bluuebunny/crossref_metadata_2025 huggingface dataset:
Diagnostic and prognostic significance of clonal T‐cell receptor beta gene rearrangements in lymph nodes of patients with mycosis fungoides
Authors: Bakels Victor , Van Oostveen Johan W. , Geerts Marie‐Louise , Gordin Roel L. J. , Walboomers Jan M. M. , Scheffer Erik , Meijer Chris J. L. M. , Willemze Rein
Published: June 2005

In this study, 25 involved and uninvolved lymph nodes from 22 patients with mycosis fungoides (MF) and seven dermatopathic lymph nodes from patients with benign skin disorders were studied for the presence of clonal T‐cell receptor beta (TCRβ) gene rearrangements by Southern blot analysis. These results were correlated with the histological classification, follow‐up data, and survival. The results of the histological classification and Southern blot analysis were concordant in 26 of 32 cases. Clonal TCRβ gene rearrangements were found in all six MF lymph nodes showing (partial) effacement of the normal lymph node architecture, but in none of the eight uninvolved dermatopathic MF lymph nodes and in none of the seven dermatopathic control lymph nodes. In addition, in 5 of 11 dermatopathic MF lymph nodes that were considered to have early involvement by MF at histological examination, clonal TCRβ gene rearrangements were detected. In the group of MF patients with dermatopathic lymphadenopathy, patients with detectable clonal T‐cell populations had a significantly shorter survival than patients without such a population (P<0.01). The results of this study indicate that within the group of dermatopathic MF lymph nodes, prognostically different groups can be distinguished and that TCRβ gene rearrangement analysis may be an important adjunct in the early diagnosis of lymph node involvement by MF.
Expression of P53 protein in normal, dysplastic, and malignant gastric mucosa: An immunohistochemical study
Authors: Joypaul B. Vickram , Newman E. Luke , Hopwood David , Grant Andrew , Qureshi Shahzad , Lane David P. , Cuschieri Alfred
Published: June 2005

Mutations in the p53 nuclear oncogene are the most frequent genetic abnormalities encountered in human malignancies. Using the polyclonal antibody CM‐1, we have examined the expression of the p53 oncoprotein immunohistochemically in archival material of normal, dysplastic, and malignant gastric mucosa. Abnormal expression of this protein was not observed in biopsies of normal gastric tissue (n=30) but was detected in 22 of the 36 gastric cancers analysed (61 per cent). Nuclear staining was diffuse in 15 of the positive cancer cases, the remaining seven showing a more varied heterogeneous staining pattern. Abnormal p53 protein was not detected in mild (n=14) or moderate (n=16) gastric dysplasia but was present in 3 out of 15 severe dysplasia cases. The results suggest that expression of the p53 oncoprotein is a common finding in gastric cancer and occurs as a late event in the malignant transformation process.
Isolation and characterization of multiple cell types from a single human colonic carcinoma: Tumourigenicity of these cell types in a xenograft system
Authors: Marsh Katrina A. , Stamp Gordon W. H. , Kirkland Susan C.
Published: June 2005

The HCA‐7 cell line was established from a moderately well differentiated mucinous adenocarcinoma of the colon, which showed histological heterogeneity. This was reflected in the morphological heterogeneity in early passages of the HCA‐7 cell line but diminished as the cells were passaged in vitro. Nine subpopulations were isolated from early passage cultures of the HCA‐7 line and maintained as cell lines. Each subpopulation demonstrated a unique set of stable biological characteristics in vitro. When established in vivo, there was a wide variation in xenograft generation time. The parent cell line gave rise to six distinct xenograft patterns, two of which had been observed in the primary carcinoma. Individual subpopulations yielded characteristic tumours which were composed of between one and four organizational patterns. Xenografts differed in both the organization of cells and the cell differentiation within the different patterns. Mucous cells were absent from some tumours while abundant in others. The subpopulations isolated from the HCA‐7 cell line provide a new and extensive model system for studying the generation and maintenance of phenotypic heterogeneity in colorectal carcinoma.
Proliferating cell nuclear antigen (PCNA) expression as a prognostic indicator for renal cell carcinoma: Comparison with tumour grade, mitotic index, and silver‐staining nucleolar organizer region numbers
Authors: Delahunt Brett , Bethwaite Peter B. , Nacey John N. , Ribas Jorge L.
Published: June 2005

Proliferating cell nuclear antigen (PCNA) expression in renal cell carcinoma (RCC) was determined by immunohistochemical staining using the PC10 clone. PCNA indices ranged from 2.4 to 53.1 per cent with mean indices of 12.6, 19.0, and 31.6 per cent for grades 1 to 3 RCC and 31.9 per cent for sarcomatoid RCC. There was a significant difference between the indices of grades 1 and 3 and grades 2 and 3 tumours and between grades 1 and 2 RCC and sarcomatoid RCC. AgNOR scores and mitotic indices were determined for each tumour and comparison of PCNA indices with mean AgNOR scores and mitotic indices showed only a weak correlation (PCNA/AGNORr= 0.406, PCNA/mitotic indexr= 0.315). Tumours were divided according to PCNA index (≤18 per cent and > 18 per cent) and there was a significant difference in survival between the two groups, for all cases, and for each of the Robson stages. Multivariate analysis using Cox's proportional hazard model showed PCNA index, tumour stage, and mean AgNOR score to be independent predictors of survival, while tumour grade and mitotic index were found to be dependent variables.
Quantitation of the renal arterial tree by fractal analysis
Authors: Cross Simon S. , Start Roger D. , Silcocks Paul B. , Bull Andrew D. , Cotton Dennis W. K. , Underwood James C. E.
Published: June 2005

To determine whether the renal arterial system has a fractal structure, the fractal dimension of renal angiograms from 52 necropsy cases was measured using an implementation of the box‐counting method on an image analysis system. The method was validated using objects with known fractal dimensions. The method was accurate with errors of less than 1.5 per cent and reproducible with initial values within 1.2 per cent of the mean of ten sets of measurements (reliability coefficient 0.968, 95 per cent confidence limits 0.911.0.984). In the 36 satisfactory angiograms the mean fractal dimension was 1.61 (SD 0.06), which was significantly greater than the topological dimension of 1 (P<0.0001), indicating that the renal arterial tree has a fractal structure. There was no significant relationship between age (P=0.494), sex (P=0.136), or systolic (P=0.069) or diastolic (P=0.990) blood pressure, but two congenitally abnormal kidneys (hypoplastic dysplasia and renal artery stenosis) had fractal dimensions at the lower end of the normal range (third percentile). Since the renal arterial tree has a fractal structure, Euclidean geometric measurements, such as area and boundary length, are invalid outside precisely defined conditions of magnification and resolution.
The earliest histopathological response to hypobaric hypoxia in rabbits in the rifugio torino (3370 M) on Monte Bianco
Authors: Smith P. , Heath D. , Williams D. , Bencini C, , Pulera N. , Giuntini C.
Published: June 2005

Twelve Dutch rabbits were kept on Monte Bianco at an altitude of 3370 m. Half of the animals were killed after 3 months, the remainder after 6 months, and a further six animals maintained at sea‐level acted as controls. The carotid bodies of all the rabbits were processed for light and electron microscopy and examined qualitatively and quantitatively. The lungs were processed for light microscopical assessment of small pulmonary arterial vessels; the thickness of the pulmonary trunks and aortas were measured; and the hearts were dissected to obtain ratios of the ventricular weight. There was a slight increase in the right ventricular weight in the hypoxic rabbits but no change in the thickness of the pulmonary trunk compared with that of the aorta. In particular, there was no hypoxic remodelling of the pulmonary vasculature such as muscularization of pulmonary arterioles or intimal longitudinal muscie in pulmonary arteries. The earliest histopathological response to hypoxia occurred in the carotid bodies in the form of an increase in the count of the dark variant of chief cell after 3 months which returned to normal after 6 months. It is concluded that the carotid body of the rabbit responds with a change in its population of dark chief cells to a level of hypoxia which is insufficient to affect the pulmonary arterioles. Changes in the cardiopulmonary system can no longer be considered to be the earliest histopathological response to hypobaric hypoxia.
Patterns of CEA‐related antigenex pression in invasive squamous carcinoma of the cervix
Authors: Sanders D. S. A. , Ferryman S. R. , Bryant F. J. , Rollason T. P.
Published: June 2005

The recognition of an adhesive role for the CEA‐related antigens emphasizes the need for clear demonstration of the changes in CEA expression and subcellular localization between normal and neoplastic tissues. Using a panel of monoclonal and polyclonal antibodies, membranous and cytoplasmic CEA expression was seen in 50 invasive cervical squamous carcinomas in four distinct patterns dependent on tumour type and differentiation. Membranous CEA expression is a marker of differentiation in squamous carcinomas and may influence tumour behaviour and hence patient survival. Strong CEA positivity was seen on the endothelium of vessels containing tumour in ten cases where vascular metastases were prominent. Staining of these ten cases revealed concomitant sialated Lewis X positivity in tumour cells with weak endothelial positivity in three cases; cervical squamous tumour cells may localize to vascular endothelium, and hence disseminate, through specific binding of CEA and/or sialated Lewis X.
Proliferative state of the urothelium with benign and atypical changes. Correlation with transferrin and epidermal growth factor receptors and blood group antigens
Authors: Limas Catherine
Published: June 2005

We attempted to investigate how the proliferative state of the urothelium correlates with the reactivity for transferrin (Tf) and epidermal grown factor (EGF) receptors and the blood group (BG) antigen. We examined morphologically normal urothelium (34 cases), benign inflammatory and reactive conditions (24 cases), and atypical changes (20 cases) without exophytic or invasive neoplasia. The Ki67 nuclear antigen was used as the proliferation index and was complemented with the in vitro BrdU incorporation assay in 32 cases. The immunohistochemical reactions for Tf and EGF receptors and for the appropriate BG antigen were scored semi‐quantitatively on frozen sections. We found that normal urothelium has very low Ki67 and BrdU indices as well as low reactivity for the two receptors and is almost invariably positive for the BG antigen. Benign conditions such as inflammation and metaplasia significantly augment the proliferation indices and the Tf receptor with little change in the EGF receptor and no significant effect on the BG antigen. Moderate atypia includes biologically heterogeneous cases which vary widely in proliferation and receptor expression. Severe atypia‐carcinoma in situ is consistently associated with markedly elevated proliferation indices, strong Tf receptor reactivity, and depressed BG antigen. The reactivity for EGF receptor is less consistently increased. Cases with a combination of strong EGF receptor reactions and absence of the expected BG antigen have a poor prognosis with progression to invasive cancer.
Ultrastructural heterogeneity in undifferentiated bronchial carcinoma
Authors: Carter Naomi , Nelson Fiona , Gosney John R.
Published: June 2005

Electron microscopy is often suggested as a useful aid to the classification of light microscopically undifferentiated bronchial malignancies, features such as dense‐core vesicles, desmosomes or tonofilaments, and microacini, allowing their designation as endocrine, squamous, or adenocarcinomas respectively. However, there is no reason to suppose that the heterogeneity of malignant bronchial tumours so often apparent by light microscopy or on immunolabelling might not occur at the ultrastructural level too. Extensive sampling of all deposits from eight subjects coming to necropsy with undifferentiated bronchial carcinoma revealed ultrastructural features of glandular and squamous differentiation to be widespread and often to occur together, although dense‐core vesicles were not seen in any of the tumours studied. Heterogeneity was present within individual tumour deposits and particularly between different deposits of those tumours which had disseminated, such that any ultrastructural diagnosis would have been significantly influenced by sampling. Such variation should be borne in mind when ultrastructural features are used to classify bronchial malignancies.
Electron microscopic study of the attachment and penetration of rabbit intestinal epithelium by Providencia alcalifaciens
Authors: Mathan Minnie M. , Mathan Vadakenadayil I. , Albert M. John
Published: June 2005

Ilea of adult rabbits with removable ileal ties (RITARD model) that developed diarrhoea and ileal loops of adult rabbits after inoculation with Providencia alcalifaciens, isolated from two patients with diarrhoea, were studied by transmission electron microscopy for enterocyte‐bacterial interactions. Two modes of entry of bacteria into epithelial cells were seen: one directly by endocytosis associated with polymerization of cytoskeletal components and the other by disruption of tight junctions with entry into and proliferation in intercellular spaces.
Antigen unmasking on formalin‐fixed, paraffin‐embedded tissue sections
Authors: Cattoretti Giorgio , Pileri Stefano , Parravicini Carlo , Becker Michael H. G. , Poggi Simonetta , Bifulco Carlo , Key Göran , D'Amato Lucia , Sabattini Elena , Feudale Elisa , Reynolds Fred , Gerdes Johannes , Rilke Franco
Published: June 2005

Enzymatic and non‐enzymatic treatments for antigen unmasking on formalin‐fixed, paraffin‐embedded, dewaxed sections were optimized and compared by the use of a panel of antibodies of diagnostic relevance (anti‐cytokeratins, vimentin. S‐100. T‐ and B‐cell receptors, Ki‐67/MIB 1, muscle actin). Non‐enzymatic unmasking was obtained by boiling the slides in a microwave oven in 0.01 M salt solution (pH 6) or in 6 M urea. Trypsin or pronase digestion was used for comparison and found to be necessary for some of the reagents. The investigation was then extended to 256 antibodies; the epitopic amino acid sequence was known for 48 of them. We found that enzymatic and non‐enzymatic antigen unmasking are not dependent on the epitope sequence, but some antigens benefit selectively from one treatment but not from the other. Denaturation of proteins is the likely mechanism which leads to immunodetection on microwave oven‐boiled slides; this suggestion is supported by the use of denaturating solutions and by the observation that endogenous enzymes were inactivated and a few antigens were no longer immunodetectable after boiling. Non‐enzymatic methods for antigen unmasking are a powerful new tool for broadening the use of antibodies for immunostaining formalin‐fixed, paraffin‐embedded sections and should be used in parallel with the traditional enzymatic methods.
Genetic alterations of the TP53 gene, p53 protein expression and hpv infection in primary cervical carcinomas
Authors: Helland Åslaug , Holm Ruth , Kristensen Gunnar , Kaern Janne , Karlsen Frank , Trope Claes , Nesland Jahn M. , Børresen Anne‐Lise
Published: June 2005

Primary cervical carcinomas from 92 patients were investigated for genetic alterations in the tumour suppressor gene TP53. Studies of allelic imbalance (AI) were performed by Southern blot analysis and by using two PCR (polymerase chain reaction) polymorphisms within the TP53 gene. AI in the tumour was observed in 22 per cent (11 of 52 informative patients) and was significantly associated with recurrence both in a univariate (P=0.013) and in a multivariate (P=0.045) analysis. The DNA samples were subjected to mutation analysis of four of the conserved domains in the TP53 gene, using PCR followed by constant denaturant gel electrophoresis (CDGE). Mutations were observed in 2 of 92 tumours (2 per cent), of which one was a silent mutation and the other a frameshift. Overexpression of the p53 protein was found by immunostaining of sections from formalin‐fixed, paraffin‐embedded material in 55 per cent (51/92) of the tumours. In 88 per cent (45/51) of these, overexpression was present in less than 5 per cent of the tumour cells. Overexpression was significantly associated with relapse‐free survival only in a univariate analysis (P=0.045). AI of the TP53 locus did not correlate with p53 expression or mutation. The important gene on 17p, responsible for the shorter disease‐free survival for patients with AI of TP53, may therefore be another gene closely linked to TP53. In addition, the 92 tumour samples were tested for the presence of human papillomavirus (HPV) types 16 and 18. Fifty‐four per cent (50/92) of the samples were positive for HPV 16 usingin situhybridization, and 93 per cent (86/92) using the PCR technique. The numbers for HPV 18 were 15 per cent (14/92) and 23 per cent (21/92), respectively. Twenty‐one per cent (19/92) were positive for both HPV 16 and HPV 18, while 4 per cent (4/92) were negative for both HPV 16 and 18. The tumour with the frameshift TP53 mutation was HPV 16‐positive, and the four samples negative for HPV 16 and 18 did not contain TP53 mutations within the conserved domains but had elevated p53 protein expression.
Immunohistochemical evidence for the expression of proliferating cell nuclear antigen (PCNA) by non‐proliferating hepatocytes adjacent to metastatic tumours and in inflammatory conditions
Authors: Harrison Rebecca F. , Reynolds Gary M. , Rowlands David C.
Published: June 2005

The possibility that proliferating cell nuclear antigen (PCNA) is expressed by non‐proliferating liver cells was investigated. Liver biopsies from 107 patients were investigated, which included histologically normal liver, metastatic tumour, and inflammatory lesions. PCNA was detected using immunohistochemical staining with the monoclonal antibody PC10. This was compared with the proportion of proliferating cells as assessed by immunostaining for the Ki‐67 antigen using the monoclonal antibody MIB 1. Most cases of histologically normal liver showed few PC10‐positive cells. PCNA‐positive hepatocytes far outnumbered those positive with MIB 1 in specimens showing metastatic tumour or an inflammatory cell infiltrate. There was no relation between the degree of PCNA overexpression and the type of tumour present or the nature of the inflammatory lesion. Other cell types, including the biliary epithelium, did not show this large difference between the proportions of PC10‐ and MIB 1‐positive cells. It is concluded that non‐proliferating hepatocytes increase their levels of PCNA in a wide variety of pathological conditions. This may be mediated by cytokines released by tumour cells or inflammatory cells.
Vascular activation in the histopathogenesis of Hodgkin's disease: Potential role of endothelial tissue factor in intravascular thrombosis and necrosis
Authors: Ruco Luigi P. , Pittiglio Marco , Dejana Elisabetta , Baroni Carlo D.
Published: June 2005

Endothelial cell activation and alterations of intravascular coagulation were investigated in 27 cases of Hodgkin's disease (HD), in five cases of anaplastic large cell lymphoma (ALCL), and in ten reactive lymph nodes. Lymph node sections were immunostained for E‐selectin, a molecule present on cytokine‐activated endothelial cells; for tissue factor (TF), a cellular initiator of the coagulation cascade; for glycoprotein (gp) II/III, a platelet‐specific antigen; and for fibrin. In HD, vascular activation was particularly prominent in the nodular sclerosis subtype, as indicated by a larger number of E‐selectin‐positive blood vessels (72 ± 49) compared with mixed cellularity (22 ± 37). High expression of E‐selectin was associated with alterations of intravascular coagulation, as indicated by immunostaining of some vascular endothelial cells for TF, by a higher incidence of intravascular thrombi, and by the extensive presence of areas of fibrin exudation and necrosis. In ALCL, the levels of endothelial cell activation and intravascular coagulation were comparable to those of HD nodular sclerosis. In reactive nodes, some E‐selectinpositive blood vessels were observed only in 3/10 cases; immunostaining for TF was not detected on endothelial cells; and alterations of intravascular coagulation were rarely observed.
Glomerular dimensions in children under 16 years of age
Authors: Moore Lynette , Williams Ruth , Staples Alan
Published: June 2005

Glomerular area, perimeter, and mean maximum glomerular diameter were assessed in post‐mortem kidney tissue from 114 children who died suddenly in accidents and who were considered to have been previously healthy. One hundred glomeruli were measured from each case using a computer‐based measurement system. Regression analysis was used to analyse glomerular size with respect to age, body weight, height, and body mass index. The estimated mean glomerular area for newborns was 7934 μm2 (SE 314), increasing 8 per cent per year to 18 219 μm2 at 15 years of age. The estimated mean maximum glomerular diameter increased from 112 μm (SE 1.9μm) for newborns, approximately 3.6μm per year to reach 167 μm at age 15. Glomerular size correlated better with age than with height, weight, or body mass index, and glomeruli in the inner cortex were significantly larger than those in the middle and outer cortex. Glomerular size was significantly greater in female children than in the male children over the age range examined.
Identification of numerical chromosome aberrations in archival tumours by in situ hybridization to routine paraffin sections: Evaluation of 23 phaeochromocytomas
Authors: Van Dekken H. , Bosman F. T. , Teijgeman R. , Vissers C. J. , Tersteeg T. A. , Kerstens H. M. J. , Vooijs G. P. , Verhofstad A. A. J.
Published: June 2005

We have applied non‐isotopic in situ hybridization (ISH) to interphase cell nuclei of 23 phaeochromocytomas (18 primary and 5 metastatic tumours) within routine paraffin‐embedded tissue sections. Each tumour was screened for numerical aberrations with a defined alphoid repetitive DNA probe set containing DNA probes specific for chromosomes 1, 7, 15, and Y. Normal adrenal medullas and other normal human cell types served as cytogenetic controls. Preservation of tissue morphology enabled targeted analysis of tumour cells. The presence of numerical chromosome changes in the tumour cells could easily be evaluated by comparing the ISH results of the DNA probes. Numerical abnormalities not previously reported in this neoplasm included overrepresentation of chromosomes 1 and 7, loss of chromosome 15, and both gain and loss of chromosome Y (P values <0.01). The percentage of aneuploid cell nuclei in a tumour correlated well with the percentage of cells in the 4C peak of flow cytometric DNA histograms from these neoplasms. We conclude that interphase ISH can be used for the identification of new and reported cytogenetic changes in tumour cell nuclei within archival tissue sections. This novel procedure also allows for retrospective analysis of previously not karyotyped material.
Loss of α6 and β4 integrin subunits coincides with loss of basement membrane components in oral squamous cell carcinomas
Authors: Downer Caroline S. , Watt Fiona M. , Speight Paul M.
Published: June 2005

In oral squamous cell carcinomas, focal or extensive loss of basement membrane components and of integrins has been reported. The purpose of this study was to investigate whether those regions of the tumour‐connective tissue interface which lack laminin and type IV collagen coincide with areas of loss of the α6 and β4 integrin subunits on basal keratinocytes. Out of a total of 15 poor and moderately or well differentiated squamous cell carcinomas, all showed some loss or fragmentation of basement membrane proteins and in 12 the loss was coincident with loss of α6 and/or β4. In three cases, there was loss of basal integrin expression in areas where the basement membrane remained intact. These results provide further evidence that loss of integrins may play an important role in tumour progression and prompt us to speculate about the sequence of events leading to tumour invasion.
TGF‐βS and TGF‐β type II receptor in human epidermis: Differential expression in acute and chronic skin wounds
Authors: Schmid Peter , Cox David , Bilbe Graeme , McMaster Gary , Morrison Chris , Stähelin Hannes , Lüscher Nicholas , Seiler Walter
Published: June 2005

Exogenously applied transforming growth factor‐beta (TGF‐β) isoforms enhance wound healing processes in animal models;1 however, little is known about the expression of endogenous TGF‐βs and TGF‐β receptors in intact human skin or during wound healing. The present study has revealed several unexpected findings by means of in situ hybridization and immunohistology techniques. In humans, TGF‐β3 is constitutively expressed in the epidermis of intact skin and in that of acute and chronic wounds—a pattern of expression closely mirrored by the TGF‐β type II receptor. Although not detected in intact skin, TGF‐β mRNA expression was observed in the regenerating epidermis of acute (thermal) wounds but was not found in chronic decubital (pressure) wounds. TGF‐β2 mRNA expression was not detected in the epidermis of any human skin or wound biopsies. From these findings we suggest that constitutive expression of TGF‐β3 is important for maintenance of epidermal differentiation and that an induction of TGF‐β1 expression is essential for re‐epithelialization of human skin wounds. Lack of TGF‐β1 expression in chronic pressure wounds may be associated with their protracted healing tendencies.
The expression of the adhesion molecules ICAM‐1, VCAM‐1, PECAM, and E‐selectin in human atherosclerosis
Authors: Davies Michael J. , Gordon J. L. , Gearing A. J. H. , Pigott R. , Woolf Neville , Katz David , Kyriakopoulos A.
Published: June 2005

The expression of PECAM, ICAM‐1, VCAM‐1, and E‐selectin was studied in 64 samples of human coronary arteries taken from 15 explanted hearts obtained within 5 min of transplantation. Normal artery (n=12), predominantly fibrous plaques (n=23), and plaques containing extracellular lipid (n=26) and three segments showing recanalization channels were studied. All endothelial cells strongly and equally expressed PECAM; positive staining was used to check that artefactual denudation of the endothelial surface had not occurred. PECAM was also present in some lipid‐filled macrophages. Normal arteries showed no VCAM‐1 staining but focal segments of the endothelium were positive for ICAM‐1 and E‐selectin. ICAM‐1 was strongly and constantly expressed by the endothelium over all types of plaques and in macrophages. E‐selectin expression was confined to endothelial cells and occurred on the surface in 35 per cent of fibrous and 22 per cent of lipid‐containing plaques. VCAM‐1 staining of surface endothelium occurred in 39 per cent of fibrous and 20 per cent of lipid‐containing plaques. A population of spindle‐shaped cells of macrophage type (positive for EMB11 antigen) expressed VCAM‐1 in lipid‐containing plaques. Adventitial vessels adjacent to plaques showed endothelial expression of ICAM‐1 and E‐selectin. VCAM‐1 staining of adventitial vessel endothelium was associated with local lymphoid aggregation. In conclusion, the expression of cell adhesion molecules is an important element in the inflammatory component of atherosclerosis and contributes to both monocyte and lymphocyte activation and recruitment from advential vessels and the arterial lumen.
Tissue iron loading and histopathological changes in hypotransferrinaemic mice
Authors: Simpson Robert J. , Konijn Abraham M. , Lombard Martin , Raja Kishor B. , Salisbury Jonathan R. , Peters Timothy J.
Published: June 2005

Tissue iron loading in hypotransferrinaemic (hpx/hpx) mice was investigated as a model for genetic (primary) haemochromatosis. Iron loading of liver preceded that in the pancreas and heart. One‐year‐old hpx/hpx mice showed iron staining in exocrine pancreas, liver parenchymal cells, and cardiac and intestinal smooth muscle cells. Iron‐loaded macrophages were observed in all these tissues. Islets of Langerhans, biliary epithelial cells, and spleen were iron‐free. The pancreas was fibrotic with massive macrophage infiltration and loss of secretory epithelium. Liver showed evidence of chronic inflammatory infiltration with increased collagen fibres in the parenchymal region but no cirrhosis. Serum aspartate aminotransferase activity and plasma glucose were increased in hpx/hpx compared with wild‐type mice. Heavy iron loading with haemosiderin deposition in the liver could be demonstrated in hpx/hpx mice from 6 weeks of age. Heterozygous hypotransferrinaemic mice showed minor increases in liver iron stores at 6–12 weeks, but not at 1 year of age. Serum ferritin levels in heterozygous mice were also increased at 6–8 weeks of age. It was concluded that 1‐year‐old hpx/hpx mice showed evidence of liver and pancreatic damage secondary to tissue iron overload. The iron loading pattern and tissue damage showed some features which were distinct from those observed in haemochromatosis.
pS2 protein in gastric carcinoma and normal gastric mucosa: Association with clinicopathological parameters and patient survival
Authors: Müller Wolfram , Borchard Franz
Published: June 2005

The presence of pS2 protein (pS2) was studied in a total of 120 consecutive patients with gastric carcinomas. This immunohistochemical study found pS2 expression in 48 per cent (n=58) of carcinomas. pS2 expression was also detected in normal gastric mucosa in 95 per cent (n=102) of specimens in upper antral mucopeptic glands and deep foveolar cells of the gastric pits but not in intestinal metaplasia. There was a significant statistical correlation between pS2 expression and extent of tumour growth (pT state) and expression of pepsinogen II by the tumours. There was no statistical correlation with clinical features such as patient age or sex or other pathological parameters (tumour stage, size, grade, and localization or growth pattern according to histological classification). There were no statistically significant differences in survival times between patients with pS2‐positive and pS2‐negative tumours. In contrast to findings concerning breast cancer, pS2 expression in gastric carcinomas has no influence on the patient's prognosis. On the other hand, strong expression of pS2 by surface epithelium of normal gastric mucosa may indicate that pS2 might play a role in physiological cell renewal of normal gastric mucosa.
Expression of p53 in oligodendrogliomas
Authors: Kros Johan M. , Godschalk Josiane J. C. J. , Krishnadath Kausilia K. , Van Eden Corbert G.
Published: June 2005

The expression of the nuclear protein p53 in oligodendrogliomas was investigated by immunohistochemistry, using a monoclonal anti‐p53 antibody (DO‐7) on formalin‐fixed, paraffin‐embedded material in 84 histologically verified cases, and compared with the histopathological grade and survival. p53‐immunoreactive cells were found in 75 per cent of the samples acquired at the first biopsy. The p53 labelling index was not related to the degree of nuclear anaplasia. Tumour cases with more than 75 per cent p53 immunostained cells had a rapidly fatal clinical course. However, no significant correlation was found between p53 labelling index and tumour grade, mitotic index, or ploidy status. In most tumour recurrences (n=25), the p53 labelling index increased or remained at the level of the first biopsy. In five cases (6 per cent), p53 was absent in the first sample as well as in the recurrence. Irrespective of the underlying aberration of either the gene or the metabolic pathway of p53, it is concluded that a high percentage (i.e., more than 75 per cent) of p53‐immunolabelled cells is predictive of an unfavourable clinical course, while a percentage lower than 75 per cent immunoreactive cells does not exclude a rapid fatal outcome.
Cytotoxicity of oxidized low‐density lipoprotein to mouse peritoneal macrophages: An ultrastructural study
Authors: Reid Vanessa C. , Mitchinson Malcolm J. , Skepper Jeremy N.
Published: June 2005

Mouse peritoneal macrophages (MPM) were incubated in culture medium containing low‐density lipoprotein (LDL), oxidized LDL (oxLDL), or as controls, for 24 h. Scanning electron microscopy of MPM exposed to oxLDL showed loss of membrane ruffles and extensive plasmalemmal ‘blebbing’. Transmission electron microscopy showed changes in up to 50 per cent of the cells, including vacuolation of the rough endoplasmic reticulum and reorganization of heterochromatin in the nuclei, characteristic of apoptosis. These changes were not seen in controls, nor in macrophages exposed to native LDL. Electron‐dense crystals were found in the cytoplasm of 1 in 50 of cells exposed to oxLDL. These were found to have a high content of calcium and phosphorus. It is proposed that oxLDL is capable of inducing apoptosis, which might explain the origin of the necrotic base of advanced atherosclerotic plaques.
An evaluation of six antibodies for immunohistochemistry of mutant p53 gene product in archival colorectal neoplasms
Authors: Baas Inge O. , Mulder Jan‐Willem R. , Offerhaus G. Johan A. , Vogelstein Bert , Hamilton Stanley R.
Published: June 2005

Immunohistochemical detection of intranuclear p53 gene product may indicate mutation of the p53 suppressor gene on chromosome 17p. We used six commercially available antibodies for p53 immunohistochemistry on 19 archival colorectal neoplasms and compared the results with the mutation status of the p53 gene and 17p allelic deletion status. By Friedman's ranking analysis, use of mouse monoclonal antibody DO7 with Target Unmasking Fluid (TUF) for antigen retrieval was the most sensitive and specific procedure (P<0·0001). Six of 7 cases with high expression (p53 Labeling Index >30 per cent using a CAS 200 image analyser) had p53 mutation. Of seven tumours without expression (LI < 1 per cent), six had no mutation and one had a truncating mutation which prohibited nuclear localization of gene product. The low expression group (1 per cent < LI <30 per cent, n = 5) consisted of three tumours without and two tumours with mutation. The sensitivity of high expression with the DO7–TUF method for p53 gene mutation was 67 per cent with specificity of 90 per cent, predictive value of a positive of 86 per cent, predictive value of a negative of 75 per cent, and efficiency of 79 per cent. This study suggests that immunohistochemistry is valuable for assessing p53 gene mutations in colorectal neoplasms, but further study is needed to elucidate the precise link between immunohistochemistry and molecular genetic alterations.
Variations in immunohistochemical detection of p53 protein overexpression in cervical carcinomas with different antibodies and methods of detection
Authors: Lambkin Helen A. , Mothersill Carmel M. , Kelehan Peter
Published: June 2005

Immunocytochemical detection of p53 protein products in paraffin sections is possible with a number of antisera, monoclonal and polyclonal. Few corroborative results amongst different laboratories have been published due to variations in the antibodies, the fixation protocols, and the immunocytochemical methods applied. Antigen unmasking methods employing microwaves or proteolytic enzymes add to the disparity in the percentage of positive cases reported. In this study, paraffin sections of 55 cases of cervical carcinoma were immunostained with monoclonal antibodies p53‐DO7 and p53‐1801 (a) without section pretreatment, (b) with pronase digestion, and (c) with microwave antigen retrieval in citric acid buffer. Specimens fixed in buffered formalin required antigen unmasking to show positive staining. Pronase digestion caused false‐negative immunostaining. Microwave pretreatment with p53‐DO7 yielded 100 per cent positivity for p53 proteins but only 7/55 cases with more than 50 per cent positive cells. Monoclonal antibody p53‐DO7 detected more positive cases than p53‐1801. Immunostaining with antibodies to p53 proteins must be interpreted cautiously as variations in fixation, antibodies, and section pretreatment will significantly affect results.
NDP‐K/nm23 expression in human breast cancer in relation to relapse, survival, and other prognostic factors: An immunohistochemical study
Authors: Sawan Ali , Lascu I. , Veron M. , Anderson J. J. , Wright C. , Horne C. H. W. , Angus B.
Published: June 2005

The nm23 gene was originally identified by Steeg et al. by screening of cDNA libraries from murine melanoma cell lines of varying metastatic potential. An inverse relationship between metastatic potential and nm23 RNA and/or protein was found in four different metastatic model systems. It was proposed that nm23 may function as a suppressor gene for tumour metastasis. It has recently been found that the sequence of nm23 and NDP‐kinase (NDP‐K) is identical. Using an immunohistochemical technique and employing a polyclonal antibody to purified NDP‐K A, we have determined NDP‐K expression in a series of 197 breast carcinomas. One hundred and sixty (81·2 per cent) of these tumours were scored positive for NDP‐K and 37 (18·8 per cent) scored negative. No relationship was found between NDP‐K/nm23 expression and patient relapse or survival. Furthermore, no relationship was found between NDP‐K/nm23 expression and a number of other prognostic factors including tumour grade, oestrogen receptor, progesterone receptor, and p53 expression. Our results contradict the hypothesis concerning the possible role of NDP‐K/nm23 as a metastatic suppressor gene in human breast cancer, but further studies using antibodies specific for NDP‐K/nm23 subtypes are clearly indicated.
Production of two novel monoclonal antibodies against human renal glomeruli and theri application to the immunohistochemical investigation of crescentic glomerulonephritis
Authors: Kohda Yukimasa , Takeya Motohiro , Arima Seiya , Arima Seiya , Nakayama Mahito , Sato Tatsuo , Takahashi Kiyoshi
Published: June 2005

Two monoclonal antiboides (MAbs) against human renal glomeruli, HEP1 and HBM1, were produced using isolated human glomeruli as an antigen. Immunohistochemical and immunoelectron microscopic analyses revealed that the recognition site of HEP1 is the cell surface of the visceral glomerularepithelial cells. HEP1 showed no reaction in the renal interstitium or in other tissues. HBM1 recognized an antigen present in the basal lamina of the renal glomeruli, Bowman's capsule, and partly in the basement membrane of renal tubules, but it was not reactive with the mesangial matrix or renal interstitium. The isotype of both MAbs was IgG1, k. The molecular weights of their corresponding antigens were 127–177 and 220kD, respectively. Besides human tissues, HEP1 also showed cross‐reactions with antigens in the visceral glomerular epithelial cells of various animals other than mice. HBM1 recognized human antigen only. Using both MAbs, eight cases of crescentic glomerulonephritis were examined immunohistochemically. As a result, it was demonstrated that the crescents may not be formed by visceral glomerular epithelial cells. From this study, these two MAbs appear to be userful markers for the evaluation of renal glomerular disorders.
Ubiquitin profile in inflammatory leukocytes and binding of ubiquitin to murine AA amyloid: Immunocytochemical and immunogold electron microscopic studies
Authors: Alizadeh‐Khiavi K. , Li W. , Chronopoulos S. , Ali‐Khan Z.
Published: June 2005

Lysosomes in activated murine monocytoid cells have been implicated in AA amyloid formation. The pathophysiology of this process is not well understood. Previous studies into the nature of the relationship between ubiquitin (UB), possessing intrinsic amyloid enhancing factor (AEF) activity; serum amyloid A (SAA), the precursor protein of AA amyloid; and activated monocytoid cells have indicated a temporal and spatial relationship between these proteins and tissue AA amyloid deposits. To extend these findings, we have examined murine peritoneal leukocytes and splenic tissues during the early amyloid deposition phase by immunocytochemical and immunogold electron microscopic methods using monospecific anti‐ubiquitin and anti‐mouse AA amyloid antibodies. We show here enrichment of endosome–lysosome‐like (EL) vesicles in the activated monocytoid cells with UB and SAA, and the presence of UB‐bound AA amyloid fibrils in the EL vesicles, perikarya, and interstitial spaces. The importance of these findings is emphasized by the fact that activated monocytoid cells, containing UB in the EL vesicles, sequester and eventually localize SAA in their EL vesicles, and that UB binds to the EL‐contained AA amyloid fibrils. These findings may also have functional consequences for studies on the role of EL and UB in amyloidogenesis.
A time sequence of vessel wall changes in an experimental model of angioplasty
Authors: More Ranjit S. , Rutty Guy , Underwood Malcolm J. , Brack Michael J. , Gershlick Anthony H.
Published: June 2005

A detailed temporal sequence of vessel wall changes in a rabbit model of balloon angioplasty, to the iliac arteries, at 30 min, 2h, day 1, day 3, day 7, day 14, 1 month, and 3 months was established using light microscopy, immunostaining for proliferating cell nuclear antigen, and electron microscopy. Complete endothelial denudation with extensive platelet and fibrin deposition was evident early. Intimal thickening, characterized by myointimal hyperplasia, was observed at day 7 and reached a maximum at 1 month. Re‐endothelization was complete by day 14. Cellular proliferation, seen early at day 1, reached a maximum at day 7 for the media and day 14 for the intima before returning to baseline levels by 1 month. This simple model of angioplasty with its reproducible myointimal hyperplasia should prove useful for assessing any potential agents for inhibiting this process.
Fractal geometric analysis of colorectal polyps
Authors: Cross Simon S. , Bury Jonathan P. , Silcocks Paul B. , Stephenson Timothy J. , Cotton Dennis W. K.
Published: June 2005

Colorectal polyps have a subjectively self‐similar structure which suggests that these structures may have fractal elements and that the fractal dimension may be a useful morphometric discriminant. The fractal dimensions of images from haematoxylin and eosin‐stained sections of 359 colorectal polyps (214 tubulovillous adenomas, 41 ‘pure’ tubular adenomas, 29 ‘pure’ villous adenomas, 68 metaplastic polyps, and 7 inflammatory polyps) were measured using a box‐counting method implemented on a microcomputer‐based image analysis system. Results were assessed using polychotomous logistic regression, confusion matrices, and kappa statistics. All examined polyps were shown to have a fractal structure in the range of scales examined. The fractal dimension was significantly different between different diagnostic categories (P<0·0001) and was a useful discriminant between these categories (kappa statistic 0·60 for the confusion matrix with size as the other variable). The fractal dimension did not shown any significant correlation with the grade of epithelial dysplasia (P>0·05). This study shows that colorectal polyps have a fractal structure over a defined range of magnification and Euclidean morphometric measurements will be invalid outside precisely defined conditions of resolution and magnification. The fractal dimension is a better way of quantitating the polyp shape and is a useful morphometric discriminant between diagnostic categories.
Bcl‐2 protein expression in carcinomas originating from the follicular epithelium of the thyroid gland
Authors: Pilotti Silvana , Collini Paola , Rilke Franco , Cattoretti Giorgio , Del Bo Romualdo , Pierotti Marco A.
Published: June 2005

The bcl‐2 product has been related to the block of programmed cell death (apoptosis) both in lymphoid and in epithelial cells. The pathological expression of bcl‐2 has been investigated mainly in haematological malignancies. Here we have investigated bcl‐2 expression in a model of epithelial tumours represented by the spectrum of carcinomas arising from the follicular epithelium of the human thyroid gland. The analysis was carried out by immunocytochemistry on archival material using monoclonal antibodies against bcl‐2 and thyroglobulin (Tg) on consecutive sections of 94 well‐differentiated carcinomas (WDCs), 19 poorly differentiated carcinomas (PDCs), and 22 undifferentiated carcinomas (UCs) of the thyroid gland. In a subset of 5 cases of UC showing a differentiated component (UC‐D), the expression of p53 protein was also investigated. As controls, fetal and adult normal thyroid glands and adenomas were analysed. bcl‐2 expression was detected in 74 of 94 cases (78·7 per cent) of WDC, 16 of 19 cases (84·2 per cent) of PDC, and 3 of 22 cases (13·6 per cent) of UC. Simultaneous expression of bcl‐2 protein and Tg was observed in 74 of 94 cases (78·7 per cent) of WDC, 13 of 19 cases (68·4 per cent) of PDC, and in no case of UC. bcl‐2 and Tg immunostaining was detected in all fetal and normal thyroid glands as well as in the adenoma specimens examined. In the subset of UC‐D, mutual exclusion of bcl‐2 and p53 expression was observed in the undifferentiated and differentiated components (p53 but not bcl‐2 expressed in the former). Our data indicate that within the spectrum of thyroid carcinomas bcl‐2 expression, similarly to Tg, is strictly correlated with cell differentiation and that its abrogation is restricted to UCs, in which alterations of the p53 gene represent a late genetic event related to tumour progression.
Classification of CEA‐related positivity in primary and metastatic malignant melanoma
Authors: Sanders D. S. A. , Evans A. T. , Allen C. A. , Bryant F. J. , Johnson G. D. , Hopkins J. , Stocks S. C. , Marsden J. R. , Kerr M. A.
Published: June 2005

Using a panel of antibodies to carcinoembryonic antigen (CEA), in paraffin‐processed biopsy material patchy, predominantly membranous positivity was seen on tumour cells in 70 per cent of cases of superficial spreading melanoma, 60 per cent of nodular melanomas, and 75 per cent of secondary deposits studied with unabsorbed polyclonal anti‐CEA only. No staining was seen using monoclonal anti‐CEAs. Localization of CEA to the cell membrane was confirmed with confocal microscopy. Immunoblotting of fresh frozen material detected CEA of around 180 kD in both primary and metastatic melanomas migrating with an apparent molecular weight of between 150 and 200 kD, indicating variable glycosylation of the protein. Recognition of an adhesive role for CEA with roles in immunolocalization and immunotherapy emphasizes the importance of more precise classification of CEA‐related positivity in human tumours.