bluuebunny/crossref_metadata_2025 huggingface dataset:
Myoendothelial contacts in arteriolosclerosis
Authors: Sosa‐melgarejo Jorge A. , Berry Colin L.
Published: June 2005

Human renal biopsies were examined electron microscopically to investigate close contacts between endothelial and smooth muscle cells in small arterioles. These myoendothelial contacts were seen in the form of cytoplasmic projections passing through fenestrae in the basal lamina. Most of these cell processes seem to arise from the endothelial cells. In the control vessels, the separation between the endothelial cells and smooth muscle cells of the tunica media is 0.09‐0.27 μm. With arteriolosclerosis there is an increasing separation between the elements of the intima and the media, from 1.0 to 2.42 μm. In spite of this increasing separation, myoendothelial contacts maintain an intercellular space of 10–15 nm, as observed in the control vessels. At 2.42 μm of separation, the amount of extracellular material accumulated is such that the cells can no longer keep in contact. Break up of the myoendothelial contacts may be responsible for impairment of communication between the tunica intima and media in the vessel wall in arteriolosclerosis.
Epstein—barr virus latent membrane protein expression by Hodgkin and Reed—Sternberg‐like cells in acute infectious mononucleosis
Authors: Isaacson Peter G. , Schmid Christina , Pan Langxing , Wotherspoon Andrew C. , Wright Dennis H.
Published: June 2005

In the light of reports of latent membrane protein (LMP) expression by Hodgkin and Reed—Sternberg (HRS) cells, paraffin sections of tonsil (two cases), lymph nodes (eight cases; three cervical, one axillary, and four inguinal) and spleen (four cases) from 14 patients with acute infectious mononucleosis (IM) have been examined for the presence of HRS‐like cells and immunostained with an antibody to LMP. Sections of the tonsils and one lymph node were also stained with a panel of antibodies which characterize HRS cells of Hodgkin's disease. The tonsils contained abundant HRS‐like cells, mainly adjacent to the crypts, which were highlighted by strong LMP expression. The immunophenotype of these cells closely, but not completely, resembled that of HRS cells of Hodgkin's disease. The lymph nodes and spleens showed the typical changes of acute IM but only few LMP‐positive HRS‐like cells were present in the cervical lymph nodes and hardly any were present in the inguinal nodes and spleen. These findings suggest that tonsillar crypt squamous epithelium may play a role in the formation of LMP‐positive HRS‐like cells; these cells could be progenitors of Hodgkin's disease HRS cells and, if so, this might explain the restricted sites of presentation of Hodgkin's disease.
Mitochondrial and chromosomal DNA alterations in human chromophobe renal cell carcinomas
Authors: Kovacs Angela , Storkel Stephan , Thoenes Wolfgang , Kovacs Gyula
Published: June 2005

Renal cell tumours are characterized by the loss of chromosome 3p and trisomy of 5q segments (common, nonpapillary renal cell carcinoma), or by trisomy of chromosomes 7 and 17 and loss of the Y chromosome (papillary renal cell carcinoma), or by random karyotype changes and mitochondrial DNA alterations (renal oncocytoma). We have studied by means of RFLP analysis the genomic and mitochondrial DNA in 11 chromophobe renal cell carcinomas, which have a unique morphology among kidney cancers. We found a loss of the constitutional heterozygosity at chromosomal regions 3p, 5q, 17p, and 17q, a combination of allclic losses that has not been found in other types of renal cell tumours. Three of the tumours showed a gross alteration in the restriction pattern of the mitochondrial DNA. A combined morphological and genetic analysis suggests that chromophobe renal cell carcinoma is a distinct entity.
A two‐colour technique for chromosome in situ hybridization in tissue sections
Authors: Lutz Steffi , Welter Cornelius , Zang Klaus D. , Seitz Gerhard , Blin Nikolaus , Urbschat Klaus
Published: June 2005

By extending non‐isotopic in situ hybridization of DNA probes (targeted to metaphase chromosomes or interphase nuclei) to hybridization using tissue sections, additional topological information on the DNA structure and specific alterations can be obtained. We have established a method for the application of two different, chromosome‐specific probes labelled with two colour dyes allowing simultaneous detection of two‐colour signals. This method was tested in and is applicable to tissue sections of various origins. To demonstrate its sensitivity, prostate carcinomas (either as cryosections or as sections from paraffin blocks) were investigated for the presence or absence of chromosomes 1 and Y. The technique presented here, comparable to immunohistochemical staining, is particularly useful for routine application in diagnostic laboratories and testing of fresh or archival material.
Seminal vesicle amyloid: The first example of exocrine cell origin of an amyloid fibril precursor
Authors: Cornwell Gibbons G. , Westermark Gunilla T. , Pitkanen Peter , Westermark Per
Published: June 2005

Amyloid fibrils have been extracted from seminal vesicles, and a dominant 14 kD amyloid fibril protein has been identified. An antiserum to this protein reacted both with amyloid and with epithelial cells in some normal seminal vesicles. These reactions were blocked with seminal vesicle secretion and seminal vesicle amyloid fibril protein, but not by degraded amyloid fibrils or fibril protein from other types of amyloid. It is concluded that seminal vesicle amyloid is derived from secretory proteins of the seminal vesicles. As such, it is the first amyloid described which appears to be the product of an exocrine secretory cell.
‘Pleural’ and pulmonary carcinosarcomas
Authors: Mayall Frederick G. , Gibbs Allen R.
Published: June 2005

The histological distinction between carcinosarcoma and mesothelioma has received little attention. The object of this study was to describe the histology and immunohistochemistry of two carcinosarcomas presenting as pleural tumours. These were compared with 12 carcinosarcomas from sites within the lung, and with the established features of mesothelioma. Histologically and immunohistochemically, these ‘pleural’ carcinosarcomas are similar to conventional pulmonary carcinosarcomas but accurate recognition is hindered by their gross appearance and biphasic histology which closely mimic mesothelioma. They may exhibit features such as neutral mucin, expression of carcinoembryonic antigen, squamous differentiation, and neuroendocrine differentiation which, when present, are evidence against mesothelioma. These tumours are rare and have passed unnoticed until now.
Flow cytometry of oesophageal mucosal biopsies; epidermal growth factor receptor, and CD15
Authors: Hopwood D. , Jankowski J. , Milne G. , Wormsley K. G.
Published: June 2005

Flow cytometry may be used to examine the properties of single or isolated cells. We have shown that it is possible to disaggregate and label oesophageal epithelial cells for two surface markers, CD15 and epidermal growth factor receptor. We have previously demonstrated these markers, in oesophageal squamous cells using immunoperoxidase techniques. These labelled disaggregated cells could then be measured by flow cytometry.
Collagen composition and ultrastructure of the so‐called amianthoid fibres in palisaded myofibroblastoma. Ultrastructural and immunohistochemical study
Authors: Skélové A. , Michal M. , Chlumské A. , Leivo I.
Published: June 2005

A very prominent feature of the recently described intranodal palisaded myofibroblastoma is the occurrence of stellate crystalline extracellular deposits known as ‘amianthoid fibres’, and thought to represent abnormally thick collagen fibrils. We showed at the ultrastructural level that these deposits are composed of fibrils with the periodicity and width typical of normal native collagen. Using antibodies directed against various extracellular components, we have determined that these structures contain type I and III collagens Our results indicate that the crystalloid deposits in intranodal palisaded myofibroblastoma called ‘amianthoid fibres’ do not contain giant collagen fibrils widely regarded as characteristic of the morphological concept ‘amianthoid’ in other locations.
Induction of emphysema and bronchial mucus cell hyperplasia by intratracheal instillation of lipopolysaccharide in the hamster
Authors: Stolk Jan , Rudolphus Arjan , Davies Philip , Osinga Don , Dijkman Joop H. , Agarwal Lilly , Keenan Kevin P. , Fletcher Dan , Kramps Johannes A.
Published: June 2005

The aim of this study was to determine whether lipopolysaccharide‐induced elastase release from recruited neutrophils in the hamster lung would induce emphysema, measured by mean linear intercept (Lm) and bronchial mucus cell hyperplasia (BMCH), scored in tissue sections stained with periodic acid‐Schiff. Lipopolysaccharide (LPS) was instilled transorally twice a week for up to 5 weeks in hamsters. At 4 weeks after seven LPS instillations, Lm amounted to 87.6 ± 1.2μm, while it was 68.3 ± 1.5 μm after seven saline instillations (P < 0.01). At 6 months after the sixth LPS instillation, the Lm of these lungs was 83.3 ± 1.6 μm, indicating irreversible tissue destruction. LPS‐treated hamsters showed marked to severe BMCH, which was most evident in large intrapulmonary airways. Instillations of highly selective inhibitor of hamster PMN elastase resulted in 50 per cent inhibition of LPS‐induced emphysema. The development of BMCH was inhibited by approximately 35 per cent by this agent.

To study the response in time of cellular infiltration after a single LPS instillation, the lungs of groups of four hamsters were lavaged at different time points. PMN recruitment showed peak values at 4 and 48 h after LPS instillation and returned to baseline values at 96 h. Simultaneous intratracheal instillation of LPS and anti‐TNFα antiserum resulted in a considerable reduction of neutrophil influx into bronchoalveolar spaces in the first 6 h after instillation.
Alteration of stromal protein and integrin expression in breast—a marker of premalignant change?
Authors: Jones J. Louise , Critchley D. R. , Walker Rosemary A.
Published: June 2005

Interactions between epithelia and the extracellular matrix are important in the medulation of cellular growth, differentiation, and motility. To investigate the possible roles of these interactions in the neoplastic process, this study examines the expression of the integrin subunits α2, α6, β1, and β4 and the stromal protein tenascin in 53 breast carcinomas, non‐involved breast tissue from 21 of these cases, and 32 normal/benign cases. Frozen tissue and an indirect immunoperoxidase technique were used throughout. Linear staining in relation to the basement membrane was seen for all integrins in the normal/benign cases. The carcinomas showed complete loss of reactivity in 65 per cent of cases for α2, 80 per cent for α6 and β4, and 90 per cent for β1. Those showing reactivity displayed a diffuse cytoplasmic type of staining. The non‐involved breast tissue showed linear basement membrane type staining with α2 and βl, but for α6 and β4 66 per cent of cases displayed reactivity identical to that of the corresponding tumour. For tenascin, band‐like staining around ducts was seen in normal/benign cases, with a diffuse coarse reactivity in all carcinomas. Most non‐involved cases stained as for normal breast. The altered α6β4 integrin staining in non‐involved tissue in cancerous breast may be an early event in the neoplastic process, and as such, may be of use as a marker of pre‐malignant change.
Proliferating cell nuclear antigen (PCNA) expression in Hodgkin's disease
Authors: Schmid Christine , Sweeney Edel , Isaacson Peter G.
Published: June 2005

Previous studies of the proliferating cell fraction in Hodgkin's disease (HD) have been directed towards the classical Hodgkin and Reed–Sternberg cells (HRS) to the exclusion of the background population and have not included cases of nodular lymphocyte predominant Hodgkin's disease (NLPHD). Using an antibody to proliferating cell nuclear antigen (PCNA), we have determined the growth fraction of HRS cells and L&H cells in paraffin sections of 15 cases of classical HD [12 nodular sclerosis (NS), 3 mixed cellularity (MC)] and eight cases of NLPHD. By double staining with anti‐PCNA and antibodies to B cells (CD20) and T cells (CD45RO), we also determined the growth fraction and immunophenotype of the background population in each case. In classical HD, 50·4 per cent of HRS cells were PCNA‐positive and judged to be proliferating, which is comparable to previous studies, while in NLPHD 76·9 per cent of L&H cells were PCNA‐positive. In both classical HD and NLPHD, the majority of PCNA‐positive cells in the background were T cells, which showed a growth fraction of 57·8 and 68·5 per cent, respectively; in comparison, only 4 per cent of B cells were PCNA‐positive in each type of HD. L&H cells are widely accepted to be B cells and there is growing evidence that HRS cells are also B cell‐derived. Our results underline a relationship between classical HD and NLPHD and suggest that the characteristic histological features of both diseases may be caused by the production and release of cytokines from altered B cells.
Mitotic indices, anti‐PCNA immunostaining, and AgNORs in thick cutaneous melanomas displaying paradoxical behaviour
Authors: Evans A. T. , Blessing K. , Orrell J. M. , Grant A.
Published: June 2005

Those melanomas which fail to behave as expected from their Breslow thickness provide interesting material for study. In an attempt to explain differences in behaviour, we have evaluated three distinct proliferative markers in 23 thick melanomas which failed to metastasize and in 20 well‐matched control tumours with documented metastasis. The test group demonstrated significantly greater numbers of mitoses when expressed as an index (mitoses per 1000 cells), whilst no difference was found when the results were expressed in terms of mitoses per unit area. Tumours showing epidermal ulceration possessed higher mitotic indices than those of non‐ulcerated lesions. High fractions of PCNA immunolabelling combined with low mitotic indices were observed frequently in the non‐metastasizing group. This result and its possible relation to survival advantage are discussed in detail. Both AgNOR numbers and patterns failed to act as prognostic variables—indeed, AgNORs failed to correiate with the other proliferative indices, suggesting that their value as a marker of tumour growth is severely limited.
Retinoblastoma and p53 gene expression related to relapse and survival in human breast cancer: An immunohistochemical study
Authors: Sawan Ali , Randall Barbara , Angus Brian , Wright Chris , Henry James A. , Ostrowski Julian , Hennessy Colm , Lennard T. W. J. , Corbett Ian , Horne C. H. W.
Published: June 2005

Inactivation of tumour suppressor genes may be an important aetiological factor in many human cancers including breast. In a study of 197 breast cancer patients, tumour tissue was snap‐frozen at the time of surgery and immunohistochemical labelling for p53 protein and retinoblastoma (Rb) gene product carried out using an indirect immunohistochemical technique. Tumours were scored by two independent observers for the intensity of nuclear staining for each antibody. Expression of p53 protein showed a significant association with a shorter time to relapse (P = 0·03) and death (P = 0·02) (log rank test). p53 expression did not correlate with nodal status but showed a significant association with high tumour grade (P = 0·001). Rb gene expression showed no relationship to relapse or survival but loss of expression showed a significant correlation with positive lymph node status. The manner by which these proteins might act to determine tumour behaviour remains to be established.
p53 Immunohistochemistry in malignant fibrous histiocytomas and other mesenchymal tumours
Authors: Soini Ylermi , Vähäkangas Kirsi , Nuorva Kyösti , Kamel Dia , Lane David P. , Pääkkö Paavo
Published: June 2005

In this study we analysed by immunohistochemistry the expression of p53 protein in 14 malignant fibrous histiocytomas (MFHs), 22 other types of sarcoma (eight leiomyosarcomas, four rhabdomyosarcomas, four liposarcomas, two fibrosarcomas, two chondrosarcomas, one malignant schwannoma, and one dermatofibrosarcoma protuberans), and 25 non‐malignant mesenchymal lesions (eight dermatofibromas, four cases of nodular fasciitis, three leiomyomas, three fibromatoses, two epithelioid leiomyomas, two neurofibromas, one schwannoma, one myositis ossificans, and one giant cell tumour of tendon sheath). Four MFHs and nine other types of sarcoma (four leiomyosarcomas, two chondrosarcomas, one liposarcoma, one fibrosarcoma, and one dermatofibrosarcoma protuberans) showed nuclear positivity for p53. Of the benign soft tissue lesions, p53 positivity was observed in two fibromatoses, one nodular fasciitis, and one dermatofibroma. The number of p53‐positive cells in these benign lesions was considerably smaller than that in most of the p53‐positive sarcomas.

The p53 positivity in MFHs and other types of sarcoma indicates that p53 gene alterations may play a part in the neoplastic transformation of these tumours. The occurrence of p53 positivity in benign mesenchymal lesions suggests that sometimes p53 protein may accumulate in cells without an associated malignancy. Because of this, p53 immunoreactivity cannot, by itself, be used as a criterion of malignancy According to our results, p53 positivity in over 1 per cent of tumour cells in mesenchymal lesions favours malignancy.
Inhibin (10.7kD prostatic peptide) in normal, hyperplastic, and malignant human endometria: An immunohistochemical study
Authors: Teni Tanuja R. , Sampat Mrudula B. , Sheth Nandini A.
Published: June 2005

The expression of inhibin, a 10.7 kD follicle‐stimulating hormone (FSH)‐suppressing prostatic peptide of 94 amino acids, was investigated in normal human endometrium, endometrial hyperplasia, and adenocarcinoma, employing the avidin‐biotin immunoperoxidase technique. The antiserum used was raised in rabbits against prostatic inhibin isolated from human seminal plasma. The study included 15 well differentiated, 32 moderately differentiated, and 21 poorly differentiated endometrial adenocarcinomas; 26 simple, five complex, and two complex atypical endometrial hyperplasias; and, for comparison, 25 normal proliferative and 30 normal secretory endometria. In malignant and hyperplastic endometrial tissues, inhibin was localized in the epithelial cytoplasm of endometrial glands while the stroma showed weak reactivity. On the other hand, inhibin was undetectable in the early proliferative phase, but was present on the luminal border of the glandular epithelium in the mid‐ and late proliferative phases. Secretory endometrium displayed strong inhibin reactivity in the cytoplasm of glandular epithelium and in the stroma. The increased inhibin reactivity in secretory endometrium as compared with the proliferative phase is indicative of a functional role for inhibin in the uterus. In addition, its localization in proliferative, hyperplastic, and malignant endometria suggests a possible regulatory role for inhibin in endometrial proliferation and growth.
Diagnostic usefulness of dipeptidyl aminopeptidase IV monoclonal antibody in paraffin‐embedded thyroid follicular tumours
Authors: Kotani T. , Asada Y. , Aratake Y. , Umeki K. , Yamamoto I. , Tokudome R. , Hirai K. , Kuma K. , Konoe K. , Araki Y. , Ohtaki S.
Published: June 2005

Monoclonal antibodies to dipeptidyl aminopeptidase IV (DAP IV, EC 3.4.14.5) were raised and selectively applied to paraffin‐embedded sections of thyroid carcinoma. Five monoclonal antibodies were found to stain paraffin sections of thyroid carcinomas. Using one of these antibodies (44‐4), we studied retrospectively aberrant expression of DAP IV in thyroid carcinoma to determine whether immunohistochemical staining with DAP IV antibody is useful in pathological diagnosis.

In almost all cases of thyroid follicular and papillary carcinoma, tumour cells were positive (99.0 per cent) with DAP IV, whereas the cases of follicular adenoma showed a low incidence (27.1 per cent) of positive staining. Follicular adenoma with incomplete capsular invasion had a higher positive incidence (50 per cent) than follicular adenoma without incomplete capsular invasion (9.6 per cent).

In positive staining cases previously diagnosed as benign tumours, 11 benign cases reacting positively with DAP IV were rediagnosed as carcinoma after re‐examinatioan of more thyroid paraffin block sections or serial sections.

These findings suggest that DAP IV monoclonal antibody is very useful in distinguishing thyroid follicular carcinoma from follicular adenoma.
In situ Hybridization demonstrates the stability of mRNA in post‐mortem rat tissues
Authors: Walker E. , McNicol A. M.
Published: June 2005

In situ hybridization was used to detect messenger RNA (mRNA) in a variety of rat tissues which were fixed in formalin either immediately after death or after a 24 h period of storage at 5°C. A synthetic polydeoxythymidine [poly d(T)] oligonucleotide probe was used to demonstrate poiyadenylated [poly (A)] mRNA in the small intestine, pancreas, liver, cerebellum, and pituitary. Of these tissues, only the liver showed a small reproducible reduction in hybridization signal following delayed fixation. Synthetic oligonucleotide probes complementary to albumin and pro‐opiomelanocortin (POMC) mRNAs were hybridized to liver and pituitary, respectively. There was no significant reduction in hybridization signal in post‐mortem tissues. The results suggest that some mRNAs may be remarkably stable under certain postmortem conditions and this should encourage the wider application of in situ hybridization techniques to post‐mortem material.
Differential expression of the mitochondrial gene cytochrome oxidase ii in benign and malignant breast tissue
Authors: Sharp Matthew G. F. , Adams Susan M. , Walker Rosemary A. , Brammar William J. , Varley Jennifer M.
Published: June 2005

Comparative analysis of expression levels of genes in benign and malignant tumours of the breast has been performed. Differential screening of cDNA libraries identified four genes of the mitochondrial genome as being expressed at different levels in the two tissues compared, but further investigations showed that only the gene encoding subunit 2 of cytochrome c oxidase (COII) is expressed at significantly higher levels in carcinomas compared with fibroadenomas. The mitochondrial genes encoding subunits 2 and 4 of NADH dehydrogenase, and subunit 6 of F0F1 ATPase, were not found to be differentially expressed in carcinomas and fibroadenomas. All four genes were expressed in the epithelium of human breast carcinomas, as shown by in situ hybridization. The expression level of the COII gene is also correlated with carcinoma grade. No gross alterations to the mitochondrial DNA from these tumours could be detected. The possible implications of these results on the behavioural differences between fibroadenomas and carcinomas are discussed.
Immunoreactivity to ubiquitin—protein conjugates is present early in the disease process in the brains of scrapie‐infected mice
Authors: Lowe James , Fergusson Jill , Kenward Nigel , Laszlo Lajos , Landon Michael , Farquhar Christine , Brown John , Hope James , Mayer R. John
Published: June 2005

Brains from mice infected with either the 87V or the ME7 strains of mouse‐passaged sheep scrapie were taken at stages during the disease process and immunostained to show the localization of ubiquitin‐protein conjugates. In both models, conjugates were seen as fine, dot‐like structures; as coarser, granular lesions within or adjacent to neurones; and in areas surrounding plaques. The dot‐like structures were visible at 28 days post‐ME7 infection and at 55 days in 87V‐infected mice. In both models, the extent of immunoreactive changes increased as the disease progressed and terminal infection was as described earlier by us (Lowe et al., J. Pathot 1990; 162: 61–66). The patterns of development of these features were distinctive in two ways: progression from region to region was observable and the density of the pathological lesions grew exponentially as the clinical symptoms appeared. The earliest pathological dot‐like structures corresponded temporally with the earliest detection of PrPsc by Western blotting, and immunogold electron microscopic investigation of the dot‐like lesions indicated that they were the multi‐vesicular, lysosome‐related, dense bodies that we have described previously in terminal disease (Laszlo et al., J Pathol 1992; 166: 333–341).

Until now, ubiquitin–protein conjugates were seen mainly in inclusion bodies associated with the terminal stages of a range of human degenerative diseases. This study establishes that ubiquitin–protein conjugates accumulate in lysosome‐related bodies very early and appear to be intimately related to the pathological processes in the animal disorders that we have studied.
KiSl—A novel monoclonal antibody which recognizes proliferating cells: Evaluation of its relationship to prognosis in mammary carcinoma
Authors: Sampson Steven A. , Kreipe Hans , Gillett Cheryl E. , Smith Paul , Chaudary Murid A. , Khan Ashraf , Wicks Kimberly , Parwaresch R. , Barnes Diana M.
Published: June 2005

Immunohistochemical staining with a novel monoclonal antibody, KiSl, which recognizes a cell cycle‐associated antigen, was investigated in 142 cases of stage I and II invasive breast carcinoma. KiS 1 staining indices were compared with disease‐free interval, overall survival, and post‐relapse survival. Using a semi‐quantitative method of assessment, we found that tumours with a high level of staining (34/142,24 per cent) had a significantly worse prognosis than those with a low level of staining (108/142, 76 per cent). Significant correlations were found between KiS1 staining and disease‐free interval (P<0·001), overall survival (P<0·001), and post‐relapse survival (P=0·008). A more time‐consuming, quantitative method of assessment gave similar results. Cox multivariate analysis showed these results to be independent of nodal status, histological type, and grade of tumour (P=0·01). We conclude that KiS1 is a valuable new antibody which affords useful prognostic information in breast carcinoma. As it can be used in formalin‐fixed, paraffinembedded material, it may be of particular use in the study of small lesions such as those identified in the Breast Cancer Screening Programme.
Isologous monoclonal antibodies can induce anti‐GBM glomerulonephritis in rats
Authors: Sado Yoshikazu , Kagawa Megumi , Rauf Syarifuddin , Naito Ichiro , Moritoh Chiharu , Okigaki Tohru
Published: June 2005

Injection of isologous monoclonal antibodies (SR2, SR3) caused anti‐glomerular basement membrane antibody‐induced glomerulonephritis (anti‐GBM nephritis) in WKY/NCrj rats. The antibodies were obtained from hybridoma cells derived from fusion of the spleen of a nephritic WKY/NCrj ral injected with rat solubilized renal basement membranes with adjuvant, and mouse SP2 myeloma cells. They belonged to the rat IgG2a subclass and bound to rat kidney in a linear pattern along the glomerular and tubular basement membranes. Histological changes in glomeruli were detected at day 1 after the injection; proteinuria with haematuria appeared on day 2; and proteinuria became severe and reached a plateau by day 5. These results demonstrate that anti‐GBM nephritis can even be induced by an isologous monoclonal antibody and that the rat IgG2a subclass is at least nephritogenic. The experimental model of anti‐GBM nephritis with isologous monoclonal antibodies makes it possible and easier to analyse further the mechanism of anti‐GBM nephritis.
Distribution and localization of Epstein–Barr virus subtypes A and B in AIDS‐related lymphomas and lymphatic tissue of HIV‐positive patients
Authors: Borisch B. , Finke J. , Hennig I. , Delacréataz F. , Schneider J. , Heitz Ph. U. , Laissue J. A.
Published: June 2005

Tissue samples of 421 HIV‐positive patients have been studied for the presence and distribution of Epstein–Barr virus (EBV) subtypes A and B. This was done by PCR, EBER in situ hybridization, and immunohistochemical detection of EBV latent membrane protein (LMP) in AIDS‐associated malignant lymphomas (16 cases) and lymphatic organs of patients without lymphoma (5 cases). Eleven cases were considered to be EBV‐positive, with type A in four, and type B virus in four other cases. In patients with malignant lymphoma, EBV was localized in tumour tissue exclusively. One patient without lymphoma presented with multiple EBV genome type B‐positive cells in all the lymphoid tissue samples examined. In HIV‐positive patients, both subtypes of EBV, A and B, may play a role in the pathogenesis of lymphoproliferative lesions.
Aberrant expression of the tumour suppressor gene p53 is very frequent in Wilms' tumours
Authors: Lemoine Nicholas R. , Hughes Christine M. , Cowell John K.
Published: June 2005

A series of 34 Wilms' tumours have been analysed for abnormal expression of the tumour suppressor gene p53 using frozen section immunohistochemistry. All tumours showed immunoreactivity with at least one of the specific antibodies used (monoclonal antibody PAb240, polyclonal antibodies CM 1 and JG8). Abnormalities of p53 expression are very frequent in this type of childhood tumour.
Autoradiographic localization of endothelin‐1 binding sites in human colonic cancer tissue
Authors: Inagaki Hiroshi , Bishop Anne E. , Eimoto Tadaaki , Polak Julia M.
Published: June 2005

Endothelin‐1, a potent vasoconstrictor, has been reported to stimulate mitogenesis in various types of normal and neoplastic cells and to be involved in neurotransmission. Recently, many human cancer cell lines, including those from the human colon, have been shown to produce endothelin. In this study, the occurrence of endothelin‐1 binding sites was investigated in human colonic cancer tissues using in vitro autoradiography. Specific [125I]endothelin‐1 binding sites were identified over tumour vessels and stromal tissues surrounding cancer cell nests. The distribution was heterogeneous, and dense silver grains were localized, especially over clusters of fibroblasts adjacent to the cancer cell nests. Endothelin binding was minimal in the cancer cells, as in the normal crypt epithelium. Quantitative analysis of the autoradiographs demonstrated high affinity (Kd = 0·50 ± 0·06 nM; mean ± SEM) binding sites, with a maximum binding capacity (Bmax) of 40 ± 3·2 amol/mm2 in the cancer tissues. Our results provide evidence that specific endothelin‐1 binding sites are expressed in the stromal tissues including tumour vessels, fibroblasts, and nerve fibres. Endothelin‐1 may play a modulatory role in blood supply, mitogenesis, and neurotransmission in a paracrine fashion through the stromal components in human colonic cancers.
The erbB‐3 Gene in human pancreatic cancer
Authors: Lemoine Nicholas R. , Lobresco Maria , Leung Hing , Barton Claire , Hughes Christine M. , Prigent Sally A. , Gullick William J. , Klöppel Günter
Published: June 2005

Abnormalities of the type 1 growth factor receptor family have been implicated in the pathogenesis of pancreatic cancer. There is evidence for a potential autocrine loop involving overexpression of the epidermal growth factor (EGF) receptor and its ligands, as well as overexpression of the erbB‐2 receptor. A third member of this receptor family, erbB‐3, has recently been recognized and found to be abnormally expressed in some types of human cancer. In this study we show that overexpression of the erbB‐3 protein occurs very frequently in carcinoma of the exocrine pancreas and also in chronic pancreatitis. We found no evidence of amplification or rearrangement of the erbB‐3 gene by Southern blot analysis of DNA from pancreatic cancer cells lines.
Production and characterization of a polyclonal antibody to the c‐erbB‐3 protein: Examination of c‐erbB‐3 protein expression in adenocarcinomas
Authors: Poller D. N. , Spendlove I. , Baker C. , Church R. , Ellis I. O. , Plowman G. D. , Mayer R. J.
Published: June 2005

A polyclonal rabbit antibody was raised to the c‐erbB‐3 protein using a synthetic peptide corresponding to amino acids 1229–1241 of the predicted protein sequence of c‐erbB‐3. In Western blot analysis this antibody detects a single band at ˜165kD in a c‐erbB‐3 transfected (293/HER‐3) human cell line. c‐erbB‐3 protein expression was then examined in a variety of adenocarcinomas. Expression of c‐erbB‐3 protein was indicated by membrane and/or cyto‐plasmic tumour cell immunoreactivity in formalin‐fixed, paraffin‐embedded tissue sections. c‐erbB‐3 protein was detected in a series of 13 out of 14 primary breast carcinomas, 3 of 5 gastric adenocarcinomas, 8 of 9 colonic adenocarcinomas, 2 of 9 prostatic adenocarcinomas, 0 of 6 renal cell carcinomas, 1 of 4 primary lung adenocarcinomas, and 5 of 7 endometrial adenocarcinomas. Immunohistochemical expression of the c‐erbB‐3 protein appears to be a relatively common event in adenocarcinomas, and further studies are now warranted to establish the role of the c‐erbB‐3 protein in neoplasia.
Detection of capsid antigen of human papillomavirus (HPV) in benign lesions of female genital tract using anti‐HPV monoclonal antibody
Authors: Iwasaki Takuya , Sata Tetsutaro , Sugase Motoyasu , Sato Yuko , Kurata Takeshi , Suzuki Kenji , Ohmoto Hiroshi , Iwamoto Seiichi , Matsukura Toshihiko
Published: June 2005

We established a murine monoclonal antibody (K1H8) to human papillomavirus (HPV) using alkaline‐disrupted virions of HPV type 1 (HPV‐l) as the immunogen. K1H8 recognized a 57 kD capsid protein of HPV‐1 and detected the antigen in paraffin sections of formalin‐fixed tissue. With K1H8, we examined immunchistochemically 68 biopsy specimens obtained from the female genital tract. The specimens were histologically condyloma acuminatum or koilocytotic lesions with or without dysplasia and each specimen was found to harbour a single type of genital HPV, such as types 6, 11, 16, 18, 31, 33, 42, 51, 52, 56, and 58, by Southern blot hybridization analysis. The antigen was localized in the nuclei and occasionally in the cytoplasm of squamous cells showing koilocytotic changes. Eighty‐four per cent of the specimens (57 cases) showed positivity for the antigen, indicating that K1H8 is a broadly‐reactive antibody to various genital HPVs. The results suggest that benign mucosal lesions of the female genital tract are more frequently associated with viral production and are a potential source of transmission.
High incidence of EBV genome in CD30‐positive non‐Hodgkin's lymphomas
Authors: Kanavaros P. , Jiwa N. M. , De Bruin P. C. , Valk P. Van Der , Noorduyn L. A. , Van Heerde P. , Gordijn R. , Horstman A. , Mullink R. , Willemze R. , Walboomers J. M. M. , Meijer C. J. L. M.
Published: June 2005

In Hodgkin's disease, Epstein–Barr virus (EBV) is found in CD30‐positive Reed–Sternberg cells. We therefore studied 60 CD30‐positive non‐Hodgkin's lymphomas (NHLs) for the presence of EBV by the polymerase chain reaction (PCR) and DNA in situ hybridization (DISH), and by immunohistochemistry for the latent EBV proteins LMP and EBNA‐2. CD30‐negative NHLs and reactive lymph nodes served as controls. The CD30‐positive cases comprised 17 anaplastic large cell lymphomas (ALCLs) (> 75 per cent CD30‐positive cells) and 43 non‐ALCLs (with 5–35 per cent CD30‐positive cells). By PCR, 40 of 60 CD30‐positive NHLs (67 per cent) were EBV‐positive; in CD30‐negative cases, 6/29 (21 per cent) were EBV‐positive, as were 12/50 (24 per cent) reactive lymph nodes. The DISH procedure demonstrated the EBV genome exclusively in the nuclei of tumour cells in 23 of the 37 PCR EBV‐positive cases that were tested. PCR‐negative cases were always DISH‐negative, as were the PCR‐positive reactive lymph nodes and CD30‐negative NHLs. Immunohistochemistry demonstrated LMP in neoplastic cells of 7/47 (15 per cent) CD30‐positive NHLs, both ALCL and non‐ALCL always in PCR EBV‐positive cases, but never in the two control groups. EBNA‐2 staining could not be detected. It is concluded that EBV is present (and transcriptionally active) in a sizeable number of NHLs and an association between the presence of the EBV genome and CD30 expression seems likely.
Changes in hepatic lymph vessels in endotoxaemia
Authors: Shibayama Yuro , Urano Toru , Nakata Katsuji
Published: June 2005

This study was undertaken into rats to investigate changes in the hepatic lymph vessels and the space of Disse in endotoxaemia and to examine their relationship with the development of endotoxin‐induced hepatic injury. Lymph stasis, namely dilatation of the lymph vessels and oedema, developed rapidly in the medium‐sized portal canals, the large portal canals, and the liver hilum after endotoxin injection, but not in the small portal canals. Such changes reached their maximum 4–8 h after endotoxin injection and had recovered markedly by 16 h after the injection. The space of Disse remained within normal limits during this period. These findings suggest that the intrahepatic lymph stasis in endotoxaemia may be caused by a reduction in the pumping activity of the extrahepatic and the intrahepatic large lymph vessels rather than by an increase of lymph formation in the liver lobules. There was no evidence suggesting a direct relationship between the disturbance of hepatic lymph flow and the development of hepatic injury in endotoxaemia.
Monoclonal antibodies against recombinant parts of the Ki‐67 antigen (MIB 1 and MIB 3) detect proliferating cells in microwave‐processed formalin‐fixed paraffin sections
Authors: Cattoretti Giorgio , Becker Michael H. G. , Key Göran , Duchrow Michael , Schlüuter Carsten , Galle Jürgen , Gerdes Johannes
Published: June 2005

The monoclonal antibody Ki‐67 reacts with a human nuclear cell proliferation‐associated antigen that is expressed in all active parts of the cell cycle. Recently we have raised monoclonal antibodies, MIB 1–3, against recombinant parts of the Ki‐67 antigen. These antibodies are true Ki‐67 equivalents, as demonstrated by immunostaining of fresh specimens, biochemistry, and molecular biological techniques. Formalin‐fixed, paraffin‐embedded sections routinely processed for immunohistochemistry failed to stain for Ki‐67 and MIB 2. Antibodies MIB 1 and MIB 3 labelled mitotic figures, while non‐mitotic proliferating cells were negative under these conditions. However, when dewaxed microwave oven‐ processed paraffin sections of formalin‐fixed tissues were used, MIB 1 and MIB 3 gave strong nuclear staining of those cells presumed to proliferate under a variety of normal and neoplastic conditions. Moreover, routine decalcification or depigmentation techniques did not alter the immunoreactivity of MIB I and MIB 3 with microwave‐processed paraffin sections. This method is highly reproducible, easy to perform at low cost, and no additional technical skill is needed because after microwave treatment just routine immunohistochemical methods are used. Since we have successfully applied this new method to sections obtained from paraffin blocks stored for a long time (in one case more than 60 years), the assessment of cell kinetics through the detection of Ki‐67 antigen is now possible on archival material collected in histopathology departments all over the world.
Epstein–Barr virus (EBV) and Hodgkin's disease in children: Incidence of EBV latent membrane protein in malignant cells
Authors: Weinreb M. , Day P. J. R. , Murray P. G. , Raafat F. , Crocker J. , Parkes S. E. , Coad N. A. G. , Jones J. T. , Mann J. R.
Published: June 2005

Previous studies have detected EBV DNA by Southern blotting or in situ hybridization in biopsy material from up to 30 per cent of adult cases of Hodgkin's disease. Here we have used monoclonal antibodies specific for the EBV latent membrane protein LMP1 to examine archival material from children with Hodgkin's disease. Material from 74 cases (54 males and 20 females) was examined and 37 (30 males and 7 females) were classified as LMPl‐positive in the malignant cells. LMP1 positivity was present in 4/13 (31 per cent) of lymphocyte predominant, 14/36 (39 per cent) of nodular sclerosis, 17/20 (85 per cent) of mixed celiularty, 1/2 (50 per cent) of lymphocyte depletion, and 1/3 (33 per cent) of unclassified subtypes. The positive cases by clinical stage were 19/22 (41 per cent), II 9/20 (45 per cent), III 11/24 (46 per cent), and IV 8/8 (100 percent). LMP1 positivity was present in 2/5 (40 percent) children aged less than 5 years, 12/ 27 (44 per cent) aged 5–10 years, and 23/42 (48 per cent) aged between 10 and 15 years. The association between EBV and Hodgkin's disease in children thus appeared to be more frequent in patients with mixed cellularity and advanced disease, but examples of EBV‐positive tumours were found in all histological subtypes, stages, and ages. Stepwise discriminant function analysis showed that clinical stage IV and mixed cellularity histology are independently associated with LMP1 positivity. These observations indicate that Hodgkin's disease in children is at least as strongly linked to EBV as is the disease in adults.
Immunoreactivity for p53 protein in malignant mesothelioma and non‐neoplastic mesothelium
Authors: Ramael Marc , Lemmens Greet , Eerdekens Christa , Buysse Corinne , Deblier Ivo , Jacobs Werner , Van Marck Eric
Published: June 2005

The presence of p53 protein in non‐neoplastic pleural mesothelium (40 cases) and in human malignant mesothelioma (36 cases) was assessed immunohistochemically using the antibodies DO7, CM‐1 and PAb240. In a quarter of the malignant mesotheliomas, there was nuclear immunoreactivity for p53 protein with both the DO7 and CM‐1 antibodies. There were no statistically significant differences between the various mesothelioma subtypes (P>0·05). No immunoreactivity was found with the PAb240 antibody, suggesting absence of mutant‐type p53 protein. Non‐ neoplastic mesothelium was not immunoreactive with any of the antibodies. We conclude that there is immunoreactivity for p53 protein in some mesotheliomas. p53 protein immunoreactivity could be used to discriminate between neoplastic and reactive mesothelium.
Dominantly inherited tubular aggregate myopathy
Authors: Cameron C. H. Stuart , Allen Ingrid V. , Patterson Victor , Avaria Maria A.
Published: June 2005

We report an unusual familial myopathy characterized morphologically by the presence of large tubular aggregates in all fibre types. Two patients, a father and daughter, presented with slowly progressive proximal weakness, limitation of eye movement, and Achilles tendon contractures. Serum creatine kinase was 5–10 times normal. Light microscopy revealed type I fibre predominance. Basophilic accumulations, which stained intensely with the NADH‐TR reaction, were present in both fibre types. Electron microscopy revealed that these consisted of tightly packed parallel tubular arrays. These varied somewhat in their ultrastructural appearance and were classified accordingly as type I, II, and III tubular structures. The tubular aggregates appear to be derived from the sarcoplasmic reticulum. This report further supports the evidence of a distinct clinico‐pathological entity of genetic origin.
Epstein–Barr virus in Reed–Sternberg G‐like cells in non Hodgkin's lymphomas
Authors: Khan G. , Norton A. J. , Slavin G.
Published: June 2005

In the course of our study on Hodgkin's disease (HD), ten cases of non‐Hodgkin's lymphomas (NHL) containing Hodgkin and Reed‐Sternberg‐like (MRS) cells were encountered. Many of these cases had initially been diagnosed as HD, but on careful review of the histology, with the aid of immunophenotyping studies, they were reclassified as NHL. The presence of Epstein–Barr virus (EBV) in these HRS‐like cells was investigated using a combination of EBER in situ hybridization (ISH) and immunostaining for the detection of EBV‐encoded latent membrane protein (LMP). HRS‐like cells in four cases (two lymphoplasmacytoid lymphomas, one Richter's transformation of lymphoplasmacytoid lymphoma, and one immunoblastic lymphoma of T‐cell type) were found to be EBV‐positive. In two of these cases, a second biopsy taken up to 10 years later also contained EBV in the HRS‐like cells. In three of the four cases, HRS‐like cells expressed the activation antigen CD30, but the expression of B‐ or T‐cell antigens was variable. All cases of T‐cell‐rich B‐cell lymphomas were negative for EBV. In conclusion, EBV may play a role in the development of HRS‐like cells i some cases of NHL. The relationship of HRS‐like cells to HRS cells of HD is discussed.
Immunohistochemical analysis of p53 protein overexpression in normal, premalignant, and malignant tissues of the cervix uteri
Authors: Holm Ruth , Skomedal Hanne , Helland åslaug , Kristensen Gunnar , Bøsrresen Anne‐Lise , Nesland Jahn M.
Published: June 2005

Two hundred and thirty‐eight cervical lesions ranging from normal to malignant were examined for overexpression of p53 protein. Whereas p53 protein was identified in 62 per cent of invasive squamous cell carcinomas, 11 per cent of invasive adenocarcinomas, and 7 per cent of squamous cell carcinomas in situ, no staining was found in adenocarcinoma in situ, dysplastic tissue, condyloma, and normal tissue. In 9 per cent of the positive cases of invasive squamous cell carcinomas. 5‐50 per cent of the tumour ceiis were immunoreactive for p53 protein, whereas the other positive specimens were characterized by only rare p53‐positive cells. We conclude that in invasive cervical carcinomas widespread overexpression of p53 protein is unusual, but occasional positive nuclei can be found frequently. Furthermore, our results indicate that altered expression of p53 protein may be involved in the progression of cervical carcinomas.
Immunocytochemical localization of c‐erbB‐2 protein in transitional cell carcinoma of the urinary bladder
Authors: Coombs L. M. , Oliver S. , Sweeney E. , Knowles M.
Published: June 2005

The level of expression and cellular localization of the c‐erbB‐2 gene product in transitional cell carcinoma of the urinary tract is controversial. Analysis of the c‐erbB‐2 gene structure and comparison of its expression in the same cells by Southern, Northern and immunoblotting, and by immunocytochemistry minimize the errors of interpretation inherent in one technique. Such a ‘correlative study’ has been performed on tumours from 82 patients, c‐erbB‐2 gene amplification was detected in 14 per cent of initial tumours and was associated with grade (.P< 0·001). Raised levels of mRNA were seen in those tumours with increased gene copy number and in 13 per cent of the remainder. Immunoblotting detected the expected 185 kD immunoreactive protein and a 155 kD piotein associated with high gene copy number. Immunocytochemistry localized c‐erbB‐2 immunoreactivity to the cell membrane and cytoplasm, and the latter predominated. Four antibodies to c‐erbB‐2 (AB‐3, 21N, pAb 1, and NCL CB11) were compared on contiguous sections of the same tumour and showed the same pattern of immunoreactivity. Similarly, analyses carried out in three independent laboratories identified the same cellular localization. Membrane and cytoplasmic immunoreactivity was demonstrated in all tumours with gene amplification or increased mRNA levels and in 40 per cent of the remaining tumours. We showed that immunocytochemistry requires careful standardization of techniques and quantitation between different groups. However, despite variations in the intensity of immunoreactivity, the total number of positive cells remained constant. Therefore quantitation must be based on the number of positive cells and, ideally, their immunoreactive content relative to normal and positive tissue controls.