bluuebunny/crossref_metadata_2025 huggingface dataset:
In situ hybridization of immunoglobulin light chain mRNA in paraffin sections using biotinylated or hapten‐labelled oligonucleotide probes
Authors: Pringle James H. , Ruprai Anita K. , Primrose Lindsay , Keyte John , Potter Linda , Close Pauline , Lauder Ian
Published: June 2005

An in situ hybridization technique has been developed for the detection of immunoglobulin light chain mRNA in routine pathology specimens. The method detects k or λ constant region sequences using a cocktail of synthetic oligonucleotide probes labelled with biotin or fluorescein 5‐isothiocyanate (FITC) reporter molecules. The probes were labelled at flanking sites chemically by primary amine directed acylation and by ‘homopolymer tailing’ with terminal deoxynucleotidyl transferase using non‐radioactive nucleotide analogues. The mRNA was unmasked in the formalin‐fixed tissue sections by digestion with varying concentrations of proteinase K, and the hybrids were demonstrated using alkaline phosphatase with either a streptavidin/biotin based four‐stage system or an anti‐FITC antibody based detection system. Alkaline phosphatase was visualized using a Fast Red naphthol‐capture method and the sections were counterstained with haematoxylin.

The results confirm that the method is specific for k or λ mRNA and show that specific mRNAs can be detected in routine formalin‐fixed sections using non‐radioactive techniques with retention of good morphology. The method reliably detects light chain mRNA in cells expressing secretory immunoglobulin. The protocol can also be applied to tissue rich in endogenous biotin by using hapten‐labelled probes.
In situ distribution of transforming growth factor α in normal human tissues and in malignant tumours of the ovary
Authors: Kommoss Friedrich , Wintzer Hanns Olof , Kleist Sabine Von , Kohler Manuela , Walker Roger , Langton Beatrice , van Tran Kim , Pfleiderer Albrecht , Bauknecht Thomas
Published: June 2005

The distribution of transforming growth factor α (TGF‐α) in human normal tissues from the uterus, Fallopian tube, ovary, small and large intestine, lung, spleen, kidney, and skin was studied by immunohistochemistry. TGF‐α was found in epidermis, bronchial epithelium, intestinal mucosa, renal tubules, endo‐ as well as in exocervical and endometrial epithelium, and in the serous epithelium of the Fallopian tube. No TGF‐α was detected in the stromal components of any of the tissues nor in any of the pre‐ and post‐menopausal ovaries studied. Twenty‐nine ovarian tumours including 23 ovarian carcinomas, one malignant mixed Muellerian tumour, two ovarian metastases of gastrointestinal carcinomas, one dysgerminoma, one sarcoma, and one fibroma were studied for TGF‐α by the same immunohistochemical method. In 25 cases, specific cytoplasmic staining for TGF‐α of epithelial tumour cells could be demonstrated. The pattern and intensity of the TGF‐α immunostain varied among the TGF‐α‐positive tumours. No TGF‐α was found by immunohistochemistry in the remaining four cases nor in the stromal tumour components of any of the lesions studied. Northern blot analysis for TGF‐α mRNA was performed on 12 of the tumours. While the immunohistochemistry and blotting results correlated well in ten cases, discordant results were obtained in two lesions.
c‐erbB‐2 oncogene expression in ovarian cancer
Authors: Haldane J. S. , Hird V. , Hughes C. M. , Gullick W. J.
Published: June 2005

One hundred and four common epithelial malignancies of the ovary were stained for c‐erbB‐2 using an affinity‐purified polyclonal antibody 21N. Twenty‐three out of 104 (22·1 per cent) showed cytoplasmic staining alone. Nine out of 104 (8·7 per cent) showed both membrane and cytoplasmic staining. In a multivariate Cox analysis with other known risk factors the relative risk for cytoplasmic staining alone was 1·456 (χ2=1·71, P>0·1) and for membrane and cytoplasmic staining 0·316 (χ2=7·95, P < 0·005). These results do not support an adverse prognostic effect of c‐erbB‐2 in our patients.
Helicobacter pylori infection of gastric mucin cell metaplasia: The duodenum revisited
Authors: Offerhaus G. Johan A. , Molyvas E. Noydas , Hoedemaeker Philip J.
Published: June 2005

The histological material of 158 Billroth II gastrectomy specimens, used for a former study that established a relationship between duodenal ulcers and the presence of gastric metaplastic epithelium in the duodenal bulb, was reinvestigated for the presence of Helicobacter pylori. The results show that in all duodenal ulcer patients with gastric mucin cell metaplasia H. pylori colonized the metaplastic epithelium accompanied by an inflammatory response. The intestinal mucosa was always negative for H. pylori. H. pylori‐positive patients also had the micro‐organism in their gastric antrum. The results further support the hypothesis that infection of gastric mucosa in the bulb by H. pylori underlies duodenal ulcer disease.
Proliferating cell nuclear antigen (PCNA) immunolocalization in paraffin sections: An index of cell proliferation with evidence of deregulated expression in some, neoplasms
Authors: Hall P. A. , Levison D. A. , Woods A. L. , Yu C. C.‐W. , Kellock D. B. , Watkins J. A. , Barnes D. M. , Gillett C. E. , Camplejohn R. , Dover R. , Waseem N. H. , Lane D. P.
Published: June 2005

Proliferating cell nuclear antigen (PCNA) is a 36 kD nuclear protein associated with the cell cycle. A monoclonal antibody, PC10, that recognizes a fixation and processing resistant epitope has been used to investigate its tissue distribution. Nuclear PCNA immunoreactivity is found in the proliferative compartment of normal tissues. PCNA immunoreactivity is induced in lectin stimulated peripheral blood mononuclear cells in parallel with bromodeoxyuridine incorporation and the number of cells with PCNA immunoreactivity is reduced by induction of differentiation in HL60 cells. In non‐Hodgkin's lymphomas a linear relation between Ki67 and PCNA staining was demonstrated. These data suggest that in normal tissues and lymphoid neoplasms, PCNA immunolocalization can be used as an index of cell proliferation. However, in some forms of neoplasia, including breast and gastric cancer and in vitro cell lines, the simple relation between PCNA expression and cell proliferation is lost. In some breast and pancreatic tumours there is apparent deregulation of PCNA with increased expression in tissues adjacent to the tumours. The over‐expression in some tumours and in adjacent morphologically normal tissue may represent autocrine or paracrine growth factor influence on PCNA gene expression.
Glutathione s‐transferase detoxication enzymes in cervical neoplasia
Authors: Carder Pauline J. , Al‐Nafussi Awatif , Rahilly Maeve , Lauder John , Harrison David J.
Published: June 2005

Altered expression of the glutathione S‐transferases (GSTs) has been implicated in the progression to tumour after exposure to carcinogens, and GST Pi has been suggested as a possible marker of preneoplasia in the cervix.

We have studied expression of the GST isoenzymes in normal cervix, non‐dysplastic cervical condylomata, cervical intraepithelial neoplasia (CIN), and invasive squamous carcinoma of the cervix using immunocytochemistry. An increase in GST Pi in CIN as compared with normal cervix was paralleled by a reduction in the expression of microsomal GST. Similar changes were seen in cervical condylomata and immature squamous metaplasia, and thus neither isoenzyme is a marker of dysplasia. Microsomal GST was expressed in only 66 per cent of cases and in 22 per cent showed strong expression in vascular endothelium.

These findings are of particular interest in view of the association between cervical carcinoma and cigarette smoking. Differences between individuals in the ability to detoxify environmental carcinogens may influence the likelihood of progression from benign proliferation to invasive malignancy.
Comparative endotoxin‐induced hepatic injury in young and aged rats
Authors: Durham Stephen K. , Brouwer Adriaan , Barelds Roel J. , Horan Michael A. , Knook Dick L.
Published: June 2005

Recent studies have demonstrated that aged rats are more susceptible to the lethal effects of endotoxin (ET) than young rats. The early (15 min to 7 h) hepatic ultrastructural and biochemical changes induced by ET in young (6 months) and aged (24 months) rats were evaluated to elucidate cell populations and/or the mechanisms that may be responsible for the previously observed differential effects. Aged rats given ET had significantly increased numbers of neutrophils in hepatic sinusoids at 30 min and thereafter as compared with ET‐treated young rats. Morphologic evidence of coagulation within hepatic sinusoids, including aggregates of fibrin enmeshed among polymorphonuclear leukocytes and platelet aggregates, was frequently observed in ET‐treated aged rats but not in ET‐treated young rats. In contrast, Kupffer cells of ET‐treated young rats frequently contained phagocytized neutrophils and platelets, whereas this phenomenon was rarely observed in Kupffer cells of ET‐treated aged rats. Hepatocellular morphologic injury was more pronounced and occurred at earlier time periods in ET‐treated aged rats, and was accompanied by significant increase in hepatic transaminases. ET‐treated aged rats had an earlier onset and greater severity of endothelial cell injury than did ET‐treated young rats. The results of this study indicate a greater aggregation of blood elements in the hepatic sinusoids of aged rats following the intravenous administration of ET, which suggests that a greater diminution in microcirculation was induced in aged rats by ET. Additionally, the increased phagocytosis of inflammatory cells by Kupffer cells of young rats may be a mechanism which affords protection against endotoxin‐induced lethality.
Nucleolar organizer regions and prognosis in renal cell carcinoma
Authors: Delahunt Brett , Ribas Jorge L. , Nancy John N. , Bethwaite Peter B.
Published: June 2005

The prognostic significance of nucleolar organizer regions (NORs) in renal cell carcinoma (RCC) was evaluated. NORs were quantified in a series of 182 cases of RCC using the silver‐colloid method. The cases were staged according to Robson's method (48 stage I, 26 stage II, 33 stage III, 75 stage IV) and mean NOR numbers for each tumour were correlated with survival over a 5‐year period. Localized tumours (stages I and II) with low NOR numbers had an almost 100 per cent 5‐year survival. Those patients with clinical evidence of metastases at presentation showed a high mortality, although those with low numbers of NORs had a significantly increased disease‐free interval. Statistical analysis using the log rank test indicated NORs to be a significant predictor of survival over the whole series (P= 0·0001) and within each of Robson's stages (P= 0·0008 stage I,P= 0·0154 stage II,P= 0·0009 stage III,P= 0·0001 stage IV). Analysis of data using Cox's proportional hazard model showed mean NOR numbers to be independent of stage as a predictor of survival.
Basaloid carcinoma of salivary glands, a variety of undifferentiated adenocarcinoma. Immunohistochemical study of intermediate filament proteins in 24 cases
Authors: Chomette G. , Auriol M. , Vaillant J. M. , Kasai T. , Okada Y. , Mori M.
Published: June 2005

Among adenoid cystic carcinomas of salivary glands (ACCs), the solid basaloid type has a poor prognosis similar to that of undifferentiated adenocarcinomas. We studied 24 cases in immunohistochemistry using antibodies reactive with keratins of various molecular weights, vimentin, S‐100 protein, and its A and B subunits. Our findings were correlated with the histological pattern and with the variable degree of differentiation of these carcinomas. In comparison with other types of ACC, intermediate filament proteins in this group were weakly expressed. The co‐expression of cytokeratin and vimentin was noted in some cases. Additional features noted were the presence of cribriform cavities associated with solid lobules and areas of necrosis giving a comedocarcinomatous pattern. In these two variants, cells characterized by the dual expression of cytokeratin and S‐100 protein were seen. In the highly malignant anaplastic variety, only a few cells were weakly positive with antisera to cytokeratin and vimentin. This group shows similarities to undifferentiated adenocarcinomas of salivary glands. Such similarities could be explained by the common origin of these tumours from intercalated ducts.
Cellular H‐ and M‐type lactate dehydrogenase (LDH) isoenzymes and tumour diagnosis—an immunohistochemical assessment
Authors: Pan Langxing , Xu Junna , Isaacson Peter G.
Published: June 2005

Cellular H‐ and M‐type lactate dehydrogenase (LDH) isoenzymes were determined immunohistochemically in a wide range of normal, inflammatory, benign neoplastic, and malignant tissues to assess the possible value of the isoenzymes in tumour diagnosis. Both types of LDH were detected almost ubiquitously in normal tissues. Increased staining intensity of LDH, especially M‐type LDH, was demonstrated in most types of malignant tumours studied. Similar increased staining intensity of the isoenzymes was also noted to parallel the proliferative activity of the cells in non‐neoplastic diseases and benign neoplasms. The results suggest that changes of cellular H‐ and M‐type LDH isoenzymes are not specific for any type of malignant tumours. The increase in cellular H‐ and M‐type LDH isoenzymes appears to be related to proliferation of cells in general.
Antipeptide antibodies against the pNR‐2 oestrogen‐regulated protein of human breast cancer cells and detection of pNR‐2 expression in normal tissues by immunohistochemistry
Authors: Piggott Nigel H. , Henry James A. , May Felicity E. B. , Westley Bruce R.
Published: June 2005

Five peptides, corresponding to regions of the predicted protein sequence of the oestrogen‐regulated pNR‐2 protein which is expressed in oestrogen‐responsive human breast cancer cells, were synthesized. Two peptides were immunogenic in rabbits and antisera against one peptide reacted with the pNR‐2 protein in sections of formalin‐fixed, paraffin‐embedded breast tumour. There was a significant correlation between the extent of pNR‐2 protein expression detected by immunohistochemistry and pNR‐2 mRNA levels determined by hybridization with a cDNA probe in a series of primary breast tumours. pNR‐2 expression was assessed immunohistochemically in a panel of normal tissues. Expression was detected in normal breast, small intestine, and stomach (body and antrum).
Oncoprotein immunoreactivity in human endocrine tumours
Authors: Roncalli M. , Springall D. R. , Varndell I. M. , Gaitonde V. V. , Hamid Q. , Ibrahim N. B. N. , Grimelius L. , Wilander E. , Polak J. M. , Coggi G.
Published: June 2005

Abnormally expressed oncogenes are implicated in neoplastic transformation. We have investigated a series of endocrine tumours using immunocytochemistry as a first screening tool to detect oncogene expression. Paraffin sections of 44 pulmonary small cell carcinomas, 15 pulmonary atypical carcinoids, 12 bronchial carcinoids, 28 medullary thyroid carcinomas, 27 phaeochromocytomas, and 17 insulinomas were immunostained with antibodies to c‐erbB‐2, c‐myc, L‐myc, and N‐myc. Diffuse immunoreactivity was detectable for c‐erbB‐2 in 8 out of 44 (18 per cent) pulmonary small cell carcinomas, 3 out of 15 (20 per cent) pulmonary atypical carcinoids, and 6 out of 27 (22 per cent) phaeochromocytomas; for c‐myc in 18 out of 44 (41 per cent) pulmonary small cell carcinomas and 5 out of 15 (33 per cent) pulmonary atypical carcinoids; for N‐myc in 6 out of 28 (21 per cent) medullary thyroid carcinomas; and for L‐myc in 4 out of 27 (15 per cent) phaeochromocytomas. There was considerable intratumoral and intertumoral heterogeneity and, in each tumour group, no relationship was found between tumour pattern, mitotic index, and oncoprotein immunoreactivity. These results suggest that oncogene products are present in a proportion of endocrine tumours, and that specific oncoproteins seem to be related to tumour type but not to other histopathological findings. Thus, oncoprotein detection may be a useful tool for identifying subsets of endocrine tumours that are not otherwise recognizable morphologically.
Enteroviruses and idiopathic dilated cardiomyopathy
Authors: Cochrane Hugh R. , May Felicity E. B. , Ashcroft Thomas , Dark John H.
Published: June 2005

Previous studies have suggested that some cases of idiopathic dilated cardiomyopathy (IDC) are due to persistent viral infection following an episode of viral myocarditis. Viral RNA sequences have recently been detected in material from patients with IDC using molecular biological techniques. We tested 40 samples from recipients' hearts explanted at cardiac transplantation for the presence of enteroviral RNA sequences, using a Northern blotting technique. Material from 19 cases of IDC and 21 cases of non‐cardiomyopathic cardiac failure was examined together with Coxsackie‐virus‐infected neonatal mouse heart as a positive control and non‐infected adult mouse heart as a negative control. A sharp band of viral RNA was detected in the positive control sample. No hybridization signal attributable to viral RNA was obtained for the negative control or for any of the test samples. We conclude that the role of enteroviruses in the pathogenesis of cardiomyopathy is not fully established and that further study is warranted.
Influence of hypobaric hypoxia in infancy on the subsequent development of vasoconstrictive pulmonary vascular disease in the Wistar albino rat
Authors: Caslin Anthony , Heath Donald , Smith Paul
Published: June 2005

A group of Wistar albino rats was injected subcutaneously with monocrotaline to induce vasoconstrictive hypertensive pulmonary vascular disease characterized by medial hypertrophy of small pulmonary arteries, the appearance of muscular pulmonary arterial vessels of arteriolar dimensions (less than 20 μm in diameter), and exudative changes in the lung parenchyma. The vascular abnormalities were quantified by measuring the percentage medial thickness of small pulmonary arteries, the number of muscular pulmonary arterial vessels below 20 μm in diameter per cm2 of lung section and by determining the smallest arterial vessels in each case showing muscularity. A second group of rats was born in a decompression chamber and kept in hypobaric hypoxia for a month of the neonatal period, developing hypoxic hypertensive pulmonary vascular disease as a consequence. The animals in this group were allowed to recover in room air for a period of 3 months and were then injected with the same dose of monocrotaline as that given to the first group. The rats previously exposed to hypoxia exhibited an exaggerated response to the alkaloid, showing in particular many more small muscular pulmonary arterial vessels which were of a smaller diameter than those found in the eupoxic rats treated with the alkaloid. The experiment demonstrates that perinatal hypoxia exaggerates the effects of agents inducing vasoconstrictive pulmonary hypertension with a shift of the segment of the pulmonary arterial tree involved to the periphery as in hypoxia. Reports of a similar phenomenon are noted as occurring in babies born at high altitude, spending their infancy there and subsequently developing primary pulmonary hypertension later in life.
Sarcomatoid tumours of lymph nodes showing follicular dendritic cell differentiation
Authors: Hollowood Kevin , Pease Christopher , Mackay Alasdair M. , Fletcher Christopher D. M.
Published: June 2005

Putative sarcomas of follicular dendritic cells are extremely uncommon and only small case numbers have been clearly documented. A major difficulty in their delineation has been the persistent controversy over the phenotype and ontogeny of normal follicular dendritic cells. Two new cases arising in adult females are presented herein, both of which had initially been mistaken for metastases from a true soft tissue sarcoma. Their morphology, immunophenotype, and ultrastructure (in one case) are described in order to aid their wider recognition. In the light of these findings, the nature and origin of follicular dendritic cells are discussed.
True histiocytic neoplasm of Langerhans' cell type
Authors: Delabie J. , De Wolf‐Peeters C. , De Vos R. , Vandenberghe E. , Kennes K. , De Jonge I. , Desmet V.
Published: June 2005

Malignant histiocytic neoplasms of dendritic cell lineage are rare and most are derived from the dendritic cells of the T‐cell and B‐cell areas of the lymph node, respectively. Only one case of a malignant histiocytic neoplasm of Langerhans' cells, the dendritic cell of the skin, has been reported.

We report the occurrence of a true histiocytic neoplasm of Langerhans' cell type in a 23‐year‐old female patient. The diagnosis of this highly aggressive neoplasm is supported by histochemical, immunohistochemical, ultrastructural, and genotypic analysis—techniques which allow differentiation from malignant melanoma and large cell anaplastic lymphoma.
Eosinophilic granuloma of the bone in Hand–Schüller‐Christian disease: Extensive in vivo eosinophil degranulation and subsequent binding of released eosinophil peroxidase (EPO) to other inflammatory cells
Authors: Zabucchi Giuliano , Soranzo Maria Rosa , Menegazzi Renzo , Cattin Luigi , Vecchio Monica , Lanza Francesco , Patriarca Pierluigi
Published: June 2005

The eosinophils from bone granuloma, bone marrow, and peripheral blood of a patient with Hand–Schüller–Christian disease (HSCD) were studied by electron microscopy and cytochemistry. Impressive eosinophil degranulation was observed. Extracellular release of eosinophil peroxidase (EPO) and EPO binding to surrounding cells were seen in the granuloma and bone marrow. Cells with peroxidase‐positive plasma membrane were also observed in peripheral blood. The pattern of eosinophil degranulation showed quite different features from those described so far. In the granuloma, the process begins with intracytoplasmic release of the granule matrix content, as revealed by both extensive extragranular accumulation of EPO and progressive decrease of the matrix electron density. Core dissolution follows thereafter, leading to complete disappearance of the granules. At the end of the process, the cells show rupture of the plasma membrane and release of their content into the surrounding environment. This pattern if secretion was also observed in blood and marrow eosinophils of the patient. In view of the previously reported findings that EPO binding to inflammatory cells influences their functions, EPO release and binding to surrounding cells in HSCD may play a role in the evolution of the inflammatory lesion in this disease.
Nucleolar ultrastructure in low‐ and high‐grade non‐Hodgkin's lymphomas
Authors: Goodlad John R. , Crocker John , Macartney James C.
Published: June 2005

Transmission electron microscopy was used to study the ultrastructure of nucleoli in seven high‐ and seven low‐grade non‐Hodgkin's lymphomas. Nucleoli with a single large fibrillar centre were predominantly seen in the low‐grade group, while the majority of nucleoli in high‐grade lesions contained several small fibrillar centres of approximately equal size. A third type of nucleolar appearance was identified, with a large central fibrillar centre surrounded by numerous small ‘satellite’ fibrillar centres. This nucleolar configuration was the least common but was more often seen in low‐grade than in high‐grade lymphomas. The mean fibrillar centre cross‐sectional area was determined for each group and measured 278 800 nm2 for low‐grade non‐Hodgkin's lymphomas and 62 507 nm2 for high‐grade lymphomas (P < 0·001). These results show that fibrillar centre size and morphology correlates with grade of non‐Hodgkin's lymphoma and may reflect differences in ribosomal deoxyribonucleic acid transcription.
A critical evaluation of AgNOR counting in benign naevi and malignant melanoma
Authors: Orrell J. M. , Evans A. T. , Grant A.
Published: June 2005

There is considerable variation in the quoted mean numbers of AgNORS per nucleus for benign melanonaevi and malignant melanomas. This is partly attributable to differnt approaches to AgNOR counting. This study summarizes our experience in devising an optimal technique for counting AgNORs. We show that it is essential to count intranucleolar AgNORs in addition to those lying outside the nucleous to obtain clear separation of naevi from melanoma. Although this seems an onerous task, we further demosnstrate that a maximum of only 30 nuclei need to be counted to obtain a mean AgNOR count per nucleus which is representative of the whole lesion. This compares with the arbitary figure of 100 nuclei chosen by most workers.

Only by optimizing and standardizing all aspects of the AgNOR technique including fixation, staining, and counting will mean AgNOR counts per nucleus become a useful quick, reproducible method which can be applied to lesions which pose diagnostic problems such as borderline melanocytic lesions.
Apoptosis induced by mild hyperthermia in human and murine tumour cell lines: A study using electron microscopy and DNA gel electrophoresis
Authors: Takano Yasuo S. , Harmon Brian V. , Kerr John F. R.
Published: June 2005

Mild hyperthermia is known to enhance apoptosis in a range of normal and neoplastic cell populations. Studies of tumours previously shown to respond to heating in this manner might be expected to provide insights not only into the mechanism of hyperthermic cell killing, but also into the apoptotic process in general. In the present study, cell death induced by 43°C heating for 30 min in two human Burkitt's lymphoma lines, BM 13674 and WW1, and in murine mastocytoma P‐815 × 2·1 was found to be exclusively apoptotic in type, identification being based on light and electron microscopic appearances and on the presence of internucleosomal cleavage of DNA into fragments that are multiples of 180–200 base pairs, which was demonstrated by agarose gel electrophoresis. The heat‐induced apoptosis was prevented by the presence of zinc sulphate, an inhibitor of the endonuclease considered to be responsible for the DNA cleavage, but was not suppressed by the protein synthesis inhibitor cycloheximide. The findings question the validity of the widely held view that active protein synthesis is an invariable prerequisite for the execution of apoptosis. It is suggested that an inositol triphosphate‐mediated increase in cytosolic Ca2+, resulting from limited membrane damage, might be the critical event responsible for activation of apoptosis by mild hyperthermia.
Morphological evidence that A‐CAM is a major intercellular adhesion molecule in human kidney
Authors: Biddlestone L. R. , Fleming S.
Published: June 2005

We have used immunocytochemistry to identify the major primary adhesion molecule of the cadherin class in human kidney. In frozen sections of kidney, A‐CAM was detected using the monoclonal antibody GC 4 on the surface of renal tubular epithelial cells. Renal tubular epithelium did not express L‐CAM. No cadherin reactivity was found on the glomerular epithelial cells. Cultured renal tubular epithelium was studied by immunofluorescence and immunogold methods. A‐CAM was found at the contact points of adjacent epithelial cells, the phenotype of which was confirmed by the demonstration of cytokeratins using the antibody CAM 5.2. The A‐CAM molecule in human kidney had an Mr of 130 kD in Western blotting experiments. These results lead us to conclude that A‐CAM is the major cadherin of adult human renal epithelium.
Control of differentiation in a rectal adenocarcinoma cell line: The role of diffusable and cell‐associated factors
Authors: Buono Raffaele Del , Pignatelli Massimo , Hall Peter A.
Published: June 2005

The rectal adenocarcinoma cell line, HRA‐19a1.1, was cultured in a three‐dimensional type I collagen gel and then xenografted in nu/nu mice to determine whether the in vivo environment could induce further morphological differentiation of the in vitro collagen gel cultures. Three phenotypic changes were observed. Type I collagen induces glandular differentiation in vitro in which well polarized cells are organized around a central lumen, as has been previously reported.1 Seven days after xenografting this structure in nu/nu mice, the glandular structures appeared to have ‘ballooned’ forming cyst‐like structures lined by a monolayer of flattened cells. There were no stromal cells associated with the graft at this stage, but with time stromal cells invaded the collagen. At points where these cells were closely associated with the HRA‐19 cells there was a marked phenotypic change, with the flattened lining cells seen at day 7 becoming columnar. By 21 days the stromal cells had replaced the collagen and the histology of the graft now resembled that of an adenocarcinoma. Placing this cell–collagen culture in a Millipore chamber prior to grafting resulted in cyst‐like structures only. Here we provide conclusive evidence that heterologous connective tissue cells can induce differentiation of a rectal adenocarcinoma cell line by a non‐ or poorly diffusiblc factors(s). Furthermore, we show that this cell–collagen xenograft method has certain advantages over conventional xenograft methods: notably, a consistent 100 per cent take rate; considerably fewer cells are required to form a tumour; and the time taken to form a tumour is dramatically reduced.
Detection of mRNA for ribosomal phosphoprotein P2 in normal colon and colonic tumours by In situ hybridization
Authors: Harvie R. , Elvin P. , McArdle C. , Morten J. E. N. , McNicol A. M.
Published: June 2005

We have localized the mRNA for the ribosomal phosphoprotein P2, a putative metastasis‐related sequence, in normal colon and colonic carcinomas by in situ hybridization, using an oligonucleotide probe end‐labelled with digoxigenin. The mRNA was identified in normal colonic epithelial cells, the intensity of the signal being greater in cells at the base of the crypts compared with those on the surface. A strong positive signal was also seen in plasma cells, in fibroblasts in granulation tissue, in ganglion cells, and in hepatocytes. A positive signal was identified in all 16 primary colonic tumours studied and in 7 hepatic metastases In contrast to previous studies based on Northern blot analysis, we were unable to demonstrate increased expression in metastases as compared with primary tumours, nor could we demonstrate any increased expression in primary tumours which were associated with distant metastases.
Gastrointestinal stromal tumours: Correlation of immunophenotype with clinicopathological features
Authors: Newman P. L. , Wadden C. , Fletcher C. D. M.
Published: June 2005

Stromal tumours of the gastrointestinal tract remain a persistent source of controversy with regard to both their proposed lines of differentiation and the difficulty in predicting their biological behaviour. We have examined 60 cases with a panel of seven antibodies directed at the identification of smooth muscular or neural differentiation. In our hands, 36 per cent of cases showed neural differentiation (although only 6.6 per cent expressed S‐100 protein); 31 per cent appeared smooth muscular; 20 per cent manifested bidirectional differentiation; and 13 per cent were negative for all the markers used. Histological appearances do not reliably reflect immunophenotype. We have attempted to correlate immunophenotype with the site of the lesions and with their clinical behaviour. Mean follow‐up of 5 years was obtained in 42 cases. Tumours with a neural phenotype have the best prognosis. Gastric tumours expressing both desmin and smooth muscle actin (in the absence of other markers) with up to 4 mitoses per 30 HPF behave in a benign fashion. Larger studies are required to substantiate the value of immunophenotyping this complex group of tumours.
Immunohistological analysis of the immunoreactivity of normal lymphoid cells and lymphomas with the monoclonal antibody OPD4
Authors: O'Grady Jane , Blaikie Andrew J. , Howie Sarah E. , Salter Donald M. , Krajewski Andrew S.
Published: June 2005

OPD4 is a recently described monoclonal antibody that recognizes a fixation‐resistant 200 kD antigen restricted to a subset of T cells. Immunolabelling with OPD4 in paraffin sections of normal lymphoid tissues and cases of malignant lymphoma was compared with that of other antibodies in common use, including the T‐cell restricted antibodies MTI and UCHL1 and the B‐cell restricted antibodies MB1, F8‐11‐13, and L26. OPD4 showed similar immunoreactivity to UCHL1 in normal tissues. OPD4 did not stain Reed‐Sternberg cells in Hodgkin's disease. In non‐Hodgkin's lymphomas, OPD4, like UCHL1, reacted with only 2/22 B‐cell lymphomas. OPD4 was, however, less useful as a marker of T‐cell lymphomas, staining only 11/32 cases, while UCHL1 stained 22/32 cases. We conclude that OPD4 is not a useful antibody for the routine diagnosis of T‐cell lymphoma.
Intercellular contacts in the media of the thoracic aorta of art fetuses treated with β‐aminopropionitrile
Authors: Sosa‐Melgarejo Jorge A. , Berry Colin L.
Published: June 2005

Beta‐aminopropionitrile (BAPN) alters the scleroproteins of the arterial media in a way which permits an increase in the stress (force/unit area) applied to smooth muscle cells at a given pressure. Pregant Sprague‐Dawley rats were treated with BAPN and the thoracic aortas of exposed fetuses were studied by electron microscopy and image analysis. Transmission electron microscopy of the media of the thoracic aorta revealed that the typs of intercellular contacts seen in the controls and in the BAPN group were intermediate junctions (IJs), nexus junctions (NJs), simple appositions (SAs), and interdigitations (Ids) as in adults vessels. The BAPN‐treated animals showed an increased density of IJs, SAs, and Ids when compared with controls but the NJs demonstrated no change. Load‐sensitive intercellular contacts have been considered to be the major route by which stress is applied across cells, and the increased number in the BAPN group probably represents a response of the vessel wall to increased tangential tension.
The vascularity of primary cutaneous melanoma
Authors: Fallowfield Mary E. , Cook Martin G.
Published: June 2005

In primary cutaneous malignant melanoma, the vascularity of the dermis immediately deep to the lesion may relate to tumour aggressiveness and to prognosis. These newly formed dermal vessels are incorporated into the melanoma to form the tumour microcirculation. We have assessed the percentage vascular volume in a series of primary melanomas in order to investigate the relationship between tumour vascularity and maximum tumour thickness.

For the 64 melanomas included in this study, there appeared to be a significant relationship between the percentage vascular volume and the maximum tumour thickness. This relationship was not influenced by the presence of necrosis, vascular invasion, regression, or lymphocytic infiltrate, nor by the growth phase of the tumour. However, the percentage vascular volume was very low in the occasional thick melanoma, at least one of which was associated with prolonged survival. It seems possible that a low tumour vascularity could correlate with a relatively favorable outcome in cutaneous melanoma.
Immunohistochemical detection of the epidermal growth factor receptor in paraffin‐embedded human tissues
Authors: Gullick William J. , Hughes Christine M. , Mellon Killian , Neal David E. , Lemoine Nicholas R.
Published: June 2005

Expression of the epidermal growth factor (EGF) receptor was evaluated by immunohistochemical staining of formalin‐fixed, paraffin‐embedded tumour tissues employing two antibodies raised to short synthetic peptides from the cytoplasmic domain of the molecule. Both antibodies gave concordant staining of a series of bladder cancers known to express or lack EGF receptors. There was no cross‐reaction with the related c‐erbB‐2 protein, which was also over‐expressed in some cases. Cancers with EGF receptor expression also expressed high levels of TGF‐alpha, a receptor agonist.
Persisten of Epstein–Barr virus in Reed–Sternberg cells throughout the course of Hodgkin's disease
Authors: Coates Philip J. , Slavin Gerard , D'Ardenne A. Jane
Published: June 2005

Non‐isotopic in situ hybridization employing digoxigenin‐labelled DNA probes has been used to localize Epstein–Barr virus (EBV) in 55 cases of Hodgkin's disease (HD). The virus was found in Reed–Sternberg (RS) and mononuclear Hodgkin's cells in nine patients (16 per cent). Further samples taken at different times from three patients also showed the presence of EBV in the malignant cell population. Estimations of the number of EBV genomes present per cell suggested wide variations between different patients, but relatively constant amounts in different samples from the same patient. These findings are compatible with a stable infection of the neoplastic cells and support the notion that EBV may play a role in the development of HD in these patients.

We also found evidence for the presence of EBV in a small percentage of non‐neoplastic cells in 8 of the 55 samples. This suggests that isolation of EBV from HD tissue does not always signify a pathogenetic role for the virus. Furthermore, it is apparent that a high percentage of HD tissues do not contain demonstrable EBV, and the virus is therefore unlikely to be a causative agent for all cases of HD.
Irreversible damage to the medullary interstitium in experimental analgesic nephropathy in F344 rats
Authors: Burrell Judith H. , Yong Jim L. C. , Macdonald Graham J.
Published: June 2005

Renal papillary necrosis (RPN) and a decreased urinary concentrating ability developed during continuous long term treatment with aspirin and paracetamol in female Fischer 344 rats. Renal structure and concentrating ability were examined after a recovery period of up to 18 weeks, when no analgesics were given, to investigate whether the analgesicinduced changes were reversible.

There was no evidence of repair to the damaged medullary interstitial matrix, or proliferation of remaining undamaged type 1 medullary interstitial cells after the recovery period following analgesic treatment. The recovery of urinary concentrating ability was related to the length of analgesic treatment and the extent of the resulting inner medullary structural damage. During the early stages of analgesic treatment, the changes in urinary concentrating ability were reversible, but after prolonged analgesic treatment, maximum urinary concentrating ability failed to recover.

This study shows that prolonged analgesic treatment in Fischer 344 rats causes progressive and irreversible damage to the interstitial matrix and type 1 interstitial cells leading to RPN. The associated urinary concentrating defect is reversible only during the early stages of structural damage to the inner medulla.
Epstein–Barr viral DNA in Hodgkin's disease: Amplification and detection using the polymerase chain reaction
Authors: Brocksmith Debra , Angel Carole A. , Pringle J. H. , Lauder I.
Published: June 2005

The presence of Epstein–Barr virus (EBV) DNA in biopsy tissues from patients with Hodgkin's disease (HD) was investigated by the polymerase chain reaction (PCR) using primers specific to a sequence within the EBV Bam H1W region. EBV genome was detected in 33 of 57 (58 per cent) cases of HD. Viral DNA was, however, also demonstrated in nine of 24 non‐Hodgkin's lymphomas, in three of nine non‐neoplastic lymph nodes and in seven of 12 normal peripheral blood samples used as controls. In all cases, the band obtained following PCR was verified using Southern blotting and hybridization with highly specific Bam H1W probes. The results suggest that the technique is sufficiently sensitive to detect EBV in persistent latent infection in B‐lymphocytes. Distinction between virus present as a possible aetiological agent of malignancy or as a latent infection is not possible when PCR is used under these conditions. The possible role of EBV as an aetiological agent of HD remains unresolved.
Characterization of integrin chains in normal and neoplastic human pancreas
Authors: Hall Peter A. , Coates Philip , Lemoine Nicholas R. , Horton Michael A.
Published: June 2005

Integrins are a complex family of non‐covalently linked heterodimeric glycoproteins which function as cell adhesion molecules, interacting with extracellular matrix molecules such as laminin, fibronectin, vitronectin, and collagen, and also having a role in intercellular adhesion. Each integrin subfamily is characterized by a common β chain associated with variable α chains. We have examined, using immunohistological methods, the expression of the VLA (very late activation) family comprising β1 in association with α1–6, and also α6 in association with β4, the LFA β chain β2, and the vitronectin receptor, in association with β1 or β5 and as the complex αvβ3. Cryostat sections of normal pancreas, pancreatic adenocarcinomas, and ampullary tumours were studied together with six pancreatic carcinoma cell lines. Normal pancreas showed expression of β1 in all parenchyma. α2 and α6 had a similar distribution whereas α3 expression was confined to ducts, including the very smallest radicles. Staining along the basement membrances of ducts was seen with β4 and the anti‐vitronection αv chain receptor antibody 13C2. Islet cells failed to stain with any antibody. No staining of epithelial components was seen with antibodies to α1, α4, α5, or to the αvβ3 form of the vitronectin receptor (β3 and αvβ3 using the antibody 23C6). Pancreatic adenocarcinomas and ampullary tumours showed expression of α2, α3, α6, β1, β4, and the vitronectin receptor (αv associated with β1 or β5). Well differentiated cell lines (BXPC‐3 and Capan‐2) showed an identical phenotype, but less well differentiated lines showed varible loss of one or more integrin chains. The functional role of integrins was quantitated by adhesion assays in which we could assess the ability of pancreatic carcinoma cells to stick to the extracellular matrix substrates fibronectin collagen, and laminin but not to fibrinogen or BSA. Binding of well differentiated cell lines to fibronectin and collagen was, in part, inhibited by exogenous RGD peptide. The distribution of integrins in normal and neoplastic cells and their functional properties were also shown to correlate well with the spatial distribution of laminin, fibronectin, and type IV collagen as shown by immunohistology in normal and neoplastic pancreas.
CD23 expression in non‐Hodgkin lymphoma: Immunohistochemical demonstration using the antibody BU38 on paraffin sections
Authors: Murray Paul G. , Janmohamed Riaz M. I. , Crocker John
Published: June 2005

The leucocyte antigen CD23 is upregulated in the early stages of B‐cell activation by Interleukin‐4 (IL‐4), and functions as an IgE receptor and lymphocyte growth factor. We have studied the expression of CD23 in 68 cases of non‐Hodgkin lymphoma (NHL) using the antibody BU38. This new antibody has the great advantage of being applicable to routinely‐processed paraffin sections. CD23 was expressed in tumour cells in 27 out of 36 cases of low grade NHL and 3 out of 32 cases of high grade NHL. Follicular dendritic cells were strongly positive and were seen in follicular lymphomas. Macrophages were also positive and were numerous in high grade lesions.
MHC antigen expression in human oral squamous carcinoma cell lines
Authors: Mutlu Serdar , Matthewst John B. , Midda Marsh , Scully Crispian , Prime Stephen S.
Published: June 2005

This study quantified the constitutive and interferon‐gamma (IFN‐γ) stimulated expression of MHC class I (HLA‐ABC and β2, microglobulin) and class I1 antigens (HLA‐DR, ‐DP, ‐DQ) on normal and malignant oral keratinocytes using radioimmunoassay and immunocytochemical techniques. Normal keratinocytes and three of four malignant cell lines (H 103, H1 57, H314) expressed MHC class I antigens constitutively; IFN‐γ increased MHC class I expression with significant changes in normals, H157 and H314. Normal keratinocytes expressed significantly more constitutive MHC class I antigens than H103 and H157 and significantly more IFN‐γ stimulated MHC class I antigens than H103, H157 and H314. MHC class II antigens predominantly were not expressed constitutively on normals, H103 and H157 but, in H314, HLA‐DR, ‐DP and ‐DQ antigens were demonstrated on 35, 11 and 5 per cent of cells, respectively, and resulted in a non‐coordinated pattern ofexpression (HLA‐DR > ‐DP = ‐DQ). IFN‐γ induced HLA‐DR on normals, H103 and H157, whilst HLA‐DP and ‐DQ remained undetectable. In H314, IFNγ enhanced HLA‐DR, ‐DP and ‐DQ (significant increase of HLA‐DQ) but the interrelationship between these antigens was maintained (HLA‐DR > ‐DP = ‐DQ). Normal keratinocytes expressed significantly more IFN‐γ stimulated HLA‐DR than H103 and H157 but significantly less HLA‐DR than H314 under similar experimental conditions. One oral malignant cell line (H191) did not express MHC class I and MHC class II antigens either constitutively or in response to IFN‐γ. The results demonstrate aberrant patterns of MHC expression (absence, enhanced, diminished) in the different malignant oral keratinocyte cell lines.
Immunocytochemistry of malignant mesothelioma: OV632 as a marker of malignant mesothelioma
Authors: Delahaye M. , Hoogsteden H. C. , Van Der Kwast Th. H.
Published: June 2005

In pleural of ascitic effusions the cytomorphological distinction of adenocarcinoma cells, reactive mesothelial cells, and malignant mesothelioma cells often causes a diagnostic dilemma. The value of immunocytochemistry was investigated on cytological smears of 24 well‐established cases of malignant mesothelioma, a selected series of 31 metastatic adenocarcinomas, and 20 smears of patients without known malignancy. In these smears we scored the immunoreactivity with a panel of four monoclonal antibodies. In addition to antibodies for epithelial membrane antigen (EMA) and carcinoembryonic antigen (CEA), the monoclonal antibody MOC31 and the ovarian carcinoma specific antibody OV632 were incorporated in the panel. With none of these four antibodies was immunostaining of reactive mesothelial cells found.

CEA‐ and MOC31‐positive tumour cells were frequent in metastatic adenocarcinomas, but occurred rarelay in malignant mesothliomas. EMA‐positive tumour cells were in all metastatic adenocarcinomas (100 per cent) and in most malignant mesotheliomas (83 per cent). In addition to the expected reactivity of OV632 with ovarian carcinomas, 22 of 24 malignant mesotheliomas contained immunopositive tumour cells, while only a small proportion of non‐ovarian adenocarcinomas reacted with this antibody.

This selective staining of malignant mesothelioma cells, but not reactive mesothelial cells, with OV632 now permits the positive identification of malignant mesothelioma cells in male patients.
New monoclonal antibodies reacting with bile ducts: Further insights into the pathogenesis of bile ductular proliferation in biliary diseases
Authors: Ayres Reuben C. S. , Hübscher Stefan G. , Shaw Jean , Garner Chris , Joplin Ruth , Williams Anne , Neuberger James M.
Published: June 2005

We have produced a range of monoclonal antibodies which stain human intrahepatic bile ducts of different sizes. Amongst 26 monoclonal antibodies produced, five clones reacted specifically with bile ducts of different sizes, of which three have been maintained in culture and their viability following freezing and thawing confirmed. Staining patterns varied between normal adult liver tissue, normal fetal liver tissue and a variety of hepatobiliary diseases. The antibodies provide further evidence of the immunological heterogeneity of the human intrahepatic biliary tree and support the hypothesis that proliferating bile ductules are derived from periseptal hepatocytes. The preparation of the antiboides, their staining reactions in normal adult, normal fetal and a variety of liver diseases are described.