of PC. In our patient group, the combination of both modalities (contrast enhanced PET/CT) yielded the best results and proved to be a useful tool for selecting candidates for peritonectomy and HIPEC.

Background & Aims: The preoperative diagnosis of cholangiocarcinoma is associated with a low sensitivity. To overcome this limitation, a new imaging modality was evaluated to detect neoplasia in vivo in the biliary tract. Methods: Fourteen patients with biliary strictures were examined. Mucosal imaging was performed with a miniaturized confocal laser scanning miniprobe introduced via the accessory channel of a cholangioscope. Thereafter, targeted biopsy specimens were taken from the same regions. Results: All strictures could be reached. Presence of irregular vessels use confocal laser microscopy enabled prediction of neoplasia with an accuracy rate of 86%, sensitivity of 83%, and specificity of 88%. The respective numbers for standard histopathology were 79%, 50%, and 100%. The mean signal-to-noise-ratio of laser microscopic images acquired from malignant strictures differed significantly from those of benign origin (1.8 ± 0.8 vs 2.6 ± 1.0; P = .005). Conclusions: Miniprobe-based confocal laser scanning microscopy considerably increases sensitivity for the detection of biliary neoplasia and therefore represents a promising diagnostic approach.

Background & Aims: Confocal fluorescence microscopy (CFM) has been mentioned to be a promising tool for in vivo histology. Recently, a portable confocal miniprobe has been developed. Our aim was to evaluate the potential benefit of CFM for detection of gastrointestinal neoplasia. Methods: A total of 47 patients with known or suspected neoplasia in the upper (n = 34) or lower gastrointestinal tract (n = 13) were examined with standard endoscopes. After mucolyis with 5–10 mL of acetic acid 1.5%, chromoendoscopy with 2–5 mL cresyl violet 0.25% was performed, with the substance also being used as a fluorophore for CFM. Real-time video sequences were recorded. Thereafter, biopsies were taken or mucosectomy/polypectomy was performed from the same examined area. All stored sequences were put into a random order and assessed by a pathologist and a gastroenterologist both blinded to any data. Results: A total of 119 CFM video sequences were recorded of 85 benign or 34 neoplastic areas. Quality of CFM images was regarded too low in 24 (pathologist) and 14 sequences (gastroenterologist). For the pathologist, accuracy of CFM detecting neoplasia was 92.6% (suitable images) and 73.9% (intention to diagnose). The respective accuracy values for the gastroenterologist were 92.4% (suitable images) and 81.5% (intention to diagnose). Agreement between CFM and histopathology was excellent (kappa values, 0.821 and 0.817). Conclusions: We have demonstrated that CFM with a miniprobe has the potential to diagnose neoplasia during ongoing endoscopy. This system has the advantage that it can be used with standard endoscopes. Further studies are warranted for validation.

Background A new portable confocal laser microscopy system has recently been developed. Objective Our purpose was to evaluate the feasibility of performing real-time microscopic imaging with a prototype of a new high-resolution miniprobe in conjunction with a video mosaicing algorithm. Design Feasibility study. Setting Tertiary referral center at a large university hospital. Patients Seven patients referred for endoscopy for various indications. Main Outcome Measurements High-resolution laser microscopy of the upper and lower GI tract was performed with standard endoscopes. Seven to 10 mL of 1% fluorescein was injected intravenously a few seconds before the procedure. No additional preparation was required. The prototype used has a lateral resolution of 1.2 μm and an axial resolution of 3 μm with a total field of 240 × 200 μm. From all stored video sequences a video mosaicing algorithm was used to combine the successive individual images, cancel motion artifacts, and reconstitute panoramas of the tissues. Results Cell-to-cell borders, single cell structures, and mucosal inflammation was readily detectable. By the use of the mosaicing algorithm, the image area could be increased 2- to 4-fold, and image definition could be further enhanced to allow finer detail visualization. Limitations Low number of patients, early feasibility study. Conclusions Our preliminary data show that high-resolution miniprobe-based confocal fluorescence microscopy in conjunction with video mosaicing has the potential to provide images similar to standard histopathologic studies. Dynamic images with a smaller field of view can be combined to reconstruct still images of high resolution covering a fairly large area.

Background: The k-Ras-protein shows in regard to its mutations status a predicitvity of the response after Cetuximab in patients with colon carcinoma. To our knowledge, not much is known, if k-RAS has also a predictivity in regard to different ways of metastasis. The goal of our investigations was the evaluation of k-Ras on primary non-metastatic colon cancer compared to lymphatic, hepatic and peritoneal metastasis. Methods: 98 patients with adenocarcinom of the colon and a postoperative pT2/3-status, who underwent surgical resection between 2006 and 2008 at the University of Tübingen had been included within the study collective. 53 patients had no metastasis (control group), while 45 patients revealed primary metastasis: lymphatic metastasis (n=20), hepatic spread (n=20) and multiple metastasize (n=5). After DNA-extraction, an amplification by ARMS-PCR was performed and the analysis of the mutation status was done by melting point analysis by Light-Cycler. Additionally sequencing of Exons 12 and 13 was performed in individual cases. Results: 29/98 (=30%) revealed a k-Ras-mutation. Out of these, 12 (=41%) had metastasis: 9 hepatic (=75%), 2 lymphatic (=17%) and 1 (=8%) multiple metastasis. Patients with k-Ras-mutation (n=29) showed a significant lower rate of lymphatic metastasis. (90% vs. 10%, p=0.031). The survival in accordance to the k-Ras mutation was not statistically significant (p=0.415). Conclusion: Patients with mutated k-Ras-gene in out patient population revealed no significant higher rate of metastasis (p=0.558), nor in regard to hepatic (p=0.090) or multiple locations (p=0.629). Our preliminary data assume, that the k-Ras mutation status has a predictive function on the rate of lymphatic metastasis in patients suffering from colon carcinoma (p=0.031). Follow-up-studies are demanded, if single mutations status might have an additional influence. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 445. doi:1538-7445.AM2012-445

Background: cMYC as well as β-Catenin are downstream-targets of the Wnt-pathway and both have an important early function within the carcinogenesis and the progression of colon cancer. Additionally cMYC was found to be associated with a worse prognosis in other neoplasms. The goal of our investigations was the evaluation of the Wnt-pathway on primary non-metastatic colon cancer compared to lymphatic, hepatic and peritoneal metastasis in regard to the expression of cMYC and β-Catenin. Methods: 98 patients with adenocarcinom of the colon and a postoperative pT2/3-status, who underwent surgical resection between 2006 and 2008 at the University of Tübingen had been included within the study collective. 53 patients had no metastasis (control group), while 45 patients revealed primary metastasis: lymphatic metastasis (n=20), hepatic spread (n=20) and multiple metastasize (n=5). cMYC and β-Catenin was coloured on a Tissue-Micro-Array by immunohistochemistry with consecutive semiquantitative analysis. Results: 17 of 98 patients (17%) had been positive for cMYC. 5 out of these 17 (29%) revelaed metastasis. 16 out of the 17 cMYC-positive patients showed a nuclear accumulation of β-Catenin (94%) (p=0.027). 40 out of 81 (49%) cMYC negative patients revealed metastasis. No significant correlation between nuclear positive cMYC and the rate or kind of metastasis was found (p=0.133). In 29 out of 81 cMYC-negative patients (36%) a strong expression of β-Catenin was found. Conclusion: cMYC is well known for a worse prognosis in case of an overexpression in patients suffering from lymphoma. A correlation between the expression of cMYC and the kind of metastasis and / or survival nor prognosis in patients with colon cancer was not able to show (p=0.211). Additonally no correlation between the nuclear exprimed β-Catenin and different ways of metastasis was found (p=0.202). Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 447. doi:1538-7445.AM2012-447

AIM: To evaluate the sensitivity of brush cytology and forceps biopsy in a homogeneous patient group with hilar cholangiocarcinoma. METHODS: Brush cytology and forceps biopsy were routinely performed in patients with suspected malignant biliary strictures. Fifty-eight consecutive patients undergoing endoscopic retrograde cholangiopancreatography (ERCP) including forceps biopsy and brush cytology in patients with hilar cholangiocarcinoma between 1995-2005. RESULTS: Positive results for malignancy were obtained in 24/58 patients (41.4%) by brush cytology and in 31/58 patients (53.4%) by forceps biopsy. The combination of both techniques brush cytology and forceps biopsy resulted only in a minor increase in diagnostic sensitivity to 60.3% (35/58 patients). In 20/58 patients (34.5%), diagnosis were obtained by both positive cytology and positive histology, in 11/58 (19%) by positive histology (negative cytology) and only 4/58 patients (6.9%) were confirmed by positive cytology (negative histology). CONCLUSION: Brush cytology and forceps biopsy have only limited sensitivity for the diagnosis of malignant hilar tumors. In our eyes, additional diagnostic techniques should be evaluated and should become routine in patients with negative cytological and histological findings.