bluuebunny/crossref_metadata_2025 huggingface dataset:
Immunocytochemical demonstration of IGF‐II immunoreactivity in human phaeochromocytoma and extra‐adrenal abdominal paraganglioma
Authors: Suzuki Toshimitsu , Watanabe Kazuo , Sugino Takashi , Tanigawa Toshitaka , Satoh Shotaro
Published: June 2005

Insulin‐like growth factor (IGF)‐II immunoreactivity was demonstrated in 14 adrenal glands, six retroperitoneal paraganglia, 18 phaeochromocytomas, and six extra‐adrenal abdominal paragangliomas through the use of a monoclonal antibody against rat IGF‐II which cross‐reacts with human IGF‐II. A subpopulation of parenchymal cells in normal adrenal medulla and in retroperitoneal paraganglia was positively immunolabelled. Eighteen cases with phaeochromocytoma including two malignant and four multiple endocrine neoplasias all contained numerous IGF‐II‐immunoreactive tumour cells. All six extra‐adrenal abdominal paragangliomas contained numerous immunoreactive tumour cells. Almost all the other normal human adult tissues examined failed to immunolabel. These results indicate that IGF‐II‐like immunoreactivity is widely expressed in normal and neoplastic human paraganglionic tissues, although its biological significance in these tissues has not been determined.
Ubiquitin: Its potential significance in murine AA amyloidogenesis
Authors: Chronopoulos Soula , Lembo Paola , Alizadeh‐khiavi K. , Ali‐khan Z.
Published: June 2005

Amyloid enhancing factor (AEF), which has recently been shown to have identity with ubiquitin (Ub), is believed to play a causative role in experimentally induced AA amyloidosis in mice. We have examined the profile of Ub in activated leukocytes and splenic reticulo‐endothelial (RE) cells and its relationship with serum amyloid A protein (SAA) and AA amyloid deposits in an alveolar hydatid cyst (AHC)‐infected mouse model of AA amyloidosis. Two monospecific antibodies, anti‐ubiquitin (RABU) and anti‐mouse AA amyloid, were used as immunological probes to localize Ub, SAA, and AA amyloid. In response to AHC infection, the dull and diffuse Ub immunoreactivity in normal mouse leukocytes and RE cells promptly changed to a discrete granular pattern suggesting an increase in the intracellular concentration of Ub and the formation of Ub‐protein conjugates. This corresponded to an elevation in SAA levels, SAA uptake by RABU‐positive phagocytic cells, co‐localization of Ub‐SAA immunoreactive splenocytes in the perifollicular areas, and deposition of Ub‐bound AA amyloid in the splenic and hepatic tissues. These results suggest that Ub‐loaded monocytoid cells may play an important role in the physiological processing of the sequestered SAA into AA amyloid. Aspects of AA amyloidogenesis are discussed in relation to other experimental models in which stress‐induced Ub—protein conjugate formation and its transport to lysosomal vesicles have been studied.
Parathyroid glands in primary hyperparathyroidism: An ultrastructural morphometric study of 25 cases
Authors: Cinti Saverio , Sbarbati Andrea , Morroni Manrico , Carboni Vittorio , Zancanaro Carlo , Cascio Vincenzo Lo , Dickersin G. Richard
Published: June 2005

In parathyroid glands removed from patients with primary hyperparathyroidism (pHPT). hyperplasias and adenomas cannot be distinguished from one another by light microscopy when only one gland is available for examination. When a second gland is available, it is necessary to establish whether it is normal, suppressed, or hyperplastic. This distinction may be difficult, and the main criterion is the amount of cytoplasmic lipid in the parenchymal cells. If the lipid is abundant, the gland is considered normal or suppressed, and if it is scanty, the gland is interpreted as hyperplastic. We have performed a morphometric ultrastructural study to test the reliability of this criterion. Twentyfive adenomatous glands removed from patients with pHPT, when compared with glands of normal size from euparathyroid patients, showed a significant increase in the parameters indicative of metabolic activity, namely the size of the Golgi apparatus, the amount of rough endoplasmic reticulum, and the length of plasmalemmas. In addition, the amount of cytoplasmic lipid was significantly reduced. Furthermore, 25 glands of normal size removed from the same patients with pHPT showed an amount of lipid similar to that of normal glands from euparathyroid patients. However, all the parameters indicative of metabolic activity were significantly higher than those in glands from euparathyroid patients and comparable to those found in adenomatous glands. These results suggest that in pHPT, normal‐size glands are as active as adenomatous glands, regardless of a higher lipid content.
Immunopathological changes in the skin following recombinant interleukin‐2 treatment
Authors: Blessing Karen , Park Kenneth G. M. , Heys Steven D. , King George , Eremin Oleg
Published: June 2005

The systemic administration of the cytokine recombinant interleukin‐2 (rIL‐2) is associated with a number of toxic effects, including dermatological complications. The clinical, histological, and immunopathological changes occurring in the skin of six patients receiving rIL‐2 for metastatic colorectal carcinoma have been studied. All of the patients developed a diffuse erythema shortly after initiation of the treatment. In one patient, generalized erythroderma and photosensitivity developed. The initial biopsy revealed patchy spongiosis, focal exocytosis, and focal basal layer damage with a perivascular chronic inflammatory cell infiltrate. With further treatment there was progressive thickening of the epidermis, Immunohistochemistry revealed changes in the expression of CD1, CD3, CD4, and CD25, particularly on perivascular cells. Additionally, there was an increase in the expression of intercellular adhesion molecule‐1 (ICAM‐1) on keratinocytes in the basal and mid‐layers of the epidermis, endothelial cells, and perivascular cells. It is hypothesized that the expression of ICAM‐1 by the epidermal keratinocytes may be the important event in initiating the cutaneous manifestations associated with rIL‐2 administration.
Production of a monoclonal antibody (IND.64) identifying a cell cycle‐associated antigen using spleen cells from nude mice bearing ichikawa tumour
Authors: Meggetto Fabienne , Saati Talal Al , Cohen‐Knafo Edith , Roubinet Francis , Selves Janick , Bouche Gérard , Key Göran , Gerdes Johannes , Delsol Georges
Published: June 2005

Using spleen cells from athymic nude mice grafted with Ichikawa tumour, we have generated the monoclonal antibody IND.64, which detects a proliferation‐associated nuclear antigen. Immunoblotting analysis with IND.64 showed a double band with apparent molecular weights of 395 and 345 kD. In normal human tissues, the antigen detected by IND.64 was expressed only by the nuclei of proliferating cells, such as germinal centre cells of reactive lymph nodes, cortical thymocytes, the basal layer of the skin, and proiiferative compartments of the stomach, small intestine, and colon. IND.64 did not react with cells known to be non‐proliferative or to show only a low turnover, such as cells of the kidney, liver, smooth muscle, cardiac muscle, and brain. The expression of this antigen during the cell cycle was determined using two approaches: IND.64 immunostaining of synchronized adult bovine aortic endothelial cells and flow cytometric analysis of double‐labelled PHA‐stimulated peripheral mononuclear blood leucocytes with a DNA marker and IND.64. The antigen recognized by IND.64 was found to appear in the late G1 phase, and persisted in phases S, G2, and M, but was absent in the G0 and early Gl phases. IND.64 was further investigated in different tumour types to evaluate the correlation between the percentage of IND.64‐positive cells (IND.64 index) and the histological grade. In non‐Hodgkin's lymphomas, an excellent correlation was found between the percentage of IND.64‐positive cells and the cytomorphological grade. In nodular sclerosis and mixed cellularity Hodgkin's disease, a high number of Reed‐Sternberg cells were positive with IND.64. The non‐lymphoid neoplasms investigated showed a variable percentage of positive cells, IND.64 appears to be a promising tissue marker to complement the evaluation of prognosis in human cancer.
Tumour angiogenesis in prostatic carcinoma with and without bone marrow metastasis: A morphometric study
Authors: Wakui Shin , Furusato Masakuni , Itoh Takaaki , Sasaki Hiroshi , Akiyama Akihito , Kinoshita Ikuhiro , Asano Koji , Tokuda Takaaki , Aizawa Shigeo , Ushigome Shinichiro
Published: June 2005

One hundred and one cases of clinical prostatic carcinoma (PCa), primary site, were analysed to define the inter‐relationship between tumour angiogenesis, histological grade, and bone marrow metastasis. Tumour angiogenesis was determined by the blood capillary density ratio (BCDR; a/b), defined as the ratio between the area of the blood capillaries (a) and the area of the tumour (b). The BCDR was evaluated by a colour image analysis system employing a computerized morphometrical method. A total of 43 cases of PCa with bone marrow metastasis (stage D2) and 58 cases of PCa without metastasis (stage B, C) were utilized. The prostatic carcinomas were classified into three groups (low, intermediate, and high) using Gleason's grading system. 1The BCDR of the primary PCa with bone marrow metastasis was similar in each of the three histologically graded scores. On the other hand, in the cases of PCa without metastasis, the BCDR of high score PCa was higher than those of the low and intermediate score PCa (U‐test; P<0·001). The BCDR of the high score PCa without metastasis was similar to that of the PCa with bone marrow metastasis. The BCDR may provide help in predicting tumour progression with regard to bone marrow metastasis of PCa with low and intermediate Gleason's scores.
L‐Cam expression in normal, premalignant, and malignant colon mucosa
Authors: Van Der Wurff Anna A. M. , Kate Joop Ten , Van Der Linden Edith P. M. , Dinjens Winand N. M. , Arends Jan‐Willem , Bosman Fred T.
Published: June 2005

L‐CAM is a cell adhesion molecule which is expressed at the intercellular borders of most epithelial cells. L‐CAM has been demonstrated to act as an invasion suppressor in carcinoma cell lines. In order to determine whether or not L‐CAM expression might distinguish between invasive and non‐invasive or metastatic and non‐metastatic colon neoplasms, we studied L‐CAM expression in normal colon mucosa, colon adenomas with various degrees of dysplasia, and colon carcinomas by immunohistochemistry, using the 6F9 monoclonal anti‐L‐CAM antibody. Normal mucosa showed evenly distributed distinct L‐CAM immunoreactivity along intercellular borders. In adenomas and carcinomas, a similar though weaker expression was observed. This pattern showed a tendency to decrease in parallel with decreasing differentiation. However, no correlation was found with Dukes stage or area within the tumour. In some carcinomas, L‐CAM was expressed at the luminal surface of the cells. In others, L‐CAM expression was not found. These results suggest that L‐CAM expression is disregulated or lost as an early event in the development of colon neoplasia and indicate that L‐CAM expression does not correlate with invasive or metastatic potential.
p53 immunostaining in the distinction between benign and malignant mesothelial proliferations using formalin‐fixed paraffin sections
Authors: Mayall Frederick G. , Goddard Heather , Gibbs Allen R.
Published: June 2005

The distinction between reactive mesothelium and mesothelioma in pleural biopsy specimens is notoriously difficult, and conventional immunohistochemical markers have provided no relief. The object of this study was to examine the frequency of immunohistochemically detectable p53 overcxpression in routinely processed, paraffin‐embedded tissue from pleural mesotheliomas and from pleura showing reactive mesothelial hyperplasia, using a polyclonal antibody to formalin‐resistant p53 epitopes, and to consider the diagnostic utility of this antibody in the distinction between mesothelioma and reactive mesothelium in pleural biopsy specimens. Immunostaining was enhanced by pepsin predigestion prior to the application of the primary antibody. Positivity occurred in 10/16 epithelial mesotheliomas, 9/19 biphasic mesotheliomas, 2/12 sarcomatous mesotheliomas but in none of 20 reactive pleura. Immunostaining was particularly intense in some of the biopsy specimens, which may be due to the rapidity with which these small pieces of tissue were fixed. In conclusion, this study suggests that p53 immunostaining can help to distinguish epithelial or biphasic mesothelioma from reactive mesothelial hyperplasia in formalin‐fixed, paraffin‐embedded pleural biopsy specimens.
Expression of the p53 protein in a spectrum of astrocytic tumours
Authors: Ellison David W. , Gatter Kevin C. , Steart Philip V. , Lane David P. , Weller Roy O.
Published: June 2005

Many human cancers are characterized by mutations of p53, a nuclear phosphoprotein which controls elements of the cell cycle. Turnover of p53 in normal cells is rapid, and the minute quantities of protein that are usually present are not detected by immunocytochemical methods. Mutations of the p53 gene in tumour cells are associated with a slower turnover and subsequent accumulation of the protein in both nucleus and cytoplasm. Genetic abnormalities of the short arm of chromosome 17, which is the site of the p53 gene locus, are a feature of astrocytic tumours. Usinga panel of five antibodies to p53 and a standard immunocytochemical method, we found detectable quantities of p53 in the cells of 3/16 diffuse astrocytomas, 8/14 anaplastic astrocytomas, and 24/34 glioblastoma multiforme. Progression of one patient's tumour from a diffuse to an anaplastic astrocytoma was characterized by the accumulation of p53. The more malignant histological features of anaplastic astrocytoma and glioblastoma multiforme appear to be reflected by a greater incidence of p53 accumulation.
Characterization of the smooth muscle cell infiltrate and associated connective matrix of lymphangiomyomatosis. Immunohistochemical and ultrastructural study of two cases
Authors: Peyrol S. , Gindre D. , Cordier J. F. , Loire R. , Grimaud J. A.
Published: June 2005

Lymphangiomyomatosis (LAM) consists of smooth muscle (SM) cell proliferation of unknown origin involving the lymph nodes and the lung interstitium. From morphological studies showing both SM differentiation of the proliferating cells and lymphatic hyperplasia, hypotheses were suggested concerning the origin of the proliferation. Two cases of LAM were investigated by electron microscopy and immunohistochemistry; tissues were obtained by lymph node and open lung biopsies. Cytoplasmic and matrix protein markers were used in order to clarify the pattern of differentiation of the proliferating cells and to characterize their connective tissue environment. The proliferating cells present ultrastructural characteristics of SM cells; they contain vimentin, desmin, and α‐SM actin and are devoid of Factor VIII, favouring a parieto‐arterial origin. The connective tissue matrix inside the infiltrate is composed of interstitial collagens and basement membrane components. At the late stage of the disease, remodelling of the interstitial matrix accompanies the infiltrate and remains perilesional.
Localization of cea messenger RNA by in situ hybridization in normal colonic mucosa and colorectal adenocarcinomas
Authors: Abbasi Tta M. , Chester Kerry A. , Macpherson Andrew J. S. , Boxer Geoffrey M. , Begent Richard H. J. , Malcolm Alan D. B.
Published: June 2005

Carcinoembryonic antigen (CEA) mRNA expression was studied in 14 cases of normal colorectal mucosa and cclonic adenocarcinomas using in situ hybridization with a 32P‐labelled cDNA probe to the unique 3‐untranslated region of CEA. This approach has the advantage that the target mRNA remains in the cell of origin, whereas there is considerable ambiguity in immunocytochemistry data for CEA because the protein is secreted. Furthermore, the specific cDNA probe overcomes potential problems of immunological cross‐reactivity with other members of the CEA family. The results demonstrated that abundant, heterogeneously distributed CEA mRNA was present in colorectal adenocarcinomas, with the highest levels in cells lining glandular structures. Parallel immunohistochemistry with anti‐CEA monoclonal antibody A5B7 showed that the regions of tumours with the highest levels of CEA mRNA also had the highest CEA protein levels, suggesting that the heterogeneous distribution reflects CEA expression rather than differential secretion of the protein. In the normal colonic mucosa, CEA mRNA expression was observed in surface epithelial cells and goblet cells of the upper crypts, with very low hybridization in the mid crypt and at the base. This crypt‐surface distribution was identical to that observed for CEA protein. In situ hybridization therefore confirms that high levels of CEA mRNA are expressed in differentiated surface epithelial cells of the normal colon.
Overexpression of p53 in Hodgkin's disease: Lack of correlation with Epstein–Barr virus infection
Authors: Niedobitek Gerald , Rowlands David C. , Young Lawrence S. , Herbst Hermann , Williams Anne , Hall Philip , Padfield James , Rooney Nick , Jones E. Lynn
Published: June 2005

Recent work has shown that p53 gene mutations are frequently found in Epstein‐Barr virus (EBV)‐positive and EBV‐negative cases of Burkitt's lymphoma but not in EBV‐associated undifferentiated nasopharyngeal carcinomas (NPCs). Similar viral gene expression patterns are observed in undifferentiated NPCs and in EBV‐positive cases of Hodgkin's disease (HD), suggesting that the contribution of the virus to the pathogenesis of these malignancies may also be similar. We have analysed 116 cases of HD for EBV association and for immunohistologically detectable overexpression of p53. p53 overexpression was detected in the tumour cell population of 37 (32 per cent) of the cases. Fifteen cases showed p53‐specific labelling of more than 40 per cent of tumour cells; in six of these, virtually all tumour cells were stained. In eight cases, between 5 and 40 per cent of tumour cells were labelled, and in another 14 cases, less than 5 per cent of tumour cells expressed detectable amounts of p53. EBV‐positive HD cases were found in all groups with different levels of p53 overexpression as well as amongst p53‐negative cases. While a more detailed analysis of the p53 gene in HD is required, these data show that overexpression of p53 in HD is heterogeneous and that there is no simple correlation between EBV infection and p53 overexpression.
Technical method a simple technique for the measurement of fractal dimensions in histopathological specimens
Authors: Sanders Harry , Crocker John
Published: June 2005

A computer program has been devised which enables a simple method for the fractal dimension analysis of histological or cytological material to be performed rapidly with relatively inexpensive hardware. The fractal dimension of a structure refers to its self‐similarity over a range of scales and appears to be well suited to histopathological preparations. Thus, the fine ‘roughness’ of object profiles can be assessed more appropriately by this technique than by means of conventional Euclidean geometry. Images of the object(s) to be investigated are firstly digitized after presentation to a high‐performance computer via a video camera or a graphics tablet system and then analysed using the computer program, which has its basis in a standard perimeter estimate method. It is proposed that such techniques may find extensive use in investigative and diagnostic histopathology.
Expression and localization of plasminogen activator inhibitor 1 mRNA in transplant kidneys
Authors: Wang Yiming , Thompson Elizabeth M. , Whawel Simon A. , Fleming Kenneth A.
Published: June 2005

Fibrinolysis in renal transplant rejection has not been systematically investigated but is known to be impaired. By an in situ hybridization technique, we have studied gene expression of plasminogen activator inhibitor 1 (PAI‐1) in human renal tissue showing severe acute vascular rejection (n=8), clinically irreversible (chronic) vascular rejection (n = 3), mild vascular rejection (n= 8), parenchymal rejection (n=4), ‘normal’ kidneys (n=6), and non‐rejecting kidneys (n=6). The results showed that in 7/8 cases showing severe acute vascular rejction and in all three chronic vascular rejection cases, PAI‐1 mRNA was expressed by endothelial cells of arterioles and arteries, and interstitial inflammatory cells but was not detectable in any other groups. The expression of PAI‐1 was frequently associated with areas of haemorrhage. Expression of PAI‐1 might be expected to promote thrombosis and ischaemia, the catastrophic consequence of severe vascular rejection. In irreversible chronic rejection, this seems to be the principal mode of action. However, our observation that there is expression of PAI‐1 in areas of haemorrhage in severe acute vascular rejection may suggest an additional potentially protective role, if it were produced as a response to haemorrhage.
Immunohistochemical demonstration of androgen receptor in breast cancer and its relationship to other prognostic factors
Authors: Isola Jorma J.
Published: June 2005

Androgen actions and androgen receptors (ARs) have been described in human breast cancer cells both in vivo and in vitro. With the use of a new monoclonal anti‐AR antibody, AR was immunohistochemically demonstrated in 76 primary breast cancers. Positive immunostaining was found in 79 per cent of tumours. Benign ductal epithelium was often AR‐positive whereas the tumour stroma lacked AR immunoreactivity. At the subcellular level, nuclear localization was evident using either cross‐linking (Zamboni's fluid) or precipitating (acetone) fixatives on frozen sections. The use of archival paraffin‐embedded tissue yielded negative results. A significant association was found between expression of AR and oestrogen receptor (ER) (P=0.0006) determined immunohistochemically on adjacent sections. Most progesterone receptor (PR)‐negative cases were also AR‐negative (P=0.02), but significant proportion (38 per cent) of AR‐positive tumours did not contain PR. Unlike ER, AR was not associated with aneuploidy or erb‐B2 oncogene overexpression, and was only marginally associated with tumour proliferation rate (S‐phase fraction by DNA flow cytometry). In conclusion, the close association of AR with ER and PR suggests that immunohistochemical determination of androgen receptors may have value as a prognostic factor and/or predictor of response to endocrine therapy.
Demonstration of coxsackie virus RNA in formalin‐fixed tissue sections from childhood myocarditis cases by in situ hybridization and the polymerase chain reaction
Authors: Hilton David A. , Variend Sadick , Pringle James H.
Published: June 2005

This is a combined study using in situ hybridization and the polymerase chain reaction to investigate the presence of Coxsackie virus RNA in formalin‐fixed tissue from cases of childhood myocarditis. Of the ten cases studied, two were positive by both methods. The virus RNA was predominantly located in areas showing an inflammatory cell infiltrate and myofibre necrosis. These findings suggest that direct lytic infection of myocytes by virus is responsible for myocaiditis in these cases, rather than an autoimmune process, which has been suggested previously. The findings in one case, where the virus showed a marked sub‐endocardial distribution, may have implications for the aetiology of endocardial fibroelastosis by confirming a viral tropism for this location. The techniques used in this study are easily repeatable and can be directly applied to look for viruses in a number of other diseases where a viral aetiology is suspected.
Protection of germinal centres from complement attack: Decay‐accelerating factor (DAF) is a constitutive protein on follicular dendritic cells. A study in reactive and neoplastic follicles
Authors: Lampert I. A. , Schofield J. B. , Amlot P. , Van Noorden Susan
Published: June 2005

The development of B‐cell memory is linked to the presence of germinal centres. This process is dependent on the presence of antigen, usually in the form of immune complexes with antibody, on the surface of the follicular dendritic cells (FDCs) that form a network in the germinal centre. The presence of immune complexes poses a constant danger of activating complement. Decay accelerating factor (DAF, CD55) and the membrane attack complex (MAC) inhibitor (CD59) are two cell proteins whose sole function is to protect cells from the action of complement, the former affecting the earlier components of the complement cascade, and the latter the terminal ones; both are bound to the cell surface via a glycosylphosphatidylinositol link. DAF but not CD59 could be demonstrated on FDCs. DAF is also present on the FDCs in follicular lymphomas despite the absence of complement (C3) in neoplastic follicles. This indicates that DAF is constitutive to FDCs but does not preclude the possibility that its expression is increased when immune complexes are deposited.
An immunohistological study of cutaneous lymphocytes in vitiligo
Authors: Badri A. M. T. Al , Todd Pamela M. , Garioch Jennifer J. , Gudgeon Janet E. , Stewart D. G. , Goudie R. B.
Published: June 2005

In a study of 49 biopsies from the margins of depigmented cutaneous lesions in 18 patients with vitiligo, highly significant overall increases were found in CD3+, CD4+, and CD8+ T cells, though cell numbers in individual cases were often within the normal range. Many of the T cells were activated (MHC class II+, interferon gamma+), of CD45RO (UCHL1+) memory subset, and many expressed the cutaneous lymphocyte‐associated antigen (HECA‐452+) typical of skin‐homing T cells. Immunohistologically, the most intense epidermal T‐cell infiltration was present within 0.6 mm of the edge of the lesion in 10 of 13 double‐stained sections with a clearly defined zone of melanocyte depletion. In 40 lesions from 17 patients seen 11–64 weeks after biopsy, no apparent association was found between T‐cell numbers and disease activity as assessed by Köbnerization of biopsy wounds or spread of depigmentation. These findings are consistent with the hypothesis that lesional T cells rather than circulating antimelanocytic antibody may be responsible for the supposedly autoimmune but characteristically patchy destruction of cutaneous melanocytes in vitiligo. Nevertheless, many of the infiltrating T cells are probably innocent bystanders attracted by upregulated cell adhesion molecules near sites of melanocyte damage.
Expression of proliferating cell nuclear antigen (PCNA) in dysplasia of the bronchial epithelium
Authors: Pendleton Neil , Dixon Glen R. , Burnett Heather E. , Occleston Nicholas L. , Myskow Michael W. , Green John A.
Published: June 2005

Proliferating cell nuclear antigen (PCNA) is expressed in cells in the cell cycle and has been studied as a marker of proliferation in lung and other tumours. We have noted immunocytochemical differences in PCNA expression between normal and neoplastic bronchial cells. As bronchial dysplasia is considered preneoplastic, we have examined PCNA expression in this condition. PCNA staining in 47 cases of bronchial dysplasia and 32 samples of normal bronchial epithelium was compared. Of the dysplasias, three were mild, 11 moderate, and 33 severe. A significant increase in PCNA counts over normal epithelium was seen only in moderate and severe dysplasias. In dysplasia, mitotic indices showed a significant positive correlation with the percentage of PCNA‐positive cells. We conclude that in moderate and severe dysplasias there is an increase in the number of cells expressing PCNA and undergoing division, indicating abnormal growth control.
Constant detection of surface and cytoplasmic immunoglobulin heavy and light chain expression in formalin‐fixed and paraffin‐embedded material
Authors: Merz Hartmut , Rickers Olaf , Schrimel Stefan , Orscheschek Karin , Feller Alfred C.
Published: June 2005

The introduction of microwave detection systems has significantly enhanced the sensitivity of immunohistochemistry in formalin‐fixed, paraffin‐embedded tissue. The combination of protease digestion and microwave treatment including the use of heavy metal salts or urea gives an Ig heavy and light chain detection system in formalin‐fixed and paraffin‐embedded material as good as that achieved using fresh frozen material, with the advantage of a well‐preserved morphology. The sensitivity of this method depends on the heavy metal salt used and its concentration. The use of urea yields equally sensitive results, while avoiding the toxicity of heavy metal salts. Heating times in the microwave oven also have an important influence on the results. The microwave‐based technique may allow for the improved accessibility of formalin‐masked antigens, as was shown here for cytoplasmic and surface immunoglobulins. It was demonstrated that formalin‐fixed specimens of Burkitt's lymphoma when subjected to the new protocol almost always showed surface Ig expression, giving results comparable to those obtained in frozen sections. Moreover, it could be shown that in cases of B‐chronic lymphocytic lymphoma (B‐CLL) or immunocytoma, surface and cytoplasmic Ig could be detected, so that the difference may be just a quantitative one and thus may no longer be safely used as a criterion in differential diagnosis. In future, the wide use of this technique will allow for the study of Ig expression at the single cell level in morphologically well‐preserved material.
Immunohistochemical identification of leucocyte populations in normal tissue and inflamed synovium of the rabbit
Authors: Wilkinson J. Michael , McDonald Gillian , Smith Simon , Galea‐lauri Joanna , Lewthwaite Joanne , Henderson Brian , Revell Peter A.
Published: June 2005

A panel of monoclonal antibodies which recognizes well‐defined markers of rabbit leucocytes has been used in an immunohistochemical survey of normal rabbit spleen, thymus, and liver, and of inflamed synovial tissue. The results demonstrate that these antibodies are well suited for the identification of leucocytes in frozen sections of rabbit tissues and that T lymphocytes, macrophages. and neutrophils can be readily distinguished. Antibodies to CD44 differentiate between medullary and cortical thymocytes, with the cortical cells being almost entirely negative for this marker. Antibodies to RLA class II show that periportal lymphocytes in the liver express RLA‐DR but not RLA‐DQ. The type A cells of the synovial lining from inflamed joint tissue are stained by antibodies that recognize macrophages in a manner similar to that found in the human synovial lining.
Monoclonal antibodies to cytoskeletal proteins: An immunohistochemical investigation of human colon cancer
Authors: Porter Rebecca M. , Holme Thomas C. , Newman E. Luke , Hopwood David , Wilkinson J. Michael , Cuschieri Alfred
Published: June 2005

Monoclonal antibodies raised to a number of microfilament‐associated proteins were shown to recognize the appropriate proteins in extracts from human colon tissue. They were then used in an immunohistochemical study of normal colonic mucosa, adenomas, and adenocarcinomas. A strong reaction was seen in stromal cells within the tumours (both adenomas and adenocarcinomas) when frozen sections were stained with antibodies to filamin and caldesmon. In addition, a similar reaction was seen in the adenocarcinomas when stained with antibodies to talin and gelsolin. We believe that immunohistochemical staining with these antibodies reveals a tumour‐induced process in the surrounding cells, possibly related to a host response to tumours.
P53 overexpression and Epstein‐Barr virus infection in undifferentiated and squamous cell nasopharyngeal carcinomas
Authors: Niedobitek Gerald , Agathanggelou Angelo , Barber Peter , Smallman Lesley A. , Jones E. Lynn , Young Lawrence S.
Published: June 2005

We have analysed 22 nasopharyngeal carcinomas (NPCs) for expression of the small nuclear Epstein‐Barr virus (EBV)‐encoded RNAs (EBERs) and for immunohistologically detectable overexpression of p53. in situ hybridization demonstrated expression of the EBERs in 13 undifferentiated NPCs while nine squamous cell NPCs were EBER‐negative. These results therefore confirm our previous DNA‐DNA in situ hybridization studies and demonstrate that in the nasopharynx EBV is exclusively associated with undifferentiated but not with squamous cell carcinomas. p53 overexpression was demonstrated by immunohistology in 5 of 9 squamous cell NPCs and in 9 of 13 undifferentiated NPCs. Thus, there appears to be no correlation of p53 overexpression with EBV infection. These results are unexpected in the light of previous studies demonstrating that the p53 gene in primary undifferentiated NPC is consistently in the wild‐type configuration. By contrast, analyses of squamous cell carcinomas of the head and neck have demonstrated that p53 overexpression in these cases is the result of p53 gene mutation. Whilst more detailed genetic analysis is required, our results suggest that mechanisms other than mutation of the p53 gene may be responsible for the stabilization of the protein in cases of undifferentiated NPC. It is tempting to speculate that an EBV‐encoded protein may be involved.
Ulcer‐associated cell lineage (‘pyloric metaplasia’) in crohn's disease: A lectin histochemical study
Authors: Roberts Ian S. D. , Stoddart Robert W.
Published: June 2005

Chronic intestinal ulceration in Crohn's disease is associated with the development of an epidermal growth factor‐secreting cell lineage, or ‘ulcer‐associated cell lineage’ (UACL).1 Expression of oligosaccharides by UACL was studied using a panel of 25 biotinylated lectins with an avidin peroxidase revealing system and compared with that of adult and fetal Brunner's glands, gastric antral mucosa, and ‘gastric metaplasia’ within the duodenum, in order to clarify further the interrelationships of these lineages. UACL was obtained from ileal resections performed for Crohn's disease.

Lectin binding of the glandular component of UACL closely resembled that of antral mucosal glands and also that of fetal and adult Brunner's glands. Lectin binding of the ductal component of immature UACL, in which a surface component had not developed, resembled that of the gland. The surface and ductal components of mature UACL showed a distinct lectin‐binding profile, which was very different from that of the gland, but closely resembled that of antral foveolar epithelium and ‘gastric metaplasia’ within the duodenum.

It is concluded that there is differentiation of UACL from the glandular to surface components and that oligosaccharide expression of the lineage reflects that of normal Brunner's gland and gastric antral mucosa.
p53 Antigen in cervical condylomata, intraepithelial neoplasia, and carcinoma: Relationship to hpv infection and integration
Authors: Cooper Kumarasen , Herrington C. Simon , Evans Mark F. , Gatter Kevin C. , McGee James O'D.
Published: June 2005

It has been proposed that wild‐type p53 cell‐regulating functions are annulled in human cervical carcinomas, either by mutations in the human papillomavirus (HPV)‐negative cases or as a consequence of their complexing with HPV E6. The aim of this study was to test this hypothesis on 39 fresh cervical biopsies by p53 immunocytochemistry (ICC) with antibody PAb 240 and with NISH (non‐isotopic in situ hybridization) and PCR (polymerase chain reaction) for HPV detection. p53 protein was present in the basal layer of pure wart virus infection; the basal to middle third of CIN (cervical intraepithelial neoplasia); in 19/22 (86 per cent) HPV‐positive cervical carcinomas, ten of which contained integrated HPV; and in 4/8 (50 per cent) HPV‐negative cervical carcinomas. Dual detection of p53 antigen and HPV 16 DNA in the same sections demonstrated either p53 protein or integrated HPV 16 alone in the majority of cells. Co‐localization of both signals was only evident in isolated cells. These data suggest that PAb 240 immunoreactivity is not mutant‐specific. They are, however, consistent with the conformation hypothesis which proposes that wild‐type p53 changes from a suppressor (PAb 240‐negative) to a promoter (PAb 240‐positive) form during cell growth response. Hence, according to this hypothesis, p53 protein expression may represent either the wild‐type promoter form or mutant p53 protein, both of which share the same conformation. This may explain co‐localization of p53 and HPV in some tumours. However, the absence of p53 protein in 50 per cent HPV‐negative squamous cell carcinomas suggests that not all HPV‐negative tumours accumulate p53 protein.
Cytochrome P450 expression is a common molecular event in soft tissue sarcomas
Authors: Murray Graeme I. , McKay Judith A. , Weaver Richard J. , Ewen Stanley W. B. , Melvin William T. , Burke M. Danny
Published: June 2005

Two major xenobiotic metabolizing sub‐families of cytochrome P450 (cytochrome P450 1A and cytochrome P450 3A) have been identified in soft tissue sarcomas. Cytochrome P450 1A was present in 70 per cent and cytochrome P450 3A was present in 78 per cent of tumours, respectively. A high proportion (86 per cent) of those tumours which contained cytochrome P450 1A or cytochrome P450 3A demonstrated co‐expression of both sub‐families. In each tumour, cytochrome P450 immunoreactivity was identified in all tumour cells and there was no intra‐tumour heterogeneity. These results indicate that expression of cytochrome P450 is a common molecular event in soft tissue sarcomas and that the presence of different sub‐families of cytochrome P450 has implications both for the pathogenesis and for the treatment of these tumours. Cytochrome P450 expression may influence the intrinsic drug resistance of these tumours and also provide a molecular target for anti‐cancer drugs which can be activated by cytochrome P450.
Demonstration of light chain mRNA in Hodgkin's disease
Authors: Hell Karen , Pringle James H. , Hansmann Martin‐L. , Lorenzen Johann , Colloby Peter , Lauder Ian , Fischer Robert
Published: June 2005

The lineage of Hodgkin and Reed‐Sternberg cells is still unclear. Detection of both immunoglobulin light chains in Hodgkin and Reed‐Sternberg cells by immunohistochemistry is a well‐known phenomenon. However, up to now, in situ hybridization techniques have failed to demonstrate light chain messenger(m) RNA in Hodgkin and Reed‐Sternberg cells. In this investigation, we have analysed 26 cases of Hodgkin's disease (nodular lymphocyte predominant Hodgkin's disease, mixed cellularity, and nodular sclerosis type) using digoxigenin‐labelled oligonucleotide probes for K and λ light chains by in situ hybridization. In nearly half of the cases of nodular lymphocyte predominant Hodgkin's disease and in one case of mixed cellularity type, mRNA for only one light chain could be clearly demonstrated in the lymphocytic and histiocytic cells, Hodgkin, and Reed‐Sternberg cells. These results support the idea that at least some cases of Hodgkin's disease are B‐cell neoplasms.
Expression of keratin mRNAS and proteins in normal salivary epithelia and pleomorphic adenomas
Authors: Su Lan , Morgan Peter R. , Harrison Dolores L. , Waseem Ahmad , Lane E. Birgitte
Published: June 2005

Control of keratin (K) gene expression may be important for cell differentiation in complex epithelia such as salivary gland. To investigate differences in distribution between keratin mRNAs and their respective proteins, a combined in situ hybridization (ISH) and immunohistochemical study was undertaken on nine normal salivary glands and seven pleomorphic adenomas. ISH employed riboprobes to K7, K8, K14, K18, and K19. Immunohistochemistry was performed on adjacent sections using monoclonal antibodies (MAbs) to the same keratins. Normal luminal cells showed abundant hybridization with probes for K7, K8, K18, and K19. Keratin 14 mRNA was present in basal and myoepithelial cells at a low level of expression. Proteins of their keratins were strongly stained. Neoplastic cells showed variable expression of mRNA and protein for K7, K8, K18, and K19. There was a high level of K14 mRNA but variable protein. The findings provide evidence that expression of these keratins in normal salivary epithelia is regulated transcriptionally and that in neoplasia this system is in considerable disarray.
Proliferative activity as revealed by Ki‐67 in uterine adenocarcinoma of endometrioid type: Comparison of tumours from patients with and without previous oestrogen therapy
Authors: Nielsen Anette Lynge , Nyholm Henrik Christian Juul
Published: June 2005

The Ki‐67 antibody recognizes a nuclear antigen related to cell proliferation, which in some studies has been shown to reflect the aggressiveness of tumours. The percentage of Ki‐67‐positive cells was estimated by immunohistochemistry on frozen tissue sections from 73 adenocarcinomas of endometrioid type (EC) (40 tumours from patients who had never received postmenopausai oestrogen treatment and 33 tumours from patients with previous postmenopausal oestrogen treatment). The Ki‐67 content was weakly but significantly (P<0.05) correlated to nuclear grade, architectural grade, and crude mitotic count, but not to stage or progesterone receptors. Ki‐67 expression in EC from patients with previous oestrogen therapy was much lower (median 10 per cent Ki‐67) than that in EC from patients who had never received oestrogen treatment (median 24 per cent Ki‐67), suggesting that a prognostic cut‐off point may be different in tumours from the two groups of patients. The mitotic count discriminated the two groups of patients to a much smaller degree. It is also shown that a quick qualitative Ki‐67 estimate can replace the time‐consuming quantitative assessment.
Comparison of three quantitation methods for PCNA immunostaining: Applicability and relation to survival in 83 astrocytic neoplasms
Authors: Haapasalo Hannu K. , Sallinen Pauli K. , Helen Pauli T. , Rantala Immo S. , Helin Heikki J. , Isola Jorma J.
Published: June 2005

Recent studies on astrocytic tumours demonstrated a close association between patient prognosis and neoplastic proliferation estimated by such methods as Ki‐67 and bromodeoxyuridine labelling. Novel monoclonal PCNA antibodies and special antigen‐retrieval techniques have the advantage of working on routinely fixed and embedded specimens and thus make the estimation of proliferation simpler. In addition to PCNA‐positive cell count expressed in percentages (PCNA‐LI), we estimated the number of PCNA‐immunopositive cells of 83 astrocytomas in two ways: (1) per mm2 of neoplastic tissue (uncorrected PCNA index); and (2) per mm2 of total neoplastic nuclear area (corrected PCNA index). Both of these methods were reproducible and showed a goad correlation with PCNA‐LI and malignancy grade (I–IV). With quantitation methods 1 and 2, the proliferative status of about 2000 cells could be estimated in about 7–10 min, whereas the PCNA count by PCNA‐LI of 200 cells took approximately the same time. The proliferation indices obtained by all three quantitation methods were highly significantly related to patient prognosis. The corrected PCNA index, having a close association with the neoplastic cellularity, even divided the glioblastoma group (grade IV) into two significantly different prognostic groups in which 56 and 17 per cent of the patients were alive after 1‐year follow‐up. The combination of PCNA immunohistochemistry and morphometry seems to give important prognostic information about astrocytomas independent of the histopathological grade.
Image analysis of Ag‐NOR proteins in transitional cell bladder cancer
Authors: Lipponen Pertti
Published: June 2005

Nuclear organizer regions (Ag‐NORs) were visualized using the standard silver‐staining technique in biopsy specimens from 112 bladder carcinomas. The area of individual Ag‐NOR dots and the area of Ag‐NOR protein/nucleus were measured using a Quantimet 570 image analyser and the results were then correlated with clinical, histological, and quantitative prognostic factors, and survival. The mean area of Ag‐NOR dots was not significantly related to any of the features analysed. The mean area of Ag‐NOR protein/nucleus was related to WHO grade (P=0.0004), papillary status (P=0.004), DNA ploidy (P=0.02), S‐phase fraction (P=0.02), mitotic frequency/mm2 of enoplastic epithelium (P=0.005), mean nuclear area (P=0.001), and SD of nuclear area (P=0.001). In the entire cohort, the mean area of Ag‐NOR protein/nucleus was related to progression with a borderline significance (P=0.09), while in survival analysis only Ag‐NOR count/nucleus had prognostic value (P=0.010). In Ta‐T1 tumours, Ag‐NOR protein area/nucleus predicted progression (T‐category) (P=0.016) and survival (P=0.014) significantly. Also, the mean area of individual Ag‐NOR dots (P=0.026) and Ag‐NOR count/nucleus (P=0.014) were related to prognosis in Ta‐T1 tumours.
Retinoblastoma and p53 tumour suppressor gene protein expression in carcinomas of the thyroid gland
Authors: Holm Ruth , Nesland Jahn M.
Published: June 2005

One hundred and thirty‐one thyroid tumours were examined immunohistochemically for expression of retinoblastoma (RB) and p53 protein. The results demonstrate that RB protein is not lost in any cases, indicating that inactivation of the RB gene is unlikely to play a central role in the pathogenesis of thyroid tumours. Eighteen of 24 (75 per cent) undifferentiated carcinomas, 6 of 32 (19 per cent) papillary carcinomas, 5 of 29 (17 per cent) follicular carcinomas, and 6 of 46 (13 per cent) medullary carcinomas showed p53 protein nuclear staining. In 46 per cent of the undifferentiated carcinomas many of the tumour cells had accumulated p53 protein, whereas in the other positive cases less than 5 per cent of the cells had increased p53 protein levels. Our results strongly suggest that p53 protein abnormalities play a crucial role in the progression of well‐differentiated to undifferentiated thyroid carcinomas.
Expression of bcl‐2 gene product in neuroblastoma
Authors: Ramani Pramila , Lu Qi‐Long
Published: June 2005

The expression of bcl‐2 is associated with inhibition of apoptosis and prolonged cell survival. The purpose of this study was to examine the immunoreactivity of neuroblastoma and ganglioneuroblastoma tissue samples to the bcl‐2 gene product in order to see if it was related to prognosis. BCL‐2 protein was detected in all the 46 formalin‐fixed, paraffin‐embedded samples from 34 patients representing all clinical stages and sites of involvement. Immuno‐positivity was observed in tumours from the primary and metastatic sites. Moreover, it was demonstrated in the pre‐chemotherapy and the post‐chemotherapy samples from six cases with stage 4 disease. It was observed in neuroblasts in various stages of differentiation. A small proportion of undifferentiated neuroblasts were negative. As BCL‐2 oncoprotein was present in all the cases irrespective of the clinical outcome, it does not appear to be one of the factors influencing prognosis.
The effect of inhibition of glutathione S‐transferase P on the growth of the jurkat human T cell line
Authors: McCaughan Frank M. , Brown Alan L. , Harrison David J.
Published: June 2005

To investigate the possible role of glutathione S‐transferase P (GSTP) in carcinogenesis and cell proliferation, ethacrynic acid (EA) was used to inhibit GSTP in the human Jurkat T cell line. At lower doses (0–30 μM), EA led to a decreased rate of proliferation as assessed by the MTT assay. This was associated with a decreased DNA S + G2/M phase population and also a dose‐dependent increase in apoptosis. At concentrations of EA >30μM, cells suffered non‐specific cytotoxic injury and underwent necrosis. The total cell number fell over the time course of the experiment. A resistant subculture of cells which proliferated in the presence of EA at 30μM was selected by continuous growth in the presence of EA. Although this had a higher basal rate of apoptosis than control cells, it also showed a significantly larger growth fraction as assessed by flow cytometry. GSTP is frequently overexpressed in human tumours and animal models of carcinogenesis, and is regarded as a marker of the ‘drug‐resistant phenotype’ of initiated cells. Our findings suggest that the role of GSTP in models of chemical carcinogenesis and in tumours may be its permissive effect on cell cycle activity and downregulation of apoptosis, thus allowing expansion of a population of initiated cells.