[Show abstract][Hide abstract] ABSTRACT: Paul Ehrlich is the founder of a number of areas in biomedical research: in the first line, immunology and chemotherapy. Aim of this historical note to the centenary of Paul Ehrlich's death is to commemorate his tribute to the establishment and development of histochemistry.
Archiv für Pathologische Anatomie und Physiologie und für Klinische Medicin 10/2014; · 2.56 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: A small group of tumors of breast and salivary glands contains squamous/epidermoid elements as a constitutive feature (e.g., squamous carcinoma, syringomatous tumors, and mucoepidermoid carcinoma). Other tumors (e.g., pleomorphic adenoma, adenomyoepithelial tumors, and adenoid cystic carcinoma) may show occasionally squamous differentiation. Furthermore, squamous metaplasia may be observed in non-neoplastic breast and salivary tissues. However, the histogenesis of these squamous differentiations is far from being understood. Based on our earlier in situ triple immunofluorescence and quantitative reverse transcription (RT)-PCR experiments for basal keratins K5/14 and p63 as well as for glandular keratins (K7/K8/18), squamous keratins (K10 and K13), and myoepithelial lineage markers (smooth muscle actin, SMA), we here traced the squamous/epidermoid differentiation lineage of 60 tumors of the breast and/or salivary glands, cultured tumor cells of 2 tumors, and of 7 squamous metaplasias of non-neoplastic breast and salivary tissues. Our results indicate that both the neoplastic lesions as well as the non-neoplastic squamous metaplasia contain p63/K5/14+ cells that differentiate toward K10/13+ squamous cells. Thus, cells with squamous/epidermoid differentiation undergo a transition from its original p63/K5/14+ precursor state to K10/13+ squamous lineage state, which can be pictured by triple-immunofluorescence experiments. Given the immunophenotypic similarity of p63/K5/14+ tumor cells to their physiological p63/K5/14+ counterparts in normal breast and salivary duct epithelium, we suggest that these cells provide an important histogenetic key to understanding the pathogenesis of squamous differentiation both in normal breast/salivary gland tissues and their corresponding tumors.
Archiv für Pathologische Anatomie und Physiologie und für Klinische Medicin 10/2014; · 2.56 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: AimsSyringomatous tumour of the nipple and low-grade adenosquamous carcinoma (LGAdSC) of the breast are regarded as distinct entities. To clear the nature of these two lesions, we compared the expression of different lineage/differentiation markers in 12 syringomatous tumours of the nipple, 9 LGAdSC, and normal breast epithelium. Methods and resultsUsing triple immunofluorescence labelling and quantitative RT-PCR studies for keratins, p63 and SMA, we demonstrated that syringomatous tumour and LGAdSC contain p63+/K5/K14+ tumour cells, K10+ squamous, and K8/18+ glandular cells with intermediary cells in both lineages. Identical p63+/K5/14+ cells were also found in the normal breast duct epithelium. Conclusions
Our data provide evidence that syringomatous tumour of the nipple and LGAdSC are identical or nearly identical lesions. They contain p63+/K5/K14+ cells as the key cells from which K10+ squamous and K8/18+ glandular lineage arise. Based on our findings in normal breast tissue and associated benign lesions, we suggest that p63+/K5/14+ cells of the normal breast duct epithelium or early related cells might play a key role in neoplastic transformation of both syringomatous tumour and LGAdSC. We propose that the differentiation patterns found in both lesions reflect the early ontogenetic stages of the normal breast epithelium.This article is protected by copyright. All rights reserved.
[Show abstract][Hide abstract] ABSTRACT: Because of the introduction of mammography screening programmes in Europe, the number of breast biopsies performed is increasing. We investigated the influence of immunohistochemistry (IHC) on the final diagnosis of breast biopsies by comparing the primary diagnoses (based on the results of haematoxylin and eosin staining only) with the final diagnoses (based on the additional information provided by IHC).
We analysed the breast biopsies which were performed at the University of Halle-Wittenberg between 2006 and 2010 and for which the pathologist requested IHC for making the final diagnosis. According to the B-categorization scheme, the primary diagnosis changed in 37 of a total of 429 biopsies (8.6%). In 18 of these biopsies (48.6%) the category changed from B1-B2 to B3-B5 or vice versa, which would imply a different work-up. Only 77% of the primary diagnoses of breast cancer in situ were confirmed.
IHC has a considerable influence on the final diagnosis of breast biopsies in several situations, including those in which the biopsied women are at risk of inadequate therapeutic intervention. The influence is particularly notable among those biopsies for which IHC is performed in order to assess the suspicion of breast cancer in situ.
[Show abstract][Hide abstract] ABSTRACT: The role of nitric oxide (NO) in the human pancreas and in pancreatitis still remains controversial. Furthermore, conflicting conclusions have been reached by different laboratories about the localization of the NO-generating enzyme (NO synthase, NOS) in the pancreas. Here, we investigated the co-expression of NOS with enzymes involved in regulation of NO signalling in the normal human pancreas and in pancreatitis. We found that the whole NO signalling machinery was up-regulated in pancreatitis, especially within the exocrine compartment. Furthermore, the exocrine parenchymal cells revealed higher levels of oxidative stress markers, nitrotyrosine and 8-hydroxyguanosine, in pancreatitis, which reflects the exceptional susceptibility of the exocrine parenchyma to oxidative stress. This study provides a direct link between oxidative stress and the enzymatic control of the NO bioavailability at the cellular level and endows with further insight into fundamental mechanisms underlying pancreatic disorders associated with disruptions in the L-arginine-NO-cGMP signalling enzyme cascade.
[Show abstract][Hide abstract] ABSTRACT: Salivary gland-like tumors of the breast show a great variety of architectural patterns and cellular differentiations such as glandular, myoepithelial, squamous, and even mesenchymal phenotypes. However, currently little is known about the evolution and cellular differentiation of these tumors. 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 construct and trace different cell lineages and define their cellular hierarchy in tumors with myoepithelial differentiation. isTILT analysis of a series of 28 breast, salivary, and lacrimal gland tumors, including pleomorphic adenomas (n=8), epithelial-myoepithelial tumors (n=9), and adenoid cystic carcinomas (n=11) revealed that all tumor types contained K5/K14-positive progenitor cells in varying frequencies from a few percent up to 15%. These K5/K14-positive tumor cells were found to differentiate to glandular- (K8/18-positive) and myoepithelial-lineage (SMA-positive)-specific cells and were also shown to generate various heterologeous cell differentiations such as squamous and mesenchymal progenies. p63 was co-expressed with K5/K14 in basal-like progenitor cells, myoepithelial, and squamous cells but not in glandular cells. Our results show that the corresponding counterpart tumors of breast and salivary/lacrimal glands have identical cellular compositions. Taken together, our isTILT and RNA-expression data indicate that look-alike tumors of the breast represent a special subgroup of basal-type tumors with benign or usually low malignant potential.Modern Pathology advance online publication, 5 April 2013; doi:10.1038/modpathol.2013.45.
[Show abstract][Hide abstract] ABSTRACT: Improvements in reagents and protocols for immunohistochemistry have led to increased sensitivity of detection systems. A significant level of signal amplification was achieved by the chain-polymer conjugate technology utilizing enzyme-labeled inert "backbone" molecule of dextran (Dako). However, the relatively large size of the dextran molecule in aqueous phase appears to create spatial hindrance compromising the penetrative ability of the detection reagent. Novel AmpliStain™ detection systems (SDT GmbH, Baesweiler, Germany) seem to overcome these constraints offering a more compact and deformable conjugate design that facilitates agile penetration through the narrowest diffusion pathways in tissue sections. Here, we compared the level of signal amplification achievable with AmpliStain™-HRP (SDT) and EnVision™+-HRP (Dako). Our results show that the AmpliStain™-HRP systems allow higher dilutions of primary antibodies in both immunohistochemistry and ELISA. Compared with EnVision™+, anti-mouse AmpliStain™ enables at least three times more sensitive detection of mouse antibodies, whereas anti-rabbit AmpliStain™ is ten times more sensitive than anti-rabbit EnVision™+.
[Show abstract][Hide abstract] ABSTRACT: BACKGROUND: As high percentage of mammographic densities complicates the assessment of imaging findings, mammographic density may influence the histopathological evaluation of core-biopsies of the breast. We measured the influence of mammographic density on the inter-observer variability of histopathological findings of breast biopsies. METHODS: Histological slides of 695 women who underwent core biopsies of the breast at University of Halle between 2006 and 2008 were evaluated in a blinded fashion by two pathologists using the five levels of the B-categorization scheme (B1-B5). To quantify mammographic density, we used a computer-based threshold method (Madena). We calculated observed and chance-corrected agreements (weighted kappa) and 95% confidence intervals (95% CI) according to four categories of mammographic density (<10%, 10<25%, 25<50%, >=50%). RESULTS: The weighted kappa decreased monotonically from 89.6% (95% CI: 85.8%, 93.3%) among women with less than 10% of mammographic density to 80.4% (95% CI: 69.9%, 90.9%) for women with more than 50% of mammographic density, respectively. Results of a kappa regression analysis showed that agreement of pathologists on clinically relevant categories (B1-B2 versus B3-B5) decreased with mammographic density. CONCLUSIONS: Mammographic density is a relevant modifier of the agreement between pathologists who assess breast biopsies using the B-categorization scheme. The influence of mammographic density on the inter-observer variability can be explained to some extent by varying prevalences of histological entities across B categories that have typically different inter-observer agreement. Women with high mammographic density are at higher risk of inter-observer variability compared to women with low mammographic density and should possibly undergo a second pathology review.
BMC Cancer 11/2012; 12(1):554. · 3.32 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Similarities in morphology and in glandular and squamous differentiation
patterns amongst syringomas of the breast nipple and of the skin suggest
a common nature, but the origin of nipple syringoma remains undefined.
Using triple immunofluorescence analysis, we found that cells
immunopositive for basal keratins K5 and 14 undergo differentiation into
glandular and squamous cell lineages. Both tumour types expressed K10,
indicative of squamous lineage, but there were specific differences in
their glandular lineage. In contrast to the breast nipple syringoma,
which expressed glandular keratins K8/18/19, syringoma of the skin only
expressed the glandular keratin K19. Therefore, syringomas of the breast
nipple and of the skin resemble glandular lineages of the breast nipple
duct or eccrine duct epithelium, respectively. From these results we
conclude that K5/14-positive cells of the breast nipple ducts are the
putative cells of origin for syringomas of the nipple, which highlights
the organotypic glandular differentiation potential.
[Show abstract][Hide abstract] ABSTRACT: It has been recommended that the histopathology results of core biopsies of the breast are categorized according to the B-categorization scheme. We measured the interobserver variability of the B-categorization of core biopsies of the breast.
Core biopsies were taken among 765 women at the University of Halle between 2006 and 2008. All histological slides were reviewed in a blinded fashion by two experienced breast pathologists. We calculated observed and chance-corrected agreements (kappa) and 95% confidence intervals (CI). The prevalence of B3-B5 biopsies was 41.6%. The observed and weighted kappa agreement of the five-level B-categorization scheme was 0.87 (95% CI: 0.84 -0.89) and 0.89 (95% CI: 0.89-0.91), respectively. The most frequent disagreement was between B2 and B3 (47 of 103 disagreements, 45.6%). Overall, 49.5% of all disagreements were clinically relevant disagreements that would imply different therapeutic strategies. Agreement was modified by referral group, Breast Imaging Reporting and Data System (BIRADS) level, radiological breast density, imaging guidance and application of immunohistological staining.
Interobserver agreement of the B-categorization scheme was high and was modified by referral status, level of radiological suspicion of breast cancer, breast density, imaging guidance of core biopsies and requirement of additional immunohistological staining.
[Show abstract][Hide abstract] ABSTRACT: The current protocols for blocking background staining in immunohistochemistry are based on conflicting reports. Background staining is thought to occur as a result of either non-specific antibody (Ab) binding to endogenous Fc receptors (FcRs) or a combination of ionic and hydrophobic interactions. In this study, cell and tissue samples were processed according to routine protocols either with or without a blocking step (goat serum or BSA). Surprisingly, no Abs in samples processed without a blocking step showed any propensity for non-specific binding leading to background staining, implying that endogenous FcRs do not retain their ability to bind the Fc portion of Abs after standard fixation. Likewise, we did not find any non-specific Ab binding ascribable to either ionic or hydrophobic interactions. We determined that traditionally used protein blocking steps are unnecessary in the immunostaining of routinely fixed cell and tissue samples.
[Show abstract][Hide abstract] ABSTRACT: To evaluate the rate, the histological spectrum and the positive predictive value (PPV) for malignancy of minimally invasive biopsies with "uncertain malignant potential (B3)" in digital mammography screening.
Consecutive data of 37,178 participants of one digital unit of the German screening program were included.
The B 3 rate was 15.1 % (148 / 979). The frequencies of lesion subtypes were as follows: atypical epithelial proliferation of ductal type (AEPDT) 35.1 % (52 / 148), radial scar (RS) 28.4 % (42 / 148), papillary lesions (PAP) 20.3 % (30 / 148), lobular carcinoma in situ 8.8 % (13 / 148), flat epithelial atypia 5.4 % (8 / 148), and mucocele-like lesions 2.0 % (3 / 148). The PPV for malignancy in surgical excisions was overall 0.28 (25 / 91); in detail 0.40 (19 / 47) for AEPDT, 0.20 (5 / 25) for RS, 0.08 (1 / 12) for PAP.
Despite a higher B 3 rate of minimally invasive biopsies with "uncertain malignant potential" in digital screening, the benign surgical biopsy rate is not disproportionally increased compared with analog screening programs. Together with defined management protocols, this results in an increased cancer detection rate per screening participant with surgical excision.
RöFo - Fortschritte auf dem Gebiet der R 04/2011; 183(8):743-8. · 2.76 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To evaluate the relevance of calcifications for invasive breast cancer detection in population-based digital mammographic screening.
This study was approved by an independent ethics committee, and no additional informed consent was required. Prospectively documented radiologic cancer features were correlated with pathologic characteristics in 241 breast malignancies diagnosed in 24067 participating women aged 50-69 years (part of the digital German Screening Program; initial screening rate, 92%; detection rate [DR], 1.0%; recall rate [RR], 7.5%). The rates of invasive cancers detected on the basis of calcifications were analyzed against pathologic tumor categories (pT categories) and histologic grades. For comparison of the study data with results of analog screening, data from the literature regarding calcification-specific RR, DR, and positive predictive value for recall (PPV(1)) were calculated.
The calcification-specific RR was 1.7% (416 of 24067). The calcification-specific DR for invasive cancer was 0.12% (29 of 24067), and the PPV(1) was 7.0% (29 of 416). Of all malignancies detected on the basis of calcification, 38% (29 of 77) were invasive. pT1 cancers showed an inverse association between tumor size and rate of detection on the basis of calcification; differences in rates among pT1 subcategories were statistically significant (P < .001). The proportion of grade 1 pT1 cancers detected on the basis of calcification (eight of 27) did not differ significantly from that of cancers detected on the basis of other radiologic features (46 of 108, P = .24). The calcification-specific invasive cancer DR was significantly higher for digital than for analog mammography.
One-third of malignancies detected on the basis of calcifications only are invasive cancers. They tend to be smaller but not less aggressive than invasive cancers detected on the basis of other features. Compared with published results of analog screening, digital screening offers the potential to increase the rate of invasive cancers detected on the basis of calcifications in population-based mammographic screening.
[Show abstract][Hide abstract] ABSTRACT: In the last decade, pioneering efforts of histochemists have led to an immense improvement in reagents and protocols. Milestones
in this development were antigen retrieval technique and signal amplification. Wide application of these techniques in pathology
and other fields of morphology has demonstrated distinct enhancement of immunostaining on archival formalin-fixed, paraffin-embedded
tissue sections for a variety of antigens. Whereas heat-induced antigen retrieval on formaldehyde-fixed and paraffin-imbedded
tissues is used in the predetection phase, signal amplification with polymeric detection systems (e.g., EnVision System from
Dako) and catalyzed reporter deposition (CARD) with tyramide are accomplished in detection and the post-detection phases respectively
(Shi et al. 2001).
[Show abstract][Hide abstract] ABSTRACT: Background staining has increased the amount of gray hair on the heads of histochemists. In most cases, background staining
is not caused by a single factor. Along with Fc receptors, frequent causes of background staining are endogenous enzyme activity,
if you use peroxidase or alkaline phosphatase as enzyme markers on your secondary antibodies, and endogenous biotin when using
a streptavidin or avidin reagents (http://www.ihcworld.com/; http://www.protocol-online.org/prot/Immunology/). When fluorescent dyes are used in experiments, autofluorescence (or natural fluorescence) of some tissue components can
cause background problems and complicate the use of fluorescence microscopy.