96. Jahrestagung der Deutschen Gesellschaft für Pathologie e. V ...

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Conclusions. The newly released 450k methylation array from Illumina provides a genome-wide representation of DNA methylation aberrations in a convenient format with direct quantification of methylation levels at each individual CpG site. However, the representation of microRNA genes and imprinted loci is quite uneven and has to be taken into account during data evaluation and derivation of any conclusion concerning these two important classes of genes. SA-P-038 Luminescent conjugated oligothiophenes: novel optical probes that detect cross beta-sheet conformation of inclusion bodies “in situ” V . Mahajan1 , T . Klingstedt2 , R . Simon2 , P . Nilsson2 , A . Thueringer1 , K . Kashofer1 , H . Denk1 , K . Zatloukal1 , J . Haybaeck1 1 2 Medical University of Graz, Institute of Pathology, Graz, Austria, Linkoping University, Sweden Aims. Mallory-Denk bodies (MDBs) are cellular hallmarks of protein aggregation diseases such as alcoholic and non-alcoholic steatohepatitis (ASH and NASH). MDBs are also seen in other liver diseases such as hepatocellular carcinoma (HCC) and alpha-1-antitrypsin deficiency. Additionally presence of Intracellular Hyaline bodies (IHBs) is also a common observation in HCC. Despite of advances in technologies highlighting protein conformation, structural characterisation of inclusion bodies remains unresolved. Therefore investigating molecular structure of the major MDB constituents keratin 8 (K8) and 18 (K18), p62 and ubiquitin may provide deeper mechanistic insights in MDB/IHB formation. Methods. Luminescent conjugated oligothiophenes (LCOs), h-HTAA, p-FTAA and PTAA contain swivelling thiophene backbones whose geometry modulates fluorescence properties. LCOs were demonstrated to specifically bind proteins with cross beta sheet conformation. In addition to the older ones the new generation of LCOs were used for in situ investigation of conformational changes in MDBs in human and murine livers. Results. LCOs demonstrated constant presence of cross beta-sheet conformation in human MDBs but not in IHBs, alpha-1-antitrypsin and ground glass inclusions. The spectral signatures collected from MDBs in ASH, NASH and HCC indicated a similar molecular structure of MDBs under the various disease conditions. MDBs induced by 3, 5-diethoxycarbonyl-1, 4-dihydrocollidine-feeding of mice revealed h-HTAA and p-FTAA binding to MDBs in all experimental stages. CHO-K1 cells transfected with various combinations of SQSTM1/p62, ubi and Krt8/ Krt18 showed that K8 was more prone than K18 to lead to generation of cross beta-sheets. Aggregates of p62 alone were reproducibly negative for LCOs. Circular dichroism analysis elucidated intrinsically different conformational nature of purified K8 and K18 polypeptides. Conclusions. The comparative higher tendency of K8 to undergo conformational changes from predominantly Alpha-helical to cross β-sheet explains its essential role in MDB formation. This elucidates why the absence of K8 prevents MDB formation whereas its excess facilitates MDB formation. The nature of keratins to acquire cross beta sheet conformation appears to be dependent on intrinsic factors but may not be the consequence of pathological conditions. With PTAA, LCOs may serve as a multicolour novel diagnostic “in situ” technology that can be applied on formalin-fixed and paraffin-embedded and fresh frozen tissues. SA-P-039 Comparison of cobas and HC2 HPV testing in a German routine laboratory H . Ikenberg1 , C . Börsch1 , B . Pittel1 , A . Xhaja1 , F . Britz2 , T . Iftner3 1 2 Cytomol, MVZ for Cytology and Molecular Biology, Frankfurt, Roche Diagnostics, Mannheim, 3University of Tübingen, Institute for Virology, Tübingen Aims. Up to now the Digene HC2 test (Qiagen, Hilden) is regarded the gold standard for human papillomavirus (HPV) DNA testing. Meanwhile several new test systems for HPV are available, among them the cobas HPV assay (Roche Diagnostics, Mannheim, Roche) which allows simultaneous genotyping for HPV 16 and 18. We compared the performance of the cobas HPV with the HC2 assay in cervical samples collected with the PreservCyt collection medium (Hologic, Frankfurt). Methods. 1781 anonymized routine specimens pretested with the HC2 test were available for analysis with cobas HPV. Cases with discrepancies between the two tests were retested with the Linear Array HPV genotyping test (LA; Roche). Partially, histologic diagnoses were available. Results. In 1566 (87.9%) of the cases HPV results were concordant. Of the 215 cases (12.1%) with discrepancies LA results are available in 214 cases. 94 cases were LA-negative: 13 of 105 cobas-pos/HC2-neg and 81 of 99 cobas-neg/HC2-pos cases. 110 cases were LA-positive: 92 of 105 cobas-pos/ HC2-neg and 18 of 99 cobas-neg/HC2-pos cases. 325 cases with histologically confirmed CIN2+ were included. In 261 out of 293 cases (89.1%) where a HC2 result was available this was positive, while 298 out of 318 of the cases (93.7%) tested with cobas HPV were positive. The rate of HPV positivity in cytologically normal cases and in HSIL was slightly higher with cobas HPV, while it was in the same range in ASC-US and in LSIL cases. With increasing severity of the cytologic findings the rate of HPV 16- and 18 positivity increased proportionally. Conclusions. In routine specimens from a German commercial laboratory the cobas HPV test showed similar performance compared to the HC2 test. Preliminary data point to a potentially higher sensitivity and specificity with cobas HPV while adding information by HPV 16 and 18 genotyping. SA-P-040 Comparison of Abbott RealTime and HC2-HPV testing in a routine laboratory H . Ikenberg1 , C . Noppen2 , M . Faber1 , A . Xhaja1 , S . Böhm3 1 2 Cytomol, MVZ for Cytology and Molecular Biology, Frankfurt, Viollier AG, Basel, Switzerland, 3University Hospital Leipzig, Dep . for Gastroenterology and Rheumatology, Leipzig Aims. The Digene Hybrid Capture2 test (HC2, Qiagen, Hilden) is the standard in routine human papillomavirus (HPV) DNA testing. Several new test systems have become commercially available in the past years, among them the automated Abbott RealTime-High-Risk-HPV assay (RealTime-HPV, Abbott Molecular, Wiesbaden). It detects 14 HR-HPV types and simultaneously differentiates between HPV16, HPV18 and 12 non-HPV16/18 HR types in a single test, while HC2 targets the same HR types, except HPV66, without typing. RealTime-HPV amplifies human β-globin in the same reaction. We evaluated RealTime-HPV versus HC2 with cervical specimens from a large German routine laboratory sampled with the PreservCyt collection medium (Hologic, Frankfurt). Methods. 505 anonymized routine specimens referred for cytology and pretested with HC2 were run with RealTime-HPV on the m2000 System (Abbott). Samples with discordant results between both assays were genotyped by Linear Array (Roche, Mannheim), targeting 37 HPV genotypes. Results were correlated with routine histology available for 280 cases (16.8%
Abstracts were LA-positive for RealTime-HPV/HC2 targeted HPV types: 27 of 33 RealTime-HPV-pos/HC2-neg and 11 of 40 RealTime-HPV-neg/HC2pos cases. 35 cases were LA-negative: 6 of 33 RealTime-HPV-pos/HC2neg and 29 of 40 RealTime-HPV-neg/HC2-pos cases. In 280 histology confirmed cases overall-agreement between RealTime-HPV and HC2 was 82.5%. Detection rates for CIN2+ and CIN3+ were 90.6% and 90.7% with RealTime-HPV and 89.3% and 88.3% with HC2, respectively. A high correlation of HPV16/18 positivity with RealTime-HPV and increasing histological severity was found. Conclusions. Clinical performance of RealTime-HPV in routine specimens was comparable to that of HC2. Discrimination of HPV 16/18 from other HR HPV types provides additional information for risk stratification. SA-P-041 FISH analysis does neither reveal genomic ETV1 amplification nor break-apart in gastrointestinal stromal tumors E . Wardelmann1 , S . Huss1 , R . Menon2 , F . Göke2 , G . Kristiansen2 , R . Buettner1 , S . Perner2 1 2 University Hospital Cologne, Institute of Pathology, Köln, University Hospital Bonn, Institute of Pathology, Bonn Aims. Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of the gastrointestinal tract. Recently the transcription factor ETV1 was shown to be highly expressed in GISTs both at transcript and protein levels. ETV1 is a member of ETS family of transcription factors. The ETS family members share an evolutionary highly conserved 80-amino-DNA binding domain and individual proteins of this family are capable of regulating gene promoter activity. Chi et al. reported that ETV1 is highly expressed in those ICCs, from which GISTs arise. Taken together, they propose that GISTs originate from ICCs with high endogenous level of ETV1 when an oncogenic activation via KIT or PDGFRA mutation occurs. However, they could not detect any genomic alteration leading to the high ETV1 expression performing FISH analysis on four GIST samples and two GIST cell lines. The present study was accomplished to evaluate in a larger cohort whether an ETV1 amplification or break apart might represent the genomic alteration causing the reported ETV1 expression. Methods. For the ETV1 amplification assay, we used the commercially available Chromosome 7 reference probe CEP 7 (D7Z1), spanning the region 7p11.1–q11.1. The target probe was located on the ETV1 locus at 7p21.2. The target probe was labelled with biotin to produce a red signal using CTD-2220I3 BAC clone. For the ETV1 break-apart assay, we used BAC clones CTD-222013 for centromeric labelling with biotin and RP-11769K2 for telomeric labelling with digoxigenin. BAC clones were obtained from Invitrogen (Invitrogen, CA, USA). Results. We performed FISH analysis on 140 GIST patients using tissue micro arrays. Neither ETV1 amplification nor break apart was detectable. Conclusions. The present study ensures that neither a genomic ETV1 amplification nor a break apart is found in GISTs. We conclude that not genetic aberrations but rather upregulation of ETV1 due to high KIT receptor levels are responsible for the high ETV1 expression in GISTs. 152 | Der Pathologe · Supplement 1 · 2012 SA-P-042 A new whole genome amplification method for studying clonal evolution patterns in malignant colorectal polyps D . Hirsch1 , J . Camps1 , S . Varma2 , R . Kemmerling3 , T . Ried1 , T . Gaiser1 1Section of Cancer Genomics, Genetics Branch, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, United States, 2HiThru Analytics, Bethesda, United States, 3University Hospital Salzburg of the Paracelsus Private Medical University, Institute of Pathology, Salzburg, Austria Aims. The transition of normal colonic epithelium to adenoma and invasive carcinoma is defined by chromosomal aberrations. To identify the genetic drivers involved in this progression in samples from individual patients, we applied array comparative genomic hybridization (aCGH) to 13 formalin-fixed paraffin-embedded (FFPE) samples of early, localized human colon adenocarcinomas arising in high-grade adenomas (so called “malignant polyps”). These lesions are small and hence the amount of DNA limited. Additionally, the quality of the DNA is compromised due to the fragmentation as a consequence of formalin fixation. To overcome these problems, we optimized a newly developed isothermal whole genome amplification system (NuGEN Ovation® WGA FFPE System). Methods. DNA was isolated from the FFPE blocks of 13 malignant polyps, each consisting of areas of adenoma and carcinoma. Starting with 100 ng of FFPE DNA, the amplification system produced 4.01±0.29 µg (mean ± standard deviation) of DNA. The samples were then analyzed using Agilent SurePrint G3 Human CGH Microarrays 4×180K. Results. Genomic imbalances were conserved in this procedure as assessed by comparing amplified and unamplified FFPE DNA using aCGH. The excellent quality of amplified DNA was further indicated by a high signal-to-noise ratio and a low derivative log2 ratio spread. Both, the amount of amplified DNA and aCGH performance were independent from the age of the FFPE block and the associated degradation of the extracted FFPE DNA. We observed losses of chromosome arms 5q and 18q in the malignant polyp samples, while the embedded early carcinomas revealed losses of 8p, 17p, and 18, and gains of 7, 8q, 13, and 20. Aberrations detected in the adenoma were invariably maintained in the embedded carcinomas. Conclusions. In conclusion, this approach demonstrates that using isothermally whole genome amplified FFPE DNA is technically suitable for aCGH. Besides demonstrating clonal origin of the adenoma and carcinoma part within a malignant polyp, the gain of chromosome arm 20q was an indicator for progression from adenoma to carcinoma in malignant polyps. SA-P-043 Dissimilarities of colorectal (CRAIT) and sinonasal adenocarcinoma (SNAIT) of intestinal type K . Donhuijsen1 , I . Kollecker1 , H . Hannig1 , H .-G . Schroeder2 1Academical Hospital Braunschweig, Department of Pathology, Braunschweig, 2Academical Hospital Braunschweig, Department of Otorhinolaryngology, Braunschweig Aims. SNAIT and CRAIT are considered to be nearly identically whereas an uninformed pathologist would diagnose an endonasal metastasis of a CRAIT instead of a primary of the inner nose. However, in a closer look are there discrepancies to detect by morphology, immunohistology and molecular results? Methods. Fifty consecutive cases of SNAIT and CRAIT were compared with regard to morphological criteria as subtype, histological inhomogeneity, mucin production, desmoplastic reaction and vascular invasion. Further the expression of CDX2, CK7, CK20, Synaptophysin, p53, Ki67, were compared and also the molecular status (30 cases) by PCR for K- RAS, b-raf, EGFR and p53. Results. SNAIT and CRAIT are histologically quite similar but not identical: adenomatous precursor lesions are absent. Tumor inhomogeneity
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Inhalt Der Pathologe · Supplement
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Inhalt Der Pathologe · Supplement
Page 6 and 7:
Editorial Liebe Kolleginnen und Kol
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Kolorektales Karzinom 2 VO-005 Tran
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Keynote Lecture VO-014 Genetic dete
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(AMACR, FASN, GOLM1, GSP-pi, ERG) t
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to assess the correct rate of R1 re
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DNA damage, and cytotoxic drugs. Au
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HCV-positive formalin-fixed and par
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well as MET activation were examine
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or BRAF mutation, c-MYC and SIRT1 e
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AG Pneumopathologie III DO-032 Remo
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immature granulopoiesis showed a st
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ned, if well-defined mantle zones,
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phomas). Most prominent gains or am
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DO-062 Tumor-associated macrophages
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DO-080 DOG1: an immunohistochemical
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AG Oralpathologie DO-087 Detection
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DO-094 Do activated fibroblasts inf
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DO-101 Histopathological analysis o
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DO-108 Identification of potential
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Conclusions. In conclusion, 454 par
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subgroup of patients with B-Raf mut
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DO-002b Impact of terminologies in
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Methods. We performed MCPyV-FISH of
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FR-013 HPV-genotype distribution in
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Methods. Three different techniques
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(i.e. investment in equipment and e
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FR-032 COLD-PCR: a powerful tool in
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dissection (MBLND) technique to imp
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SO-005 Prevalence of mutations in s
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SO-011 Methylation profiling and in
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more effective than SAHA alone and
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SO-022 Specialized pathology review
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SO-027 Ki-67 in mitotic score group
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nificantly associated with advanced
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liver did not correlate with the mu
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AG Paidopathologie II SO-046 Overex
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Results. Histomorphology of the ova
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p42 and p27 have not yet been inves
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SO-068 Recent advances in understan
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SO-075 A novel, dual role of CCN3 i
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Pancreatic Adenocarcinoma SG-137 Se
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sease and 30 colon cancer serum sam
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Conclusions. Although CRC is charac
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differentiated and TNM stage III or
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pressed moderate levels of the prot
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FR-P-037 MALDI imaging reveals COX7
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in the context of therapy response
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on primary cells of wild-type and c
Page 102 and 103: chemotherapy was recommended and co
Page 104 and 105: aims are twofold: 1) to verify the
Page 106 and 107: Results. Her2/neu status in TMAs an
Page 108 and 109: prognostic impact was found in rect
Page 110 and 111: provided information on survival ti
Page 112 and 113: Methods. Biopsy specimens from 430
Page 114 and 115: the OS rate was 53% for patients wi
Page 116 and 117: FR-P-098 Immunohistological stainin
Page 118 and 119: FR-P-104 Histopathological evaluati
Page 120 and 121: within the earliest morphologic det
Page 122 and 123: Conclusions. In primary imaging a g
Page 124 and 125: fibrotic neointma development as we
Page 126 and 127: FR-P-129 Alpha-1-antitrypsin-PiZ-an
Page 128 and 129: FR-P-136 The expression of central
Page 130 and 131: cosa and in the periarterial tissue
Page 132 and 133: FR-P-149 Combination of Castleman d
Page 134 and 135: or without different concentrations
Page 136 and 137: Results. In 18 cases (79%) the caus
Page 138 and 139: FR-P-168 Autopsy findings in a 2-ye
Page 140 and 141: FR-P-174 MPGN-like glomerulonephrit
Page 142 and 143: Poster: Gynäkopathologie und Mamma
Page 144 and 145: SA-P-011 Genetic aberrations of pre
Page 146 and 147: Mechanismen, die eine Progression d
Page 148 and 149: internet server. A network of inter
Page 150 and 151: Methods. HE-gefärbte Schnittpräpa
Page 154 and 155: and desmoplastic reaction are diffe
Page 156 and 157: one patient revealed more than 9 mi
Page 158 and 159: situation, the V600E mutation in th
Page 160 and 161: SA-P-064 Evolution of molecular pat
Page 162 and 163: nic markers such as myf4 and MyoD1
Page 164 and 165: Conclusions. To our knowledge, this
Page 166 and 167: SA-P-085 Expression of the eukaryot
Page 168 and 169: Results. The papillary RCC type II
Page 170 and 171: SA-P-098 Male infertility: assessme
Page 172 and 173: SA-P-104 Process oriented scientifi
Page 174 and 175: Conclusions. The IBDW offers a uniq
Page 176 and 177: L Lasitschka F. DO-046 Lehmann A. F
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