Noninvasive diagnosis of urothelial cancer in urine using DNA hypermethylation signatures-Gender matters.

Urothelial cancer (UCa) is the most predominant cancer of the urinary tract and noninvasive diagnosis using hypermethylation signatures in urinary cells is promising. Here, we assess gender differences in a newly identified set of methylation biomarkers. UCa-associated hypermethylated sites were identified in urine of a male screening cohort (n = 24) applying Infinium-450K-methylation arrays and verified in two separate mixed-gender study groups (n = 617 in total) using mass spectrometry as an independent technique. Additionally, tissue samples (n = 56) of mixed-gender UCa and urological controls (UCt) were analyzed. The hypermethylation signature of UCa in urine was specific and sensitive across all stages and grades of UCa and independent on hematuria. Individual CpG sensitivities reached up to 81.3% at 95% specificity. Albeit similar methylation differences in tissue of both genders, differences were less pronounced in urine from women, most likely due to the frequent presence of squamous epithelial cells and leukocytes. Increased repression of methylation levels was observed at leukocyte counts ≥500/µl urine which was apparent in 30% of female and 7% of male UCa cases, further confirming the significance of the relative amounts of cancerous and noncancerous cells in urine. Our study shows that gender difference is a most relevant issue when evaluating the performance of urinary biomarkers in cancer diagnostics. In case of UCa, the clinical benefits of methylation signatures to male patients may outweigh those in females due to the general composition of women's urine. Accordingly, these markers offer a diagnostic option specifically in males to decrease the number of invasive cystoscopies.

Impact of real-time elastography on magnetic resonance imaging/ultrasound fusion guided biopsy in patients with prior negative prostate biopsies.

PURPOSE: The fusion of multiparametric resonance imaging and ultrasound has been proven capable of detecting prostate cancer in different biopsy settings. The addition of real-time elastography promises to increase the precision of the outcome of targeted biopsies. We investigated whether real-time elastography improves magnetic resonance imaging/transrectal ultrasound fusion targeted biopsy in patients after previous negative biopsies. MATERIALS AND METHODS: Prospectively 121 men underwent 3T magnetic resonance imaging. Using magnetic resonance imaging/real-time elastography fusion every suspicious lesion was characterized according to its tissue density and sampled by 2 fusion guided targeted biopsies. Additionally, all patients underwent 12-core systematic biopsy. The detection rate of clinically significant and insignificant cancers was compared between targeted und systematic biopsies. The accuracy to predict high grade prostate cancer was evaluated for with the PI-RADS scoring system and compared to the magnetic resonance imaging/real-time elastography fusion score. RESULTS: Overall prostate cancer was detected in 52 patients (43%). Targeted fusion guided biopsy revealed prostate cancer in 32 men (26.4%) and systematic biopsy in 46 (38%). The proportion of clinically significant cancers was higher for targeted biopsy (90.6%) compared to systematic biopsy (73.9%). The detection rate per core was higher for targeted biopsies (14.7%) compared to systematic biopsies (6.5%, p <0.001). The prediction of biopsy result according to magnetic resonance imaging/real-time elastography fusion was better (AUC 0.86) than magnetic resonance imaging alone (AUC 0.79). Sensitivity and specificity for magnetic resonance imaging/real-time elastography fusion was 77.8% and 77.3% vs 74.1% and 62.9% for magnetic resonance imaging. CONCLUSIONS: Magnetic resonance imaging/transrectal ultrasound fusion enhances the likelihood of detecting clinically significant cancers in a repeat biopsy setting. Adding real-time elastography to magnetic resonance imaging supports the characterization of cancer suspicious lesions.

Automated quantification of FISH signals in urinary cells enables the assessment of chromosomal aberration patterns characteristic for bladder cancer.

Targeting the centromeres of chromosomes 3, 7, 17 (CEP3, 7, 17) and the 9p21-locus (LSI9p21) for diagnosing bladder cancer (BC) is time- and cost-intensive and requires a manual investigation of the sample by a well-trained investigator thus overall limiting its use in clinical diagnostics and large-scaled epidemiological studies. Here we introduce a new computer-assisted FISH spot analysis tool enabling an automated, objective and quantitative assessment of FISH patterns in the urinary sediment. Utilizing a controllable microscope workstation, the microscope software Scan^R was programmed to allow automatic batch-scanning of up to 32 samples and identifying quadruple FISH signals in DAPI-scanned nuclei of urinary sediments. The assay allowed a time- and cost-efficient, automated and objective assessment of CEP3, 7 and 17 FISH signals and facilitated the quantification of nuclei harboring specific FISH patterns in all cells of the urinary sediment. To explore the diagnostic capability of the developed tool, we analyzed the abundance of 51 different FISH patterns in a pilot set of urine specimens from 14 patients with BC and 21 population controls (PC). Herein, the results of the fully automated approach yielded a high degree of conformity when compared to those obtained by an expert-guided re-evaluation of archived scans. The best cancer-identifying pattern was characterized by a concurrent gain of CEP3, 7 and 17. Overall, our automated analysis refines current FISH protocols and encourages its use to establish reliable diagnostic cutoffs in future large-scale studies with well-characterized specimens-collectives.

Monte Carlo-based parametrization of the lateral dose spread for clinical treatment planning of scanned proton and carbon ion beams.

Ion beam therapy using state-of-the-art pencil-beam scanning offers unprecedented tumour-dose conformality with superior sparing of healthy tissue and critical organs compared to conventional radiation modalities for external treatment of deep-seated tumours. For inverse plan optimization, the commonly employed analytical treatment-planning systems (TPSs) have to meet reasonable compromises in the accuracy of the pencil-beam modelling to ensure good performances in clinically tolerable execution times. In particular, the complex lateral spreading of ion beams in air and in the traversed tissue is typically approximated with ideal Gaussian-shaped distributions, enabling straightforward superimposition of several scattering contributions. This work presents the double Gaussian parametrization of scanned proton and carbon ion beams in water that has been introduced in an upgraded version of the worldwide first commercial ion TPS for clinical use at the Heidelberg Ion Beam Therapy Center (HIT). First, the Monte Carlo results obtained from a detailed implementation of the HIT beamline have been validated against available experimental data. Then, for generating the TPS lateral parametrization, radial beam broadening has been calculated in a water target placed at a representative position after scattering in the beamline elements and air for 20 initial beam energies for each ion species. The simulated profiles were finally fitted with an idealized double Gaussian distribution that did not perfectly describe the nature of the data, thus requiring a careful choice of the fitting conditions. The obtained parametrization is in clinical use not only at the HIT center, but also at the Centro Nazionale di Adroterapia Oncologica.

Implementation and initial clinical experience of offline PET/CT-based verification of scanned carbon ion treatment.

BACKGROUND AND PURPOSE: We report on the implementation of offline PET/CT-based treatment verification at the Heidelberg Ion Beam Therapy Centre (HIT) and present first clinical cases for post-activation measurements after scanned carbon ion irradiation. Key ingredient of this in-vivo treatment verification is the comparison of irradiation-induced patient activation measured by a PET scanner with a prediction simulated by means of Monte Carlo techniques. MATERIAL AND METHODS: At HIT, a commercial full-ring PET/CT scanner has been installed in close vicinity to the treatment rooms. After selected irradiation fractions, the patient either walks to the scanner for acquisition of the activation data or is transported using a shuttle system. The expected activity distribution is obtained from the production of ß(+)-active isotopes simulated by the FLUKA code on the basis of the patient-specific treatment plan, post-processed considering the time course of the respective treatment fraction, the estimated biological washout of the induced activity and a simplified model of the imaging process. RESULTS: We present four patients with different indications of head, head/neck, liver and pelvic tumours. A clear correlation between the measured PET signal and the simulated activity pattern is observed for all patients, thus supporting a proper treatment delivery. In the case of a pelvic tumour patient it was possible to detect minor treatment delivery inaccuracies. CONCLUSIONS: The initial clinical experience proves the feasibility of the implemented strategy for offline confirmation of scanned carbon ion irradiation and therefore constitutes a first step towards a comprehensive PET/CT-based treatment verification in the clinical routine at HIT.

Intraoperative frozen section of the prostate decreases positive margin rate while ensuring nerve sparing procedure during radical prostatectomy.

PURPOSE: We evaluated whether intraoperative frozen section analysis of the prostate surface might provide significant information to ensure nerve sparing and minimize the positive margin rate. MATERIALS AND METHODS: In 236 patients treated with radical prostatectomy between June 2011 and September 2012 whole surface frozen section analysis of the removed prostate was done intraoperatively. The apex and base were circumferentially dissected as well as the whole posterolateral tissue corresponding to the neurovascular bundles. Multiple perpendicular sections were cut systematically for frozen section analysis. Pathology results were reported to navigate the procedure. RESULTS: Frozen section analysis identified positive surgical margins in 22% of cases, including the neurovascular bundles in 56.9%, apex in 34.5% and base in 8.6%. Of positive frozen section cases 92.3% could be converted to negative status, while 7.7% remained positive. The final positive margin rate in the total cohort was 3%, including a false-negative frozen section rate of 1.6%. In 14.8% of cases the initial nerve sparing plan was changed intraoperatively due to the positive frozen section and the secondary resected specimen detected cancer in 25%. Final pathology results showed Gleason upgrading or up-staging in 40.7% of cases compared to preoperative variables. When comparing patients with positive vs negative frozen sections, preoperative variables did not significantly differ, while postoperatively pathological stage, tumor volume, operative time and final margin status differed significantly. Of patients with exclusively unilateral positive biopsies 13% had a positive surgical margin intraoperatively on the opposite, biopsy negative side. CONCLUSIONS: The surface frozen section technique is associated with a low false-negative surgical margin rate. It might allow for safer preservation of functional anatomical structures in misclassified patients or even patients at higher preoperative risk.

Multiparametric ultrasound of the prostate: adding contrast enhanced ultrasound to real-time elastography to detect histopathologically confirmed cancer.

PURPOSE: We prospectively assessed whether a combined approach of real-time elastography and contrast enhanced ultrasound would improve prostate cancer visualization. MATERIAL AND METHODS: Between June 2011 and January 2012, 100 patients with biopsy proven prostate cancer underwent preoperative transrectal multiparametric ultrasound combining real-time elastography and contrast enhanced ultrasound. After initial elastographic screening for suspicious lesions, defined as blue areas with decreased tissue strain, each lesion was allocated to the corresponding prostate sector. The target lesion was defined as the largest cancer suspicious area. Perfusion was monitored after intravenous injection of contrast agent. Target lesions were examined for hypoperfusion, normoperfusion or hyperperfusion. Imaging results were correlated with final pathological evaluation on whole mount slides after radical prostatectomy. RESULTS: Of 100 patients 86 were eligible for final analysis. Real-time elastography detected prostate cancer with 49% sensitivity and 73.6% specificity. Histopathology confirmed malignancy in 56 of the 86 target lesions (65.1%). Of these 56 lesions 52 (92.9%) showed suspicious perfusion, including hypoperfusion in 48.2% and hyperperfusion in 48.2%, while only 4 (7.1%) showed normal perfusion patterns (p = 0.001). The multiparametric approach decreased the false-positive value of real-time elastography alone from 34.9% to 10.3% and improved the positive predictive value of cancer detection from 65.1% to 89.7%. CONCLUSIONS: Perfusion patterns of prostate cancer suspicious elastographic lesions are heterogeneous. However, the combined approach of real-time elastography and contrast enhanced ultrasound in this pilot study significantly decreased false-positive results and improved the positive predictive value of correctly identifying histopathological cancer.

The impact of real-time elastography guiding a systematic prostate biopsy to improve cancer detection rate: a prospective study of 353 patients.

PURPOSE: We evaluated whether real-time elastography guided biopsy improves prostate cancer detection compared to conventional systematic gray scale ultrasound guidance. MATERIALS AND METHODS: A total of 353 consecutive patients suspicious for prostate cancer were prospectively randomized for real-time elastography (178) or gray scale ultrasound (175). Each patient enrolled in the study underwent a 10-core prostate biopsy. Six lateral prostate sectors (base, mid, apex) were scanned for cancer suspicious areas, defined as stiffer blue lesions using real-time elastography and hypoechoic lesions using gray scale ultrasound. Suspicious areas were sampled by a single targeted biopsy and considered representative of a defined prostate sector. If real-time elastography or gray scale ultrasound did not visualize a suspicious area in a sector, the biopsy core was taken systematically. Imaging findings were correlated with histopathological reports. Real-time elastography and gray scale ultrasound cases were compared in terms of cancer detection rate and imaging guidance accuracy. RESULTS: Characteristics of patients undergoing real-time elastography and gray scale ultrasound, including age, prostate specific antigen, prostate volume and digital rectal examination, were not significantly different (p>0.05). Prostate cancer was detected in 160 of 353 patients (45.3%). The prostate cancer detection rate was significantly higher in patients who underwent biopsy with the real-time elastography guided approach compared to the gray scale ultrasound guided biopsy at 51.1% (91 of 178) vs 39.4% (69 of 175) (p=0.027). Overall sensitivity and specificity to detect prostate cancer was 60.8% and 68.4% for real-time elastography vs 15% and 92.3% for gray scale ultrasound, respectively. CONCLUSIONS: Sensitivity to visualize and detect prostate cancer improved using real-time elastography in addition to gray scale ultrasound during prostate biopsy. Overall sensitivity did not reach levels to omit a systematic biopsy approach.

Comprehensive report on prostate cancer misclassification by 16 currently used low-risk and active surveillance criteria.

UNLABELLED: What's known on the subject? and What does the study add? Prostate cancer characterisation, based on laboratory findings, clinical examination and histopathological cancer features that are used to define selection criteria for AS, is not ideal. Consequently, a panel of strict or more lenient criteria to select patients for AS have been published. Studies investigating the relationship between pretreatment variables and final pathology have been done in the past showing the risk of cancer misclassification for some criteria. No study has presented an overview of cancer selection using a panel of 16 currently used AS criteria that is presented in the present study. In an exactly defined cohort after radical prostatectomy, each set of criteria was used as a diagnostic test to separate between patients with more favourable (pT2, no Gleason upgrade between biopsy grading and final pathology) and unfavourable cancer features (pT3, pN+, Gleason upgrade). To the best of our knowledge a comparison of test quality criteria for AS criteria given by sensitivity, specificity, positive and negative predictive value and likelihood ratio has not yet been reported. Moreover, we showed that tumour characterisation, by a formally sufficient 12-core biopsy, in the present dataset harboured a risk of ≈20% that unfavourable cancer features were missed regardless of whether strict or more lenient selection criteria for AS were chosen. OBJECTIVE: To evaluate final histopathological features among men diagnosed with prostate cancer eligible for low-risk (LR) or active surveillance (AS) criteria. PATIENTS AND METHODS: Retrospective application of 16 definitions for AS or LR prostate cancer to a contemporary (January 2008 to March 2011) open retropubic radical prostatectomy (RRP) series of 1745 patients. EXCLUSION CRITERIA: neoadjuvant hormones, radiotherapy, inadequate histopathological reports, <10 biopsy cores. Report on the number of men with insignificant tumours (defined as: ≤pT2, Gleason score ≤6, tumour volume <0.5 mL) and men who had unfavourable tumour characteristics on final pathology (defined as: extracapsular extension or seminal vesicle invasion or lymph node metastasis or Gleason upgrading). Sensitivity, specificity, positive predictive value (PPV) and negative predictive values (NPV) were calculated. RESULTS: Eligibility of patients in the final study cohort (n = 1070) varied from 5.1% to 92.7% depending on the AS or LR criteria used. Final pathology revealed 77 insignificant cancers and 578 patients who had unfavourable histopathological criteria. The detection rate for insignificant cancers on final pathology was variable ranging from 7.8% to 28.3% depending on the AS- or LR-prediction tool used; unfavourable tumour characteristics were found in up to 33.5% on final pathology. The sensitivity, specificity, PPV and NPV were 8.5-97.9%, 24.7-97.8%, 67.7-89.1% and 45.3-78.2%, respectively. The likelihood ratio to correctly identify a patient with LR disease on final pathology ranged from 1.3 to 8. CONCLUSIONS: AS or LR criteria have a significant risk of cancer misclassification. Better prediction tools are needed to improve these criteria. Re-biopsy might improve safety and should be considered more frequently in patients who opt for AS.

Comparison of a lightweight polypropylene mesh (Optilene® LP) and a large-pore knitted PTFE mesh (GORE® INFINIT® mesh)--Biocompatibility in a standardized endoscopic extraperitoneal hernia model.

PURPOSE: The use of a mesh with good biocompatibility properties is of decisive importance for the avoidance of recurrences and chronic pain in endoscopic hernia repair surgery. As we know from numerous experiments and clinical experience, large-pore, lightweight polypropylene meshes possess the best biocompatibility. However, large-pore meshes of different polymers may be used as well and might be an alternative solution. METHODS: Utilizing a totally extraperitoneal technique in an established animal model, 20 domestic pigs were implanted with either a lightweight large-pore polypropylene (PP) mesh (Optilene® LP) or a medium-weight large-pore knitted polytetrafluorethylene (PTFE) mesh (GORE® INFINIT® mesh). After 94 days, the pigs were sacrificed and postmortem diagnostic laparoscopy was performed, followed by explantation of the specimens for macroscopic, histological and immunohistochemical evaluation. RESULTS: The mean mesh shrinkage rate was 14.2% for Optilene® LP vs. 24.7% for INFINIT® mesh (p = 0.017). The partial volume of the inflammatory cells was 11.2% for Optilene® LP vs. 13.9% for INFINIT (n.s.). CD68 was significantly higher for INFINIT (11.8% vs. 5.6%, p = 0.007). The markers of cell turnover, namely Ki67 and the apoptotic index, were comparable at 6.4% vs. 12.4% (n.s.) and 1.6% vs. 2.0% (n.s.). In the extracellular matrix, TGF-ß was 35.4% for Optilene® LP and 31.0% for INFINIT® (n.s.). Collagen I (pos/300 µm) deposits were 117.8 and 114.9, respectively. CONCLUSION: In our experimental examinations, Optilene® LP and INFINIT® showed a comparable biocompatibility in terms of chronic inflammatory reaction; however, the shrinkage rate was significantly higher for INFINIT® after 3 months. The higher shrinkage rate of INFINIT® should be taken into account when choosing the mesh size for an adequate hernia overlap.

Comparison of real-time elastography with grey-scale ultrasonography for detection of organ-confined prostate cancer and extra capsular extension: a prospective analysis using whole mount sections after radical prostatectomy.

OBJECTIVE: •To evaluate whether transrectal real-time elastography (RTE) improves the detection of intraprostatic prostate cancer (PCa) lesions and extracapsular extension (ECE) compared with conventional grey-scale ultrasonography (GSU). PATIENTS AND METHODS: •In total, 229 patients with biopsy-proven PCa were prospectively screened for cancer-suspicious areas and ECE using GSU and RTE. •The largest tumour focus detected by RTE was defined as the index lesion. •The prostate gland was stratified into six sectors on GSU and RTE, which were compared with histopathological whole mount sections after radical prostatectomy. RESULTS: •Histopathologically, PCa was confirmed in 894 out of 1374 (61.8%) evaluated sectors and ECE was identified in 47 (21%) patients. •Of these 894 sectors, RTE correctly detected 594 (66.4%) and GSU 215 (24.0%) cancer suspicious lesions. •Sensitivity was 51% and specificity 72% using RTE compared to 18% and 90% for GSU. •RTE identified the largest side specific tumour focus in 68% of patients. •ECE was identified with a sensitivity of 38% and specificity of 96% using RTE compared to 15% and 97% using GSU. CONCLUSIONS: •Compared with GSU, RTE provides a statistically significant improvement in detection of PCa lesions and ECE. •RTE enhances GSU, although improvement is still needed to achieve a clinically meaningful sensitivity.

Open radical retropubic prostatectomy gives favourable surgical and functional outcomes after transurethral resection of the prostate.

OBJECTIVES: To assess the peri- and postoperative outcome of patients treated with open radical retropubic prostatectomy (RRP) for prostate cancer and who had previously undergone transurethral resection of the prostate (TURP). PATIENTS AND METHODS: Prospectively collected data from a consecutive series of 1760 patients who had RRP between July 2003 and June 2007 at our institution were used to retrospectively match 62 cases (with previous TURP) with the same number of controls (without previous TURP). Matching variables were patient age, body mass index, prostate volume, preoperative total prostate-specific antigen (PSA) level, Gleason score, pathological stage, and intraoperative nerve-sparing procedure. Complete 1-year follow-up data were available for all patients. All collected data on surgery and perioperative complications were analysed. Functional outcome data at the 1-year follow-up were evaluated by applying an institutional questionnaire. Sexual function was assessed using the abbreviated International Index of Erectile Function-5 questionnaire, and urinary control was evaluated by defining complete urinary control as no pad usage. RESULTS: The rate of complete urinary control rate in cases and controls was similar (81% vs 82%). When nerves were spared, 60% (15/25) of patients in either group were capable of sexual intercourse. The overall positive surgical margin rate was insignificantly higher in cases (19% vs 13, P>0.05). After 1 year of follow-up the biochemical recurrence rate (PSA>0.04 ng/mL) did not differ significantly in patients who had RRP after TURP vs RRP alone (six of 62, 10%, vs five of 62, 8%; P=0.77). CONCLUSIONS: RRP for prostate cancer in patients who have had previous TURP does not result in a higher perioperative complication rate, or a worse functional outcome.

[Breast-conserving treatment of breast cancer after augmentation with injected hydrophilic polyacrylamide gel and review of the literature]. / Brusterhaltende Therapie des Mammakarzinoms nach Augmentation mit injiziertem hydrophilem Polyacrylamidgel und Literaturübersicht.

We report on a 33-year-old female patient with invasive ductal breast cancer. Despite breast augmentation with injected hydrophilic polyacrylamide gel in her history, she was successfully treated with breast-conserving therapy. The widespread migration of the gel conglomerates first complicated diagnostic imaging, surgical treatment and tumour aftercare. Removing the gel proved a difficult task. Nevertheless, the gel was macroscopically totally removed allowing a breast-conserving therapy. Wound healing took place without complications. After adjuvant chemotherapy, radiotherapy and hormonal therapy, the patient stays tumour free with a satisfactory cosmetic result.

The use of composite meshes in laparoscopic repair of abdominal wall hernias: are there differences in biocompatibily?: experimental results obtained in a laparoscopic porcine model.

INTRODUCTION: In recent years, laparoscopic repair of abdominal wall hernias has become increasingly established in routine clinical practice thanks to the myriad advantages it confers. Apart from the risk of intestinal damage following adhesiolysis, to date no information is available on the best way of preventing the formation of new adhesions in the vicinity of the implanted meshes. Numerous experimental investigations, mainly conducted on an open small-animal model, have demonstrated the advantages of coating meshes, inter alia with absorbable materials, compared with uncoated polypropylene meshes. In our established laparoscopic porcine model we set about investigating three of these meshes, which are already available on the market. MATERIALS AND METHODS: In total, 18 domestic pigs underwent laparoscopic surgery and three different composite meshes were tested in each case on six animals (Dynamesh IPOM, Proceed, Parietene Composite). At 4 months, postmortem diagnostic laparoscopy was carried out, followed by full-wall excision of the specimens. Planimetric analysis was conducted to investigate the size of the entire surface area and the extent of adhesions. Histological investigations were performed on five sections for each specimen. These focused on the partial volumes of inflammatory cells, the proliferation marker Ki67, apoptotic index, inflammatory cell marker CD68 and transforming growth factor beta (TGF-beta) as a marker of the extracellular matrix. RESULTS: A similar value of 14% was obtained for shrinkage of Dynamesh IPOM and Parietene Composite, while Proceed showed a 25% reduction in its surface area. Markedly lower values of 12.8% were obtained for Parietene Composite in respect of adhesions to the greater omentum, compared with 31.7% for Proceed and 33.2% for Dynamesh IPOM (p = 0.01). Overall, Parietene Composite performed best in the histological and immunhistochemistry tests. CONCLUSIONS: On the whole, all composite meshes showed evidence of good biocompatibility. However, none of the coatings was completely able to prevent adhesions. Coating of polypropylene meshes with collagen appears to confer significant advantages compared with other coatings.

Cryopreserved human amniotic membrane for soft tissue repair in rats.

Fresh amniotic membrane has been used in medicine since 1910. The reconstruction of immunologic privileged ocular surfaces with cryopreserved amniotic membrane was introduced in the 1990s. The aim of this study was to analyze the use of cryopreserved human amniotic membrane (HAM) as a surgical patch in immunologic unprivileged anatomic sites. In part I of the investigation, the abdominal wall muscle of 36 rats was covered with mono- and multilayered HAM. After 3, 14, and 28 days, respectively, these grafts were evaluated macro- and microscopically. Multilayer samples displayed slower degradation and less inflammation compared with monolayer coverage. In part II of the study, abdominal wall closure with multilayer HAM and with polypropylene mesh was conducted in 20 rats. All rats showed sufficient closure after 21 days, but significantly lower intraabdominal adhesion formation was observed in the HAM rats. The results of this study might pave the way for the use of cryopreserved HAM as graft material in reconstructive surgery.

Does the additional application of a polylactide film (SurgiWrap) to a lightweight mesh (TiMesh) reduce adhesions after laparoscopic intraperitoneal implantation procedures? Experimental results obtained with the laparoscopic porcine model.

BACKGROUND: Intraperitoneal repair of incisional hernias using a mesh makes particular demands on the material used. In addition to good integration of the mesh on the parietal side, adhesions to the visceral peritoneum must be avoided. Large-pore, light-weight meshes induce fewer adhesions than heavy-weight polypropylene meshes. Although numerous adhesion-barrier substances for use in combination with a polypropylene mesh have been tested already, mostly in open small animal models, unequivocal benefits have been identified to date in only a few of the experiments. METHODS: Using the laparoscopic intraperitoneal onlay mesh technique, six pigs were implanted with either a lightweight polypropylene mesh (TiMesh light) or TiMesh plus an adhesion-barrier film made of polylactide (SurgiWrap). After 3 months, the animals underwent a postmortem laparoscopy, and specimens were obtained for planimetric and histologic investigations. RESULTS: No adhesions to intestinal structures were found in any of the animals. Adhesions between the greater omentum and the mesh did not differ significantly between the TiMesh (32%) and SurgiWrap (33.5%) groups. The shrinkage of the mesh's surface area was comparable between the two groups (18% vs. 21%). Histology showed pronounced inflammatory reaction and bridging of scar tissue between the filaments with the use of SurgiWrap versus TiMesh light without film. However, immunohistochemical investigations examining the partial volume of the inflammatory cells, the proliferation marker Ki67, and the apoptotic index at the interface of the filaments all failed to show any significant differences. CONCLUSION: To avoid adhesions, it is essential that the acute and chronic inflammatory reaction to the implanted material be as small as possible. This requirement is met specifically by the lightweight polypropylene mesh TiMesh light. The additional application of a slowly absorbable adhesion-barrier film made of polylactide (SurgiWrap) does not appear to confer any further benefit.

[The leishmaniasis - a parasitel infection as differential diagnosis of malignant tumours of oral mucosa. A case report and review of literature]. / Die Leishmaniasis - eine parasitäre Erkrankung als Differenzialdiagnose maligner Mundschleimhauterkrankungen. Fallbericht und Literaturübersicht.

Although parasitel infections in northern Europe are rare, it must be considered as differential diagnosis of malignant tumours of mucous membrane. With increasing tourisms in endemic areas, infections with parasite pathogen are spreading in non-endemic areas as well. In this case a mucous membrane malignancy with clinical feature of ulcer on unusual location was imitated. In this reported case the patient suffers with hepatitis c, causing cirrhosis of the liver and making a liver transplantation necessary. In this patient a history of a leishmaniosis which had been treated successful by the tropical institute is reported, but because of a new actually leishmaniosis-infection a liver transplantation is contraindicated. Under oral therapy with Miltefosin (IMPADIVO) a remission was successful. The leishmaniosis is a classical tropical disease. WHO reported a morbidity of nearly 12 million people in 88 countries around the world especially in tropical areas. Repeatedly infections in northern Europe caused by the phlebotonus-sandflies are described. Therefore leishmaniosis must be considered as differential diagnosis in suspect lesions of mucous membrane.

Investigating the accuracy of the FLUKA code for transport of therapeutic ion beams in matter.

In-beam positron emission tomography (PET) is currently used for monitoring the dose delivery at the heavy ion therapy facility at GSI Darmstadt. The method is based on the fact that carbon ions produce positron emitting isotopes in fragmentation reactions with the atomic nuclei of the tissue. The relation between dose and beta(+)-activity is not straightforward. Hence it is not possible to infer the delivered dose directly from the PET distribution. To overcome this problem and enable therapy monitoring, beta(+)-distributions are simulated on the basis of the treatment plan and compared with the measured ones. Following the positive clinical impact, it is planned to apply the method at future ion therapy facilities, where beams from protons up to oxygen nuclei will be available. A simulation code capable of handling all these ions and predicting the irradiation-induced beta(+)-activity distributions is desirable. An established and general purpose radiation transport code is preferred. FLUKA is a candidate for such a code. For application to in-beam PET therapy monitoring, the code has to model with high accuracy both the electromagnetic and nuclear interactions responsible for dose deposition and beta(+)-activity production, respectively. In this work, the electromagnetic interaction in FLUKA was adjusted to reproduce the same particle range as from the experimentally validated treatment planning software TRiP, used at GSI. Furthermore, projectile fragmentation spectra in water targets have been studied in comparison to available experimental data. Finally, cross sections for the production of the most abundant fragments have been calculated and compared to values found in the literature.

SOCS-3 is frequently methylated in head and neck squamous cell carcinoma and its precursor lesions and causes growth inhibition.

The suppressors of cytokine signaling (SOCS) are inhibitors of cytokine signaling that function via the Janus kinase (JAK)/signal transducers and activators of transcription (STAT) pathway. Recently, methylation of SOCS-1 and SOCS-3 has been implicated in the tumorigenesis of liver and lung cancer. This study was performed to elucidate the role of SOCS-1 and SOCS-3 in squamous cell carcinoma of the head and neck (HNSCC) and its precursor lesions. HNSCC of 94 patients and corresponding normal mucosa, lymph node metastases as well as 16 high- and 21 low-grade squamous cell dysplasias were studied by using methylation-specific PCR (MSP) for the SOCS-1 and SOCS-3 promoter after microdissection. The presence of SOCS-3 mRNA transcripts was confirmed by semiquantitative real-time PCR, and the SOCS-3 protein was analysed immunohistochemically. SOCS-3 hypermethylation was found in 85/94 HNSCC (90%) and in 10/16 high-grade and 9/21 low-grade dysplasias (63 and 43%, respectively). SOCS-1 promoter hypermethylation was detected in 10/94 HNSCC samples (11%) and in 2/16 high-grade and 1/21 low-grade dysplasias (13 and 5%, respectively). Lymph node metastases exhibited an identical methylation status as the primary tumors. Methylation of the SOCS-3 promoter correlated with downregulation of SOCS-3 transcripts and protein expression in these tumors and various cell lines. In the cell lines tested, SOCS-3 and SOCS-1 transcripts increased upon treatment with the demethylation compound 5-aza-2-deoxycytidine (5-AZA-DC). Overexpression of wild-type SOCS-3 in carcinoma cells with methylated SOCS-3 resulted in the induction of apoptosis and growth suppression as well as downregulation of STAT3, bcl-2 as well as bcl-xL. Our data suggest that promoter methylation and subsequent transcript downregulation of SOCS-3 transcripts and, to a much lesser extent, SOCS-1 are involved in the multistep carcinogenesis of HNSCC. During its involvement in tumor growth, restoration of SOCS-3 may hold treatment potential for HNSCC.

Mutations of BRAF and RAS are rare events in germ cell tumours.

The BRAF gene, one of the human isoforms of RAF, is activated by oncogenic Ras, leading to cooperative effects in cells responding to growth factor signals. Recently, somatic missense mutations in the BRAF gene have been detected in a variety of human tumors. We have studied male germ cell tumours (GCT) for probable mutations of the BRAF and Ras oncogene. Microsatellite instability (MSI) was analysed using mono- or di-nucleotide marker. Mutational analysis of 62 GCT (30 seminomas and 32 nonseminomas) was performed after microdissection of the different tumour components. The expression of Erk1/2, an important downstream point of convergence in the Ras-RAF-MEK-Erk pathway was assessed immunohistochemically. Activating BRAF missense mutations were identified in 3 out of 32 cases of nonseminomas (9%) but not in seminomas. The mutations were 1796T>A mutations and were found within the embryonic carcinoma component of these tumors. Two out of 30 seminomas (7%) and 3 out of 32 nonseminomas (9%) exhibited KRAS gene mutations. MSI was observed in 4 out 62 tumours (7%) [1 seminoma and 3 nonseminomas (embryonal carcinoma)]. All of the microsatellite instable embryonal carcinomas had a mutated BRAF gene. All 5 GCT with RAS mutations had an intact BRAF gene. We identified constitutively activated Erk in almost all tumours tested. Our data indicate that BRAF gene mutations are a rare event in GCT and are independent of KRAS mutations. In embryonal carcinomas, BRAF mutations may be linked to the proficiency of these tumours in repairing mismatched bases in DNA. The finding of activated Erk suggests a causative role for MAPK activation in GCT independent of activating BRAF or RAS mutations.