TY  - JOUR
AB  - Tissues used for clinical diagnostics are mostly formalin-fixed and paraffin-embedded (FFPE) which provides many advantages. However, the quality of the obtained nucleic acids (NA) is reduced and this turns out to be a challenge for further molecular analyses. Although the spectrum of analyses of NA extracted from FFPE tissue has increased, the standard operating procedures for NA isolation from old tissue blocks still need to be improved. Here, we compared the efficiency of different NA extraction methods, using FFPE tissues of variable age and origin, with respect to downstream analyses. Our study showed that the phenol-chloroform isoamyl alcohol (PCI) and the commercial Qiagen protocol yielded samples with highest purity. The PCI protocol delivered the longest amplicons even from samples from the 1970s. We developed a short (1 h) tissue lysis procedure that turned out to be highly time- and cost-effective when DNA quality was tested using single and multiplex PCR. Compared to a 1-day lysis-protocol, the amplicons were only 100 bp shorter. In addition, single-copy genes used in daily routine were successfully amplified from long-term stored FFPE samples following 1-h tissue-lysis. The RNA integrity numbers (RIN) determined on RNA isolated from FFPE tissues indicated degraded RNA; however, all RINs were above the generally agreed threshold of 1.4. We showed that, depending on the purpose of the analysis, NA retrieved from FFPE tissues older than 40 years may be successfully used for molecular analysis.
AU  - Ludyga, N.
AU  - Grünwald, B.
AU  - Azimzadeh, O.
AU  - Englert, S.
AU  - Höfler, H.
AU  - Tapio, S.
AU  - Aubele, M.
C1  - 7247
C2  - 29592
SP  - 131-140
TI  - Nucleic acids from long-term preserved FFPE tissues are suitable for downstream analyses.
JO  - Virchows Arch.
VL  - 460
IS  - 2
PB  - Springer
PY  - 2012
SN  - 0042-6423
ER  - 

TY  - JOUR
AB  - no Abstract
AU  - Straub, B.K.*
AU  - Esposito, I.
AU  - Gotthardt, D.*
AU  - Radeleff, B.*
AU  - Antolovic, D.*
AU  - Flechtenmacher, C.*
AU  - Schirmacher, P.*
C1  - 6508
C2  - 28833
SP  - 761-765
TI  - IgG4-associated cholangitis with cholangiocarcinoma.
JO  - Virchows Arch.
VL  - 458
IS  - 6
PB  - Springer
PY  - 2011
SN  - 0042-6423
ER  - 

TY  - JOUR
AB  - Epithelial ovarian cancer is a highly metastatic disease and the leading cause of death among cancer of the female genital tract. Abnormal epidermal growth factor receptor (EGFR) signalling has been shown to be involved in epithelial-mesenchymal transition (EMT), an early step during metastasis. Additionally, over-expression of the E-cadherin repressor Snail, a key regulator of EMT, has previously been found to be associated with unfavourable prognostic features. Thus, the aim of our study was to elucidate the role of EGFR-dependent signalling pathways for Snail expression in ovarian cancer. For this purpose, we analysed 25 formalin-fixed and paraffin-embedded (FFPE) primary tumours and their corresponding metastases for the expression of 25 signalling pathway molecules by reverse phase protein arrays. We found a significant correlation of Snail with EGFR((Tyr1086)) and p38 MAPK((Thr180/Tyr182)) in primary ovarian carcinoma and with EGFR((Tyr1086)) in their corresponding metastasis. Additionally, we showed that high expression levels of Snail in primary tumours combined with high expression levels of the phosphorylated p38 MAPK((Thr180/Tyr182)) in metastasis lead to an increased risk for death in ovarian carcinoma patients. Thus, for future combinatorial cancer therapy, drug combinations that best target the deregulated protein network in each individual patient should be selected.
AU  - Hipp, S.*
AU  - Berg, D.*
AU  - Ergin, B.*
AU  - Schuster, T.*
AU  - Hapfelmeier, A.*
AU  - Walch, A.K.
AU  - Avril, S.*
AU  - Schmalfeldt, B.*
AU  - Höfler, H.
AU  - Becker, K.F.*
C1  - 2163
C2  - 27713
CY  - Berlin [u.a.]
SP  - 705-713
TI  - Interaction of Snail and p38 mitogen-activated protein kinase results in shorter overall survival of ovarian cancer patients.
JO  - Virchows Arch.
VL  - 457
IS  - 6
PB  - Springer
PY  - 2010
SN  - 0042-6423
ER  - 

TY  - JOUR
AB  - Trastuzumab-based therapy has been shown to confer overall survival benefit in HER2-positive patients with advanced gastric cancer in a large multicentric trial (ToGA study). Subgroup analysis identified adenocarcinomas of the stomach and gastroesophageal (GE) junction with overexpression of HER2 according to immunohistochemistry (IHC) as potential responders. Due to recent approval of trastuzumab for HER2 positive metastatic gastric and GE-junction cancer in Europe (EMEA) HER2 diagnostics is now mandatory with IHC being the primary test followed by fluorescence in situ hybridization (FISH) in IHC2+ cases. However, in order to not miss patients potentially responding to targeted therapy determination of a HER2-positive status for gastric cancer required modification of scoring as had been proposed in a pre-ToGA study. To validate this new HER2 status testing procedure in terms of inter-laboratory and inter-observer consensus for IHC scoring a series of 547 gastric cancer tissue samples on a tissue microarray (TMA) was used. In the first step, 30 representative cores were used to identify specific IHC HER2 scoring issues among eight French and German laboratories, while in the second step the full set of 547 cores was used to determine IHC HER2 intensity and area score concordance between six German pathologists. Specific issues relating to discordance were identified and recommendations formulated which proved to be effective to reliably determine HER2 status in a prospective test series of 447 diagnostic gastric cancer specimens.
AU  - Rüschoff, J.*
AU  - Dietel, M.*
AU  - Baretton, G.*
AU  - Arbogast, S.*
AU  - Walch, A.K.
AU  - Monges, G.*
AU  - Chenard, M.-P.*
AU  - Penault-Llorca, F.*
AU  - Nagelmeier, I.*
AU  - Schlake, W.*
AU  - Höfler, H.*
AU  - Kreipe, H.H.*
C1  - 2207
C2  - 27560
SP  - 299-307
TI  - HER2 diagnostics in gastric cancer-guideline validation and development of standardized immunohistochemical testing.
JO  - Virchows Arch.
VL  - 457
IS  - 3
PB  - Springer
PY  - 2010
SN  - 0042-6423
ER  - 

TY  - JOUR
AB  - In a previous retrospective study, we demonstrated the prognostic value of protein tyrosine kinase 6 (PTK6) protein expression in breast carcinomas. Here, we analyzed PTK6 gene amplification using fluorescence in situ hybridization technique in a cohort of 426 invasive breast carcinomas and compared it with PTK6 expression level as well as with the clinical outcome of patients. Forty-five percent of tumors show increased PTK6 gene copy numbers when compared to normal tissue. Most of these, however, were related to chromosome 20 polysomy (30%), while gene amplification accounted for only 15%. Only "low level" amplification of the PTK6 gene, with up to eight signals per nucleus, was found. The PTK6 cytogenetic status (normal, gene amplification, polysomy 20) was not associated with histopathological parameters or with the protein expression of HER receptors. No statistical association was identified between PTK6 gene status and expression level. Further, the PTK6 gene status does not influence the disease-free survival of patients at a parts per thousand yen240 months. Based on these results, we state that the PTK6 overexpression is not essentially attributed to gene amplification, and the PTK6 protein expression-but not gene status-is of prognostic value in breast carcinomas. PTK6 protein overexpression may result from polysomy 20 in a minority of the tumors. In a marked proportion of tumors, however, the overexpression is likely to be caused by posttranscriptional regulation mechanisms.
AU  - Aubele, M.
AU  - Vidojkovic, S.
AU  - Braselmann, H.
AU  - Ritterswürden, D.
AU  - Auer, G.*
AU  - Atkinson, M.J.
AU  - Tapio, S.
AU  - Höfler, H.
AU  - Rauser, S.
AU  - Bartlett, J.M.*
C1  - 1841
C2  - 26764
CY  - Heidelberg
SP  - 117-123
TI  - Overexpression of PTK6 (breast tumor kinase) protein - a prognostic factor for long-term breast cancer survival - is not due to gene amplification.
JO  - Virchows Arch.
VL  - 455
IS  - 2
PB  - Springer
PY  - 2009
SN  - 0042-6423
ER  - 

TY  - JOUR
AB  - Proteomics raises high expectations in finding novel and reliable biomarkers for diagnosis, prognosis and therapy prediction. The goal of the 2-day workshop "Protein analysis of tissues-current views and clinical perspectives" was to bring together scientists from multiple areas of protein research interested in tissue analysis.
AU  - Becker, K.F.*
AU  - Walch, A.K.
AU  - Ueffing, M.
C1  - 1391
C2  - 26740
SP  - 191-192
TI  - Protein analysis of tissues - current views and clinical perspectives.
JO  - Virchows Arch.
VL  - 455
IS  - 2
PY  - 2009
SN  - 0042-6423
ER  - 

TY  - JOUR
AB  - Secondary sclerosing cholangitis (SSC) is a chronic cholestatic disorder caused by mechanical, infectious, toxic, or ischemic factors. A new variant of SSC occurring after long-term treatment in intensive care units (ICU) has been recently described and characterized from the clinical point of view. The aim of this study was the histomorphological characterization of ICU-treatment-related SSC (ICU-SSC) and the definition of histological changes occurring over time based on the morphological findings. Liver biopsies of ten patients affected by ICU-SSC obtained at different time points (1.5 to 57 months) after the initial injury were analyzed. The main morphological alterations included degenerative changes of portal bile ducts, portal edema, inflammation, and fibrosis as well as biliary interface activity and bilirubinostasis. Perivenular necroses and bile infarcts were found in eight and six patients, respectively. Bile duct loss was not observed. No correlation between morphological features of biopsies and liver chemistry tests or outcome could be established. Based on the morphological observation, a possible disease-progression model starting with an initial damage of portal bile ducts (primary insult) with associated portal/periportal changes (inflammation, ductular reaction) and resulting in secondary parenchymal changes is proposed.
AU  - Esposito, I.
AU  - Kubisova, A.*
AU  - Stiehl, A.*
AU  - Kulaksiz, H.*
AU  - Schirmacher, P.*
C1  - 2656
C2  - 25668
SP  - 339-345
TI  - Secondary sclerosing cholangitis after intensive care unit treatment: Clues to the histopathological differential diagnosis.
JO  - Virchows Arch.
VL  - 453
IS  - 4
PB  - Springer
PY  - 2008
SN  - 0042-6423
ER  - 

TY  - JOUR
AB  - Pheochromocytomas are neuroendocrine tumors arising in the neural crest-derived chromaffin cells of the adrenal gland or in extra-adrenal sympathetic ganglia (paragangliomas). In a rat model of multiple endocrine neoplasia (MEN), absence of functional p27Kip1 protein predisposes to pheochromocytoma and paraganglioma development. As no data is available regarding the involvement of p27Kip1 in human pheochromocytoma and/or paraganglioma, we set out to determine the expression pattern of p27Kip1 in those tumor types. A panel of 25 pheochromocytomas and 23 paragangliomas was collected. Two pheochromocytomas were from MEN2 patients. The paragangliomas included 15 tumors that developed at the carotid bifurcation, three in the jugulo-tympanic area, and five at other sites. Except for the MEN2 cases, all others were apparently sporadic. Immunohistochemistry for p27Kip1 and the proliferation marker Ki67 was performed. We found that p27Kip1 expression is reduced/lost in 56% of pheochromocytomas, but only in 18.1% of paragangliomas. Downregulation of p27Kip1 was not associated with increased proliferation. Cases showing reduced/lost p27Kip1 expression were screened for the presence of somatic mutations in CDKN1B (p27Kip1) and for allelic imbalance at the p27Kip1 locus. Three cases had allelic imbalance but none had mutations. In conclusion, pheochromocytomas display extreme reduction/loss of p27Kip1 expression at high frequency.
AU  - Pellegata, N.S.
AU  - Quintanilla-Martinez, L.
AU  - Keller, G.*
AU  - Liyanarachchi, S.*
AU  - Höfler, H.
AU  - Atkinson, M.J.
AU  - Fend, F.*
C1  - 4333
C2  - 24640
SP  - 37-46
TI  - Human pheochromocytomas show reduced p27Kip1 expression that is not associated with somatic gene mutations and rarely with deletions.
JO  - Virchows Arch.
VL  - 451
IS  - 1
PB  - Springer
PY  - 2007
SN  - 0042-6423
ER  - 

TY  - JOUR
AU  - Rosivatz, E.*
AU  - Becker, K.-F.*
AU  - Kremmer, E.
AU  - Schott, C.*
AU  - Blechschmidt, K.*
AU  - Höfler, H.
AU  - Sarbia, M.*
C1  - 4010
C2  - 23662
SP  - 277-287
TI  - Expression and nuclear localization of Snail, an E-cadherin repressor, in adenocarcinomas of the upper gastrointestinal tract.
JO  - Virchows Arch.
VL  - 448
PY  - 2006
SN  - 0042-6423
ER  - 

TY  - JOUR
AU  - Fend, F.*
AU  - Bock, O.*
AU  - Kremer, M.*
AU  - Specht, K.*
AU  - Quintanilla-Martinez, L.
C1  - 4311
C2  - 23199
SP  - 909-919
TI  - Ancillary techniques in bone marrow pathology: Molecular diagnostic on bone marrow trephine biopsies.
JO  - Virchows Arch.
VL  - 447
PY  - 2005
SN  - 0042-6423
ER  - 

TY  - JOUR
AU  - Kayser, G.*
AU  - Gerlach, U.*
AU  - Walch, A.K.
AU  - Nitschke, R.*
AU  - Haxelmans, S.*
AU  - Kayser, K.*
AU  - Hopt, R.*
AU  - Werner, M.*
AU  - Lassmann, S.*
C1  - 5360
C2  - 22858
SP  - 61-65
TI  - Numerical and structural centrosome aberrations are on early and stable event in the adenoma-carcinoma sequence of colorectal carcinomas.
JO  - Virchows Arch.
VL  - 447
PY  - 2005
SN  - 0042-6423
ER  - 

TY  - JOUR
AU  - Kremer, M.*
AU  - Quintanilla-Martinez, L.
AU  - Nährig, J.*
AU  - Schilling, Ch. von*
AU  - Fend, F.*
C1  - 4312
C2  - 23200
SP  - 920-937
TI  - Immunohistochemistry in bone marrow pathology: A useful adjunct for morphologic diagnosis.
JO  - Virchows Arch.
VL  - 447
PY  - 2005
SN  - 0042-6423
ER  - 

TY  - JOUR
AB  - Adenocarcinoma of the gastroesophageal junction is rapidly rising in incidence. It has been proposed that these tumors be classified as three different types: distal esophageal (AEG I), cardia (AEG II), and subcardia (AEG III). Using comparative genomic hybridization (CGH) analysis, one recent study reported that the 14q chromosomal arm showed a significantly higher rate of deletion in esophageal than in cardia adenocarcinoma. Using a microsatellite analysis technique, we analyzed this area and regions in the vicinity of the APC, DCC, and p53 genes. Tumor and normal tissues were microdissected from 54 cases (27 AEG I and 27 AEG III). DNA was extracted and then amplified using seven fluorescent-labeled microsatellite markers, one pair each on 5q, 18q, and 17p and four on 14q. The results were analyzed for loss of heterozygosity (LOH) and microsatellite instability (MSI). LOH varied from 20% to 30% at each locus except for the 17p locus, where it was slightly above 50% in both groups. No significant differences in LOH or MSI were found between the esophageal and gastric tumors, including the 14q chromosomal arm. These results fail to confirm the finding that abnormalities on the 14q chromosomal arm distinguish between distal esophageal and proximal gastric tumors.
AU  - Yanagi, M.*
AU  - Keller, G.*
AU  - Müller, J.*
AU  - Walch, A.K.
AU  - Werner, M.*
AU  - Stein, H.J.*
AU  - Siewert, J.R.*
AU  - Höfler, H.
C1  - 21559
C2  - 19684
SP  - 605-610
TI  - Comparison of loss of heterozygosity and microsatellite instability in adenocarcinomas of the distal esophagus and proximal stomach.
JO  - Virchows Arch.
VL  - 437
IS  - 6
PY  - 2000
SN  - 0042-6423
ER  - 

TY  - JOUR
AB  - Epithelial proliferation in the ventral surface of mouse tongue follows a pronounced circadian rhythm with a peak in mitotic activity at 10.00 a.m., preceded by a wave of DNA synthesis 8 h earlier. Nearly all cells (85%) pass through G2 and mitosis immediately after the S-phase; they subsequently divide again, usually after 2 or 3 days, indicating cohorts of cells with different G1-duration. The fraction of all nucleated cells comprised in one daily proliferation wave is about 20% indicating a turnover time of the nucleated cell compartment of about 5 days. Cytotoxic injury by a single radiation dose of 20 Gy causes a steep decrease in cell counts, leading to complete denudation after 9-13 days. The difference between the latent period before ulceration and the tissue turnover time is explained by a marked proliferative activity of the doomed cells. The mitotic index increases steeply after day 1 to three times the control level, but most mitotic figures display gross abnormalities such as multipolar spindles or chromosome clumping. As a consequence cells with abnormal or multiple nuclei appear in the basal layers 3 days post irradiation and subsequently migrate to the upper layers. After denudation the epithelium rapidly becomes restored, with a phase of transient hyperplasia on days 13-14. Normal architecture is regained by day 15. Over the whole healing period the mitotic index remains at a high level, with most of the mitoses appearing histologically normal.
AU  - Dörr, W.
AU  - Kummermehr, J.C.
C1  - 40726
C2  - 40183
SP  - 287-294
TI  - Proliferation kinetics of mouse tongue epithelium under normal conditions and following single dose irradiation.
JO  - Virchows Arch.
VL  - 60
IS  - 5
PY  - 1991
SN  - 0042-6423
ER  - 

TY  - JOUR
AB  - Crystalline arrays of coated parallel tubules (CPT) were observed by electron microscopy within dilated cisternae of the rough endoplasmic reticulum of pleomorphic mononuclear cells in a human osteosarcoma. The wall of the peculiar intracisternal tubules consisted of an electron-dense thin membrane-like envelope which appeared to be composed of granular subunits. The electron-lucent tubular core together with the limiting envelope was approximately 15 nm in diameter. A coat of fuzzy material, approximately 10 nm thick, tightly surrounded the membrane-like wall of the tubules. Cross sections of accumulations of CPT showed the tubules to be arranged in a hexagonal crystalline array. The nature and significance of the intracisternal CPT are unknown.
AU  - Marquart, K.H.
C1  - 42331
C2  - 38692
SP  - 309-313
TI  - Intracisternal crystalline arrays of coated parallel tubules in cells of a human osteosarcoma.
JO  - Virchows Arch.
VL  - 391
IS  - 3
PY  - 1981
SN  - 0042-6423
ER  - 

TY  - JOUR
AB  - Prenatal X-irradiation of mice in the late organogenesis stage either with a fractionated or a single exposure dose (3 × 160 R or 200 R) leads to remarkable, previously undescribed malformations of the skull. These malformations range from mild hyperostotic nodule formation in about 90% of the offspring to excessive formation of desmal bony tissues, which extend deep into the forebrain and are thus only detectable in histological sections. Metaplastic and hyperplastic formation of cartilage in all the neurocranial regions is observed in about 10% of the offspring. The pathogenesis of these overgrowth phenomena is presumably related to a growth disturbance of both the mesenchymal skull primordium and the brain. While malformation of the latter leads to a decrease of intracranial pressure and consequently to altered growth activity of the skull sutures, the reparative and proliferative capacities of the mesenchyme are also stimulated, in a hyperplastic direction, by X-irradiation.
AU  - Schmahl, W.G.
AU  - Meyer, I.
AU  - Kriegel, H.
AU  - Tempel, K.H.*
C1  - 41641
C2  - 35777
SP  - 173-184
TI  - Cartilaginous metaplasia and overgrowth of neurocranium skull after X-irradiation in utero.
JO  - Virchows Arch.
VL  - 384
IS  - 2
PY  - 1979
SN  - 0042-6423
ER  -