bluuebunny/crossref_metadata_2025 huggingface dataset:
Paraffin wax‐embedded material as a source of DNA for the detection of somatic genetic changes
Authors: Sundaresan V. , Ganly P. , Hasleton P. , Bleehen N. M. , Rabbitts P.
Published: June 2005

Loss of genetic material, corresponding to chromosomal deletions, has been detected in a wide range of tumours and may indicate the position of a tumour suppressor gene. In order to identify the position of such a gene more precisely, many tumour samples must be studied until a minimum consensus deletion is characterized. This process is particularly necessary for lung tumours in which the deletion in chromosome 3, seen with such high frequencies in all histological subtypes, is almost always large. We have recently described the use of the polymerase chain reaction (PCR) for restriction fragment length polymorphism (RFLP) analysis of DNA isolated from small bronchial biopsies of lung tumours. In this study we adapted this technique to allow genotyping of DNA isolated from paraffin wax‐embedded material (PWEM) microdissected from glass slides. We have investigated 12 lung tumours at polymorphic loci on chromosome 3 and showed allelic loss in all samples. In adapting PCR–RFLP analysis for DNA isolated from PWEM, we have concentrated on those approaches which might be adaptable to routine clinical practice. Somatic genetic changes are now being identified in many tumour types, and this information is expected to be of diagnostic and prognostic significance.
Detection of calcitonin and calcitonin gene‐related peptide mRNA in human medullary thyroid carcinoma. A retrospective study
Authors: Denijn Maryléne , De Weger Roel A. , Van Unnik Jan A. M. , Otter Wim Den , Lips Cees J. M.
Published: June 2005

In Situ hybridization finds many applications in modern pathology. In many cases, special attention is paid to the processing of the tissues prior to in situ hybridization. In order to investigate the value of RNA in situ hybridization (RISH) in retrospective studies, we performed RISH for calcitonin and calcitonin gene‐related peptide (CGRP)‐I and HomRNA in eight medullary thyroid carcinomas processed in 1981–1983. RISH was successful with radioactive calcitonin and CGRP‐1 probes. With biotinylated probes, only calcitcnin‐specific probes gave adequate results. The concentrations of CGRP mRNA were probably too low to be detected by non‐radioactive RISH. The results of RISH were correlated with the immunohistochemical localization of the polypeptides. The results matched in all cases except one, where hybridization for calcitonin mRNA was found, but no immunoreactive calcitonin polypeptide. We con‐clude that RISH can be successfully used for retrospective analysis, even after long storage of tissue embedded in poraffin.
In situ correlation of synthesis and storage of parathormone in parathyroid gland disease
Authors: Kendall C. H. , Potter Linda , Brown R. , Jasani B. , Pringle J. H. , Lauder I.
Published: June 2005

The distribution and expression of parathyroid hormone (PTH) were investigated in normal and abnormal parathyroid tissue. PTH was detected using a monoclonal antibody with specificity for the 44–68 region of the PTH molecule. Prominent reactivity for PTH was seen in normal parathyroid with a granular pattern of staining. Active parathyroid tissue (adenoma and hyperplasia) showed much less reactivity for PTH, although there was prominent reactivity in the normal tissue around adenomas. Comparison of expression of PTH with that of parathormone mRNA showed a reciprocal pauern in normal tissue and, to a less marked extent, in abnormal tissue. Parathyroid carcinoma in particular had coinciding areas of PTH and PTH mRNA expression. Oxyphil cells had little or no PTH expression, except in the associated ‘colloid’ in some cases. The findings indicate an inverse relationship between storage and cellular synthesis of PTH, this being more marked in physiological than in pathological conditions of the parathyroid.
Localization of plasminogen activator inhibitor‐1 production in inflamed appendix by in situ mRNA hybridization
Authors: Whawell Simon A. , Wang Yiming , Fleming Kenneth A. , Thompson Elizabeth M. , Thompson Jeremy N
Published: June 2005

The peritoneum has been shown to possess fibrinolytic activity which is thought to play a role in the prevention of intra‐abdominal adhesion formation. Recently inflamed peritoneal tissue has been shown to have reduced fibrinolytic activity secondary to increased levels of plasminogen activator inhibitor‐1 (PAI‐1). The aim of this study was to localize the production of PAI‐1 in appendix tissue using in situ mRNA hybridization. Sections of normal and inflamed appendix were hybridized with a digoxigenin‐labelled cDNA probe. PAI‐1 production was localized to both mcsothelium and serosal blood vessel endothelium in all inflamed appendix samples. Cell identities were confirmed using immunohistochemistry directed against mesothelial and endothlial cell markers. Staining was not seen on sections of normal appendix or on negative control slides of inflamed appendix (hybridization with plasmid DNA, PAI‐1 probe following ribonuclease digestion). The identification of the cells expressing the PAI‐1 gene in peritoneum increases our understanding of the pathophysiological changes in fibrinolytic activity which occur in inflammation and may lead to adhesion formation.
Gastric intestinal metaplasia type III cases are classified as low‐grade dysplasia on the basis of morphometry
Authors: Tosi Piero , Filipe M. Isabel , Luzi Pietro , Miracco Clelia , Santopietro Rosa , Lio Roberto , Sporza Vincenzo , Barbini Paolo
Published: June 2005

The aim of this study was to try to place gastric intestinal metaplasia, type III (type III IM) in the stepwise chain of events from atrophic gastritis to cancer. A number of dysplastic, periulcer regenerative, and type III IM lesions were qualitatively diagnosed (and graded) blindly and independently by several pathologists. These lesions were further analysed by means of quantitative parameters, with the aim of differentiating dysplastic from regenerative changes. Inconsistencies between the qualitative and quantitative classification (about 7 per cent of cases) were eliminated and homogeneous groups (low‐grade dysplasia, high‐grade dysplasia, regenerative changes) were obtained. These cases were taken as the gold standard against which type III IM was compared. The results indicate that the great majority (91·4 per cent) of cases of type III IM fulfil the nuclear and architectural criteria for low‐grade dysplasia.
Growth factors: Regulation of normal and neoplastic growth
Authors: Pusztai L. , Lewis C. E. , Lorenzen J. , McGee J. O'D
Published: June 2005

This paper presents an overview of current trends in growth factor research. The first part of the review considers the current classification of growth factors and their receptors. A model of cell proliferation regulation by growth factors is then presented. The final section reviews the latest concepts of the involvement of growth factors in the development of neoplasia.
Proliferation and differentiation of tumour cells from B‐cell lymphoma of mucosa‐associated lymphoid tissue in vitro
Authors: Hussell Tracy , Isaacson Peter G. , Spencer Jo
Published: June 2005

Several characteristics of lymphomas of mucosa‐associated lymphoid tissue (MALT type) suggest that they are antigen‐dependent and that their growth parodies a normal immune response. We have previously shown that three cases of low‐grade B‐cell MALT‐type lymphoma recognize autoantigens. In this study, we investigated the response of three low‐grade and one high‐grade case of MALT‐type lymphoma to anti‐idiotypic antibody as a model of antigen binding either alone or as a co‐stimulus with B‐cell mitogens. We also studied the response of tumour cells to interleukin‐6 (IL‐6), which induces differentiation to immunoglobulin‐producing cells in many systems. Of the four cases studied, one low‐grade case showed markedly enhanced proliferation in response to anti‐idiotype alone. This could not be increased by the addition of mitogens. In the remaining two low‐grade cases, mitogen responsiveness was observed which was affected by anti‐idiotype either by an enhancement or by a reduction in the proliferative response. The high‐grade case failed to respond to the stimuli studied. No response to IL‐6 was observed. This study supports the suggestion that antigen may affect the pathogenesis of low‐grade tumours of MALT type.
Bronchus‐associated lymphoid tissue (BALT) in human fetal and infant lung
Authors: Gould Stephen J. , Isaacson Peter G.
Published: June 2005

Bronchus‐associated lymphoid tissue (BALT) has been defined as the organized lymphoid tissue of the lung. Although well described in a variety of animal species, documentation of its presence and development in human lung is limited. Because the tissue to volume ratio in adult lungs is so low, a systematic search for BALT would involve so many sections as to be impractical. In this study, therefore, we have studied post‐mortem specimens of fetal (n=102) and infant (n=17) lungs, which have a much higher tissue to volume ratio. Fetal death was due to various causes but all but two infants died from sudden infant death syndrome. In the fetal lungs, the presence of BALT was almost invariably associated with chorioamnionitis or intrauterine pneumonia, being present in 24 of 51 of these cases (47 per cent). The earliest ill‐defined lymphoid aggregate was seen at 16 weeks' gestation, while lymphoepithelium, a hallmark of mucosaassociated lymphoid tissue, could be identified at 20 weeks. In 51 fetuses without infection, BALT was found in only five cases (10 per cent). BALT was identified in 13/17 (77 per cent) of infant lungs and well‐developed lymphoepithelium was evident in four cases. This study shows that BALT may be present in the human fetal and infant lung, but that its appearance is probably dependent on antigenic stimulation.
Integrin expression in normal, hyperplastic, dysplastic, and malignant oral epithelium
Authors: Jones Judith , Sugiyama Masaru , Watt Fiona M. , Speight Paul M.
Published: June 2005

We have examined the distribution of a range of integrin subunits in normal and lesional oral mucosa. The α2, α3, α6, β1β, and β4 subunits were highly expressed in normal epithelium, and there was weaker, more variable expression of α5 and αv. Expression of all subunits was highest in the basal layer of normal epithelium, but extensive staining above the basal layer was also observed, particularly in the floor of the mouth and the lateral margin of the tongue. In dysplastic lesions and hyperplastic epithelium adjacent to ulcers, suprabasal staining was even more pronounced. Staining patterns in squamous cell carcinomas showed considerable variation, both within and between individual tumours: in some areas there was staining reminiscent of normal epithelium, but uniform staining throughout tumour islands, and patchy and variable cytoplasmic and pericellular staining were also seen. Thirteen out of 17 carcinomas showed some loss of integrin expression: six out of ten moderately well differentiated tumours and all the poorly differentiated tumours. Focal loss of a6 and β4 was most commonly observed, but loss of α2 and β3 also occurred. Since integrins regulate not only keratinocyte adhesion, but also the initiation of terminal differentiation, the changes in integrin expression that we have observed may have significance for the behaviour of individual tumours.
Adenoid cystic carcinoma: A study of nucleolar organizer regions (AgNOR) counts and their relation to prognosis
Authors: Fonseca Isabel , Soares Jorge
Published: June 2005

We studied the AgNOR counts in 30 cases of adenoid cystic carcinoma from salivary glands (n = 18) and non‐salivary sites (n=12) in an attempt to correlate them with the evolution of the disease. AgNOR counts per nucleus varied between 1·96 and 6·12 (mean value 4·2 ± 0·99) as compared with 1·21 ± 1·4 in normal salivary tissue. There was no significant difference between cases that had an unfavourable clinical outcome (recurrence, metastases, and/or died of the disease) and cases without disease complications (4·31 vs. 4·03 AgNORs per nucleus). No difference was found between tumours located at salivary and non‐salivary sites or between major and minor salivary glands. AgNOR counts also did not correlate with the grade of differentiation of the neoplasms. In adenoid cystic carcinoma, AgNOR counts do not seem to be a prognostic indicator, in contrast to the usefulness of this method in other tumour types.
Differentiation‐associated changes in mucin glycoprotein antigenicity in mucosa adjacent to rare gastrointestinal tract tumours of non‐mucosal origin
Authors: Pilbrow Simon J. , Hertzog Paul J. , Linnane Anthony W.
Published: June 2005

We have investigated changes in mucin antigenicity and morphology in the perineoplastic mucosa adjacent to rare, predominantly non‐mucosal gastrointestinal (GI) tumours. Twenty‐nine tumours of small and large intestine, including primary mesenchymal and ectodermal tumours, were examined immunohistochemically using 11 monoclonal antibodies (MAbs) raised against SIMA and LIMA (small and large intestinal mucin antigens). Non‐epithelial GI tumours were essentially non‐reactive, while adjacent mucosa showed altered mucin expression and morphology, in particular, features of transitional mucosa (TM). Combinations of different SIMA epitopes were detected adjacent to all colorectal tumours, and, similarly, LIMA epitopes adjacent to small intestinal tumours. Specific patterns adjacent to certain tumours may reflect influences of factors produced by individual tumours on mucin composition. Altered antigenicity and morphology in TM thus appear to be reactive changes in response to a wide range of GI tumours, presumably as a consequence of factors secreted by the tumour and/or a host response to the tumour.
Early increase precedes a depletion of VIP and PGP‐9.5 in the skin of insulin‐dependent diabetics—correlation between quantitative immunohistochemistry and clinical assessment of peripheral neuropathy
Authors: Properzi Giuliana , Villa Sandro Franca , Poccia Gianfranco , Aloisi Paolo , Gu Xu‐Hong , Terenghi Giorgio , Polak Julia M.
Published: June 2005

Diabetic neuropathy affects both sensory and autonomic peripheral nerve fibres. Vasoactive intestinal polypeptide (VIP) is present in autonomic fibres which modulate sweat secretion, while calcitonin gene‐related peptide (CGRP) is localized to cutaneous sensory fibres. In this study, immunohistochemistry and image analysis were used to assess changes of VIP and CGRP, and of the pan‐neuronal marker protein gene‐product (PGP)‐9.5, in skin biopsies of 18 patients affected by type 1 diabetes (age range 18–46 years) and from seven aged‐matched controls. Patients were divided into three groups: group 1 (n=6), with diabetes for 6 months to 3 years; group 2 (n=5), with the disease for 5–10 years; and group 3 (n=7), with diabetes for more than 10 years. VIP immunoreactivity (IR) and PGP‐9.5‐IR were significantly reduced around sweat glands (P <0.005) in groups 2 and 3. Epidermal CGRP‐IR and PGP‐9.5‐IR were significantly reduced in group 3 (P <0.05). Twenty‐eight per cent (5/18) of all patients showed high VIP‐IR around sweat glands (>95 per cent confidence limits of controls) and all of these patients had diabetes for less than 3 years. Conversely, 55 per cent (10/18) of patients had low VIP‐IR (<5 per cent confidence limit of controls). The latter, compared with the former, showed a significantly longer duration of diabetes (Fisher exact test P=0·002), presence of clinical autonomic neuropathy (Fisher exact test P=0.04), and a reduced sural nerve conduction velocity (Fisher exact test P=0.04). These results suggest that quantitative immunohistochemical analysis of peptide‐containing cutaneous nerves allows an objective evaluation of nerve fibre alterations at early stages of diabetes than is currently possible with neurophysiological functional tests.
Detection of monoclonality in low‐grade B‐cell lymphomas using the polymerase chain reaction is dependent on primer selection and lymphoma type
Authors: Diss Tim C. , Peng Huaizheng , Wotherspoon Andrew C. , Isaacson Peter G. , Pan Langxing
Published: June 2005

Detection of B‐cell monoclonality using the polymerase chain reaction (PCR) promises the quick and cost‐effective separation of monoclonal from polyclonal B‐cell disease. However, the efficiency of the method has yet to be fully assessed, particularly with regard to disease type and selection of PCR primers.

We have evaluated two approaches based on amplification of the immunoglobulin heavy chain gene using frame work 2 (Fr2) and framework 3 (Fr3) region primers. Frozen tissue samples from 94 cases of low‐grade B‐cell lymphoma were investigated, all of which had previously been shown to be monoclonal by Southern blot analysis. Using a Fr2 primer, we were able to show monoclonality in 85 per cent of cases; with Fr3, 80 per cent of cases; and using both techniques in separate reactions, 90 per cent of cases.

Thus, a significant false‐negative rate exists with either primer which can be reduced by using both. We also found a difference in the efficiency of detection in different types of lymphoma; only 87 percent of mucosa‐associated lymphomas and centroblastic/centrocytic lymphomas were shown to be monoclonal, whereas all of the other lymphoma types tested were positive using one or both methods.

We conclude that PCR detection of B‐cell monoclonality allows rapid analysis of tissue samples, including paraffin‐processed material. False‐negative results which occur in some types of lymphoma can be reduced by the use of two or more primer combinations.
Gene rearrangement of bcl‐1 and bcl‐2 is confined to distinct subgroups of high‐grade malignant B‐cell lymphomas
Authors: Ott M. Michaela , Dahlheimer Kathrin , Müller‐Hermelink Hans K. , Feller Alfred C.
Published: June 2005

The occurrence of bcl‐1 and bcl‐2 gene rearrangements was investigated in 37 cases of high‐grade B‐cell lymphomas. Bcl‐2 rearrangement was detectable only in single cases of primary centroblastic lymphoma with a follicular growth pattern, whereas secondary centroblastic lymphomas evolving from a centroblastic–centrocytic lymphoma were positive in up to 60 per cent of the cases analysed. Bcl‐1 rearrangement was found only in one case of immunoblastic B‐cell lymphoma with a history of a pre‐existing lymphoplasmacytoid immunocytoma. It is concluded that there may be a subgroup of centroblastic lymphomas with a biology similar to that of centroblastic–centrocytic lymphomas. The detection of bcl‐1 rearrangement in high‐grade lymphomas may indicate a secondary high‐grade lymphoma.
Establishment and characterization of a new human clear‐cell sarcoma cell‐line, HS‐MM
Authors: Sonobe Hiroshi , Furihata Mutsuo , Iwata Jun , Ohtsuki Yuji , Mizobuchi Hiroo , Yamamoto Hiroshi , Kumano Osamu
Published: June 2005

We have established and morphologically characterized a new human clear‐cell sarcoma cell‐line, HS‐MM, from the pleural effusion of a 39‐year‐old man with pulmonary metastasis derived from the primary popliteal tumour. The HS‐MM cells were round or spindle‐shaped, with round nuclei containing extremely prominent nucleoli. Light microscopically, the heterotransplanted nude mouse tumours showed essentially the same features as those of the original sarcoma, revealing an alveolar pattern of tumour cells with abundant clear cytoplasm. Both in vitro and in vivo, the cells reacted with anti‐S‐100 protein and melanoma‐specific HMB 45 antibodies by immunohistochemistry. Ultrastructurally, the cells contained round euchromatin‐rich nuclei with large nuclcoli revealing conspicuous nucleolonema, and a large amount of glycogen and a few lysosomal dense bodies, but no premelanosomes in their cytoplasm. The HS‐MM cell line was thus fully proven to exhibit the unique characteristics of a clear cell sarcoma both in vitro and in vivo, being also compatible with an amelanotic melanoma. This cell line will therefore be extremely useful for clinicopathological and histogenetic studies on clear cell sarcomas.
Expression of the pro‐opiomelanocortin gene in lung neuroendocrine tumours: In situ hybridization and immunohistochemical studies
Authors: Black M. , Carey F. A. , Farquharson M. A. , Murray G. D. , McNicol A. M.
Published: June 2005

Neuroendocrine tumours of the lung may be associated with the ectopic adrenocorticotrophin (ACTH) syndrome and may synthesize and secrete ACTH‐related peptides in the absence of the syndrome. However, immunocytochemical analysis may not confirm these biochemical findings, particularly in small cell carcinoma, which is poorly granulated. To investigate further the morphological evidence for expression of the pro‐opiomelanocortin (POMC) gene in neuroendocrine lung tumours, we have examined a series of 46 small cell carcinomas and 13 carcinoid tumours of the lung by in situ hybridization for POMC mRNA using a digoxigenin‐labelled oligoprobe. We have compared the findings with the immunocytochemical detection of ACTH and β‐endorphin. In situ hybridization was positive in 15 of 46 small cell carcinomas (33 per cent) and in 8 of 13 carcinoid tumours (62 per cent). Immunocytochemical staining was positive in only one carcinoid tumour. These in situ hybridization studies have corroborated biochemical data indicating POMC gene expression in a high proportion of lung neuroendocrine tumours. This suggests that the low levels of expression detected by immunocytochemistry may be due to low levels of hormone storage. Multivariate analysis showed a weak negative association between POMC expression and survival in small cell carcinomas, although this did not reach statistical significance.
Demonstration of pro‐opiomelanocortin mRNA in pituitary adenomas and para‐adenomatous gland in cushing's disease and Nelson's syndrome
Authors: Fehn Marita , Farquharson Maura A. , Sautner Doris , Saeger Wolfgang , Lüdecke Dieter K. , McNicol Anne M.
Published: June 2005

Pro‐opiomelanocortin (POMC) mRNA was demonstrated in pituitary adenomas from 16 patients with Cushing's disease and 10 with Nelson's syndrome. The intensity of signal was significantly greater in Nelson's syndrome than in Cushing's disease and there was a trend towards a greater proportion of positive cells. This probably reflects inhibition of POMC gene expression by the high circulating levels of cortisol in Cushing's disease. In the para‐adenomatous gland, the intensity of signal was variable in cells showing Crooke's hyaline change, ranging from negative to strongly positive, in keeping with the functional heterogeneity of corticotrophs. In one case, junctional corticotrophs were present and these were more intensely stained than anterior lobe corticotrophs in the same gland. This supports the concept that these cells are subject to different regulatory influences from corticotrophs in the anterior lobe. Whether this is related to differences in embryological origins or to local factors is at present unclear.
Manganese superoxide dismutase content and localization in human thyroid tumours
Authors: Nishida Shozo , Akai Fumiharu , Iwasaki Hiromitsu , Hosokawa Kouichi , Kusunoki Takashi , Suzuki Keiichiro , Taniguchl Naoyuki , Hashimoto Shigeo , Tamura Takanori T.
Published: June 2005

Manganese‐containing superoxide dismutase (Mn‐SOD) content and its immunohistochemical localization in human thyroid tumours and some other thyroid diseases were examined and compared with adjacent normal thyroid tissue. Enzyme‐linked immunosorbent assay (ELISA) was used in this study for the measurement of Mn‐SOD. The content of Mn‐SOD tended to increase in diffuse hyperplasia, adenomatous goitre, and foliicular adenoma. In papillary carcinoma, it was significantly higher than in adjacent normal thyroid tissue. Foliicular carcinoma also revealed a markedly high Mn‐SOD conient. In the immunohistochemicai study, adjaceni normal thyroid tissue showed granular positive staining of Mn‐SOD in the cytoplasm. An increase of Mn‐SOD was observed in the papillary proliferative lesion of diffuse hyperplasia and in the follicles adjacent to lymphoid tissue in chronic thyroiditis with hypothyroidism. Strong positive staining of Mn‐SOD was observed in papillary and foliicular carcinomas, whereas in anaplastic carcinoma staining was markedly less intense. These results indicate that the Mn‐SOD content varies according to the degree of differentiation of thyroid carcinomas.
Expression of large intestinal mucin antigen (LIMA) epitopes in the normal and neoplastic gastrointestinal tract
Authors: Pilbrow Simon J. , Hertzog Paul J. , Pingzower Gideon D. , Linnane Anthony W.
Published: June 2005

This study has identified the expression in normal and neoplastic gastrointestinal (GI) tract of epitopes on the colonic mucin LIMA (large intestinal mucin antigen), which are unique markers of normal colonic differentiation. Six anti‐LIMA monoclonal antibodies (MAbs) (22D4, 9B5, 2C3, 23B2, 46A2, and 10B3) were studied immunohistochemically in normal GI tract, colorectal adenomas, and colorectal and gastric cancers. All MAbs showed specificities consistent with distinct epitopes, five of which were neuraminidase‐resistant and four periodate‐sensitive. Each reacted with mucin in 60–100 per cent normal colons—MAbs 10B3 an 23B2 also with small intestinal mucin—but none with gastric mucin. Five MAbs showed crypt and regional gradients in normal colon. MAbs, 22D4, 9B5, and 2C3 showing a hierarchy of reactivities in the crypt. Individual adenomas showed decreasing goblet cell (GC) LIMA expression with increasing size. However, 30 per cent of familial adenomatous polyposis (FAP) patients had generalized background losses of 9B5 and 2C3 GC reactivity, retaining 22D4, whilst 44 per cent of non‐FAP patients lost 22D4 GC reactivity, regaining 9B5 and 2C3—evidence for polymorphism of mucin expression. All colorectal cancers expressed LIMA epitopes (frequently weaker than normal), and three MAbs (22D4, 9B5, and 2C3) showed deeper than normal staining in adjacent crypts. Eighty‐five per cent of gastric cancers also expressed LIMA epitopes.
Expression of the nuclear membrane protein statin in cycling cells
Authors: Ansari Bijan , Dover Robin , Gillmore Caroline P. , Hall Peter A.
Published: June 2005

Statin is a 57 kD protein previously reported to be expressed by cells in G0. We have studied the detailed distribution of statin immunoreactivity in normal human and rat tissues, and correlated this with investigation of in vitro model systems. By laser confocal microscopy, statin immunoreactivity is localized to the nuclear membrane. In contrast to previous reports, using in vitro model systems we found that statin was also expressed by replicating cells as judged by both co‐localization with [3H]thymidine‐labelled and Ki67‐labelled cells. Furthermore, in a nude mouse xenograft model the number of statin‐labelled cells exceeded the number of quiescent cells as assessed by both fraction of labelled mitosis methods and labelling with [3H]thymidine and Ki67. We conclude that although there is an association between expression of the 57 kD nuclear membrane protein statin and growth arrest, this is not absolute and it is expressed in a sub‐population of cycling cells. The properties of statin closely resemble those of nuclear lamins, members of the intermediate filament family.
p53 Immunoreactivity in inflammatory and neoplastic diseases of the uterine cervix
Authors: Bosari Silvano , Roncalli Massimo , Viale Giuseppe , Bossi Paola , Coggi Guido
Published: June 2005

Immunoreactivity for the tumour suppressor gene product p53 is commonly found in many different human malignancies and few premalignant lesions. Data on cervical neoplasms, however, are still lacking. We retrospectively investigated p53 immunoreactivity in 92 lesions of the uterine cervix, including 44 cases of chronic cervicitis, 29 squamous intraepithelial lesions (SILs), and 19 invasive carcinomas. p53 immunoreactivity confined to the basal cell layer, was detected in 74 per cent of cases showing chronic cervicitis and in all cases with low‐grade SILs. Conversely, suprabasal and/or diffuse p53 immunoreactivity was exclusively demonstrated in 25 per cent of high‐grade SILs and in 74 per cent of invasive carcinomas. The results of this investigation document a high prevalence of p53‐immunoreactive malignant tumours of the uterine cervix. In high‐grade SILs, p53‐immunoreactive cells paralleled the height of involvement by dysplastic changes within the squamous epithelium. A prolonged half‐life of the protein is the most likely explanation for the occurrence of p53 immunoreactivity in neoplastic cells. The unexpected finding of p53‐immunoreactive cells in inflammatory lesions, though possibly related to an increased proliferation rate of the basal cell compartment, requires further study and underlines the need for a careful approach to p53 immunocytochemistry.
BCL‐2 expression in adult and embryonic non‐haematopoietic tissues
Authors: Lu Qi‐Long , Poulsom Richard , Wong Leslie , Hanby Andrew M.
Published: June 2005

The B‐cell leukaemia/lymphoma‐2 (bcl‐2) proto‐oncogene is unusual as its product appears to provide survival advantage to B cells by blocking apoptosis. In this study, the expression of bcl‐2 has been examined in normal nonhaematopoietic tissues, embryos, and psoriatic skin by immunohistochemical staining. Bcl‐2 protein expression is mainly observed in cell populations with a long life and/or proliferating ability such as duct cells in exocrine glands, basal keratinocytes, cells at the bottom of colon crypts, and neurons. In the skin of both adult and embryo and also embryonic kidney and cartilage, bcl‐2 expression was observed in cells which were undergoing morphological transition from undifferentiated stem cells to committed precursor cells. The finding of bcl‐2 expression in the terminal differentiated syncytial trophoblast, but not cytotrophoblast, and in some cells responsive to hormone stimulation such as in the endometrium and myometrium suggests that the gene expression may be related too hormone responsiveness. As no bci‐2 localization was seen in the benign hyperproliferative skin condition psoriasis, this does not suggest a straightforward link to proliferation. These observations support the view that the bcl‐2 gene may have an important role in cell development, maturation, and the path to terminal differentiation.
TGF alpha and TGF beta expression in mammary carcinoma
Authors: Dublin Edwin A. , Barnes Diana M. , Wang Dennis Y. , King Roger J. B. , Levison David A.
Published: June 2005

TGF alpha and beta expression was examined using rabbit polyclonal antibodies and immunohistochemistry on a series of 195 breast carcinomas. TGF alpha immunoreactivity was observed in all but nine of the tumours, with over 50 per cent staining strongly. The polyclonal TGF alpha antibody (CIM1), when compared with a commercially available mouse monoclonal TGF alpha antibody used on the same sections, gave a good correlation (r=0.52, P<0.001). Both TGF alpha antibodies produced a granular cytoplasmic staining pattern, that with CIM 1 being coarser, suggestive of binding to an aggregated protein or organelle. Eighty‐one per cent of tumours stained with the TGF beta antibody, 35 per cent strongly. There was significant co‐expression of TGF alpha and TGF beta (P<0.001). However, they were not found to be useful prognostic indicators, lacking any significant correlation with histological classification, tumour size, nodal status, oestrogen receptor status, S‐phase fraction, or overall survival over a 9–12 year period. The expression of these growth factors in most breast carcinomas suggests that they have important biological roles, but the exact nature of these roles remains unclear at the moment.
Modulation of cell surface componts of human breast cancer cells by retinoic acid: Enhanced HLA‐DR Expression
Authors: Gardner F. J. E. , Walker Rosemary A.
Published: June 2005

The effect of the differentiating agent all‐trans‐retinoic acid on the expression of components of the milk fat globule membrane and HLA‐DR by breast cancer cell lines has been examined. Effects on proliferation were also considered, to determine whether any cell surface changes were related to or independent of proliferation effects. No significant differences were observed in the expression of components detected by the milk fat globule membrane antibodies HMFG1 and HMFG2 over 8 days culture with 10‐7‐10‐9 M retinoic acid for the cell lines MCF‐7, T‐47 D, ZR‐75, MDA‐MB‐231, and BT‐20. In contrast, there was enhanced expression of HLA‐DR by two oestrogen receptor‐positive cell lines T‐47 D and ZR‐75 and the oestrogen receptor‐negative line MDA‐MB‐231, with differing sensitivities. These effects were independent of inhibition of proliferation, which was only observed for oestrogen receptor‐positive cell lines and for different durations of exposure. The finding of enhanced HLA‐DR expression after retinoic acid treatment has not previously been reported and is of interest regarding clinical potential for the induction of tumour immunity.
Evidence of perforin‐mediated cardiac myocyte injury in acute murine myocarditis caused by coxsackie virus B3
Authors: Seko Yoshinori , Shinkai Yoichi , Kawasaki Akemi , Yagita Hideo , Okumura Ko , Yazaki Yoshio
Published: June 2005

We have recently demonstrated that killer cells expressing a cytolytic factor, perforin, infiltrate the hearts of mice with acute viral myocarditis and may play an important role in the mechanism of myocardial damage. To clarify the mechanism of in vivo cardiac myocyte injury mediated by perforin, we investigated the release of perforin molecules from killer cells by immunoelectron microscopy and examined the circular lesions formed by perforin on the membrane of cardiac myocytes. We found that there was massive release of perforin molecules from the killer cells directly onto the surface of the cardiac myocytes. Furthermore, electron microscopy of ultrathin ventricular sections treated with trypsin revealed numerous circular lesions with the characteristics of perforin pores, in the membranes of cardiac myocytes. These findings provide the first direct evidence that killer cells injure cardiac myocytes by releasing perforin and may play a critical role in the myocardial damage occurring in acute viral myocarditis.
Immunohistochemical distribution of heart‐type fatty acid‐binding protein immunoreactivity in normal human tissues and in acute myocardial infarct
Authors: Watanabe Kazuo , Wakabayashi Hironao , Veerkamp J. H. , Ono Teruo , Suzuki Toshimitsu
Published: June 2005

The cellular distribution of heart‐type fatty acid‐binding protein (H‐FABP) immunoreactivity was examined in normal human tissues using a polyclonal antibody against human H‐FABP. Immunoreactivity was detected in cardio‐myocytes of both ventricles and atria as well as in all striated muscles investigated. In addition, staining was frequently observed in parietal cells of the stomach, renal epithelial cells, acinar and ductal cells of the breast, ductal cells of the salivary gland, corpus luteum, and Leydig cells of the testis. Adipocytes and vascular endothelial cells were positive but other tissues and cells examined were negative. Old infarcts of the heart replaced by fibrous connective tissues were not labelled. Necrotic cardiomyocytes and morphologically normal cardiomyocytes in acute ischaemic lesions 1 h after onset showed reduced or no H‐FABP immunoreactivity. Thus, decreased immunoreactivity for H‐FABP may be a good histological marker of damaged cardiomyocytes.
p53 and c‐erbB‐2 protein expression in adenocarcinomas and epithelial dysplasias of the gall bladder
Authors: Kamel Dia , Pääkkö Paavo , Nuorva Kyösti , Vähäkangas Kirsi , Soini Ylermi
Published: June 2005

In this study we investigated immunohistochemically the expression of p53 and c‐erbB‐2 proteins in 30 gall bladder adenocarcinomas, one carcinoma in situ, eight gall bladder epithelial dysplasias, and four cases of chronic cholecystitis. p53 expression could be found in 14 (47 per cent) adenocarcinomas and in two out of eight epithelial dysplasias. There were significantly more p53‐positive grade II‐III tumours than grade I tumours (P=0.032 according to Fisher's exact probability test). c‐erbB‐2 expression was found in three (10 per cent) adenocarcinomas, but all dysplasias were c‐erbB‐2‐negative. All three c‐erbB‐2‐positive cases were also p53‐positive. The results indicate that p53 mutations and c‐erbB‐2 gene alterations play a role in the neoplastic transformation of gall bladder epithelial cells. Co‐expression of p53 and c‐erbB‐2 suggests that alterations of these genes might act in concert in the neoplastic transformation. The occurrence of p53 expression in gall bladder dysplasias suggests that p53 mutations could be an early event in the evolution of some gall bladder carcinomas, as has been suggested for some other types of tumours, such as lung squamous cell carcinomas.
Carcinoembryonic antigen (CEA) in benign and malignant epithelium of the gall bladder, extrahepatic bile ducts, and ampulla of vater
Authors: Maxwell Perry , Davis Richard I. , Sloan James M.
Published: June 2005

Carcinoembryonic antigen (CEA) has been reported in benign and malignant epithelium of the gall bladder, although cross‐reactivity with CEA‐related antigens cannot be excluded. We have investigated the immunoreactivity of three monoclonal and two polyclonal antisera to CEA in benign and malignant epithelium of the gall bladder, extrahepatic bile ducts, and ampulla of Vater. The results varied with the antisera, due to cross‐reactivity with CEA‐related antigens and according to the epitope specificities. Little CEA was found in benign epithelium using monoclonal antisera or following absorption of a polyclonal antiserum from Dako with human liver powder. The latter was the most reliable at detecting malignant epithelium of the gall bladder and extrahepatic bile ducts. All antisera showed equal immunoreactivity in tumours of the ampulla of Vater. The differences seen in the immunoreactivities between tumours of the upper and lower extrahepatic biliary tract probably indicate a variation in the occurrence of CEA epitopes.
Lymphoma in the BB/E rat: C‐myc translocation identified
Authors: Meehan C. J. , Krajewski A. S. , Butcher G. W. , Smith W. , Baird J. D.
Published: June 2005

In a prospective necropsy study involving 257 animals in the BB rat colony in Edinburgh, there was an increased incidence of lymphoma (average 10.9 per cent in all three subgroups: 8 per cent in diabetic, 20 per cent in non‐diabetic diabetes‐prone, and 3.2 per cent in diabetes‐resistant rats). The incidence was significantly increased (P<0.05) in the non‐diabetic diabetes‐prone subgroup. These results differ markedly from previous results and indicate that the relationship between lymphoma and diabetes is more complex than previously suggested. All the lymphomas bar one involved the ileocaecal nodes and were classified as immunoblastic lymphomas of B‐cell origin. There was a striking resemblance both in tissue distribution and in histological classification to the lymphoma seen in the established B‐cell lymphoma model, the LOUVAIN rat. Southern blot analysis carried out on the BB rat lymphomas revealed a translocation of variable length involving the c‐myc oncogene. Such a translocation has not been demonstrated in the BB rat before.
Absence of Epstein—Barr viral RNA in lymphomatoid papulosis
Authors: Kadin Marshall E. , Vonderheid Eric C. , Weiss Lawrence M.
Published: June 2005

Epstein‐Barr virus (EBV) recently has been implicated in the pathogenesis of Hodgkin's disease (HD). Lymphomatoid papulosis (LyP) is a premalignant cutaneous lymphoproliferative disorder which shares several characteristics with HD. The hypothesis has been made that EBV may be associated with the pathogenesis of LyP. We therefore examined 17 skin biopsy specimens and two lymph nodes from nine patients with LyP for EBV RNA using the highly sensitive and specific EBER method. In all specimens, the large atypical cells were negative for EBV while poly T studies confirmed the presence of adequate RNA for detection of EBER. The negative EBER results were confirmed in seven LyP patients whose biopsies were also stained for latent membrane protein (LMP‐1). Interestingly, one patient with clonally related LyP and HD had no EBV RNA detected in any specimen. We conclude that EBV is unlikely to be an important aetiological agent in LyP. If confirmed in other patients, HD associated with LyP may have a different aetiology from HD arising de novo.
Identical expression of ELAM‐1, VCAM‐1, and ICAM‐1 in sarcoidosis and usual interstitial pneumonitis
Authors: Van Dinther‐janssen A. C. H. M. , Van Maarsseveen T. C. M. Th. , Eckert H. , Newman W. , Meijer C. J. L. M.
Published: June 2005

Extravasation of leucocytes in tissues is mediated by leucocyte—endothelial cell interactions in which adhesion molecules play an important role. Until now, two pathways have been unravelled, i.e., the LFA‐1/ICAM‐1 and the VLA‐4/VCAM‐1 pathways. ELAM‐1 has been shown to be involved in granulocyte accumulation and recently also in lymphocyte migration. The role of HECA‐452 is under investigation. In this study we have investigated the expression of the above‐mentioned adhesion molecules in lung tissue of patients with pulmonary sarcoidosis and usual interstitial pneumonitis (UIP), and in mediastinal lymph nodes of patients with sarcoidosis. ICAM‐1 (CD54) was broadly distributed on the endothelium of all the vessels found in sarcoidosis and UIP. VCAM‐1 was present on the endothelium of the venules, capillaries, and arterioles in both sarcoidosis and UIP. ELAM‐1 reacted with endothelial cells lining venules and capillaries in chronic progressive sarcoidosis and in the active phase of UIP but not in the stationary phases of both diseases. HECA‐452 activity could be detected only on high endothelial venules within sarcoid lymph nodes. In lung tissues, macrophages bearing the ICAM‐1 antigen were present in sarcoid tissue but not in the interstitium and alveolar space of UIP. LFA‐1 (CD11a/CD18) and VLA‐4 (CD49d/CD29) were present on all leucocytes found but seemed to be more highly expressed on lymphocytes in sarcoidosis. These findings suggest that the LFA‐1/ICAM‐1 and VLA‐4/VCAM‐1 pathways are involved in leucocyte migration in both types of lung disease, while in the active phases of sarcoidosis and UIP, ELAM‐1 is also involved.
The diagnostic implications of variable cytokeratin expression in mesotheliomas
Authors: Mayall Frederick G. , Goddard Heather , Gibbs Allen R.
Published: June 2005

Immunostaining for cytokeratin has a well‐established diagnostic application in distinguishing sarcomatous mesotheliomas from sarcomas based on the premise that cytokeratin expression is common in mesotheliomas but very rare in sarcomas. However, the frequency of cytokeratin detection is highly dependent on the choice of antibody. The aim of this study was to examine the distribution of simple cytokeratins and stratified cytokeratins in 45 mesotheliomas of various types and to assess the diagnostic utility of a simple cytokeratin antibody, a stratified cytokeratin antibody, and a broad spectrum cytokeratin antibody with the aim of establishing the superiority of one for routine diagnostic use. Particular attention was paid to the potential utility of these antibodies in biopsy specimens. The broad spectrum cytokeratin antibody performed better than both the simple cytokeratin antibody (paired t‐test: P < 0.01) and the stratified cytokeratin antibody (P < 0.01) as a diagnostic marker of mesothelioma and should be used in preference to the other two antibodies, particularly when considering small biopsy specimens.
Immune complex degradation by cultured rat mesangial cells
Authors: Furness Peter N.
Published: June 2005

The binding and degradation of radiolabelled immune complexes by cultured rat glomerular mesangial cells were measured and compared with the binding and degradation by thioglycollate‐elicited rat peritoneal macrophages. Mesangial cells are generally considered to be a modified pericyte with smooth muscle‐like properties, but they were able to bind and degrade soluble immune complexes at rates comparable to those of the macrophages. In a second study, the ability of cultured mesangial cells to bind and degrade immune complexes of varying molecular weight was assessed. Very large, insoluble complexes were found to bind to mesangial cells more avidly than small soluble complexes, but unlike the small complexes, the large complexes did not appear to undergo degradation. These findings support a role for the intrinsic mesangial cell in the elimination of small soluble immune complexes as they arrive in the glomerulus. They also provide a possible explanation for the paradox that large immune complexes—i.e., electron‐dense deposits—can persist in the mesangium next to the intrinsic mesangial cells without being rapidly destroyed.
DNA cytometric and interphase cytogenetic analyses of paraffin‐embedded hydatidiform moles and hydropic abortions
Authors: Van De Kaa Christina A. , Hanselaar Antonius G. J. M. , Hopman ANTON H. N. , Nelson Karin A. M. , Peperkamp Antonius R. , Gemmink Johanna H. , Beck Johan L. M. , De Wilde Peter C. M. , Ramaekers Frans C. S. , Vooijs G. Peter
Published: June 2005

The combined application of DNA cytometric and interphase cytogenetic analyses was used to find objective criteria for the differential diagnosis of complete hydatidiform mole, partial hydatidiform mole and hydropic abortion. DNA ploidy and G0/G1 exceeding rates were determined using image and flow cytometric analyses on paraffin‐embedded tissues of 166 cases: 71 cases of complete mole, 20 cases of partial mole, and 75 cases of abortions. To determine the existence and histological distribution of cell subpopulations with numerical chromosome aberrations, interphase cytogenetic analysis using probes specific for chromosomes 1, X, and Y was applied to paraffin tissue sections of 23 cases: 12 cases of complete mole, 3 cases of partial mole, and 8 cases of abortions. In contrast to previously reported findings that complete moles are diploid, the results of this study showed that complete moles are DNA‐polyploid (96 per cent), with high G0/G1 exceeding rates and a high frequency of numerical chromosomal aberrations in the trophoblast hyperplasia. The majority of the partial moles were DNA‐triploid (55 per cent). This study, however, also showed the presence of DNA‐polyploid partial moles (30 per cent). Abortions were DNA‐diploid (60 per cent) or DNA‐triploid (39 per cent). DNA cytometric analysis, especially image DNA cytometric analysis with determination of the G0/G1 exceeding rate, and interphase cytogenetic analysis provide objective measurements which are contributory in the differential diagnosis between complete mole, partial mole, and hydropic abortion.
T‐cell lymphoma developing in Hodgkin's disease: Evidence for two clones
Authors: Wlodarska Iwona , Delabie Jan , De Wolf‐peeters Chris , Mecucci Cristina , Stul Michel , Verhoef Gregor , Cassiman Jean‐Jacques , Berghe Herman Van Den
Published: June 2005

A case of peripheral T‐cell lymphoma (PTL) occurring in a patient with Hodgkin's disease (HD) in relapse is described. The second neoplasm developed 25 months after the diagnosis of HD. Cytogenetic analysis on the lymph node biopsy at the time of diagnosis of PTL revealed the co‐existence of two distinct, abnormal cell clones. The first clone was characterized by a reciprocal translocation t(5;7)(q13;q35) involving 7q35, namely the TCR‐β gene, as expected in T‐cell lymphomas. The second cell clone carried trisomies for chromosomes 2, 5, 7, and 14. By immunophenotypic and molccular analysis as well as by in situ hybridization, it was possible to prove that the malignant T‐cells and the Reed‐Sternberg cells corresponded to different cell clones, one carrying the structural chromosome abnormalities and one carrying the numerical chromosome anomalies. These results indicate that the present case represented a true composite lymphoma.