Пауль Эрлих является основателем ряда важных направлений в биомедицинских исследованиях, среди ко... more Пауль Эрлих является основателем ряда важных направлений в биомедицинских исследованиях, среди которых на первом месте-иммунология и химиотерапия. Целью данного исторического экскурса к столетию со дня смерти П. Эрлиха является описание его роли в создании и развитии гистохимии. Ключевые слова: Пауль Эрлих, гистохимия, иммунология.
Extraocular sebaceous carcinoma (ESC) is a rare appendiceal skin tumor. In contrast to ocular seb... more Extraocular sebaceous carcinoma (ESC) is a rare appendiceal skin tumor. In contrast to ocular sebaceous carcinoma, information about the exact cellular architecture of these lesions is scarce and the histogenesis of ESC is unknown. Here, we extend our previous study and investigate 28 extraocular carcinomas in comparison to 54 benign sebaceous tumors and 8 cases of normal sebaceous glands using a broad spectrum of antibodies against p63, several keratins, adipophilin, EMA, Ki67, androgen receptor, and mismatch repair proteins. This observational study demonstrates that p63- and K5/14-positive basaloid cells are key cells in normal sebaceous gland and in all sebaceous tumors and that these basaloid cells give rise to EMA+, adipophilin+ sebocytes, and K5/14+, K7±, K10± ductal structures. Finally, about half of ESC is associated with superficial in situ neoplasia, which provides evidence that at least part of these carcinomas arises from flat superficial in situ carcinoma. In contrast to the normal sebaceous gland, about half of all sebaceous tumors lack keratin K7. MMR protein IHC-profiles role will be discussed.
Background/Aim: Polymorphous adenocarcinoma (PAC) is a low-grade salivary gland malignancy in con... more Background/Aim: Polymorphous adenocarcinoma (PAC) is a low-grade salivary gland malignancy in contrast to variants with papillary (PAP) or cribriform (CASG) architecture and confers the second most common malignancy of minor salivary glands. Our study aimed to identify prognostic factors and to evaluate histomorphological and molecular diagnostic criteria of PACs. Patients and Methods: A series of 155 PACs, including 10 PAPs and 12 CASGs from the population-based Cancer Registry of North Rhine-Westphalia (LKR-NRW) and the Hamburg Salivary Gland Reference Centre (HRC) were analyzed. Results: One fifth of the tumors were located in the major salivary glands and PACS/CASGS invariably lacked p40 expression. Fifty-two percent of PACs showed a PRKD1 E710D mutation. Ordinary PACs had a disease-specific 10-year survival probability of 97% compared to 90% when combining PAPs and CASGs. T-stage at diagnosis was a prognostic factor with 98% for stages T1/T2 versus 75% for T3/T4. Conclusion: Diagnostic algorithms for the PAC/CASG spectrum of tumors need to be improved and should include molecular markers.
Adenoid cystic carcinoma (ACC) is an aggressive head and neck malignancy characterized by a t (6;... more Adenoid cystic carcinoma (ACC) is an aggressive head and neck malignancy characterized by a t (6;9) translocation resulting in an MYB–NFIB gene fusion or, more rarely, an MYBL1 fusion. The true frequency and clinical significance of these alterations are still unclear. Here, we have used tissue microarrays and analyzed 391 ACCs and 647 non-ACC salivary neoplasms to study the prevalence, expression, and clinical significance of MYB/MYBL1 alterations by FISH and immunohistochemistry. Alterations of MYB or MYBL1 were found in 78% of the cases, of which 62% had MYB alterations and 16% had MYBL1 rearrangements. Overexpression of MYB/MYBL1 oncoproteins was detected in 93% of the cases. MYB split signal, seen in 39% of the cases, was specific for ACC and not encountered in non-ACC salivary tumors. Loss of the 3′-part of MYB was enriched in grade 3 tumors and was a significant independent prognostic biomarker for overall survival in multivariate analyses. We hypothesize that loss of the 3′-...
Objectives: Information of the clinicopathological characteristics and outcome data of patients w... more Objectives: Information of the clinicopathological characteristics and outcome data of patients with adenosquamous carcinoma of the pancreas (ASCAP) remains limited. This study's aim is to describe the clinical, pathological, and molecular characteristics of 25 resected ASCAPs. Methods: Of all 25 cases, patient characteristics, follow-up data, and pathological/immunohistological features were reviewed and analyzed. Results: In this 3-institutional retrospective analysis of 562 pancreatic cancer patients, we identified 25 cases with histologically confirmed ASCAP (4.4%). Follow-up was available in 21 ASCAP and 50 pancreatic ductal adenocarcinoma control patients with a median overall survival of 8.2 and 21 months, respectively. Age, tumor size, localization in the tail, lymph node status, and resection margin seem to be the most significant factors of survival in our ASCAP cohort. In contrast to pancreatic ductal adenocarcinoma, positive expression of p63, keratins K5/14, and the epidermal growth factor receptor are a robust marker profile of these tumors. Conclusions: Adenosquamous carcinoma of the pancreas comprises a group of neoplasms in which stage and adverse morphological features contribute to its bad prognosis. Further work must be pursued to improve detection and treatment options to reduce mortality. Specifically, differences in biology might become a target for the development of possible therapies.
Biomedical Journal of Scientific & Technical Research, 2018
The investigation conforms with Guide for the Care and Use of Laboratory Animals published by the... more The investigation conforms with Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85-23, revised 1996) and was per
Background: Salivary gland-like tumors are currently subdivided into those with and those without... more Background: Salivary gland-like tumors are currently subdivided into those with and those without myoepithelial differentiation. Furthermore, they show a wide range of differentiation patterns including squamous differentiation. Many authors believe that the myoepithelial cell plays a major role in this context. However, currently little is known about the evolution and cellular differentiation of these tumors. Material and Methods: For that reason, we performed an in situ Triple Immunofluorescence Lineage/Differentiation Tracing (isTILT) and qRT-PCR study of basal (K5/K14), glandular (K7/K8/18), and epidermal-specific squamous (K10) keratins, p63, and smooth muscle actin (SMA; myoepithelial marker) with the aim to trace different cell lineages and define their cellular hierarchy in tumors. We investigated 50 cases of salivary gland-like tumors of the breast (11 Adenoid-cystic carcinomas, 9 (adeno-)myoepithelial tumors, 8 adeno-squamous carcinomas including 12 syringomas of the nipp...
Nitric oxide (NO) is generated by a family of enzymes termed NO synthases (NOS) that convert L-ar... more Nitric oxide (NO) is generated by a family of enzymes termed NO synthases (NOS) that convert L-arginine to NO and citrulline. The role of NO as an important biological mediator and recognition of the pathophysiological significance of superoxides/NO interaction has led to an intensive research and development of therapies based on the interception of the NO signaling cascade in the pancreatitis course. However, the presence and localization of the NO-generating enzymes in various organs including pancreas are subject to controversy. We assumed that this controversy might reflect rather the diversity of experimental approaches and an insufficient sensitivity of the methods used. Applying tyramide signal amplification (TSA) immunohistochemical technology, we were able detect all three NOS isoforms both in exocrine and endocrine compartments and in the vasculature in the normal pancreas and in pancreatitis. This also allowed us to demonstrate that oxidative stress runs ahead of NOS up-regulation, which implies that the NO enhancement in the course of pancreatitis is likely to be an adaptive mechanism aimed at maintaining the homeostatic cellular level of the bioactive NO. The aims of this minireview are to describe normal intrapancreatic NO pathways and the role of NO in the pancreatitis course.
PURPOSE: Telomerase has been detected in a majority of human malignant tumors, making telomerase ... more PURPOSE: Telomerase has been detected in a majority of human malignant tumors, making telomerase activity (TA) one key difference between mortal and immortal cells. In this study, we evaluated in blind-trial fashion the association of TA with cytologic and final clinical/pathologic diagnosis in fine-needle aspirates (FNAs) of breast lesions. MATERIALS AND METHODS: In 172 FNAs, including 80 samples that were cytologically malignant, 18 that were atypical but not diagnostic for malignancy, and 74 that were cytologically benign, TA was determined by a modified nonradioactive telomeric repeat amplification protocol (TRAP) assay. Final diagnosis was made by pathologic examination of follow-up surgical material available for all the cytologically malignant samples, a majority of the cytologically atypical samples, and a portion of the cytologically benign samples. RESULTS: TA was detected in 85 of 172 samples. Comparison of the cytologic and histologic diagnoseswith TA showed that 80 of 8...
Breast biology and pathology are currently shaped by the two-cell concept that recognizes only gl... more Breast biology and pathology are currently shaped by the two-cell concept that recognizes only glandular and myoepithelial cells. In the present study, we have visualized a previously unidentified cell population within the epithelial compartment of the breast, which displays the phenotypic characteristics of a committed stem cell. Immunofluorescence double labeling with digital image processing and Western blotting were applied to normal breast tissue as well as to noninvasive and invasive breast cancers using antibodies to basal cytokeratin 5 (Ck5), glandular cytokeratins 8/18 (Ck8/18/19), and smooth muscle ␣-actin (SMA) as markers for myoepithelial cells (SMA). A distinct population of cells was identified that expressed Ck5 in the absence of Ck8/18/19 or SMA. These cells differentiate toward glandular epithelial or myoepithelial Ck5-negative end cells passing through either Ck5/Ck8/18/19 or Ck5/SMA-positive intermediates. Our experiments clearly demonstrate a precursor or committed stem cell function of the Ck5-positive cell that is responsible for regeneration of the human adult breast epithelium. However, the observation that the vast majority of breast cancers display the glandular epithelial immunophenotype strongly suggests that the neoplastic cells derive from a late stage of the glandular epithelial differentiation pathway. The significance of this new cell biological model is that it might serve as a tool to unravel the regulatory mechanisms that govern regeneration and abnormal proliferation of breast epithelium at the cellular level.
Background: As an exocrine gland, the breast can manifest tumors with similar morphological featu... more Background: As an exocrine gland, the breast can manifest tumors with similar morphological features as the salivary gland. Defining the cellular identity of such tumours is one of the keys to a better understanding how these tumors evolve and grow. Material and Methods: We analyzed corresponding counterpart tumors such as pleomorphic adenoma, basal-cell adenoma, adenoid cystic carcinoma, adenosquamous carcinoma, and adenomyoepithelial tumors. With basal keratins, K5 and K14, as a reference, we have used antibodies against glandular keratins 8/18, myoepithelial markers (SMA, SMMHC, CD10) and functional markers such as Ki67 to investigate their expression in breast and salivary gland tumors. We used modern triple immunofluorescence labeling for simultaneously detecting these proteins in tumour cells with the obj ective to define different cell lineages and their differentiation status. The data were corroborated by real time PCR analysis of cRNAs for these proteins. Drawing parallels between tumors of the breast and salivary gland must help to unravel the nature of these lesions in these two organs. Results: All salivary gland tumors were found to contain progenitor cells with expression of basal keratins 5/14. They also showed an orderly sequential expression of basal keratins 5/14 and lineage specific proteins K8/18 for the glandular differentiation, myoepithelial markers for the myoepithelial differentiation, and K10 and vimentin for heterologeous squamous and mesenchymal differentiation, respectively, indicating the evolution of lineage specific cells from K5/14+ precursor cells. Normal breast and salivary gland epithelium showed striking similarities in their cellular composition, as both glands contain K5/14-positive progenitor cells which differentiate to glandular (K8/18+) or myoepithelial (SMA+;SMMHC+, CD10+) cells via intermediary cells, with a sequential expression of basal and lineage specific proteins. However, there are striking differences in the proliferative activity of different subtypes of cells — for instance, of precursor cells and differentiating glandular cells in both organs, which might be the source of differences in incidence of these tumors in both glands. Discussion: In this study we show that: (i) with few exceptions, salivary gland-like tumours represent K5/14- positive tumors with a glandular and/or myoepithelial differentiation and/or with a heterologeous (for example squamous) differentiation; (ii) the corresponding counterpart tumors both of breast and salivary gland are identical. As the constituent cells of all these tumors mimic the cellular composition and differentiation capabilities of progenitor cells both of breast and salivary gland epithelium, we believe that these tumors may originate from immature, probably pluripotent K5/14+ progenitor cells. Differences in proliferative activity of the various subtypes of cells in both organs might be an explanation for their rather different incidences. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-01-07.
A general problem in immunocytochemistry is the development of a reliable multiple immunolabeling... more A general problem in immunocytochemistry is the development of a reliable multiple immunolabeling method with primary antibodies originating from the same host species. Here, we briefly outline different approaches intended to close this technological gap and focus on multiple immunolabeling with monoclonal primary antibodies. To this end, we generated a basic universal protocol for the use of secondary antibodies selectively recognizing different isotypes/subclasses of monoclonal primary antibodies. This approach is widely applicable and offers a simple procedure for simultaneously detecting two or more antigens.
Cytokeratin 5/6 immunohistochemistry assists the differential diagnosis of atypical proliferation... more Cytokeratin 5/6 immunohistochemistry assists the differential diagnosis of atypical proliferations of the breast Aims: This study was performed to determine the diagnostic value of keratin 5/6 (CK 5/6) immunophenotyping on routinely processed breast tissues. Methods and results: Six hundred and ninety-nine breast lesions, including normal tissues as well as benign and malignant lesions in 321 formalin-fixed, paraffinembedded samples from 158 different patients were investigated immunohistochemically, following wet autoclave pre-treatment for antigen retrieval. In normal breast tissues, both myoepithelial and luminal epithelial cells expressed CK 5/6 in varying amounts. While myoepithelial immunoreactivity was most pronounced in the duct system, luminal epithelial immunoreactivity was strongest in the terminal duct lobular units. In ductal hyperplasias (DH), luminal epithelial cells predominantly revealed CK 5/6 immunoreaction. In contrast, neoplastic epithelial cells in atypical ductal and lobular hyperplasias (ADH and ALH) lacked such an expression, whereas in ductal in-situ carcinomas (DCIS) and in infiltrating ductal carcinomas 3.7% and 7.7%, of the cases respectively, showed positive immunostaining for CK 5/6. Conclusions: Immunophenotyping of keratin 5/6 expression can be helpful in the diagnosis of atypical hyperplasias and in-situ carcinomas of the breast. It is particularly valuable in the differential diagnosis between benign and atypical proliferative lesions. Keywords: benign breast disease, breast cancer, cytokeratin, ductal carcinoma in-situ, immunohistochemistry, premalignant breast disease Recently, a new antibody (anti-cytokeratin no. 5/6, Boehringer, Mannheim, Germany) has been advocated for immunohistochemistry in formalin-fixed, paraffinembedded tissues. This antibody had originally been intended for the differential diagnosis of squamous cell carcinoma and adenocarcinoma. It has also been shown to
European Journal of Surgical Oncology (EJSO), 2007
Introduction: Tumor volume is one of the best documented prognostic factors for prostate cancer. ... more Introduction: Tumor volume is one of the best documented prognostic factors for prostate cancer. There are several methods to gain this important parameter but unfortunately most of the clinicians in the world do not get this information in their routine practice from the pathologist. We developed a standardized method to handle radical prostatectomy specimens including a special form of mapping in order to document relevant morphological data. The aim of this study was to investigate if our model of mapping prostate cancer, which we use in routine practice, may serve for visual estimation of tumor volume. Methods: We estimated the tumor volume of prostate cancer by visual estimation of 350 maps of radical prostatectomy specimens and correlated these data with established prognostic parameters and clinical outcome. Results: Significant correlations between tumor volumes, as obtained from our mapping, and known prognostic parameters such as preoperative serum levels of prostatic specific antigen, loss of differentiation, histological grade, lymph node metastasis, and margins were found. In a multivariate analysis, only Gleason score and tumor stage were shown to be independent prognostic parameters. Discussion: We demonstrate that mapping of prostate cancer is more than a simple method of documentation but may serve as a method for visual estimation of tumor volume of prostate cancer after radical prostatectomy. This method can further be used for a visual documentation of the tumor stage independent of changes in the TNM classification. The method is inexpensive and practicable and can therefore be applied in routine surgical pathology.
The incidence of Ewing's tumors (ETs) is lower in Asians or African-Americans tha... more The incidence of Ewing's tumors (ETs) is lower in Asians or African-Americans than in Caucasians. Japanese ETs were available for analysis of chromosomal aberrations by comparative genomic hybridization (n = 16) and for expression of chimeric EWS transcripts by reverse-transcriptase polymerase chain reaction (n = 11). These results in Japanese patients were compared with those of 62 ETs in European Caucasian patients registered in the European Intergroup Cooperative Ewing's Sarcoma Study. Japanese patients with ET had lower overall survival (P = 0.0446) and relapse-free survival (P = 0.0371) compared with European Caucasian patients. Ten of 11 Japanese ETs and 31 of 62 European Caucasian ETs had type I (EWS exon 7 to FLI1 exon 6) fusion transcripts. In Japanese ETs, the median numbers of chromosomal aberrations were 2.0 and 6.0 in 11 primary tumors and five relapsed tumors, respectively. In European Caucasian ETs, the median number of changes were 2.5 and 5.0 in 52 primary and 10 relapsed tumors, respectively. Frequent gains were 8q (38%), 8p (31%) and 12q (25%) in Japanese ETs and 8q (52%), 8p (48%) and 12q (19%) in European Caucasian ETs. Frequent losses were 19q (44%), 19p (38%) and 17p (25%) in Japanese ETs and 16q (21%), 19q (18%) and 17p (15%) in European Caucasian ETs. The incidence of losses of 19p (P = 0.0215) and 19q (P = 0.0277) were significantly higher in Japanese ETs than in European Caucasian ETs. An amplification (1p33-p34) was observed in only one Japanese ET. Japanese patients with ET in this study had a worse prognosis than European Caucasian patients. In molecular genetic analyses, Japanese ETs had a higher frequency of loss of chromosome 19 than European Caucasian ETs. Different genetic aberrations may explain the different incidences and prognoses of ET between Caucasian and Japanese patients.
Mast cells are ubiquitous throughout the human tissues and play an essential role in physiology a... more Mast cells are ubiquitous throughout the human tissues and play an essential role in physiology and pathology. For evaluation of patients with pathological conditions, mast cells were primarily detected using metachromatic staining with toluidine blue. In the last decades, the staining arsenal of pathologists was enriched with enzyme histochemical and immunohistochemical methods, and it was established that depending on species and tissue localization mast cells are not similar both in appearance and function. The aim of this study was to characterize different mast cell populations using the up-to-date methods of their identification. We compared standard metachromatic method for mast cells with enzyme histochemical detection of chloroacetyl esterase and with immunohistochemical detection of tryptase and chymase in human and rodent tissues. Combination of these methods allowed us to assay quantitatively mast cell populations in different organs of humans and rodents. Furthermore, w...
Пауль Эрлих является основателем ряда важных направлений в биомедицинских исследованиях, среди ко... more Пауль Эрлих является основателем ряда важных направлений в биомедицинских исследованиях, среди которых на первом месте-иммунология и химиотерапия. Целью данного исторического экскурса к столетию со дня смерти П. Эрлиха является описание его роли в создании и развитии гистохимии. Ключевые слова: Пауль Эрлих, гистохимия, иммунология.
Extraocular sebaceous carcinoma (ESC) is a rare appendiceal skin tumor. In contrast to ocular seb... more Extraocular sebaceous carcinoma (ESC) is a rare appendiceal skin tumor. In contrast to ocular sebaceous carcinoma, information about the exact cellular architecture of these lesions is scarce and the histogenesis of ESC is unknown. Here, we extend our previous study and investigate 28 extraocular carcinomas in comparison to 54 benign sebaceous tumors and 8 cases of normal sebaceous glands using a broad spectrum of antibodies against p63, several keratins, adipophilin, EMA, Ki67, androgen receptor, and mismatch repair proteins. This observational study demonstrates that p63- and K5/14-positive basaloid cells are key cells in normal sebaceous gland and in all sebaceous tumors and that these basaloid cells give rise to EMA+, adipophilin+ sebocytes, and K5/14+, K7±, K10± ductal structures. Finally, about half of ESC is associated with superficial in situ neoplasia, which provides evidence that at least part of these carcinomas arises from flat superficial in situ carcinoma. In contrast to the normal sebaceous gland, about half of all sebaceous tumors lack keratin K7. MMR protein IHC-profiles role will be discussed.
Background/Aim: Polymorphous adenocarcinoma (PAC) is a low-grade salivary gland malignancy in con... more Background/Aim: Polymorphous adenocarcinoma (PAC) is a low-grade salivary gland malignancy in contrast to variants with papillary (PAP) or cribriform (CASG) architecture and confers the second most common malignancy of minor salivary glands. Our study aimed to identify prognostic factors and to evaluate histomorphological and molecular diagnostic criteria of PACs. Patients and Methods: A series of 155 PACs, including 10 PAPs and 12 CASGs from the population-based Cancer Registry of North Rhine-Westphalia (LKR-NRW) and the Hamburg Salivary Gland Reference Centre (HRC) were analyzed. Results: One fifth of the tumors were located in the major salivary glands and PACS/CASGS invariably lacked p40 expression. Fifty-two percent of PACs showed a PRKD1 E710D mutation. Ordinary PACs had a disease-specific 10-year survival probability of 97% compared to 90% when combining PAPs and CASGs. T-stage at diagnosis was a prognostic factor with 98% for stages T1/T2 versus 75% for T3/T4. Conclusion: Diagnostic algorithms for the PAC/CASG spectrum of tumors need to be improved and should include molecular markers.
Adenoid cystic carcinoma (ACC) is an aggressive head and neck malignancy characterized by a t (6;... more Adenoid cystic carcinoma (ACC) is an aggressive head and neck malignancy characterized by a t (6;9) translocation resulting in an MYB–NFIB gene fusion or, more rarely, an MYBL1 fusion. The true frequency and clinical significance of these alterations are still unclear. Here, we have used tissue microarrays and analyzed 391 ACCs and 647 non-ACC salivary neoplasms to study the prevalence, expression, and clinical significance of MYB/MYBL1 alterations by FISH and immunohistochemistry. Alterations of MYB or MYBL1 were found in 78% of the cases, of which 62% had MYB alterations and 16% had MYBL1 rearrangements. Overexpression of MYB/MYBL1 oncoproteins was detected in 93% of the cases. MYB split signal, seen in 39% of the cases, was specific for ACC and not encountered in non-ACC salivary tumors. Loss of the 3′-part of MYB was enriched in grade 3 tumors and was a significant independent prognostic biomarker for overall survival in multivariate analyses. We hypothesize that loss of the 3′-...
Objectives: Information of the clinicopathological characteristics and outcome data of patients w... more Objectives: Information of the clinicopathological characteristics and outcome data of patients with adenosquamous carcinoma of the pancreas (ASCAP) remains limited. This study's aim is to describe the clinical, pathological, and molecular characteristics of 25 resected ASCAPs. Methods: Of all 25 cases, patient characteristics, follow-up data, and pathological/immunohistological features were reviewed and analyzed. Results: In this 3-institutional retrospective analysis of 562 pancreatic cancer patients, we identified 25 cases with histologically confirmed ASCAP (4.4%). Follow-up was available in 21 ASCAP and 50 pancreatic ductal adenocarcinoma control patients with a median overall survival of 8.2 and 21 months, respectively. Age, tumor size, localization in the tail, lymph node status, and resection margin seem to be the most significant factors of survival in our ASCAP cohort. In contrast to pancreatic ductal adenocarcinoma, positive expression of p63, keratins K5/14, and the epidermal growth factor receptor are a robust marker profile of these tumors. Conclusions: Adenosquamous carcinoma of the pancreas comprises a group of neoplasms in which stage and adverse morphological features contribute to its bad prognosis. Further work must be pursued to improve detection and treatment options to reduce mortality. Specifically, differences in biology might become a target for the development of possible therapies.
Biomedical Journal of Scientific & Technical Research, 2018
The investigation conforms with Guide for the Care and Use of Laboratory Animals published by the... more The investigation conforms with Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85-23, revised 1996) and was per
Background: Salivary gland-like tumors are currently subdivided into those with and those without... more Background: Salivary gland-like tumors are currently subdivided into those with and those without myoepithelial differentiation. Furthermore, they show a wide range of differentiation patterns including squamous differentiation. Many authors believe that the myoepithelial cell plays a major role in this context. However, currently little is known about the evolution and cellular differentiation of these tumors. Material and Methods: For that reason, we performed an in situ Triple Immunofluorescence Lineage/Differentiation Tracing (isTILT) and qRT-PCR study of basal (K5/K14), glandular (K7/K8/18), and epidermal-specific squamous (K10) keratins, p63, and smooth muscle actin (SMA; myoepithelial marker) with the aim to trace different cell lineages and define their cellular hierarchy in tumors. We investigated 50 cases of salivary gland-like tumors of the breast (11 Adenoid-cystic carcinomas, 9 (adeno-)myoepithelial tumors, 8 adeno-squamous carcinomas including 12 syringomas of the nipp...
Nitric oxide (NO) is generated by a family of enzymes termed NO synthases (NOS) that convert L-ar... more Nitric oxide (NO) is generated by a family of enzymes termed NO synthases (NOS) that convert L-arginine to NO and citrulline. The role of NO as an important biological mediator and recognition of the pathophysiological significance of superoxides/NO interaction has led to an intensive research and development of therapies based on the interception of the NO signaling cascade in the pancreatitis course. However, the presence and localization of the NO-generating enzymes in various organs including pancreas are subject to controversy. We assumed that this controversy might reflect rather the diversity of experimental approaches and an insufficient sensitivity of the methods used. Applying tyramide signal amplification (TSA) immunohistochemical technology, we were able detect all three NOS isoforms both in exocrine and endocrine compartments and in the vasculature in the normal pancreas and in pancreatitis. This also allowed us to demonstrate that oxidative stress runs ahead of NOS up-regulation, which implies that the NO enhancement in the course of pancreatitis is likely to be an adaptive mechanism aimed at maintaining the homeostatic cellular level of the bioactive NO. The aims of this minireview are to describe normal intrapancreatic NO pathways and the role of NO in the pancreatitis course.
PURPOSE: Telomerase has been detected in a majority of human malignant tumors, making telomerase ... more PURPOSE: Telomerase has been detected in a majority of human malignant tumors, making telomerase activity (TA) one key difference between mortal and immortal cells. In this study, we evaluated in blind-trial fashion the association of TA with cytologic and final clinical/pathologic diagnosis in fine-needle aspirates (FNAs) of breast lesions. MATERIALS AND METHODS: In 172 FNAs, including 80 samples that were cytologically malignant, 18 that were atypical but not diagnostic for malignancy, and 74 that were cytologically benign, TA was determined by a modified nonradioactive telomeric repeat amplification protocol (TRAP) assay. Final diagnosis was made by pathologic examination of follow-up surgical material available for all the cytologically malignant samples, a majority of the cytologically atypical samples, and a portion of the cytologically benign samples. RESULTS: TA was detected in 85 of 172 samples. Comparison of the cytologic and histologic diagnoseswith TA showed that 80 of 8...
Breast biology and pathology are currently shaped by the two-cell concept that recognizes only gl... more Breast biology and pathology are currently shaped by the two-cell concept that recognizes only glandular and myoepithelial cells. In the present study, we have visualized a previously unidentified cell population within the epithelial compartment of the breast, which displays the phenotypic characteristics of a committed stem cell. Immunofluorescence double labeling with digital image processing and Western blotting were applied to normal breast tissue as well as to noninvasive and invasive breast cancers using antibodies to basal cytokeratin 5 (Ck5), glandular cytokeratins 8/18 (Ck8/18/19), and smooth muscle ␣-actin (SMA) as markers for myoepithelial cells (SMA). A distinct population of cells was identified that expressed Ck5 in the absence of Ck8/18/19 or SMA. These cells differentiate toward glandular epithelial or myoepithelial Ck5-negative end cells passing through either Ck5/Ck8/18/19 or Ck5/SMA-positive intermediates. Our experiments clearly demonstrate a precursor or committed stem cell function of the Ck5-positive cell that is responsible for regeneration of the human adult breast epithelium. However, the observation that the vast majority of breast cancers display the glandular epithelial immunophenotype strongly suggests that the neoplastic cells derive from a late stage of the glandular epithelial differentiation pathway. The significance of this new cell biological model is that it might serve as a tool to unravel the regulatory mechanisms that govern regeneration and abnormal proliferation of breast epithelium at the cellular level.
Background: As an exocrine gland, the breast can manifest tumors with similar morphological featu... more Background: As an exocrine gland, the breast can manifest tumors with similar morphological features as the salivary gland. Defining the cellular identity of such tumours is one of the keys to a better understanding how these tumors evolve and grow. Material and Methods: We analyzed corresponding counterpart tumors such as pleomorphic adenoma, basal-cell adenoma, adenoid cystic carcinoma, adenosquamous carcinoma, and adenomyoepithelial tumors. With basal keratins, K5 and K14, as a reference, we have used antibodies against glandular keratins 8/18, myoepithelial markers (SMA, SMMHC, CD10) and functional markers such as Ki67 to investigate their expression in breast and salivary gland tumors. We used modern triple immunofluorescence labeling for simultaneously detecting these proteins in tumour cells with the obj ective to define different cell lineages and their differentiation status. The data were corroborated by real time PCR analysis of cRNAs for these proteins. Drawing parallels between tumors of the breast and salivary gland must help to unravel the nature of these lesions in these two organs. Results: All salivary gland tumors were found to contain progenitor cells with expression of basal keratins 5/14. They also showed an orderly sequential expression of basal keratins 5/14 and lineage specific proteins K8/18 for the glandular differentiation, myoepithelial markers for the myoepithelial differentiation, and K10 and vimentin for heterologeous squamous and mesenchymal differentiation, respectively, indicating the evolution of lineage specific cells from K5/14+ precursor cells. Normal breast and salivary gland epithelium showed striking similarities in their cellular composition, as both glands contain K5/14-positive progenitor cells which differentiate to glandular (K8/18+) or myoepithelial (SMA+;SMMHC+, CD10+) cells via intermediary cells, with a sequential expression of basal and lineage specific proteins. However, there are striking differences in the proliferative activity of different subtypes of cells — for instance, of precursor cells and differentiating glandular cells in both organs, which might be the source of differences in incidence of these tumors in both glands. Discussion: In this study we show that: (i) with few exceptions, salivary gland-like tumours represent K5/14- positive tumors with a glandular and/or myoepithelial differentiation and/or with a heterologeous (for example squamous) differentiation; (ii) the corresponding counterpart tumors both of breast and salivary gland are identical. As the constituent cells of all these tumors mimic the cellular composition and differentiation capabilities of progenitor cells both of breast and salivary gland epithelium, we believe that these tumors may originate from immature, probably pluripotent K5/14+ progenitor cells. Differences in proliferative activity of the various subtypes of cells in both organs might be an explanation for their rather different incidences. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-01-07.
A general problem in immunocytochemistry is the development of a reliable multiple immunolabeling... more A general problem in immunocytochemistry is the development of a reliable multiple immunolabeling method with primary antibodies originating from the same host species. Here, we briefly outline different approaches intended to close this technological gap and focus on multiple immunolabeling with monoclonal primary antibodies. To this end, we generated a basic universal protocol for the use of secondary antibodies selectively recognizing different isotypes/subclasses of monoclonal primary antibodies. This approach is widely applicable and offers a simple procedure for simultaneously detecting two or more antigens.
Cytokeratin 5/6 immunohistochemistry assists the differential diagnosis of atypical proliferation... more Cytokeratin 5/6 immunohistochemistry assists the differential diagnosis of atypical proliferations of the breast Aims: This study was performed to determine the diagnostic value of keratin 5/6 (CK 5/6) immunophenotyping on routinely processed breast tissues. Methods and results: Six hundred and ninety-nine breast lesions, including normal tissues as well as benign and malignant lesions in 321 formalin-fixed, paraffinembedded samples from 158 different patients were investigated immunohistochemically, following wet autoclave pre-treatment for antigen retrieval. In normal breast tissues, both myoepithelial and luminal epithelial cells expressed CK 5/6 in varying amounts. While myoepithelial immunoreactivity was most pronounced in the duct system, luminal epithelial immunoreactivity was strongest in the terminal duct lobular units. In ductal hyperplasias (DH), luminal epithelial cells predominantly revealed CK 5/6 immunoreaction. In contrast, neoplastic epithelial cells in atypical ductal and lobular hyperplasias (ADH and ALH) lacked such an expression, whereas in ductal in-situ carcinomas (DCIS) and in infiltrating ductal carcinomas 3.7% and 7.7%, of the cases respectively, showed positive immunostaining for CK 5/6. Conclusions: Immunophenotyping of keratin 5/6 expression can be helpful in the diagnosis of atypical hyperplasias and in-situ carcinomas of the breast. It is particularly valuable in the differential diagnosis between benign and atypical proliferative lesions. Keywords: benign breast disease, breast cancer, cytokeratin, ductal carcinoma in-situ, immunohistochemistry, premalignant breast disease Recently, a new antibody (anti-cytokeratin no. 5/6, Boehringer, Mannheim, Germany) has been advocated for immunohistochemistry in formalin-fixed, paraffinembedded tissues. This antibody had originally been intended for the differential diagnosis of squamous cell carcinoma and adenocarcinoma. It has also been shown to
European Journal of Surgical Oncology (EJSO), 2007
Introduction: Tumor volume is one of the best documented prognostic factors for prostate cancer. ... more Introduction: Tumor volume is one of the best documented prognostic factors for prostate cancer. There are several methods to gain this important parameter but unfortunately most of the clinicians in the world do not get this information in their routine practice from the pathologist. We developed a standardized method to handle radical prostatectomy specimens including a special form of mapping in order to document relevant morphological data. The aim of this study was to investigate if our model of mapping prostate cancer, which we use in routine practice, may serve for visual estimation of tumor volume. Methods: We estimated the tumor volume of prostate cancer by visual estimation of 350 maps of radical prostatectomy specimens and correlated these data with established prognostic parameters and clinical outcome. Results: Significant correlations between tumor volumes, as obtained from our mapping, and known prognostic parameters such as preoperative serum levels of prostatic specific antigen, loss of differentiation, histological grade, lymph node metastasis, and margins were found. In a multivariate analysis, only Gleason score and tumor stage were shown to be independent prognostic parameters. Discussion: We demonstrate that mapping of prostate cancer is more than a simple method of documentation but may serve as a method for visual estimation of tumor volume of prostate cancer after radical prostatectomy. This method can further be used for a visual documentation of the tumor stage independent of changes in the TNM classification. The method is inexpensive and practicable and can therefore be applied in routine surgical pathology.
The incidence of Ewing's tumors (ETs) is lower in Asians or African-Americans tha... more The incidence of Ewing's tumors (ETs) is lower in Asians or African-Americans than in Caucasians. Japanese ETs were available for analysis of chromosomal aberrations by comparative genomic hybridization (n = 16) and for expression of chimeric EWS transcripts by reverse-transcriptase polymerase chain reaction (n = 11). These results in Japanese patients were compared with those of 62 ETs in European Caucasian patients registered in the European Intergroup Cooperative Ewing's Sarcoma Study. Japanese patients with ET had lower overall survival (P = 0.0446) and relapse-free survival (P = 0.0371) compared with European Caucasian patients. Ten of 11 Japanese ETs and 31 of 62 European Caucasian ETs had type I (EWS exon 7 to FLI1 exon 6) fusion transcripts. In Japanese ETs, the median numbers of chromosomal aberrations were 2.0 and 6.0 in 11 primary tumors and five relapsed tumors, respectively. In European Caucasian ETs, the median number of changes were 2.5 and 5.0 in 52 primary and 10 relapsed tumors, respectively. Frequent gains were 8q (38%), 8p (31%) and 12q (25%) in Japanese ETs and 8q (52%), 8p (48%) and 12q (19%) in European Caucasian ETs. Frequent losses were 19q (44%), 19p (38%) and 17p (25%) in Japanese ETs and 16q (21%), 19q (18%) and 17p (15%) in European Caucasian ETs. The incidence of losses of 19p (P = 0.0215) and 19q (P = 0.0277) were significantly higher in Japanese ETs than in European Caucasian ETs. An amplification (1p33-p34) was observed in only one Japanese ET. Japanese patients with ET in this study had a worse prognosis than European Caucasian patients. In molecular genetic analyses, Japanese ETs had a higher frequency of loss of chromosome 19 than European Caucasian ETs. Different genetic aberrations may explain the different incidences and prognoses of ET between Caucasian and Japanese patients.
Mast cells are ubiquitous throughout the human tissues and play an essential role in physiology a... more Mast cells are ubiquitous throughout the human tissues and play an essential role in physiology and pathology. For evaluation of patients with pathological conditions, mast cells were primarily detected using metachromatic staining with toluidine blue. In the last decades, the staining arsenal of pathologists was enriched with enzyme histochemical and immunohistochemical methods, and it was established that depending on species and tissue localization mast cells are not similar both in appearance and function. The aim of this study was to characterize different mast cell populations using the up-to-date methods of their identification. We compared standard metachromatic method for mast cells with enzyme histochemical detection of chloroacetyl esterase and with immunohistochemical detection of tryptase and chymase in human and rodent tissues. Combination of these methods allowed us to assay quantitatively mast cell populations in different organs of humans and rodents. Furthermore, w...
Uploads
Papers by Werner Boecker