bluuebunny/crossref_metadata_2025 huggingface dataset:
Ki‐S1 and PCNA expression in erythroid precursors and megakaryocytes—A comparative study on proliferative and endoreduplicative activity in reactive and neoplastic bone marrow lesions
Authors: Thiele Juergen , Bertsch Hans Peter , Kracht Lutz Walter , Anwander Thomas , Zimmer Joerg Detlef , Kreipe Hans , Fischer Robert
Published: June 2005

The monoclonal antibody Ki‐S1 reacts with a cell proliferation‐associated nuclear antigen which is expressed in the G1 through G2/M phases of the cell cycle and is resistant to formalin fixation. We have studied Ki‐S1 and PCNA (PC10) immunostaining of erythroid precursors (proliferative activity) and megakaryocytes (endoreduplicative activity) in bone marrow trephine biopsies in a variety of reactive and neoplastic lesions using double immunohis‐tochemistry to identify both cell lineages. A significant increase in Ki‐S1 labelling compared with PCNA positivity was found in all conditions studied. In particular, specimens derived from secondary polycythaemia (SP), polycythaemia vera (P. vera), and primary osteomyelofibrosis (OMF), and from splenic tissue with myeloid metaplasia (MM), revealed a disproportionally high labelling index of erythropoiesis, which was not present in chronic myelogenous leukaemia (CML), AIDS, and autoimmune (idiopathic) thrombocytopenia (ITP). Enhancement of Ki‐S1 (PCNA) staining in SP and P. vera is in keeping with the relevant increase in erythroid precursor proliferation, but in OMF and MM there is overexpression of both proliferation markers, possibly due to secondary folic acid deficiency, which is known to cause a block in the S‐phase of the cell cycle. A significant correlation was observed between the sizes of megakaryocytes and their nuclei with Ki‐S1 (and also PCNA) staining. Ki‐S1 (and PCNA) labelling of predominantly smaller elements of this lineage supports a hypothesis that the phases of the cell cycle have different durations in the various steps of polyploidization, with a prolongation of G1/G2 at higher ploidy levels. A considerable number of large to giant (hyperpolyploid) megakaryocytes may have reached the end stage of endoreduplicative activity (Ki‐S1‐/PCNA‐negative GO phase). A comparative evaluation of these two proliferation markers adds significantly to our understanding of cell kinetics in erythro‐ and megakaryo‐poiesis, and sheds some light on the biological behaviour of these cell lineages in various disorders.
Cell proliferation correlates with prognosis in acinic cell carcinomas of salivary gland origin. Immunohistochemical study of 30 cases using the MIB 1 antibody in formalin‐fixed paraffin sections
Authors: Skalova Alena , Leivo Ilmo , Von Boguslawsky Kristina , Saksela Eero
Published: June 2005

Immunohistochemical staining with the MIB 1 antibody was used to assess cell proliferation in 30 cases of acinic cell carcinoma of salivary glands. Until now, no prognostic factors have been available for these rare tumours. The MIB 1 monoclonal antibody recognizes the Ki‐67 antigen in formalin‐fixed, paraffin‐embedded tissues. A MIB 1 index was developed as a means of expressing the percentage of MIB 1‐positive tumour cell nuclei, and the tumours were scored without prior information of clinical behaviour. The staining results were then compared with the clinical outcome of the patients. All eight patients who developed tumour recurrences had MIB 1 indices higher than 5 per cent. Tumour recurrences could be predicted even in cases of bland morphology and low mitotic rate. Three patients died of their recurrent tumours, and had MIB 1 indices of 56.2, 12.7, 7.8 per cent in their primary tumours. Five of seven patients with MIB 1 indices higher than 10 per cent had unfavourable outcomes. None of the 17 patients with MIB 1 indices lower than 5 per cent developed recurrences during follow‐up periods up to 30 years. The present results indicate that MIB 1 staining appears to be a significant prognostic factor in acinic cell carcinomas of salivary gland origin.
Optimization of immunohistochemical detection of ERBB2 in human breast cancer: Impact of fixation
Authors: Penault‐Llorca Frédérique , Adelaïde José , Houvenaeghel Gilles , Hassoun Jacques , Birnbaum Daniel , Jacquemier Jocelyne
Published: June 2005

In an attempt to standardize the immunohistochemical detection of ERBB2, we evaluated six normal breast samples and 58 breast carcinomas for (1) the effect of four different fixatives (Bouin's fluid, buffered formalin, alcoholic formalin, and methacarn), and (2) the sensitivity and specificity of eight commercially available antibodies. ERBB2 gene copy numbers and RNA transcripts were measured by Southern and Northern blot analysis in all samples. The BT‐474 and MCF7 cell lines were processed in the same way as the tissue samples and used as references of activation or the normal state of the ERBB2 gene, respectively. Complete concordance between immunohistochemistry and molecular biology was observed in 43/58 cases (74 per cent). However, fixation and processing protocols significantly affected the reactivity of the antigenic determinants. The most consistent results were obtained with NCLCB11 on frozen sections and with Tab250 on paraffin‐embedded sections. For prospective studies, the use of alcoholic formalin in association with a highly specific antibody (Tab250, 9G6, NCLCB11, or OA‐11‐854) gave the best results. Methacarn appears to be a reliable fixative, but was studied in only 28 cases. For retrospective studies, the most important parameter to take into account is the sensitivity of the antibody. Thus, with Bcuin s fixation, 3B5 was the only reliable antibody, whereas with buffered formalin, Tab250 gave the most consistent results. This study will help inter‐laboratory comparisons, and in determining the prognostic significance of ERBB2 expression in breast carcinomas using a standardized methodology.
Epstein‐Barr virus in nasal T‐cell lymphomas (polymorphic reticulosis/midline malignant reticulosis) in Western China
Authors: Van Gorp Joost , Weiping Liu , Jacobse Karen , Yong‐Huei Liu , Feng‐Yuan Li , de Weger Roel A. , Li Gandi
Published: June 2005

Polymorphic reticulosis (PR) or midline malignant reticulosis (MMR) is considered to be malignant, or at least pre‐malignant T‐cell proliferations of the nose or midline area. Recent reports of small series of nasal T‐cell lymphomas have shown a strong association with Epstein‐Barr virus (EBV). Furthermore, a peculiar phenotype is described, with expression of CD56 and not of CD3, suggesting a possible origin from natural killer (NK) cells. We have analysed a series of 38 cases of PR/MMR for the presence of EBV by in situ hybridization (ISH) of the EBV‐encoded RNAs 1 and 2 (EBER). Twenty cases were tested for expression of EBV‐encoded latent membrane protein 1 (LMP‐1). Special attention was also paid to the expression of CD3 and the NK cell‐related marker CD56. Thirty‐two cases (84 per cent) showed positive EBER ISH. In 5 of 20 cases, LMP‐1 expression was detected. In three cases, a few scattered cells were positive, and in two cases, LMP‐1 was detected in clusters of atypical cell. Most of the neoplasms showed expression of CD3 (89 per cent) and in 27 cases (71 per cent), CD56 was detected. These results are consistent with an aetiopathogenetic role for EBV in most, but not all, cases of PR/MMR. Our findings are less supportive of a major role for LMP‐1 in tumour genesis. CD3 expression in most of the cases of PR/MMR underlines the T‐cell origin of these neoplasms, often with aberrant expression of CD56.
Expression of the trefoil peptides pS2 and human spasmolytic polypeptide (hSP) in Barrett's metaplasia and the native oesophageal epithelium: Delineation of epithelial phenotype
Authors: Hanby Andrew M. , Jankowski Janusz A. Z. , Elia George , Poulsom Richard , Wright Nicholas A.
Published: June 2005

The distribution of the two trefoil peptides pS2 and human spasmolytic polypeptide (hSP) and their mRNAs was investigated in 90 selected oesophageal biopsies, 23 of which contained epithelium with wholly gastric cardiac‐type morphology, 52 specialized (intestinal)‐type metaplasia, and 15 non‐metaplastic oesophageal epithelium. No fundic‐type epithelium was represented. The cardiac‐type epithelium resembled true gastric antral epithelium, with hSP and pS2 mRNA localization to the superficial/foveolar compartment and hSP mRNA alone in deeper glands. The pattern of peptide distribution was broadly in line with the mRNA, but hSP peptide was generally not demonstrable in the surface epithelium. pS2 immunostaining was diffusely cytoplasmic, whereas in specialized‐type mucosa it was aggregated and cytoplasmic hSP mRNA was demonstrable in surface epithelium of incomplete‐ but not complete‐type intestinal (specialized) metaplasia. Deep glands with morphological features of pyloric glands/ulcer‐associated cell lineage (UACL) typically contained only hSP peptide and its mRNA. In three biopsies containing specialized epithelium, small foci morphologically identical to true small intestinal surface epithelium were seen in which only pS2 mRNA and peptide were found and then only in rare goblet cells. Neither squamous epithelium nor oesophageal glands contained demonstrable pS2/hSP mRNA or peptide. The function of these proteins in the metaplastic mucosa of Barrett's oesophagus is unknown but they may reflect an epithelium responding to repeated insult. Their localization facilitates definition of the disparate epithelium types seen in this portion of the gut, with both native gastric and intestinal epithelia, and may help to pinpoint high‐risk epithelium in Barrett's oesophagus, a potentially preneoplastic condition.
DNA in situ hybridization as a diagnostic tool in the discrimination of melanoma and spitz naevus
Authors: de wit Peter E. J. , Kerstens Harold M. J. , Poddighe Pino J. , Van Muijen Goos N. P. , Ruiter Dirk J.
Published: June 2005

As the clinical and histological differential diagnosis between Spitz naevus and cutaneous melanoma may be very difficult, we have investigated whether DNA in situ hybridization may be helpful in resolving this problem. To this end, routinely‐processed paraffin sections of 15 typical Spitz naevi, 15 typical nodular melanomas, and five cases originally misdiagnosed as Spitz naevi but which later metastasized and were reclassified as melanoma were analysed using a method previously described (De Wit et al., J Invest Dermatol 1992; 98: 450–458). Microscopical semi‐quantitative evaluation revealed that the number of nuclei with supernumerary aberrations of the centromere region of chromosome 1, suggestive of aneuploidy, was significantly different in Spitz naevi and nodular melanoma. The mean number of aberrant nuclei per high power field was 0.41 and 4.01, respectively (P=0.0001). On applying the results of the typical lesions to the equivocal, originally misdiagnosed lesions, three out of five could be identified as melanoma. These results suggest that the application of DNA in situ hybridization may contribute to the positive identification of histologically equivocal pigmented lesions. The advantages of this technique are that it is cheap, requires little tissue, and can be applied on routinely‐processed paraffin sections.
Borrelia burgdorferi‐induced ultrastructural alterations in human phacgocytes: A clue to pathogenicity?
Authors: Riting Michael G. , Häpl Thomas , Krause Andreas , Kressel Michael , Groscurth Peter , Burmester Gerd R.
Published: June 2005

A chronic infection with the spirochaete Borrelia burgdorferi typically results in a multistage, multistem illness. Thus, Lyme borreliosis may provide an interesting model to study the pathomechanisms of microbial persistence. In the present investivation, human peripheral blood monocytes, polymorphonuclear leukocytes, and synovial macrophages were incubated with B. burgdorferi and examined by light and electron microscopy. It was found that incubation with the spirochaetes induced distinct features in the phagocytes. Features which may be related to the pathogenesis of Lyme disease included the segmental uptake of spirochates with leaky lysosomes, the invagination of large membrane areas, the extra‐lysosomal degradation of internalized B. burgdorferi cells and, finally, the formation of mononuclear synctial cells and homotypic cell clusters. Feautres of unknown relevance were the occurrence of two types of cytoplasmic inclusion bodies and exocytic vesicles. These novel findings suggest that reactive alterations of the phagocytes may contribute to the pathogenesis of Lyme borreliosis, which could help to focus future histopathological studies. Moreover, these results may provide new insights into the pathogenesis of other infectious diseases characterized similarly by microbial persistence.
The ulceration‐associated cell lineage (UACL) reiterates the Brunner's gland differentiation programme but acquires the proliferative organization of the gastric gland
Authors: Ahnen Dennis J. , Poulsom Richard , Stamp Gordon W. H. , Elia George , Pike Christine , Jeffery Rosemary , Longcroft Janet , Rio Marie‐Christine , Chambon Pierre , Wright Nicholas A.
Published: June 2005

The ulceration‐associated cell lineage (UACL) develops in the human gastrointestinal mucosa after ulceration; it grows out from the bases of adjacent crypts and ramifies in the lamina propria to form a new gland, finally giving rise to a duct by which the glandular secretion and indeed cells are carried to the surface. Using immunocytochemistry and in situ hybridization with 35S‐labelled riboprobes, we have defined the pattern of trefoil peptide gene expression (pS2; human spasmolytic polypeptide, hSP), epidermal growth factor/urogastrone (EGF/URO), and the distribution of cell proliferation during the development of the UACL, as indicated by immunostaining for proliferating cell nuclear antigen (PCNA). Our studies reveal that the morphogenesis of the UACL shows a marked morphological resemblance to developing Brunner's glands; the pattern of trefoil peptide gene expression during UACL development is also very similar. However; trefoil peptide gene expression in the mature UACL complex is unique amongst gastrointestinal cells. The mature UACL shows a distinctive proliferative organization: while the early buds and glands are non‐proliferative, apparently being fed by cells from the parent crypts, a definitive proliferative zone develops within the duct. This, of course, corresponds to the location of the gastric gland proliferative zone. We propose that while the UACL shows novel features, it shares its differentiation programme with Brunner's glands, but its pattern of cell renewal eventually is that of the gastric gland.
Expression of pS2 protein in transitional cell bladder tumours
Authors: Lipponen P. K. , Eskelinen M. J.
Published: June 2005

A series of 201 bladder cancer biopsy specimens was analysed immunohistochemically for the expression of pS2 protein. Altogether, 61 per cent of the tumours were pS2‐negative; in 16 per cent less than 1 per cent and in 23 per cent of cases more than 1 per cent of cells were pS2‐positive. Normal transitional epithelium was negative for pS2. The fraction of positive cells was higher in poorly differentiated non‐papillary tumours and in invasive tumours with pelvic lymph‐node (P=0.05) and distant metastasis (P=0.10). pS2 expression was not related to sex, while patients aged 60–70 years had low fractions of pS2‐positive cells (P=0.03). DNA ploidy, S‐phase fraction, mitotic index, morphometric nuclear features, and expression of c‐erbB‐2, p53, and epidermal growth factor receptor were independent of expression of pS2. Tumours expressing pS2 in over 10 per cent of cells had a lower survival probability (P=0.0486). The results show that pS2 is expressed in 40 per cent of transitional cell bladder tumours, but that this marker has no clinical significance over established prognostic factors.
Site of production of IGF1 in the normal and stimulated mouse thyroid
Authors: Thomas Geraldine A. , Davies Howard G. , Williams E. Dillwyn
Published: June 2005

Insulin‐like growth factor 1 (IGF1) has emerged as an essential factor in the follicular cell growth response in vitro to TSH, although its source within the thyroid in vivo is not clear. We have studied the localisation of IGF1 mRNA by in situ hybridization using digoxigenin labelled oligoprobes in tissue sections of mouse thyroid. Our results show that in the thyroid IGF1 mRNA is predominantly present in follicular cells and C cells rather than the stroma. Follicular cell levels are higher during postnatal thyroid growth and during the growth response to goitrogen administration, but decrease in the mature animal. This decrease in production is limited to the follicular cells, as IGF1 mRNA is still easily demonstrable in C cells and in the parathyroid. Immunocytochemistry for IGF1 peptide shows a weak and variable follicular cell content in both juvenile and mature mice, but a more uniform distribution during growth in response to a goitrogenic stimulus. These studies show that the follicular cells are the main source of IGF1 in the thyroid, and suggest that the role of IGF1 in follicular cell growth is as an autocrine factor.
Brief, high‐temperature heat denaturation (pressure cooking): A simple and effective method of antigen retrieval for routinely processed tissues
Authors: Norton Andrew J. , Jordan Suzanne , Yeomans Patricia
Published: June 2005

The merits of microwave‐based antigen retrieval methods for diagnostic immunohistochemistry have recently been emphasized. As such technology has drawbacks for busy routine laboratories, we have investigated an alternative method of heat‐mediated antigen retrieval (HMAR), using a domestic pressure cooker as a source of superheating. A simple protocol, with superheating in 0.01 M citrate buffer (pH 6.0) for 2 min, gave results comparable to those obtainable in a microwave oven. The range of antigens retrieved and the antigens for which heating was ineffective matched the published microwave experience. The advantages included the speed of treatment, the reproducibility of results with large batches of slides, the ability to use metal slide racks, and economy of time and equipment costs.
Human non‐small cell lung cancer: P53 protein accumulation is an early event and persists during metastatic progression
Authors: Fontanini Gabriella , Vignati Silvana , Bigini Daniela , Basolo Fulvio , Pingitore Raffaele , Bevilacqua Generoso , Merlo Giorgio R. , Ribecchini Alessandro , Angeletti Carlo Alberto
Published: June 2005

Mutations in the p53 tumour suppressor gene, with consequent accumulation of the p53 protein, are frequently observed in non‐small cell lung cancer (NSCLC). Little is known, however, about the timing of their appearance or their maintenance through cancer progression and metastatic spread. We have examined the normal epithelium and a panel of bronchial lesions, including dysplastic, neoplastic, and metastatic leisons, for p53 immunoreactivity and for expression of proliferating cell nuclear antigen (PCNA). No p53 immunoreactivity was found in normal and hyperplastic epithelium, nor in squamous metaplastic lesions. Twenty out of 30 invasive tumours and 13 out of 17 in situ carcinomas adjacent to an invasive tumour showed p53 immunoreactivity. There was a strict correlation between the level of p53 expression in the non‐invasive and the invasive components of the tumours. Five out of eight pairs of primary tumours and matching metastases expressed p53, at identical levels in both compartments. These data indicate that p53 overexpression can occur in the earliest recognized phase of NSCLC and that the alteration is maintained during progression from in situ to invasive carcinoma and metastatic spread. PCNA expression increased from early to advanced phases of NSCLC. High PCNA immunoreactivity was observed in tumours expressing high p53 levels. A significant association was observed for PCNA expression between preinvasive and invasive lesions.
Testicular germ cell tumours of childhood in Denmark, 1943‐1989: Incidence and evaluation of histology using immunohistochemical techniques
Authors: Visfeldt Jakob , Jørgensen Niels , Müler Jørn , SkakkebæK Niels E. , Møller Henrik
Published: June 2005

In the Danish Cancer Registry, 72 testicular tumours in boys younger than 15 years of age were recorded during the period 1943‐1989 and material from 34 of these was retrieved from Danish departments of pathology. The histological types were evaluated and the role of immunohistochemical staining was analysed. The survival of the patients was correlated with the histological diagnoses, and changes in the incidence of testicular cancers in childhood were analysed. Twenty‐nine of the 34 patients had germ cell tumours, which fell into two groups (infantile and pubertal) with distinct differences. The tumours of infancy usually presented before the age of 3 years and were either pure yolk sac tumours or teratomas. The tumours of puberty showed no morphological or immunohistochemical differences from adult germ cell tumours. In the infantile group, immunohistochemical staining confirmed the morphological evaluation but was not necessary for diagnosis. Patients in the infantile group seemed to have a better prognosis than adult patients, only one patient dying from his disease, whereas the pubertal patients seemed to have a prognosis similar to that of adult patients. The incidence of infantile testicular cancer in Denmark appears to have increased at almost the same rate as that observed in adult men, but due to the small numbers in infancy, this cannot be statistically substantiated. We conclude that the testicular germ cell tumours of infancy and puberty may arise from different precursor cells, but both groups seem to arise prenatally.
Decreased expression of cellular markers in Epstein‐Barr virus‐positive Hodgkin's disease
Authors: Bai M. C. , Jiwa N. M. , Horstman A. , Vos W. , Kluin PH. H. , Valk P. Van Der , Mullink H. , Walboomers J. M. M. , Meijer C. J. L. M.
Published: June 2005

Epstein‐Barr virus (EBV) has been demonstrated in the Reed–Stenberg cells and their mononuclear variants (Hodgkin cells; H‐RS cells) in a substantial number of Hodgkin's disease (HD) cases. Moreover, EBV can modulate both in vivo and in vitro the expression of several cellular genes, including lymphoid differentiation markers. Therefore we investigated, in 64 cases of HD, the relationship between the presence of EBV and the expression of lymphoid (CD45RB), T‐ (CD3, CD45RO), B‐ (CD20, MB2 antigen, CDw75), and myeloid‐cell lineage markers (CD15), and of activation markers (CD30, EMA, and the 115D8 antigen) on the H‐RS cells. EBV‐positive cases, as demonstrated by the presence of EBER‐1 and ‐2 RNA and LMP‐1 protein expression, showed a significant reduction in the expression on H‐RS cells of T‐cell lineage (CD3, p<0·02), B‐cell lineage (CD20, P<0·005), AND ACTIVATION MARKERS (ema; P<0·002 and the 115D8 antigen; P<0·001) as compared with EBV‐negative cases. No differences were found in the expression of CD15, CD30, CD45RO, CD45RB CDw75, or the MB2 antigen on H‐RS cells in EBV‐positive and EBV‐negative HD cases. Interestingly, in 11 cases of EBV‐negative HD, B‐as well as T‐cell lineage markers could be found on some H‐RS cells. These data suggest that EBV in H‐RS cells is able to down‐regulate the expression of T‐ (CD3) and B‐ (CD20) cell lineage markers and lymphoid activation markers (EMA and the 115D8 antigen). For this reason, the origin of H‐RS cells in HD, as studies by immunohistochemistry, cannot be discussed without taking into account the presence of EBV.
Langerhans' cell histiocytosis cells are activated langerhans' cells
Authors: Emile Jean‐François , Fraitag Sylvie , Leborgne Michèle , de Prost Yves , Brousse Nicole
Published: June 2005

Langerhans' cell histiocytosis (LCH) is characterized by the presence of large mononucleated cells, associated with inflammatory cells. The Langerhans' cell (LC) lineage of the mononucleated cells is suggested by the presence of Birbeck granules and the expression of CD1a. We investigated the presence of 14 markers expressed by normal LCs in vitro. Nine skin and one lymph node frozen biopsies of LCH children were analysed by in situ immunohistochemistry. The data were compared with six skin and five lymph node frozen biopsies. LCH cells of the ten samples were positivie for all 14 LC markers. We observed three different groups of markers, according to the respective staining of normal LCs and LCH cells. Group 1 included DR, DQ, CD1a, CD1c, and ICAM‐3. Markers of group 1 were present on the majority of both normal LCs and LCH cells. Group 2 included CD1b, CD4, LFA‐1, LFA‐3, CD32, and CD68. Markers of group 2 were detected on the majority of LCH cells, but only on a fraction of normal LCs. Group 3 included CD11b, CD24, and B7/BB1. Markers of this group were detected on LCH cells, but not on normal LCs. This in situ immunohistochemical study confirms that LCH cells belong to the LC lineage. The different clinical LCH syndromes had the same immunohistochemical staining. The expression of some markers of groups 2 and 3 is known to be related to the activation of LCs in vitro. Our study suggests that LCH cells are activated LCs.
Assessment of epidermal dendritic cell markers and T‐lymphocytes in psoriasis
Authors: Jones J. Louise , Berth‐Jones John , Fletcher Alan , Hutchinson Peter E.
Published: June 2005

Epidermal dendritic and T‐cell counts have been performed in lesional and non‐lesional skin from 35 psoriatic patients. The aims were to investigate absolute changes and interrelationships between these cellular elements in psoriasis and to explain apparent discrepancies between these results and reports in the literature. In non‐lesional skin, the most frequently expressed dendritic cell marker was CD1a. HLA‐DR+ and α‐mannosidase+ dendritic cells were approximately 50 per cent and S100+ cells were 25 per cent as frequent. T‐lymphocytes were rare, CD4+ cells predominating. In lesional psoriatic epidermis, there was a definite increase (approximately two‐fold) in the absolute number of CD1a+ dendritic cells. This differs from the conclusions from the majority of previous studies. However, when cell counts were expressed per unit area of vertical section, there was a decrease in CD1a+ cells in lesional skin, which is an explanation for this discrepancy. There was a greater increase in absolute HLA‐DR+ cell counts, so that the numbers of cells expressing CD1a and HLA‐DR were similar in lesional skin. S100 expression increased proportionately with CD1a+, but there was no absolute increase in α‐mannosidase+ cells, which might represent a separate sub‐population of dendritic cells. The greatest cellular increase was in T‐lymphocytes, particularly CD8+. In lesional skin, direct correlations have been demonstrated between epidermal thickness, HLA‐DR+ dendritic cells and T‐lymphocytes, particularly CD8+ cells. We would suggest that the present method of quantification is of value for the analysis of absolute changes in epidermal infiltrates, particularly psoriasis, and could be applied to other epidermal pathologies.
Expression of integrins and CD44 isoforms in non‐Hodgkin's lymphomas: CD44 variant isoforms are preferentially expressed in high‐grade malignant lymphomas
Authors: Terpe Hans‐Joachim , Koopmann Rita , Imhof Beat A. , Günthert Ursula
Published: June 2005

Low‐ and high‐grade malignant non‐Hodgkin's lymphomas have been investigated by immunohistochemistry for their expression of various integrins and CD44 isoforms. Comparison with the expression patterns obtained in non‐malignant adult lymph nodes revealed the following differences: α6 and β4 integrins were expressed in several high‐grade malignant lymphomas to a lower degree than in both the low‐grade malignant lymphomas and the normal lymph nodes; all other integrins (α2, α4, α5, αv, β1, β2, β3, and β7) did not exhibit significant differences in the expression levels between malignant and non‐malignant tissues. The standard isoform of CD44 (CD44s) was highly expressed in all lymphoid tissues. Using CD44 exon‐specific monoclonal antibodies, CD44 variant isoforms were not detected in non‐malignant lymph nodes and were detected only rarely in low‐grade malignant lymphomas. In contrast, high‐grade malignant lymphomas expressed several CD44 variant isoforms, which included the products from the variant exons 3v, 6v, and 9v, but not 4v. Specifically, detection of exon 3v and 6v products indicates a more aggressive phenotype.
Expression of E‐cadherin (E‐CD) as related to other prognostic factors and survival in breast cancer
Authors: Lipponen P. , Saarelainen E. , Ji H. , Aaltomaa S. , Syrjänen K.
Published: June 2005

A series of 208 breast cancer biopsies were analysed immunohistochemically for expression of E‐cadherin (E‐CD). Altogether, 72 per cent of the tumours showed E‐CD positivity in over 50 per cent of cells, the staining being heterogeneous in nearly all tumours. In only 16 per cent of ductal carcinomas was positive staining seen in less than 1 per cent of cells. Expression of E‐CD was not related to tumour diameter, nodal status, metastasis at diagnosis, histological grade, DNA ploidy, S‐phase fraction, nuclear area, mitotic frequency, or PR content. There was a significant relationship between expression of E‐CD, histological type (P=0.01), the proportion of intraductal growth (P=0.008), the density of tumour‐infiltrating lymphocytes (P=0.0007), ER content (P=0.012), and morphometric nuclear factors (P=0.02). Expression of E‐CD showed a weak association with a high survival probability (P=0.02), while the relation to recurrence‐free survival was not significant (P=0.11). In axillary lymph node‐negative tumours, E‐CD expression was not related significantly to survival (P=0.11) or to recurrence‐free survival (P=0.06). In multivariate analysis, E‐CD expression had no independent prognostic value, while the axillary lymph node status, tumour diameter, patient age, and mitotic frequency were independent prognostic factors. The results indicate that E‐CD expression is related to several histological features in breast cancer, but has no independent prognostic value over standard prognostic factors.
Demonstration of skin‐derived antileukoproteinase (skalp) and its target enzyme human leukocyte elastase in squamous cell carcinoma
Authors: Alkemade Hans A. C. , van Vlijmen‐Willems Ivonne M. J. J. , van Haelst Urbaan J. G. M. , van de Kerkhof Peter C. M. , Schalkwijk Joost
Published: June 2005

Skin‐derived antileukoproteinase (SKALP), also known as elafin, is a strong and specific inhibitor of elastase and proteinase 3. SKALP is not present in normal epidermis, but is expressed by epidermal keratinocytes under hyperproliferative conditions such as psoriasis, wound healing, and in cell culture. In human epidermal tumours, SKALP is differentially expressed and restricted to tumours with distinct squamous differentiation. We have studied the presence of both SKALP and one of its known target enzymes, leukocyte elastase, in 41 squamous cell carcinomas of the skin. SKALP expression correlated with the degree of differentiation: strong expression was seen in well‐differentiated cells and expression was absent in poorly differentiated tumour cells. Most of the squamous cell carcinomas showed elastase‐positive cells in the tumour stroma and also within the tumour cell nests. SKALP may interfere with the proteoloytic activity of infiltrating inflammatory cells or with hitherto unknown proteinases from the tumour cells. We hypothesize that in squamous cell carcinoma progressive loss of SKALP expression could facilitate tumour spread.
In situ localization of type III and type IV collagen‐expressing cells in human diabetic nephropathy
Authors: Razzaque Mohammed S. , Koji Takehiko , Taguchi Takashi , Harada Takashi , Nakane Paul K.
Published: June 2005

Nodular intercapillary glomerulosclerosis is the most typical lesion of diabetic nephropathy (DN) and is characterized by increased extracellular matrix (ECM) and amorphous masses of mesangial matrix. The local exaggeration of these deposits results in the formation in the typical diabetic nodule. To clarify the composition of the ECM of sclerotic lesions in DN, we investigated the distribution of type III and type IV collagens and their mRNAs by immunohistochemistry and in situ hybridization, respectively. In normal renal tissues, there was no intraglomerular immunostaining for type III collagen, while strongly positive staining was found in the extraglomerular interstitum. Positive immunostaining for type IV collagen was also present in the mesangium, glomerular basement membrane (GBM), Bowman's capsule, and the vascular pole of the normal glomerulus. In DN, the nodular lesions were negative for type III collagen and strongly positive for type IV collagen. On the other hand, in the late stage of global sclerosis, both type III and type IV collagens were diffusely present in the sclerotic matrix. To determine the origins of these type III and type IV collagens in the sclerotic matrix, in situ hybridization was performed, utilizing thymine‐thymine (T‐T) dimerized synthetic oligonucleotides complementary to either proα(III) chain or pro α1 (IV) chain mRNAs as probes. The signals were detected by enzyme immunohistochemistry using an anti‐T‐T antibody. Intraglomerular cells (glomerular epithelial and mesangil cells) containing type III collagen mRNA were found in DN with sclerotic lesions, but not in normal glomeruli. At this stage of sclerosis, intraglomerular cells (mainly glomerular epithelial cells and infrequently mesangial cells) were positive for type IV collagen mRNA, but there were few positive cells in globally sclerotic glomeruli. This study provides evidence that both type III and type IV collagens are synthesized by intraglomerular cells during sclerosis and become significant constituents of the sclerotic matrix in DN.
p53 Expression, mutation, and allelic deletion in ovarian cancer
Authors: McManus D. T. , Yap E. P. H. , Maxwell P. , Russell S. E. H. , Toner P. G. , McGee J. O'D.
Published: June 2005

The aim of this investigation was to study the prevalence of p53 gene mutations and allelic deletions in ovarian cancer and the relationship between these events and p53 expression. Genomic p53 was amplified by the polymerase chain reaction (PCR) from paraffin‐embedded sections of tumour and non‐tumour tissue. Exons 5–9 were screened for mutations using non‐isotopic single‐strand conformation polymorphism (SSCP) analysis. Allelic deletions at a sequence polymorphism in exon 4 were studied for loss of heterozygosity (LOH) by restriction analysis and by SSCP. p53 expression was detected by immunohistochemistry with the p53 antibodies CM1 and Do7. Of 44 cases, ten (23 per cent), including a mucinous tumour of low malignant potential, showed mutations. There was a statistically significant correlation between p53 mutation and expression (P<0·01) but seven cases showed discordance. Of 18 informative cases, eight (44 per cent) demonstrated LOH. Mutations were identified in three of the informative cases, two of which also had LOH. The remaining case showed mutations in two amplicons. p53 dysfunction, as indicated by mutation, deletion, or increased expression, is common in ovarian cancer. There is an association between these molecular events but the exact mechanisms of p53 inactivation remain to be elucidated.
HLA‐DR antigen expression and lymphocyte subsets in transitional cell carcinoma of the urinary bladder. An immunohistological study on frozen sections
Authors: Ioachim‐Velogianni Elli , Stavropoulos Nickolaos E. , Kitsiou Evangelia , Stefanaki Stella , Agnantis Niki J.
Published: June 2005

Lymphocyte subpopulations (B cells, CD4, CD8), interleukin‐20 receptors (IL‐2), monocytes/macrophages (Leu M5), and HLA‐DR antigen expression were studied immunohistochemically on frozen sections from 38 bladder cancer specimens. T cells predominated over B cells in all tumours. CD4‐positive lymphocytes predominated over CD8 in the stroma (CD4/CD8: 1·35/1), while in epithelial tumour cells CD8 was the prominent subpopulation (CD8/CD4: 1·75/1). Aberrant HLA‐DR expression was found in 21·05 per cent of bladder tumours. A strong correlation between CD4 and CD8 population densities and macrophages with the other subpopulations was noticed. In HLA‐DR‐positive tumours, there was no correlation of the percentage of positive cells with CD4‐ and CD8‐positive lymphocyte populations. Various parameters including IL‐2 receptors, B cells, CD8‐ and CD4‐positive cells, and macrophages did not differ significantly between the groups of tumours expressing and not expressing HLA‐DR antigen. There were no statistically significant differences in the population densities of B cells, CD8‐ or CD4‐positive cells, IL‐2 receptor, monocytes/macrophages, and HLA‐DR antigen expression among various clinicopathological parameters, including growth pattern, histological grade and clinical stage or patient's age and sex. These findings suggest that in transitional cell carcinoma of the urinary bladder, HLA‐DR antigen expression is independent of lymphocyte subpopulations. It is therefore possible that HLA‐DR expression by tumour cells reflect the existence of separate HLA‐DR‐positive or HLA‐DR‐negative tumour clones.
Proliferative activity in benign and neoplastic prostatic epithelium
Authors: Limas Catherine , Frizelle Sandra P.
Published: June 2005

The increasing availability of means for the early detection of prostatic cancer has brought under scrutiny the criteria used for prognosis and emphasized our limitations in understanding what determines the rate of progression in these cancers. The rate of cancer cell proliferation has been under intense investigation, which, however, has yielded conflicting results. In this study we evaluated the proliferative activity of benign and neoplastic prostatic epithelium, using various existing methodologies. We first analysed the variability introduced by the methodological approach and then attempted to demonstrate whether determination of the proliferative capacity had any clinical consequence that complemented the histological grading. Tissue samples from patients, 88 with cancer and 46 with benign prostatic pathology, were studied using in vitro bromodeoxyuridine (BrdU) incorporation as well as Ki67 and the proliferating cell nuclear antigen (PCNA) to estimate the proliferative activity. Increased proliferation was found consistently in inflammation and metaplasia, but not in hyperplasia. In contrast, cancers showed marked variability. Although average proliferation indices increased with grade, there was a wide scatter of values. Correlation was stronger with stage, but also depended on the methodology. Bromodeoxyuridine indices over 10 per mille had a positive predictive value of 79 per cent for cancers extending beyond the prostatic capsule and may prove particularly helpful for evaluating patients with grade 7 cancer. This observation is significant, since grade 7 cancers are the most frequent and the least predictable.
Possible role of tissue‐bound calcium ions in citrate‐mediated high‐temperature antigen retrieval
Authors: Morgan J. M. , Navabi H. , Schmid K. W. , Jasani B.
Published: June 2005

High‐temperature preheating of sections in the presence of a salt (e.g., citrate) or a protein denaturant (e.g., urea) solution has been shown recently to provide a reliable alternative to tissue proteolysis for antigen retrieval from formaldehyde‐fixed, paraffin‐embedded tissues. However, the underlying mechanism of action of this form of pretreatment remains highly speculative. In this study, we show that calcium chelating agents such EDTA and EGTA are more effective than citrate in the retrieval of a citrate‐sensitive nuclear antigen, Ki‐67. Also, sodium carbonate and another calcium precipitating agent are both able to effect antigen retrieval at high temperatures. The overall data therefore suggest that either the chelation or the precipitation of tissue‐bound calcium ions, and perhaps also other divalent metal cations, is a critical step in salt‐mediated antigen retrieval. As a corollary, it is suggested that tight complexing of calcium ions or other divalent metal cations with proteins during formaldehyde tissue fixation is responsible for the masking of certain antigens.
Molecular mechanisms of drug resistance in tumours
Authors: Harrison David J.
Published: June 2005

Despite advances in the design and use of chemotherapeutic drugs, the majority of human cancers are resistant to therapy at presentation or become resistant after an initial partial response. This suggests that resistance may be inherent in a tumour cell or may evolve under the selection pressure of drug administration. A number of possible molecular explanations for drug resistance exist. There may be exclusion of drug from the cell, failure to activate the prodrug to its active form, increased detoxification, alteration in the drug target, enahanced repair capability of the cell after injury, or failure to engage an appropriate response, leading to apoptosis in the damaged cell. Many of these factors may co‐exist in vivo in human tumours; some are a feature of cell lineage whilst others appear de novo during disease progression. Modulation of these mechanisms has been of some value in laboratory studies but widespread clinical application and benefit remain some way off.
Differentiation of a human rhabdomyosarcoma cell line after antineoplastic drug treatment
Authors: Melguizo C. , Prados J. , Aneiros J. , Fernandez J. E. , Velez C. , Aranega A.
Published: June 2005

The feasibility of treating solid tumours with differentiation therapy using antineoplastic drugs is currently being investigated, but the emergence of multidrug resistance remains the major limitation to this therapeutic approach. A rhabdomyosarcoma cell line resistant to actinomycin D (RD‐DAC) has been used as an in vitro model to investigate, with light and electron microscopy, the degree of differentiation in multidrug‐resistant cells. The parental cell line (RD), derived from a human embryonic‐type rhabdomyosarcoma, is undifferentiated, with no evidence of specific ultrastructural markers. Examination of resistant cells by transmission electron microscopy revealed myofilaments arranged parallel to the long axis of the cell, which was considered clear evidence of myogenic differentiation. These observations suggest that actinomycin D, the drug of choice in the treatment of rhabdomyosarcoma, induces differentiation in the cell line RD. It is postulated that multidrug resistance can interfere with cellular differentiation.
mdm2 gene alterations and mdm2 protein expression in breast carcinomas
Authors: Marchetti Antonio , Buttitta Fiamma , Girlando Salvatore , Palma Paolo Dalla , Pellegrini Silvia , Fina Paolo , Doglioni Claudio , Bevilacqua Generoso , Barbareschi Mattia
Published: June 2005

This study investigates the mdm2 gene status and expression in 66 surgically resected human breast carcinomas, with correlations with clinico‐pathological and biological data (histological type, grading, steroid receptor status, p53 expression, proliferative activity). Four (7.7 per cent) out of 52 informative cases bear mdm2 gene amplification (four‐ to ten‐fold) and 8 (15.4 per cent) of 52 cases showed borderline amplification (three‐fold). Nine (13.6 per cent) out of 66 cases showed strong mdm2 nuclear immunoreactivity. Twenty‐seven (40.9 per cent) cases showed isolated mdm2 reactive nuclei. All cases with clear amplification showed a high percentage of mdm2 immunoreactive nuclei. The relationship between gene amplification and mdm2 protein expression is highly significant (P<0.0001). No association was observed between mdm2 gene amplification and any of the considered clinico‐pathological and biological parameters, while mdm2 immunoreactivity showed a significant association only with oestrogen receptor immunoreactivity (P=0.009). p53 expression was associated neither with mdm2 gene amplification nor with mdm2 immunoreactivity. It could be tempting to hypothesize that the evaluation of the combined mdm2/p53 immunohistochemical phenotype in human breast carcinoma could give us better prognostic information than the evaluation of the expression of the p53 protein alone.
Epstein‐Barr virus association with early cancers found together with gastric medullary carcinomas demonstrating lymphoid infiltration
Authors: Takano Yasuo , Kato Yo
Published: June 2005

Seven early gastric cancers obtained from patients also demonstrating Epstein‐Barr virus (EBV)‐positive gastric medullary carcinoma with lymphoid infiltration were investigated using a combined polymerase chain reaction (PCR) and in situ hybridization (ISH) approach. Sharing the same background mucosa as gastric medullary cancers, they comprised four intramucosal carcinomas, predominantly well‐differentiated adenocarcinomas, and three submucosal carcinomas, histologically showing mixtures of well and poorly differentiated adenocarcinoma. In the three cases of submucosal carcinoma, the presence of EBV was proven by means of both PCR and ISH. However, not all cancer cells were positive for EBV on the basis of ISH examination, in contrast to the large series of gastric carcinoma with lymphoid infiltration previously investigated. All four mucosal carcinomas were EBV‐negative. Lymphocyte‐determined membrane antigen (LYDMA) monoclonality, performed by PCR, and latent membrane protein‐1 (LMP‐1) and Epstein‐Barr virus (EBNA2) expression, assessed immunohistochemically, were negative in all seven cases. The results suggest that EBV becomes associated with gastric medullary carcinoma with lymphoid infiltration (GMCL) at a relatively early stage of the disease, shortly after the tumour has initially progressed to an invasive form, and plays some role in the manifestation as GMCL.
Expression of VE (vascular endothelial)‐cadherin and other endothelial‐specific markers in haemangiomas
Authors: Martìn‐padura Inés , de Castellarnau Conxita , Uccini Stefania , Pilozzi Emanuela , Natali Pier Giorgio , Nicotra Maria Rita , Ughi Francesca , Azzolini Cesare , Dejana Elisabetta , Ruco Luigi
Published: June 2005

Haemangiomas are vascular tumours characterized by rapid growth and increased endothelial turnover. VE‐cadherin is a recently discovered endothelial cell‐specific cadherin located at intercellular junctions. In different types of epithelial tumours, cadherin expression is inversely correlated with invasiveness and metastatic dissemination. In this immunohistochemical study, VE‐cadherin expression has been analysed in different types of haemangioma. VE‐cadherin is highly expressed in endothelial cells of haemangiomas and is decreased, but still detectable, in some cases of haemangionendothelioma and angionsarcoma. The antigenic profile of most haemangioma cells was similar to that of normal endothelium. CD31, CD34, ICAM‐1, von Willebrand factor, and VLA integrins were expressed in haemangioma endothelium; in addition, the major components of vascular basement membrane, namely fibronectin, collagen type IV, and laminin, were correctly expressed and organized. Surprisingly, a marked reactivity for the M form of laminin (merosin) was detected in the basement membranes of two juvenile capillary haemangiomas. Overall, this study shows that, with the exception of angiosarcoma and haemangionendothelioma, vascular tumours maintain most of the differentiation characteristics of normal endothelium. This encourages speculation that in these pathologies, abnormal endothelial proliferation is more related to the release of local factors than to an altered endothelial phenotype.