Obschon die endosonografisch geführte Feinnadelbiopsie (EUS-FNP) vor mehr als 22 Jahren in die klinische Medizin eingeführt wurde und heute aus der Routinediagnostik nicht mehr wegzudenken ist, werden weiterhin viele Detailfragen kontrovers beurteilt. Es gibt zahlreiche, unterschiedliche Punktionstechniken, mit denen versucht wird, möglichst viel Gewebe aus endosonografisch erkannten Zielläsionen zu gewinnen. Außerdem spielen Fragen wie die nach der "besten" Punktionsnadel, der "besten" Gewebeanalysemethode und der optimalen Ausbildungszielplanung für Endosonografiker eine wichtige Rolle. Ziel ist die weitere Verbesserung der mittlerweile fast flächendeckend angebotenen, minimalinvasiven EUS-Methode. Die vorliegende Übersicht geht einigen wichtigen Fragen nach und versucht, diese anhand der gegenwärtig erhältlichen "Evidenzlage" zu charakterisieren und zu bewerten. Dabei wird der hohe Stellenwert der Methodik im klinischen Alltag erneut deutlich. Die Bewertung einiger methodischer und technischer Schritte bietet auch kleine Überraschungen. Mit dem Studium dieser aktuellen Bestandsaufnahme und Zukunftsaussichten lassen sich die Feinnadelpunktionsergebnisse in jeder Endoskopieeinheit dauerhaft verbessern.

Endosonography with fine-needle aspiration biopsy (EUS-FNA) has become a widespreadly available clinical tool to diagnose numerous different lesions in humans. EUS-FNA is frequently used for tissue-based diagnoses such as lymphatic diseases (ranging from tuberculosis / sarcoidosis to malignant lymphoma) or solid tumors (such as pancreatic carcinoma, neuroendocrine tumors, sub-epithelial gastrointestinal tumors and others). Outcomes of EUS-FNA results, however, vary which is caused by several different factors ranging from experience of the endoscopist over technical factors such as use of stylet or suction for puncture through the skills of the cyto-pathologist who takes care of the specimen obtained by EUS-FNA. Though introduced since more than 20 years ago EUS-FNA has still not yet been perfectionized and several issues remain controversial among endoscopist. These issues include needle size and type (FNA versus TNB needles), use of a stylet and suction for FNA sampling, pure cytologic assessment versus cyto-histologic techniques, grading of the investigator´s and pathologist´s experience and improvement of EUS training for novices. In this report we briefly review the actual literature and summarize the available evidence on some controversely discussed issues. The results support the view that use of a stylet rarely aids to increase the amount of tissue obtained during EUS-FNA, whereas use of suction can be helpful in certain situations. Novel cutting needles may potentially improve number and size of core biopsies that can be rendered for special histologic tissue processing techniques. An in-room-cytopathologist not necessarily improves outcome of EUS-FNA results but may have a role during build-up of EUS units to become more successful. EUS-FNA education requires skilled endoscopists on both sides and can presumably be improved by objective testing of practical expertise by peer review and introducing objective sampling parameters. Novel techniques and equipment are about to evolve in the near future.

Objective: To identify and image protein biomarker candidates in the synovial tissue of patients with rheumatoid arthritis (RA) and patients with osteoarthritis (OA).Methods: A novel matrix-assisted laser desorption/ionization (MALDI) imaging mass spectrometry (IMS) technique was applied to the analysis of synovial tissue. Patients were classified according to the American College of Rheumatology (ACR) criteria for RA. Frozen sections were stained to obtain morphological data. Serial sections were desiccated, and spotted with matrix for MALDI analysis. Ions generated by laser irradiation of the tissue were separated in time, based on their m/z ratio, and were subsequently detected. IMS was used in a 'profiling' mode to detect discrete spots for rapid evaluation of proteomic patterns in various tissue compartments. Photomicrographs of the stained tissue images were reviewed by a pathologist. Areas of interest (10 discrete areas/compartment) were marked digitally and the histology-annotated images were merged to form a photomicrograph of the section taken before the MALDI measurement. Pixel coordinates of these areas were transferred to a robotic spotter, the matrix was spotted, and the coordinates of the spots were transferred to a mass spectrometer for spectral acquisition. The data generated were then subjected to biocomputation analysis to reveal the biomarker candidates.Results: Several peaks (m/z) consistent in mass with calgranulins, defensins, and thymosins were detected and their distribution in various synovial compartments (synovial lining and sublining layer) was demonstrated.Conclusion: MALDI IMS is a powerful tool for the rapid detection of numerous proteins (in situ proteomics) and was applied here for the analysis of the distribution of proteins in synovial tissue sections.

OBJECTIVES:With advances in laparoscopic surgery, laparoscopic deroofing has gained wide acceptance in the surgical community to treat symptomatic non-parasitic hepatic cysts. Published non-surgical data still favour aspiration and sclerotherapy as treatment in these cases, though morbidity is higher and recurrence rates are not acceptable. We reviewed all patients that had been treated by laparoscopic deroofing in our department over a period of 6 years in order to find out if the surgical approach should be considered the standard treatment.MATERIALS AND METHODS:Over a 6 year period, 34 laparoscopic deroofings were performed in 21 patients with symptomatic cysts. Patients underwent laparoscopy followed by radical deroofing using an Ultracision device.RESULTS:All cases were completed laparoscopically, no intraoperative adverse events were recorded. Mean operation time was 101 ± 22.7 min. The mean size of treated cysts was 9.7 ± 2.18 cm. Follow up showed only one symptomatic recurrence (3.3%) two months after surgery.CONCLUSION:Laparoscopic deroofing of hepatic cysts is a safe and effective treatment option. Recurrence rates are unprecedentedly low. Our data suggest that the risk of operation is justified and that the method is superior to sclerotherapy.

Ascites is a common clinical symptom in liver cirrhosis, inflammatory disorders of the abdomen and a major late manifestation of metastatic malignancies. Standard cytopathological techniques and immunocytochemistry have specificities and sensitivities of approximately 95 and 60%, respectively for the presence of tumor cells. Development of faster and more accurate screening methods would be of great clinical utility. In this work we examined differential analysis of the unbound proteins in the supernatant of ascites fluid by Protein-Chip SELDI mass spectrometry. There were 21 tumor cell-positive and 34 tumor cell-negative samples. We used principal component analysis coupled with linear regression applied to the mass spectra of the samples to distinguish between the sample groups. Two sample sets for statistical analysis were created after randomization, a training set with 37 samples and a validation set with 18 samples resulting in a specificity of 93% and a sensitivity of 83% on the training set. The validation set yielded a specificity and sensitivity of 75%. This study suggests that SELDI-TOF mass spectrometry appears to have great potential as a surrogate diagnostic tool to evaluate effusion specimens.

Einleitung: In Anbetracht der Fortschritte in der laparaskopischen Chirurgie innerhalb der letzten Dekade hat die laparoskopische Entdachung von non-parasitären, symptomatischen Leberzysten eine weite Akzeptanz in chirurgischen Kliniken gefunden.

Der Anstieg der Leberwerte AST und ALT in der 2. Schwangerschaftshälfte muss an das HELLP-Syndrom denken lassen, da dieses eine der schwerwiegendsten schwangerschaftsspezifischen Erkrankungen darstellt. Differentialdiagnostisch kommen neben der akuten Hepatitis insbesondere die Schwangerschaftsbedingte Cholestase in betracht. Die Diagnose HELLP-Syndrom lässt sich schließlich oft erst durch den Verlauf der Laborparameter mit der typischen Konstellation der Thrombozytopenie und dem Abfall des Haptoglobins, sowie der präeklamptischen Symptomatik stellen, die aber auch fehlen kann.

Endometriose ist eine der häufigsten Erkrankung bei Frauen im Reproduktionsalter, die verschiedenste Erscheinungsformen zeigen kann und für die Diagnostik eine schwierig fassbare Entität darstellt. Bislang wurde eine Reihe von Screening-Markern für Endometriose vorgestellt, um eine Verkürzung des Intervalls zwischen Beginn der Symptome und Diagnose und Therapie der Erkrankung zu erreichen.

Einleitung: in 1–2% muss bei hypertensiven Schwangerschaftserkrankungen mit schweren Verlaufsformen gerechnet werden. Unverändert bestehen erhebliche Schwierigkeiten hinsichtlich der frühen Diagnosestellung bei der Präeklampsie bzw. dem HELLP-Syndrom. Unter der Vorstellung, dass sich alle krankheitsassoziierten Veränderungen auf der Proteomebene manifestieren, haben wir Serumproben von Patientinnen mit Präeklampsie, HELLP-Syndrom und unkomplizierten Schwangerschaften massenspektrometrisch analysiert.

Hintergrund: Endometriose stellt neben Malignomen des weiblichen Genitale die häufigste Ursache für organdestruktives Wachstum dar und ist gekennzeichnet durch das Vorkommen von endometrialem Gewebe außerhalb des Cavum uteri. Für die Etablierung von Endometrioseherden und der weiteren Proliferation der Läsionen spielt neben verschiedenen Adhäsionsmolekülen, wie ICAM und VCAM, die lokale Expression von gewebedegradierenden Matrix-Metalloproteinasen (MMPs), im besonderen MMP-2, MMP-9 und MMP-11, eine zentrale Rolle.

Pouchitis after restorative proctocolectomy for ulcerative colitis is usually of ill-defined etiology and is encountered with sclerosing cholangitis, bacterial overgrowth, and ischemia. Recently, appendiceal involvement, ileitis, and fissures in the colectomy specimen have been associated with short- and long-term development of pouchitis. To corroborate these recent findings, the histology of 40 colectomies (70% males; mean age 46.3 years, age range 20-70 years; mean follow-up period 3.7 years, range 1-13 years) with yearly follow-up biopsies was correlated with pouchitis and clinical symptoms. Appendicitis, fissures, and ileitis were present in 47, 45 and 5% of the patients, respectively. Pouchitis in patients with appendicitis or with fissures was noted in 44 and 50% at first biopsy and in 70 and 58% during follow-up (p = NS). Of the patients without appendicitis or without fissures, 33 and 33% demonstrated pouchitis at the first biopsy and 30 and 55% during follow-up (p = NS). Clinico-histological correlation revealed normal/near-normal biopsies with the lowest clinical severity score in 77% and with the highest clinical score in 43% (p < 0.025). The histological findings of appendiceal involvement, fissuring ulcers, and ileitis in colectomies for ulcerative colitis do not correlate with the finding of pouchitis in early or late pouch biopsies. A high clinical suspicion score is frequently not correlated with significant inflammation of the pouch.

PURPOSE:Our study was designed to evaluate promoter CpG island hypermethylation in the diagnosis and prognosis of prostate cancer.EXPERIMENTAL DESIGN:Primary prostate cancers from 53 patients, pelvic lymph nodes, noncancerous prostate tissues, and prostate cell lines were analyzed. Real-time methylation-specific PCR was used to identify CpG island hypermethylation at five promising gene loci (i.e., GSTP1, APC, PTGS2, MDR1, and RASSF1a).RESULTS:At three gene loci (GSTP1, APC, and PTGS1) and CpG island, hypermethylation was highly prevalent in prostate cancers (71-91%), and analysis of receiver operator curves showed that hypermethylation at these three gene loci can distinguish between prostate cancer and noncancerous prostatic tissue (i.e., benign hyperplasia) with a sensitivity of 71.1% to 96.2% and a specificity of 92.9% to 100%. Using sensitive SYBR green methylation-specific PCR technology, we observed a respective 28% and 71% hypermethylation rate at the RASSF1a and MDR1 loci in benign prostate hyperplasia, which may represent early nonaggressive carcinogenesis. Methylation characteristics in prostate cancer metastases (i.e., pelvic lymph nodes) were comparable to the respective primary cancer. Statistical analysis showed no correlation between the methylation status of a single gene locus and clinicopathologic variables (e.g., preoperative prostate specific antigen levels, Gleason score, capsular penetration, involvement of seminal vesicle, and age). In contrast, the methylation of two (GSTP1/APC; GSTP1/PTGS2) or three (GSTP1/APC/PTGS2) gene loci correlated with prognostic indicators (i.e., pathologic stage, extraprostatic extension, and Gleason score, but not with prostate specific antigen levels).CONCLUSIONS:Our data suggest that the evaluation of DNA hypermethylation at three gene loci (i.e., GSTP1, APC, and PTGS2) is of diagnostic and prognostic value in prostate cancer.

Background: The t(2;5)(p23;q35) translocation is associated with a high percentage of anaplastic large-cell lymphomas (ALCL) of T- or null-cell phenotype. The translocation produces an 80 kDa hyperphosphorylated chimeric protein (p80) derived from the fusion of the anaplastic lymphoma kinase (ALK) with nucleophosmin (NPM). The NPM-ALK chimeric protein is an activated tyrosine kinase that has been shown to be a potent oncogene and presumably plays a causative role in lymphomagenesis. Materials and Methods: A transgenic mouse line was generated, where the human NPM-ALK cDNA is driven by the lck promoter conferring transgene expression to early T-cells. Results: Mice rapidly developed large cell lymphoblastic lymphomas with a median latency of 8 weeks, primarily involving the thymus, with lymph node as well as histologically evident extranodal organ infiltration by large tumor cells. Conclusion: The transgenic approach described provides direct evidence for the strong transforming potential of NPM-ALK in T-cells and furthermore represents a system for the analysis of the oncogenic events mediated by NPM-ALK in vivo, which might be instrumental in the development of tyrosine kinase inhibitor therapies of potential clinical use.