bluuebunny/crossref_metadata_2025 huggingface dataset:
p53 expression in Barrett's oesophagus, dysplasia, and adenocarcinoma using antibody DO‐7
Authors: Symmans Pennie J. , Linehan Jacqueline M. , Brito Maria J. , Filipe M. Isabel
Published: June 2005

Barrett's oesophagus has a well‐recognized association with oesophageal adenocarcinoma, with phenotypic progression through dysplasia to malignancy. The nuclear phosphoprotein p53 is a putative tumour suppressor with mutations resulting in both loss of negative growth regulatory function and possible gain of oncogene function. Many mutant forms have a prolonged half‐life and are demonstrable with immunohistochemical techniques. We examined 62 endoscopic oesophageal biopsies and 36 oesophageal resections for p53 overexpression using the monoclonal antibody DO‐7 on paraffin‐embedded tissue. The series included 40 cases of Barrett's metaplasia, 13 cases of dysplasia, and 81 cases of adenocarcinoma. None of the cases of metaplasia was p53‐positive, compared with 4/13 cases of dysplasia and 52/81 cases of adenocarcinoma. There was no association between the degree of dysplasia and p53 expression, although a trend emerged of increasing p53 expression with higher tumour grade. We conclude that p53 overexpression is frequent in oesophageal adenocarcinoma and may be related to tumour grade. p53 overexpression is not restricted to neoplastic lesions and mutation of this tumour suppressor may occur early in the malignant progression of Barrett's oesophagus.
Computer‐assisted chromatin texture characterization of Feulgen‐stained nuclei in a series of 331 transitional bladder cell carcinomas
Authors: Van Velthoven Roland , Petein Michel , Zlotta Alexandre , Oosterlinck Willem J. , van der Meijden Adrian , Zandona Cassio , Roels Hendrik , Pasteels Jean‐Lambert , Schulman Claude , Kiss Robert
Published: June 2005

The chromatin patterns of Feulgen‐stained nuclei in a series of six specimens of normal mucosa and 331 transitional bladder carcinomas, including 293 superficial (Ta and T1) and 38 invasive (T2–T4) cases, were quantitatively described by means of eight parameters relating to densitometric, run‐length distribution, and co‐occurrence matrix features. The results show that the chromatin texture of the superficial lesions was markedly different from that of the invasive tumours, which exhibited a distinctly more dense and heterogeneous chromatin pattern. The data also show that the increasing level of malignancy, as revealed by the increasing clinical stage, was accompanied by an increase in the overall chromatin condensation level. Only some areas of the nucleus actually increased in density; other pale areas appeared concomitantly with these increasingly denser chromatin areas. This chromatin density increase corresponded to a marked increase in the frequency of small dense chromatin clumps; these joined together into very large dense chromatin clumps, which were distributed more and more heterogeneously in the nucleus as the clinical stage of the tumour increased.
Non‐enzymatic retrieval of antigen permits staining of follicle centre cells by the rabbit polyclonal antibody to protein gene product 9.5
Authors: Langlois N. E. I. , King G. , Herriot R. , Thompson W. D.
Published: June 2005

The rabbit polyclonal antibody to protein gene product 9.5 (PGP9.5) will detect the L1 isoenzyme of ubiquitin carboxy‐terminal hydrolase (UCH), which is a marker for neurones and neuroendocrine tissue. We re‐evaluated this antibody using the technique of non‐enzymatic antigen retrieval (boiling sections in citrate buffer, heated by microwave oven) followed by streptavidin‐biotin‐peroxidase staining. Due to the fortuitous choice of appendix as positive control material containing small nerves, we found strong, repeatable cytoplasmic and nuclear staining of lymphoid follicle centre cells in addition to neural tissue. This effect could be repeated on other lymphoid tissues and was not dependent on microwave heating, but did require boiling in an ionic buffer solution. Staining was also observed with a fresh batch of antibody and with four of the five different batches of antibody which were supplied to us. This pattern was not obtained in fresh tissue, in fixed material following trypsinization, or by increasing the primary antibody concentration. We suggest that the boiling of sections in citrate buffer is exposing an epitope for the anti‐PGP9.5 antibody which is inaccessible in the native or fixed state and therefore we would recommend retesting of antibody specificity following non‐enzymatic retrieval of antigen.
Alpha‐B crystallin in the normal human myocardium and cardiac conducting system
Authors: Leach I. H. , Tsang M. L. , Church R. J. , Lowe J.
Published: June 2005

The alpha crystallins are major protein components of the ocular lens and show both structural and functional homology with the family of small heat shock proteins. αB crystallin is also present in various extra‐lenticular tissues, with a high concentration in cardiac muscle. In this study, the myocardium and conducting system from 15 adult and 25 fetal and infant hearts were examined by immunohistochemistry using a previously characterized antiserum to αB crystallin. Contractile myocardium showed moderate staining, with particular localization to Z bands and intercalated discs. Fibres of the sino‐atrial and atrio‐ventricular nodes and His bundle showed less intense staining than contractile fibres, whereas fibres of the left and right bundle branches showed more intense staining. This distribution is similar to that previously demonstrated for the intermediate filament desmin. This observation, together with currently available evidence, suggests that cardiac αB crystallin may play a role in protecting the cytoskeleton during cell stress. For practical purposes, immunostaining with αB crystallin greatly facilitates the identification of cardiac conducting fibres.
Human glomerular growth during childhood: A morphometric study
Authors: Akaoka K. , White R. H. R. , Raafat F.
Published: June 2005

Morphometric measurements of glomerular size were made on renal biopsy specimens obtained from 16 children with minimal change nephrotic syndrome (MCNS) and 16 with isolated recurrent haematuria (RH), whose ages ranged from 1.8 to 15.2 years. Glomeruli were normal on light and electron microscopy, and immunofluorescence was essentially negative. Tracings were made of the outlines of glomerular capsules, tufts, and individual capillary lumens; using computerized digitometry, the mean areas of each item were determined. The number of capillary lumens per glomerulus was counted automatically. From these data, the mean mesangial area was also calculated. Mesangial cells were counted in at least 100 distal mesangial areas per biopsy. The mean glomerular capsular area (GCA) correlated well with the mean glomerular tuft area (GTA) but frequent capsular artefacts rendered the former measurement less suitable for comparison with diseased glomeruli. The mean GTA correlated significantly with age and body surface area in both MCNS and RH, as did the number of capillary lumens per glomerulus, but the mean capillary lumina area showed no such correlations. The mean mesangial area increased with age in proportion to GTA, whereas mesangial cell counts remained normal. These findings indicate that glomerular growth during childhood is proportional to body growth, and are consistent with capillary subdivision as the mechanism of enlargement.
An in vitro model of urothelial regeneration: Effects of growth factors and extracellular matrix proteins
Authors: Rebel Johanna M. , de Boer Willem I. , Thijssen Cornelia D. , Vermey Marcel , Zwarthoff Ellen C. , Van der Kwast Theodorus H.
Published: June 2005

Although the cellular turnover of resting urothelium is very low, its regenerative capacity is known to be outstanding. In organotypic mouse urothelial cultures closely mimicking the differentiation and multilayering of normal urothelium, we examined the cell biological mechanisms underlying urothelial regeneration and the specific role of growth factors and several extracellular matrix (ECM) components. Exposure to epidermal growth factor (EGF) and acidic fibroblast growth factor (aFGF) and culture on laminin resulted in enhanced expansion of the urothelium. Microscopy and assessment of proliferative activity revealed that enhanced urothelial expansion due to EGF could be attributed to increased proliferative activity and an increase in cell numbers, whereas aFGF‐stimulated expansion must be considered the consequence of increased cellularity and migration. Laminin‐enhanced urothelial expansion was shown to be the result of spreading of the entire urothelial organotypic culture. This was associated with a considerable decrease in the number of cell layers. A synergistic effect of growth factors and laminin was not found. This organotypic urothelial cell culture model seems to be very useful in studying strategies to improve urothelial regeneration.
Alcoholic muscle disease: Features and mechanisms
Authors: Preedy Victor R. , Salisbury Jonathan R. , Peters Timothy J.
Published: June 2005

Approximately 50 per cent of all chronic alcohol misusers have alcoholic muscle disease. Chronic alcoholic skeletal muscle myopathy is characterized by a selective atrophy of type II fibres, so that up to 20 per cent of the entire skeletal musculature is lost. The pathogenetic mechanism for the myopathy is currently unknown but a model has been described in which various anatomically‐distinct skeletal muscles are employed to reflect type I and II fibres, i.e. the soleus and plantaris, respectively. In chronic studies, rats were fed nutritionally complete liquid diets containing either ethanol or glucose (controls) for up to 6 weeks. In acute studies, rats were given single boluses of ethanol and rates of protein synthesis were examined at 2.5 h. The results show that the myopathy is due to defective skeletal muscle protein synthesis. The information gained from these studies enhances our understanding of skeletal muscle diseases characterized by preferential effects on anaerobic fibres and should be applicable to disease processes in other toxic or metabolic myopathies.
Dermal vascularity in lentigo maligna
Authors: Trotter Martin J. , Tron Victor A.
Published: June 2005

Lentigo maligna (LM) may represent a tumour arrested in an in situ phase, lacking the angiogenic capacity and underlying dermal neovascularization required for invasive growth. The acquisition of an angiogenic phenotype might be associated with the development of lentigo maligna melanoma (LMM). To investigate this thesis, sections of formalin‐fixed, paraffin‐embedded tissue from 15 LMMs, and 11 LM excision specimens were stained with the vascular endothelial marker Ulex europaeus agglutinin I. Dermal vessels were counted and vascular morphometry was performed. In specimens in which LMM was present, dermal vascularity was significantly increased in LM compared with normal skin. The most significant increases were found for dermal vascular density (39 per cent increase, P=0.008) and for total vessel surface area (62 per cent increase, P=0.005). However, when no LMM was present, the vascular density underlying LM (79 ± 9 vessels/mm2) did not differ significantly from that of adjacent normal skin (67 + 6 vessels/mm2), although focal ‘hot spots’ of increased vascularity were present. We conclude that increased dermal vascularity is present beneath in situ LM and that this increased vascular density is closely associated with the presence of invasive LMM in the same specimen.
Production of two monoclonal antibodies (FB1 AND FB21) useful for the identification of human B lymphocytes in formalin‐fixed, paraffin‐embedded tissues
Authors: Nozawa Yoshihiro , Abe Masafumi , Ohno Hitoshi , Fukuhara Shirou , Wakasa Haruki
Published: June 2005

Two monoclonal antibodies (FBI and FB21) reactive in formalin‐fixed, paraffin‐embedded tissue sections are reported in this paper. FB1 and FB21 recognize a cytoplasmic antigen and a surface antigen of B cells, respectively. FBI reacts with mantle zone (MZ) B cells, germinal centre (GC) cells, and marginal zone (MrZ) B cells, but not with T cells in lymphoid tissues. FB21 reacts with MZ B cells, GC cells in lymphoid tissues, and T cells of peripheral blood, but not with MrZ B cells in the spleen. Neither monoclonal antibody (MoAb) reacts with monocytes, granulocytes, or plasma cells. FB1 reacted with all the B‐cell lymphomas tested and with CD20‐positive Reed‐Sternberg cells in two of five cases of Hodgkin's disease, but not with multiple myelomas or T‐cell lymphomas. FB21 reacted with B‐cell lymphoma in 20 of 22 cases, but not with multiple myelomas, T‐cell lymphomas, or Reed‐Sternberg cells of Hodgkin's disease. Immunoprecipitation studies revealed that FB1 recognizes the same two polypeptide chains that are recognized by L26 and is a member of the CD20 antibody cluster. FB21 was thought to recognize a sialic acid‐dependent carbohydrate epitope and this was confirmed at the Fifth International Conference on Human Leukocyte Differentiation Antigens (Boston, 1993) FB21 did not react with splenic MrZ B cells and was different from the pan B markers reported previously [CD20 (L26), CD45RA (MB1), and CD74 (LN‐2)]. FB21 recognizes a subset of B cells and appears to be closely related to CD75/76 antibodies. FB1 and FB21 are useful MoAbs for the diagnosis and analysis of B‐cell lymphomas.
Demonstration of URR‐duplication variants of human papillomavirus type 6 in paraffin‐embedded tissue sections of one condyloma acuminatum and one buschke‐loewenstein tumour
Authors: Albert Rüubben , Spelten Bettina , Albrecht Jörg , Grußendorf‐Conen Elke‐Ingrid
Published: June 2005

Human papillomavirus type 6 (HPV 6) induces condylomata acuminata and laryngeal papillomas. Occasionally, HPV 6 may also be found in low‐grade verrucous carcinomas. In some tumours, genetic analysis revealed the presence of HPV 6 variants with rearrangements, mostly DNA duplications, within the upstream regulatory region (URR). In this study, we analysed 98 formalin‐fixed, paraffin‐embedded condylomata acuminata obtained from 54 patients for the presence of URR‐duplication variants of HPV 6. HPV 6 DNA could be amplified by polymerase chain reaction (PCR) from 40 samples. One condyloma acuminatum contained a HPV 6 genome with rearranged URR. Analysis using restriction enzyme cleavage suggested a DNA duplication within the URR of approximately 200 bp, spanning the Hpa II site at nt 7863 and the Dde I site at nt 7843 but not involving the Rsa I site at nt 7633. In addition, we analysed the distribution of the already characterized URR‐duplication variant HPV 6ACIB within different paraffin‐embedded tissue sections of a Buschke‐Loewenstein tumour. No correlation could be demonstrated between the presence of the rearranged genome and malignant histological changes. This result and the demonstration of an URR‐duplication variant in a typical condyloma acuminatum suggest that duplications within the URR of HPV 6 are not directly related to malignant progression of HPV 6‐induced tumours.
Mucosa‐associated lymphoid tissue (MALT) in the human conjunctiva
Authors: Wotherspoon Andrew C. , Isaacson Peter G. , Hardman‐Lea Simon
Published: June 2005

The recent description of conjunctival lymphomas with features of lymphomas that arise in mucosa‐associated lymphoid tissue (MALT) infers the presence of MALT in the human conjunctiva. Some authors suggest that organized MALT is found in the normal human conjunctiva, but others disagree. In order to resolve this problem, we have examined the conjunctival fornix from unselected autopsies with no prior history of conjunctival or ocular disease for the presence of organized MALT. In only 31 per cent of cases was organized lymphoid tissue found with the morphological and immunophenotypic features of MALT. This finding suggests that MALT is not found in the normal human conjunctiva but is acquired during life in a proportion of apparently asymptomatic individuals.
Splenic interleukin 1 gene expression is associated with accumulation of macrophages and oxygen radical production in Plasmodium vinckei malaria
Authors: Prada Javier , Neifer Stefan , Muller Simone , Rudolph Roland , Bienzle Ulrich , Kremsner Peter G.
Published: June 2005

High levels of interleukin 1 alpha (IL‐1 alpha) were detected in vitro, in murine peritoneal macrophages stimulated with Plasmodium vinckei exogenous antigens, and in vivo, in sera of P. vinckei‐parasitized mice. Moreover, high production of IL‐1 alpha mRNA could be detected by in situ hybridization analysis in spleen sections of mice during the course of P. vinckei malaria. The observed IL‐1 alpha gene expression in the spleen was associated with the accumulation of F4/80+ macrophages in the red pulp and in the marginal zone of follicles, as well as with the relative proportions of Mac‐1+ cells in the spleen and the capacity of spleen cells to produce reactive oxygen intermediates during murine malaria.
Immunohistochemical localization of glutathione s‐transferases in sarcomas
Authors: McKay Judith A. , Murray Graeme I. , Ewen Stanley W. B. , Melvin William T. , Burke M. Danny
Published: June 2005

The glutathione S‐transferases (GSTs) are a multi‐gene family of enzymes involved in detoxifying electrophilic compounds and the expression of these enzymes in tumours has been proposed as one important mechanism of anti‐cancer drug resistance. In this study, the localization of the major cytoplasmic forms of GST has been studied in soft tissue sarcomas by immunohistochemistry. The alpha, mu, and pi forms of GST were identified in 59, 68, and 51 per cent of tumours, respectively. In addition, GST pi immunoreactivity was consistently identified in fibroblasts in adjacent non‐neoplastic tissue. The presence of specific forms of GST in soft tissue sarcomas may contribute to the drug resistance frequently observed in these tumours.
Detection of T‐cell receptor β chain mRNA in frozen and paraffin‐embedded biopsy tissue using digoxigenin‐labelled oligonucleotide probes in situ
Authors: Hodges Elizabeth , Howell William M. , Tyacke Sean R. , Wong Raymond , Cawley Michael I. D. , Smith John L.
Published: June 2005

In situ hybridization techniques using a cocktail of digoxigenin‐labelled T‐cell receptor (TcR) constant (C) region β oligonucleotide probes were used to detect TcRβ mRNA in frozen and paraffin‐embedded tissue sections. The specificity of the Cβ cocktail was confirmed by Northern blot analysis. The TcR Cβ cocktail successfully hybridized to T cells in frozen and paraffin‐embedded tissue obtained from patients with inflammatory arthropathies, B‐ and T‐cell non‐Hodgkin's lymphoma (NHL), and reactive tonsillitis, and showed staining patterns comparable to those obtained by conventional immunohistological detection of T cells. This is the first report of in situ studies using labelled TcR Cβ oligonucleotide probes and may indicate the feasibility of investigating clonal T‐cell populations using digoxigenin‐labelled clonospecific probes in clinical samples in situ.
Racial variation in the O‐acetylation phenotype of human colonic mucosa
Authors: Campbell Fiona , Appleton Mark A. C. , Fuller Clare E. , Greeff Michael P. , Hallgrimsson Jonas , Katoh Ryohei , Ng O. L. Irene , Satir Ali , Williams Geraint T. , Williams E. Dillwyn
Published: June 2005

O‐acetylated and non‐O‐acetylated sialoglycoproteins can be distinguished by the mPAS (mild periodic acid‐Schiff) histochemical technique. Individual adults show one of three different patterns of staining of large intestinal mucosa: uniformly mPAS‐positive, uniformly mPAS‐negative, or mPAS‐negative with scattered mPAS‐positive crypts. To test our hypothesis that these variations are the results of a single autosomal gene (oat) polymorphism, we have studied the frequency of the three patterns of staining in a total of 435 adult colon specimens from six geographically separate populations: British, South African blacks, Icelanders, Japanese, Hong Kong Chinese, and Bahrainis. The distribution of the three types of staining fell into two groups. In Japanese and Chinese, uniformly mPAS‐positive cases were much more frequent than uniformly mPAS‐negative cases; this distribution differed significantly (X2, P<0·001) from that in non‐Sino‐Japanese, where the uniformly mPAS‐positive phenotype was much less frequently found than the uniformly mPAS‐negative phenotype. In neither of the groups did the frequency of the three phenotypes differ significantly from that predicted for a single gene polymorphism by the Hardy‐Weinberg law. The variation in staining patterns between populations is consistent with variation in frequency of a single polymorphic autosomal gene (oat) controlling O‐acetylation of sialic acid, probably by an O‐acetyl transferase enzyme. Loss of function mutation in the high acetylator gene (oata) in a colonic crypt stem cell in heterozygous individuals would account for the scattered discordant crypts. Gene frequencies for a variety of enzymes differ between the Sino‐Japanese and non‐Sino‐Japanese races. This newly described gene polymorphism may be related to differential susceptibility to organisms binding specifically to either O‐acetylated or non‐O‐acetylated sialoglycoproteins, or to differential enteric colonization by bacterial flora that vary in their relative secretion of sialidases and sialate O‐acetyl esterase.
Human colonic stem cell mutation frequency with and without irradiation
Authors: Campbell Fiona , Fuller Clare E. , Williams Geraint T. , Williams E. Dillwyn
Published: June 2005

Mild periodic acid‐Schiff (mPAS) staining distinguishes O‐acetylated from non‐O‐acetylated sialoglycoproteins. In human colonic mucosa, individuals possess one of three phenotypes: uniformly mPAS‐positive (non‐O‐acetylated), uniformly mPAS‐negative (O‐acetylated), and negative with infrequent scattered positive crypts. This is due to a polymorphism in a single autosomal gene (oat). Discordant crypts have not been found in children's colons, suggesting that they result from somatic mutation in heterozygous individuals. We now present evidence to confirm this based on a study of radiation‐induced changes. Comparison of mPAS staining of large intestinal mucosa from patients given radiation 4 weeks before surgery for carcinoma of the rectum with matched controls receiving surgery alone showed a similar phenotype distribution, but informative irradiated patients showed an increased frequency of discordant crypts (irradiated vs. non‐irradiated 14·5 ± 8·2 × 10−4 vs. 6·1 ± 4·2 × 10−4). When these were classified as wholly or partially involved by the aberrant phenotype, the increase was most marked in partially involved crypts (7·5 ± 4·5 × 10−4 vs. 0·3 ± 0·5 × 10−4, Mann‐Whitney U, P<0·005). Two patients receiving radiotherapy many years before colectomy showed a very high total discordant crypt frequency but relatively few partially affected crypts. Studies of somatic mutation in colonic or small intestinal crypts following a single dose of mutagen in mice have shown early partial crypt involvement by the mutated phenotype and later complete crypt involvement. The demonstration in man that recent radiation greatly increases the frequency of partially involved discordant crypts while long‐term radiation greatly increases the frequency of wholly involved discordant crypts confirms that the discordant crypts result from stem cell mutation in individuals heterozygous for a polymorphic gene controlling O‐acetylation of sialoglycoproteins. They show that the mPAS technique can be used to identify and quantify colonic stem cell somatic mutation in man.
Detection of the c‐met proto‐oncogene product in normal skin and tumours of melanocytic origin
Authors: Saitoh Kazuya , Takahashi Hiroyuki , Sawada Norimasa , Parsons Peter G.
Published: June 2005

The proto‐oncogene c‐met product (c‐MET) is a receptor tyrosine kinase and functions as a receptor for hepatocyte growth factor (HGF). Although the function of c‐MET has yet to be fully clarified, HGF stimulates the phosphorylation of tyrosyl residues on c‐MET and triggers the signal transduction pathways, resulting in a contribution to the malignant progression of melanonocytes with synergic factors such as basic fibroblast growth factor and mast cell growth factor. Using immunohistochemical methods, we have studied the localization of c‐MET in normal skin and various melanocytic tumours, c‐MET was detected in keratinocytes, melanocytes, sebaceous cells, and other cells of the skin. In particular, basal melanocytes almost always showed nuclear labelling. Melanocytic naevi generally revealed predominantly nuclear staining of cells in the epidermis, whereas only a few cases showed a distinct cytoplasmic localization of c‐MET in dermal naevus cells. The distribution pattern of c‐MET in melanoma cells was basically similar to that of benign lesions, although the numbers tested were small. Cultured human melanoma cells also showed predominantly nuclear labelling, but were unresponsive to exogenous c‐MET ligand HGF. Treatment with the glucosidase inhibitor castanospermine caused accumulation of protein at 220 kD, without diminishing the amount of normally‐processed 190‐kD c‐MET. Although there was no significant difference in c‐MET distribution between benign and malignant melanocytic lesions, it is suggested that malignant transformation of melanocytes may be associated with loss of response to HGF or other growth‐regulating factors.
Wet autoclave pretreatment for antigen retrieval in diagnostic immunohistochemistry
Authors: Bankfalvi Agnes , Navabi Hossein , Bier Bert , Böcker Werner , Jasani Bharat , Schmid Kurt W.
Published: June 2005

Interest has recently been shown in adapting the microwave oven heating technique for antigen retrieval to routine diagnostic immunocytochemical practice. Although it has proved effective as a specialist method for individual antigen localization in many laboratories, it has certain drawbacks which have hampered its wider routine application. These include the need to monitor the sections during the microwave treatment to prevent damage or drying, the limited number of sections that can be accommodated in the microwave oven, and the inevitable alteration in nuclear morphology induced by the microwaves. In order to obviate these difficulties, we have modified the wet autoclave method of Shin et al. (Lab Invest 1991; 64: 693–702) as a routine technique for retrieval of a variety of cell surface, cytoplasmic, and nuclear antigens in formalin‐fixed and paraffin‐embedded tissue. The technique produces even enhancement of several refractory antigens in anatomically different sites and has the potential to handle reliably up to 200 sections at a time without significant damage to the section or to nuclear morphology.
Loss of membranous E‐cadherin expression in pancreatic cancer: Correlation with lymph node metastasis, high grade, and advanced stage
Authors: Pignatelli Massimo , Ansari Tareq W. , Gunter Pat , Liu Dan , Hirano Shinji , Takeichi Masatoshi , Klöppel Günter , Lemoine Nicholas R.
Published: June 2005

Epithelial cadherin (E‐cadherin) is a Ca2+‐dependent cell‐cell adhesion molecule that connects cells via homotypic interactions. Its function is critical in the induction and maintenance of cell polarity and differentiation, and its loss of downregulation is associated with an invasive and poorly differentiated phenotype in colon and other tumours. We have used an avidin‐biotin immunoperoxidase technique to localize E‐cadherin in microwave‐treated, paraffin‐embedded sections from 36 patients with pancreatic adenocarcinomas. E‐cadherin was expressed by normal ductal and acinar cells with typical membranous staining at the intercellular junctions. Loss of normal surface E‐cadherin expression was found in 19/36 (53 per cent) tumours compared to the adjacent normal ductal cells. Abnormal E‐cadherin expression was found more frequently in poorly differentiated (grade III) (6/7, 86 per cent) than in well‐differentiated tumours (grade I) (4/14, 28 per cent) (P=0·012). Membranous E‐cadherin expression was also lost more frequently in primary tumours with lymph node (stage III) (14/23, 61 per cent) and distant metastasis (stage IV) (2/2, 100 per cent) compared with 3/11 (27 per cent) lymph node‐negative tumours (stage I) (P=0·043). In conclusions, our data indicate that loss of membranous E‐cadherin expression is associated with high grade and advanced stage in pancreatic cancer.
Atypical alveolar hyperplasia: Relationship with pulmonary adenocarcinoma, p53, and c‐erbB‐2 expression
Authors: Kerr Keith M. , Carey Francis A. , King George , Lamb David
Published: June 2005

Atypical alveolar hyperplasia (AAH) has recently been described in human lungs in association with primary lung cancer, particularly adenocarcinoma. Unlike proximal bronchogenic carcinoma, peripheral (parenchymal) adenocarcinoma of the lung does not have a well‐recognized progenitor lesion. Epidemiological, morphometric, and cytofluorometric data in the literature suggest that AAH is a candidate premalignant entity. In this study, 97 AAH lesions were found in lungs resected from 29 patients (1–13 lesions per case, mean 3·5) being treated for presumed carcinoma (25/29 had adenocarcinoma). From a study case‐load of 285 adenocarcinoma‐bearing lungs, the AAH incidence was 8·8 per cent. Sections of 67 AAH lesions from 19 patients were stained using monoclonal antibodies against Ki67 (MIB1), p53 (DO7), and c‐erbB‐2 (NCL‐CB11). Ki67 was expressed in up to 10 per cent of AAH nuclei. Thirty‐nine lesions (58 per cent) showed stainable p53 protein, while five (7 per cent) expressed membrane c‐erbB‐2 oncoprotein. These latter five lesions were all strongly positive for p53, and both p53 and c‐erbB‐2 staining was associated with increased cellular crowding and pleomorphism in AAH. These data demonstrate that AAH exhibits some genetic changes associated with malignancy and thereby support the hypothesis that AAH is premalignant.
Incidence of latent infection of Epstein–Barr virus in lung cancers—an analysis of EBER1 expression in lung cancers by in situ hybridization
Authors: Kasai Kiyoshi , Sato Yuichi , Kameya Toru , Inoue Hayato , Yoshimura Hirokuni , Kon Shinichiro , Kikuchi Kokichi
Published: June 2005

To evaluate the presence of Epstein–Barr virus (EBV) in lung cancers of Japanese patients, 81 lung cancers were examined using a highly sensitive in situ hybridization (ISH) method, employing an antisense oligonucleotide probe for EBV‐encoded small nuclear RNA‐1 (EBER). EBER1 expression was demonstrated in one poorly differentiated squamous cell carcinoma associated with marked lymphoid stroma (PDSCC‐LS), two well differentiated adenocarcinomas, and two moderately differentiated squamous cell carcinomas, but was not detectable in other lung cancers, including small cell carcinomas. Unlike lymphoepithelioma‐like undifferentiated carcinoma (LELC) of the lung, the PDSCC‐LS consisted of poorly differentiated cells with distinct cell borders and nuclei with a coarse chromatin pattern and some prominent nucleoli. Most of the cancer cells expressed intense EBER1 signals. Although small to moderate numbers of cells positive for EBER1 were present in two adenocarcinomas and two squamous cell carcinomas, EBER1 signals varied in intensity and number in these four cases. Although polymerase chain reaction (PCR) and Southern blot hybridization with a 32P‐labelled probe internal to the primers were conducted to detect the EBV genome in 24 lung cancers, including five EBER1‐positive cases, the genome was found to be positive in the five cases with EBER1‐positive staining, including the PDSCC‐LS, two adenocarcinomas and two squamous cell carcinomas, but not in the other cases. This study indicates that the morphological features of EBV‐associated lung cancers are not restricted to the typical LELC type.
Prognostication of astrocytoma patient survival by Ki‐67 (MIB‐1), PCNA, and S‐phase fraction using archival paraffin‐embedded samples
Authors: Sallinen Pauli K. , Haapasalo Hannu K. , Visakorpi Tapio , Helén Pauli T. , Rantala Immo S. , Isola Jorma J. , Helin Heikki J.
Published: June 2005

The prognostic power of three proliferation estimation methods, Ki‐67 (MIB‐1) and PCNA immunohistochemistry, and flow cytometry (S‐phase and S+G2/M fractions, respectively), were evaluated in 50 cases of astrocytoma. Each proliferation index showed a strong association with the grade of malignancy (grades I‐IV). The MIB‐1 labelling index (LI) provided additional information, as it could be used for the discrimination of grade II and grade III astrocytomas (P=0·0357). All three proliferation estimation methods also had strong prognostic potential (MIB‐1 LI: P<0·0001; PCNA Li: P<0·0001; S‐phase: P=0·0004; S+G2/M: P=0·0124). According to the receiver operating characteristics (ROC) curve, the MIB‐1 LI showed generally the best sensitivity and specificity in placing the patients correctly into groups of survivors and non‐survivors, which was further confirmed in the multivariate analysis. Only 4 per cent of the patients having high MIB‐1 scores (>15·3 per cent) were alive after 2‐years' follow‐up. In contrast, 72 per cent of patients with tumours of low proliferation activity survived. It appears that Ki‐67 (MIB‐1) immunolabelling using archival paraffin‐embedded samples is of value in predicting prognosis in astrocytic tumours.
Characterization of a subtype of primary osteoclastoma: Extracellular calcium but not calcitonin inhibits aggressive HLA‐DR‐positive osteoclastoma possessing ‘functional’ calcitonin receptors
Authors: Rathod Hersha , Malcolm Archibald J. , Gillespie James I. , Berry Vanita , Pooley Joseph , Piggott Nigel H. , Datta Harish K.
Published: June 2005

We report here a case of primary osteoclastoma that despite possessing HLA‐DR‐positive status and ‘functional’ calcitonin receptors, exhibited aggressive in vitro and in vivo bone resorptive activity. In the osteoclast bone slice assay employing scanning electron microscopy, the giant cell‐mediated bone resorption was uninhibited by salmon calcitonin (10 nM) and significantly inhibited by raised extracellular calcium (20 mM). In Fura‐2AM based microspectrofluorimetric assays, the presence of the ‘functional’ calcitonin receptors was ascertained by a rise in intracellular calcium induced by calcitonin and high extracellular calcium. These findings provide evidence for a hitherto unrecognized subtype of giant cells that have HLA‐DR‐positive status, exhibit avid bone resorptive activity, but remain insensitive to calcitonin despite possessing calcitonin receptors.
Intrahepatic expression of hepatitis C virus antigens in chronic liver disease
Authors: Nouri‐Aria Kayhan T. , Sallie Richard , Mizokami Masashi , Portmann Bernard C. , Williams Roger
Published: June 2005

Localization of hepatitis C virus (HCV) antigens was studied in fresh frozen and formalin‐fixed, paraffinembedded liver tissue by immunoperoxidase using monoclonal antibodies to nucleocapsid protein and polyclonal human immunoglobulin G purified from plasma containing antibodies to structural and non‐structural antigens of hepatitis C virus. The results observed using monoclonal antibody to HCV core were similar to those of polyclonal IgG against HCV antigens in the majority of cases and both correlated well with HCV status as defined by ‘nested’ polymerase chain reaction. HCV antigens were detected in both hepatocytes and mononuclear cells. Using polyclonal human IgG, a small proportion of biliary epithelial cells were also positive in 6/29 patients. In most of the specimens examined, relatively few cells (1–5 per cent) were found to be positive for HCV antigens. The cryostat sections, using polyclonal IgG against HCV antigens, exhibited greater immunohistochemical staining, suggesting that the fixation and processing of the tissue may be a major factor in the conservation and the outcome of HCV antigen(s) findings. However, the results using monoclonal antibodies may reflect the specificity of antigen expression.
Is the carbohydrate sialosyl‐Tn a marker for altered, non‐malignant activity in squamous epithelium in the head and neck region?
Authors: Bryne M. , Gravdahl C. , Koppang H. S. , Kjærheim Å. , Dabelsteen E.
Published: June 2005

Cell surface carbohydrates are involved in many cell functions such as cellular differentiation, adhesion, and invasion. A carbohydrate, sialosyl‐Tn (STn), is expressed in many human carcinomas but generally not in normal epithelia. In the oral mucosa, however, STn had recently been observed on basal cell in some lesions with epithelial hyperplasia and dysplasia. The aim of this study was to carry out a systematic investigation of STn expression on epithelial basal cells in hyperplastic, ‘borderline’ malignant, and malignant head and neck lesions, to see if the expression of STn is associated specifically with hyperplastic conditions. Using the primary monoclonal antibody TKH2, normal controls did not reveal STn. STn was detected on probably post‐mitotic basal cells in hyperplastic head and neck lesions and on basal cells adjacent to cancers, but not within the carcinomas. A Ki67 antibody reacted with basal cells in other locations. The most highly differentiated lesions, such as focal epithelial hyperplasia and verrucous hyperplasia, revealed a high percentage (86 per cent in both cases) of STn reactivity. The least‐differentiated verrucous carcinomas (VCs) and keratoacanthomas (KAs) did not express STn, in contrast to the highly differentiated VCs and KAs. These findings indicate that STn‐negative cases may have a greater malignant potential that the STn‐positive cases. In conclusion, STn expressed on basal cells is possibly a marker for non‐malignant conditions with altered basal cell activity and for highly differentiated verrucous carcinomas.
Human papillomavirus DNA in oesophageal carcinomas in South Africa
Authors: Cooper Kumarasen , Taylor Louise , Govind Sanjay
Published: June 2005

Using morphological criteria, the presence of human papillomavirus (HPV) in oesophageal carcinomas has been inferred in patients from Finland and South Africa. However, studies to demonstrate the viral antigen in tissue sections of these tumours have proved disappointing. This study investigates 48 archival oesophageal carcinoma biopsies from South Africa for the presence of HPV DNA using non‐isotopic in situ hybridization (NISH) with HPV DNA probes to HPV 6, 11, 16, 18, 31, and 33. HPV DNA sequences were detected in 25/48 (52 per cent) oesophageal cancers. HPV 16 was present in 84 per cent of the HPV‐positive cancers. A NISH type 2 signal pattern (punctate/dot) was present in all HPV‐positive tumours. This signal pattern was previously shown to represent integrated HPV DNA within host chromosome. Integrated HPV DNA in oesophageal cancers has also been demonstrated in patients from China and Japan. In addition, the prevalence of HPV DNA in oesophageal cancers from high‐risk countries like South Africa (52 per cent) and China (49 per cent) would appear to be consistent.
17q allele loss is associated with lymph node metastasis in locally aggressive human colorectal cancer
Authors: Purdie Colin A. , Piris Juan , Bird Colin C. , Wyllie Andrew H.
Published: June 2005

A consecutive series of 87 colorectal tumours were studied for loss of a polymorphic probe on chromosome 17q and of the 64 informative cases, 13 (20 per cent) showed loss of heterozygosity (LOH). Examples of LOH were found in carcinomas of all stages and in a large non‐invasive adenoma. There was no correlation between 17q LOH and patient age, sex, standard clinicopathological variables (differentiation and nature of tumour margin), DNA ploidy, or tumour site, nor was 17q LOH associated with 17p LOH defined at four loci adjacent to p53. However, comparison of Dukes' B and C carcinomas revealed that tumours which had metastasized to regional lymph nodes at the time of primary surgery were significantly more likely to have lost this 17q allele. Clinical follow‐up of this cohort of patients showed no significant difference in survival between patients whose tumours had lost or retained 17q. Thus, we conclude that 17q allele loss is associated with lymph node metastasis in locally aggressive colorectal tumours but probably not with blood‐borne metastasis.
Chromosome 1 aneusomy, identified by interphase cytogenetics, in mammographically detected ductal carcinoma in situ of the breast
Authors: Harrison Michele , Magee Hilary M. , O'loughlin John , Gorey Thomas F. , Dervan Peter A.
Published: June 2005

Because of the relative rarity of ductal carcinoma in situ (DCIS) in the premammographic screening era, the unavailability of adequate fresh tissue for culture, and the lack of cytogenetic expertise in most pathology departments, there is little information on karyotypic abnormalities in DCIS. The purpose of this study was to investigate the frequency of chromosome 1 aneusomy in DCIS, using interphase cytogenetic techniques, and to correlate the findings with nuclear grade. Twenty‐one cases of DCIS, identified in a mammographically screened population, were studied by in situ hybridization. Chromosome 1 numbers were identified in interphase nucler in conventional histology sections, using a specific centromeric probe (pUC 1.77). In each case, 100 tumour nuclei were compared with 100 normal nuclei. Eighteen of 21 (86 per cent) cases were aneusomic for chromosome 2. This included 15 of 16 (94 per cent) pure comedo or predominantly comedo DCIS. Fifteen of 16 (94 per cent) DCIS with grade 3 nuclei and 3 of 5 (60 per cent) cases with grade 2 nuclei were aneusomic. One case with grade 3 nuclei (a comedo carcinoma) was negative. We conclude that chromosome 1 aneusomy precedes invasion and is a relatively consistent occurrence in those DCIS with high nuclear grade.
Upregulation of integrin expression in benign vulvar warts
Authors: Williams A. T. , Sexton C. J. , Hanna N. F. , Leigh I. M.
Published: June 2005

Benign anogenital warts demonstrate features both of hyperproliferation and of aberrant differentation associated with human papillomavirus (HPV) infection. The expression of integrins has been examined in normal vulvar epithelium and benign vulvar warts to determine whether HPV infection is associated with changes in integrin expression, either during terminal differentiation or among the proliferating cell population. The expression of integrins in normal vulvar epithelium was similar to that seen in the epidermis, with little expression of α1 and expression of α2, α3, and β1 at the periphery of basal cells. In vulvar wart epithelium, α1 was not significantly expressed, but upregulation of α2, α3, and β1 was seen at the periphery of cells throughout the basal and spinous layers. In addition, α2 staining was greater at foci along the basal layer of normal vulva, whilst such heterogeneity was not detected in wart sections. Expression of β4 and αv was confined to the basal and lateral aspects of normal basal cells, but expressed at the periphery of basal cells and to a lesser extent in epibasal and spinous layers of warts. The observed differences in integrin expression are consistent with those reported in other hyperproliferative epithelia.
Mononuclear cell thyroiditis in rats with graft‐versus‐host disease
Authors: Fujiwara Kouji , Peszkowski Michael J. , Larsson Åke , Yamasaki Akira , Toda Shuji , Watanabe Teruo
Published: June 2005

Rats with graft‐versus‐host disease (GVHD), induced by injecting spleen cells of parental strain rats (Brown Norway) into non‐irradiated (Brown Norway × Lewis) F1 hybrid rats, develop thyroiditis. This is characterized by mononuclear cell infiltration with destruction of the epithelium and a significant reduction in serum thyroid hormone levels. Immunohistochemically, the mononuclear cells consisted mainly of CD8‐positive cells and macrophages. These findings provide evidence that the thyroid gland can now be enumerated as one of the target organs during GVHD. The CD8‐positive cells may serve as important effector cells in lesion development, either by direct cytotoxicity or by supporting the cytotoxic potential of macrophages.
Low incidence of mbr bcl‐2/JH fusion genes in Hodgkin's disease
Authors: Kneba Michael , Eick Stefan , Herbst Hermann , Pott Christiane , Bolz Ingrid , Dallenbach Friederike , Hiddemann Wolfgang , Stein Harald
Published: June 2005

Lymph node biopsies from 140 cases of Hodgkin's disease (HD) and from 30 non‐malignant lesions were screened for the presence of t(14;18) translocations involving the major breakpoint region (mbr) of the bcl‐2 gene and the joining region (J)H of the immunoglobulin heavy chain gene, using a polymerase chain reaction (PCR) assay with subsequent nucleotide sequencing of amplified bcl‐2/JH junctional regions. Expression of the bcl‐2 protein within the Hodgkin and Reed‐Sternberg (HRS) cells was investigated in 86 cases of HD by immunohistochemistry on cryostat or paraffin sections. Although bcl‐2 expression could be found in a proportion of neoplastic cells in up to one‐third of HD cases, the frequency of t(14;18) gene fusions detected by PCR was low. We identified such gene fusions in only 3 out of 140 (2 per cent) HD cases, one biopsy of which presented with four clonally distinct bcl‐2/JH sequences. No t(14;18) was found in any of 30 reactive lymph node lesions. All fusion gene sequences were unique regarding the localization of the chromosome 14 and 18 breakpoints and the extranucleotide N‐insertions. None of these gene fusions conformed to t(14;18) breakpoint sequences previously characterized in our laboratories. Our findings point to a mere coincidence in some cases of HD lesions and cells carrying a t(14;18) in the same biopsy and argue against a significant role of bcl‐2 in the pathogenesis of HD.
Immunohistochemical localization of thrombomodulin in normal human skin and skin tumours
Authors: Jackson D. E. , Mitchell C. A. , Bird P. , Salem H. H. , Hayman J. A.
Published: June 2005

Thrombomodulin (TM) expression has been investigated in sections of normal human skin, in cultured normal human keratinocytes, and in a variety of skin tumours. TM was present in squamous epithelial cells in the spinous layer of normal epidermis and in the outer root sheath of hair follicles, but was absent in the cells of the basal layer. It appeared to be predominantly localized to the cell membrane and the intercellular bridges in these areas. Cultured normal human keratinocytes demonstrated functionally active constitutive TM expression on their cell surface. Immunoperoxidase staining of skin tumours using anti‐human TM antibodies demonstrated a typical cell membrane positivity in tumours with squamous or hair follicle differentiation. Basal cell carcinomas showed TM expression only in areas where incomplete squamoid metaplasia occurred. Sweat gland tumours and lesions of the melanogenic system failed to express TM. The localization of TM by immunostaining in various benign and malignant skin tumours typically correlated with their normal skin element of origin. The physiological significance of TM expression in the epidermis is currently undefined.
An evaluation of the staining of lymphomas and normal tissues by the rabbit polyclonal antibody to protein gene product 9.5 following non‐enzymatic retrieval of antigen
Authors: Langlois N. E. I. , King G. , Herriot R. , Thompson W. D.
Published: June 2005

We have previously described the staining of normal follicle centre lymphoid cells by the rabbit polyclonal antibody to protein gene product 9.5 (PGP9.5), following an antigen unmasking step employing heat pretreatment. Applying this finding to a range of lymphomas to determine whether this phenomenon could have a role in identifying lymphomas of follicle centre origin revealed no relationship between type or grade of lymphoma and staining of neoplastic cells. Testing a range of normal tissues following antigen retrieval displayed increased sensitivity and a greater range of tissue positivity. Immunoblots of gel electrophoresed tonsil and brain extract were performed. Although subjecting these blots to antigen unmasking increased sensitivity, no novel epitopes, in terms of additional bands, appeared. This suggested that antibody specificity remained unaltered.