bluuebunny/crossref_metadata_2025 huggingface dataset:
Papillomavirus screening in cervical cell samples using dual‐label dot‐blot analysis
Authors: Potter Colin G. , Cooper Kum , Stickland Julia E. , Ramshaw Anna L. , O' McGee James D.
Published: June 2005

The presence of human papillomavirus (HPV) in cervical cells is closely related to the development of cervical carcinoma. Detection of virus may be by Southern blot, dot blot or the highly sensitive polymerase chain reaction. Whatever method is employed, there are problems of false negatives due to poor clinical samples in which the DNA may be degraded or is absent altogether. Here we describe a new method of dual labelling for dot blots using a 32P‐labelled probe for HPV and a 35S‐labelled probe for human actin genes. The samples were counted on a Betaplate™ flat‐bed scintillation counter and the data analysed to separate the activities of the two isotopes. The counts from the actin probe show whether human DNA is present or not and false negatives from this cause may thereby be eliminated. The counts due to HPV when compared with those for actin give a quantitative measure of HPV abundance for the particular sample and this may have clinical relevance.
Three‐dimensional reconstruction of human embryonic notochords: Clue to the pathogenesis of chordoma
Authors: Salisbury Jonathan R. , Deverell Mark H. , Cookson M. John , Whimster William F.
Published: June 2005

Three‐dimensional reconstruction experiments performed on serial sections of human embryos showed that the anatomy of the caudal and rostral ends of the notochord was complex. Forking of the ends, with separate fragments of chordal tissue, was demonstrated and these provide a way by which notochordal cell rests could be left behind in the basicranial and sacral regions when the notochord involutes elsewhere. Assuming the histogenesis of chordomas from notochordal cell rests, this would furnish an explanation for the observed skeletal distribution of chordomas.
Clinical necropsy rates during the 1980s: The continued decline
Authors: Start R. D. , McCulloch T. A. , Benbow E. W. , Lauder I. , Underwood J. C. E.
Published: June 2005

The trends in necropsy rates during the 1980s in three groups of British teaching hospitals in Leicester, Manchester, and Sheffield were compared in a retrospective study. The clinical necropsy rates declined in all three cities: in Leicester, from 16 to 10 per cent; in Manchester, from 14 to 8 per cent; and in Sheffield from 18 to 11 per cent. The medico‐legal and overall necropsy rates showed variable trends between the cities. Specific events and changes in organization during the review period were correlated with the changing trends. The observed trends emphasize the continued decline in clinical necropsy rates over the last decade and illustrate the importance of monitoring differential necropsy rates.
Apoptotic bodies in a murine model of alcoholic liver disease: Reversibility of ethanol‐induced changes
Authors: Goldin R. D. , Hunt N. C. , Clark J. , Wickramasinghe S. N.
Published: June 2005

The aim of this study was to investigate the effects of ethanol exposure on the location and number of apoptotic bodies in murine liver. Livers from 21 control mice were compared with those of 41 animals exposed to ethanol vapour by inhalation for up to 23 days. In the control livers, apoptotic bodies were identified in association with 36 per cent of the terminal hepatic venules examined and 88 per cent of these bodies were located in the first two rows of perivenular hepatocytes. By comparison, in the ethanol‐exposed group, apoptotic bodies were observed in association with 72 per cent of the terminal hepatic venules examined and more were found further from them. This increase in the number of apoptotic bodies was shown to depend on the duration of ethanol exposure and, furthermore, the effects of ethanol were shown to be completely reversed by a period of abstinence. These results indicate that ethanol exposure has time‐dependent, reversible effects on both the location and the number of apoptotic bodies in murine liver. This extends knowledge gained from previous work on rat livers.
Immunohistochemical detection of cd1a antigen in formalin‐fixed and paraffin‐embedded tissue sections with monoclonal antibody 010
Authors: Krenàcs Làszlò , Tiszalvicz Làszlò , Krenàcs Tibor , Boumsell Laurence
Published: June 2005

The immunoreactivity of a CD1a monoclonal antibody (MAb), denoted 010, was investigated by means of the streptavidin‐biotin‐peroxidase method in formalin‐fixed and paraffin‐embedded tissues from 47 cases. The samples comprised reactive lymphoid proliferations of skin, tonsil, and lymph node including dermatopathic lymphadenopathy and Langerhans' cell histiocytosis, Hodgkin's and non‐Hodgkin's lymphomas, and thymomas. Interdigitating and dermal dendritic cells, veiled cells, Langerhans' cells, and also cortical thymocytes and their neoplastic counterparts displayed immunostaining with MAb 010 in paraffin sections. These results are identical to previous ones reported for other CD1a MAbs in fresh or frozen specimens. The findings suggest that the binding site of 010 is a fixation‐resistant epitope of CD1a antigen which has not been previously identified.
Expression of the pS2 peptide in primary breast carcinomas: Comparison of membrane and cytoplasmic staining patterns
Authors: Dookeran K. A. , Rye P. D. , Dearing S. J. , Walker R. A.
Published: June 2005

The pS2 protein is oestrogen‐regulated in breast cancer cell lines. Previous studies have shown a relationship to oestrogen receptor in primary breast carcinomas. This study examined 178 breast carcinomas for pS2 using immunohistochemistry. A high frequency (77 per cent) of positive tumours was found, using a 10 per cent cut‐off point to define a positive tumour. There was no relationship with menopausal status or node status, a significant association with differentiation, a weak association with oestrogen receptor, and no association with progesterone receptor or overall survival. Two patterns of cellular localization were observed: cytoplasmic and membrane. The former showed a stronger relationship with oestrogen receptor status, although there were oestrogen receptornegative tumours with marked pS2 staining. Membrane staining showed a stronger relationship with differentiation, with a staining pattern similar to that observed for milk fat globule membrane. The staining patterns observed may support a role for pS2 in a secretory mechanism. However, the expression and function of pS2 in breast carcinomas emain complex, and are not simply related to oestrogen regulation.
Parathyroid hormone‐related protein (PTHrP) gene expression in solid tumours associated with normocalcaemia and hypercalcaemia
Authors: Dunne Fidelma P. , Lee Susan , Ratcliffe Wendy A. , Hutchesson Andrew C. , Bundred Nigel J. , Heath David A.
Published: June 2005

Parathyroid hormone‐related protein (PTHrP) is an important humoral factor in the syndrome of humoral hypercalcaemia of malignancy (HHM) and its importance is evident by the many studies examining either PTHrP mRNA expression, intracellular peptide, or circulating PTHrP levels in patients with malignancy. However, the relationship between PTHrP mRNA expression, intracellular localization of peptide, and circulating PTHrP levels in the same group of patients with malignancy has not been examined. This study was carried out to explore this relationship in a group of patients with solid tumours associated with either normocalcaemia or hypercalcaemia. PTHrP mRNA and peptide were localized by in situ hybridization and immunohistochemistry respectively in 26 squamous carcinomas and 15 adenocarcinomas from patients who were either hypercalcaemic or normocalcaemic. Plasma PTHrP1–86 and serum PTH1–84 concentrations were measured by two‐site immunoradiometric assays. PTHrP mRNA and peptide were localized in 11 (100 per cent) and 10 (91 per cent) of 11 squamous tumours from hypercalcaemic patients, all of whom had detectable circulating PTHrP levels, and in 14 (97 per cent) and 11 (73 per cent) respectively of 15 squamous tumours from normocalcaemic patients. PTHrP mRNA and peptide were localized in only two (28 per cent) and four (57 per cent) respectively of seven adenocarcinomas associated with hypercalcaemia. Since the majority of squamous tumours synthesized PTHrP irrespective of the calcium status of the patient, this suggests that the clinical expression of tumour‐derived PTH‐like bioactivity may depend on the rate of secretion of PTHrP rather than gene expression, and that the bioactivity of secreted PTHrP may be modulated by post‐translational processing.
Thymic extracellular matrix in human malnutrition
Authors: Lyra Jurandy S. P. Ocampo , Madi Kalil , Maeda Cristina T. , Savino Wilson
Published: June 2005

Previous studies have shown that malnutrition severely affects both lymphoid and epithelial components of the thymus. Yet, few data are available concerning the extracellular matrix (ECM) of the thymic microenvironment in malnutrition. We studied by histological, ultrastructural, and immunohistochemical means thymuses obtained in necropsies from 19 malnourished children. We observed a consistent increase in the intralobular ECM‐containing network which could be ascertained histologically by the dense reticulin staining. This abnormally dense ECM network contained fibronectin, laminin, and type IV collagen. Importantly, the enhancement of thymic ECM in malnourished individuals positively correlated with the degree of thymocyte depletion. This correlation may represent a cause‐effect relationship in which the contact of thymocytes with abnormally high amounts of thymic ECM triggers and/or enhances programmed cell death.
Role of respiratory viral infection in sids: Detection of viral nucleic acid by in situ hybridization
Authors: An Shu F. , Gould Stephen , Keeling Jean W. , Fleming Kenneth A.
Published: June 2005

There is considerable evidence suggesting that respiratory viral infection is involved in the genesis of the sudden infant death syndrome (SIDS), with rates of about 20 per cent of SIDS victims compared to about 13 per cent of controls. Since the techniques used previously are prone to under‐reporting from autopsy material, non‐isotopic in situ hybridization (NISH) has been used to detect viral nucleic acid in lung in SIDS. Forty‐five SIDS cases (30 males) were examined (age range 3 weeks–14 months, mean age 3·9 months). Thirty non‐SIDS cases (15 males) were also examined (age range 5 weeks–24 months, mean age 9·0 months). Eleven of 45 (24·4 per cent) SIDS cases were positive by NISH compared to 1 of 30 (3·3 per cent) non‐SIDS cases (P=0·012). There were eight cases of adenovirus type 5, two cases of respiratory syncytial virus (RSV), and one case of parainfluenza virus type 2. The one positive control case was adenovirus type 5. Only lung parenchyma was examined here. Additional examination of the upper respiratory tract may increase the number of positive cases.
Expression of hepatocyte growth factor mRNA during oval cell activation in the rat liver
Authors: Alison Malcolm R. , Poulsom Richard , Jeffery Rosemary , Anilkumar T. V. , Jagoe Roger , Sarraf Catherine E.
Published: June 2005

The customary wave of hepatocyte regeneration which occurs in the rat liver after two‐thirds partial hepatectomy can be abolished by oral administration of the carcinogen 2‐acetylaminofluorene. Instead, regeneration is achieved through the proliferation and differentiation of potential stem cells (oval cells) which appear to emanate from the portal space. Ultrastructural studies have illustrated the undifferentiated nature of these cells in the first 3 days after resection, but very rapidly they acquire features of small hepatocytes or biliary epithelia. Oval cell progeny can form either cohesive columns of cells within sinusoids which may later differentiate into new hepatic plates, or single cells that can insinuate within existing plates. Using a 35S antisense riboprobe to hepatocyte growth factor (HGF) mRNA, the synthesis of HGF mRNA was observed in sinusoid‐lining cells. There were few HGF mRNA‐expressing cells in the liver removed at resection, but numbers steadily increased in the remnant over the next 7 days. In particular, an almost nine‐fold increase in the density of HGF mRNA‐producing cells occurred in the periportal areas, resulting in approximately double the density present within the centrilobular parenchyma. The superabundance of HGF‐producing cells in the immediate vicinity of oval cell proliferation and differentiation strongly suggests that this growth factor is involved in all aspects of stem cell behaviour—proliferation, migration, and differentiation, through a paracrine mechanism.
The role of apoptosis in growing and stationary rat ascites hepatoma, Yoshida AH‐130
Authors: Tessitore Luciana , Costelli Paola , Sacchi Cristina , Piacentini Mauro , Baccino Francesco M.
Published: June 2005

The occurence of apoptosis and its contribution to growth‐phase transitions in the rat ascites hepatoma Yoshida AH‐130 have been evaluated. Apoptosis was observed by light microscopy in both exponentially‐growing and stationary tumours as characteristic chromatin condensation and compacting of the cytoplasm, but the frequency of apoptotic bodies (apoptotic index) was four‐fold higher in stationary than in logarithmic‐growing tumours. Apoptosing cells exhibited strong immunocytochemical reactivity for ‘tissue’ transglutaminase, and transglutaminase activity was higher in stationary tumour cells. A ladder pattern of DNA fragmentation was revealed by agarose gel electrophoresis which was more pronounced in stationary tumours. The apoptotic bodies were either free or contained in vacuoles within otherwise normal tumour cells, suggesting active engulfment of dead cells by viable homologous cells. Such ‘homophagic’ disposal of the apoptotic bodies probably accounted for a component of the enhanced cell protein degradation previously observed in stationary AH‐130 tumours (Tessitore L, Bonelli G, Cecchini G, Amenta JS, Baccino FM, Arch Biochem Biophys 1987; 255: 372–384). The data indicate that, in concurrence with changes in the cell proliferation rate, modulations of apoptosis play a substantial role in determining the net growth rate of such an anaplastic tumour as the Yoshida AH‐130 hepatoma.
Studies of c‐jun oncogene expression in human breast using a new monoclonal antibody, NCL‐DK4
Authors: Tiniakos Dina G. , Scott Lynn E. , Corbett Ian P. , Piggott Nigel H. , Horne Charles H. W.
Published: June 2005

A new monoclonal antibody to human c‐jun oncoprotein, designated NCL‐DK4, has been produced. NCL‐DK4 has been proved to be highly effective for use on formalin‐fixed, paraffin‐embedded tissues, enabling the study of c‐jun expression at a cellular level in both normal and neoplastic human tissues. The expression of c‐jun oncogene has been examined in normal, benign, and malignant breast tissues, and c‐jun‐specific immunoreactivity in carcinomas has been related to histological type, tumour grade, c‐erbB‐2, oestrogen receptor, progesterone receptor, and epidermal growth factor receptor expression. Normal and benign breast tissues showed c‐jun‐specific immunostaining, which was weaker and in fewer cells compared with the c‐jun immunoreactivity observed in breast carcinomas. No relationship was found between the degree of immunostaining and the extent of proliferative changes in benign breast tissues. Ninety per cent of all breast carcinomas studied showed c‐jun‐specific nuclear staining. There were no statistically significant differences in the intensity of c‐jun immunoreactivity among grade I, II, and III infiltrating ductal carcinomas. There was no significant relationship between c‐jun oncoprotein expression and c‐erbB‐2, oestrogen, progesterone, and epidermal growth factor receptor immunoreactivity.
Salivary duct carcinoma—a highly aggressive salivary gland tumour with overexpression of c‐erB‐2
Authors: Hellquist Henrik B. , Karlsson Mats G. , Nilsson Christer
Published: June 2005

The clinicopathological and immunocytochemical features of nine cases of salivary duct carcinoma are described. This relatively rare tumour, which only recently has been widely recognized as a separate entity, is highly maligant and caused the death in eight of the patients. The tumour cells are arranged in cribriform and solid growth patterns, where the solid tumour nests frequently have comedo necrosis, and a fibrous, often sclerotic, stroma is present. The infiltrating desmoplasmic component and the diffuse invasive growth into adjacent adipose parotid tissue have similarities to ductal breast carcinoma. Immunocytochemical investigationof salivary duct carcinoma showed constant overexpression of c‐erbB‐2 as detected by membrane accentuation, and high proliferative activity as detected by nuclear positivity for MIB 1 (Ki‐67). Changes in the expression of p53 and retinoblastoma gene product do not constitute a constant event event in salivary duct carcinoma. A few of the tumours showed scattered cells with distinct nuclear positivity for both progesterone and oestrogen receptors. We emphasize that this highly malignant salivary gland tumour has a characteristic morphology, may not be as rare as previously considered, and that prompt and aggressive therapy is needed.
Primary cutaneous T‐cell lymphoma: Clinicopathological features and prognostic parameters of 35 cases other than mycosis fungoides and cd30‐positive large cell lymphoma
Authors: Beljaards Rob C. , Meijer Chris J. L. M. , Van Der Putte Sebastiaan C. J. , Hollema Harry , Geerts Marie‐Louise , Bezemer P. Dick , Willemze Rein
Published: June 2005

Within the group of primary cutaneous T‐cell lymphomas (CTCLs), mycosis fungoides (MF), Sézary's syndrome (SS), and CD30‐positive lymphomas have been delineated as clinicopathological entities. Primary CTCLs that do not belong to one of these entities represent a heterogeneous and ill‐defined group of neoplasms. This paper describes the clinical and histological features of 35 of such cases. The object of t his study was to define prognostic parameters for this group of primary CTCLs. Using a slightly modified version of the updated Kiel classification, a subdivision was made into CTCL, pleomorphic, small cell type (n = 3); plemorphic, medium‐sized cell type (n = 6); pleomorphic, large cell type (n = 18); and immunoblastic lymphomas (n = 8). Altogether, these lymphomas had a poor prognosis with estimated 2‐ and 4‐year survival rates of 53 and 22 per cent, respectively. Patients with pleomorphic, small and medium‐sized cell lymphomas (n = 9) proved to have a significantly better survival that those with pleomkorphic, large cell lymphomas (P = 0·032) and immunoblastic lymphomas (P = 0·008). Primary cutaneous immunoblastic lymphomas had the worst prognosis with an estimated 2‐year survival rate of 14 per cent. Other parameters including age (P = 0·345), sex (P = 0·345), sex (P = 0·662), extent of skin lesions at presentation (P = 0·0854), and mode of initial treatment (P = 0·609) had no significant effect on the survival time. The results of this study suggest that primary CTCLs other than classical MF, SS, and CD30‐positive lymphomas have a poor prognosis in most cases, and that the current classification may be a useful means of predicting the clinical behaviour in these lymphomas.
Morphological changes of neural and vascular peptides in human skin suction blister injury
Authors: Gu X. H. , Terenghi G. , Purkis P. E. , Price D. A. , Leigh I. M. , Polak J. M.
Published: June 2005

Suction blister injury is an experimental model for the investigation of the possible derangement of dermal/epidermal interaction in injury. An extensive fibre network can be stained in skin using antisera to the pan‐neuronal marker protein gene product 9.5 (PGP) and the sensory neuropeptide calcitonin gene‐related peptide (CGRP), while microvessels are identifiable with antisera to the endothelial marker von Willebrand's factor (vWf) and the peptide endothelin (ET). To investigate the possible involvement of superficial cutaneous innervation and microcirculation during the repair process in injury, human skin biopsies taken at differnt times after suction blister injury were investigated by quantitative immunohistochemistry. Neural and endothelial changes were seen in both edge and blister areas. PGP‐and CGRP‐immunoreactive nerves showed an increase in both areas compared with control skin up to 6 h after injury, followed by a decrease which lasted until 72 h. This was followed by a gradual increase of both nerve types starting from the blister edge and lasting up to 8 days after injury when the values were similar to controls. Similarly, in the blister area of the skin, vWF‐immunoreactive capillaries showed statistically significant increases between 0 and 6 h after injury, followed by a decrease at 12 and 18 h which was maintaine dup to 72h. ET‐1 immunoreactivity showed a similar, although more variable, pattern of changes. At the blister edge from 23 h onwards, both vWf and ET‐1 immunoreactivities showed a second increase from the edge of the blister spreading towards the centre of the blister ahead of the nerve increase. This lasted up to 8 days, when vWf immunoreactivity showed a statistically significant increase compared with control skin. Generally the numerical increase observed at early time points was accompanied by a strong staining intensity, which reverted to a normal staining intensity at later time points. These results suggest that there is a reactive process for all immunoreactive elements in the early stages of injury, followed by changes indicative of neuronal and endothelial damage (depletion phase) and the subsequent repair process. The repair started with re‐epidermalization from the blister edge, followed by revascularization and, lastly, by reinnervation of the tissue. These results indicate a close relationship between epidermis, blood vessels, and nerve fibres during the healing process which follows suction blister injury.
Whipple's disease: A histological, immunocytochemical, and electron microscopic study of the small intestinal epithelium
Authors: Ectors Nadine L. , Geboes Karel J. , de Vos Rita M. , Heidbuchel Hein P. , Rutgeerts Paul J. , Desmet Valeer J. , Vantrappen Gaston R.
Published: June 2005

At endoscopy, the duodenum in Whipple's disease frequently appears abnormal and some clinical features such as gastrointestinal blood loss and anaemia suggest epithelial damage. However, the intestinal epithelial cells themselves appear to be normal on light and electron microscopy. The aims of this study were to analyse in detail the cytological changes in epithelial cells over time and in response to therapy in biopsies obtained from 20 patients, to investigate the functional repercussion on digestive enzymes such as lactase, and to assess the expression by the epithelial cells of MHC antigens. Cytological changes were minimal at both the light‐ and the electron‐microscopic level and MHC class I expression was preserved. However, changes indicative of functional deficits were demonstrated. Lactase and MHC class II expression were reduced or even absent. Antibiotic therapy resulted in normalization within 3–6 months. These findings are consistent with the clinical evolution and are of interest with regard to the importance of the immune response in aetiopathogenesis.
Attitudes of senior pathologists towards the autopsy
Authors: Start R. D. , McCulloch T. A. , Silcocks P. B. , Cotton D. W. K.
Published: June 2005

The attitudes of 205 consultant British pathologists in four regions were assessed by a postal questionnaire in which they were asked to indicate their level of agreement with 15 statements relating to autopsies. A total of 144 pathologists completed the questionnaire (response rate 70 per cent). Senior pathologists strongly agreed with statements relating to the importance of autopsies within pathology workloads, medical audit, and accreditation for training posts. There was strong support for the attendance of clinicians at autopsy demonstrations and for the suggestion that material from medico‐legal autopsies should be made available for teaching and research. There was strong disagreement with the suggestions that advances in diagnostic techniques have diminished the role of autopsies, that performing autopsies does not further pathologists' education, that the cost of autopsies may not be justifiable within a limited budget, and that the autopsy should no longer be part of the MRCPath examination. These results are discussed in the context of the current status of the autopsy in general.
The observation of angiogenin and basic fibroblast growth factor gene expression in human colonic adenocarcinomas, gastric adenocarcinomas, and hepatocellular carcinomas
Authors: Li Dechun , Bell Julie , Brown Alex , Berry Colin L.
Published: June 2005

Using digoxigenin‐labelled, synthetic oligonucleotide probe cocktails of angiogenin and bFGF genes, the expression of the two genes was observed by in situ hybridization in ten colonic adenocarcinomas, seven gastric adenocarcinomas, and four hepatocellular carcinomas. The angiogenin gene was expressed in eight of the ten cases of colonic adenocarcinoma and in all of the four cases where dysplastic glands were found. Angiogenin expression was evident in four of the seven cases of gastric adenocarcinoma. bFGF expression was detected in only five of the seven gastric carcinoma cases. The mRNAs for angiogenin and bFGF were mainly cytoplasmic in distribution and were only occasionally seen in the nuclei of the positive cells. Neither the angiogenin gene mRNA nor the bFGF mRNA was expressed in the four cases of hepatocellular carcinoma. It is postulated that the angiogenin gene may play an important role in angiogenesis in colonic adenocarcinomas; in gastric cancers, both angiogenin and bFGF were involved in this process. For hepatocellular carcinomas, neither angiogenin nor bFGF production appeared to be related to angiogenesis.
L‐CAM expression in lymph node and liver metastases of colorectal carcinomas
Authors: Van Der Wurff Anna A. M. , Arends Jan‐Willem , Van Der Linden Edith P. M. , Kate Joop Ten , Bosman Fred T.
Published: June 2005

L‐CAM, also known as E‐cadherin, is a cell adhesion molecule expressed on the plasma membranes of epithelial cells at the intercellular interface. From in vitro gene transfection experiments the idea has been conceived that loss of L‐CAM expression might be related to the invasive capacity as well as metastatic potential of tumour cells. In several tumours a relation between the grade of differentiation and L‐CAM expression has been noticed: loss of differentiation appears to be associated with loss of L‐CAM immunoreactivity. Also, in lymph node metastases of poorly differentiated carcinomas loss of L‐CAM expression was demonstrated. In this study we describe L‐CAM expression in lymphogenous and haematogenous metastases of large bowel adenocarcinomas, using an indirect immunoperoxidase method with the monoclonal anti‐L‐CAM antibody 6F9. All the metastases studied—lymphogenous as well as haematogenous—demonstrated L‐CAM immunoreactivity in a pattern comparable to that of primary tumours. Intratumour heterogeneity in expression was noted, with normal intercellular, apical (non‐functional), and focally negative areas in the same tumour. The data indicate that primary tumours and their metastases do not differ strikingly in their pattern of L‐CAM expression. This would be consistent with transient rather than constitutive down‐regulation of L‐CAM in invasive and metastatic cancer cells.
The prognostic significance of immunohistochemically detected lymph node micrometastases in colorectal carcinoma
Authors: Jeffers Michael D. , O'Dowd Gerard M. , Mulcahy Hugh , Stagg Mary , O'Donoghue Diarmuid P. , Toner Mary
Published: June 2005

Micrometastases have been detected by immunocytochemical means in the lymph nodes of patients with otherwise node‐negative cancer of the colon and rectum. This study examines the incidence and prognostic significance of nodal micrometastases in Dukes' B carcinoma. Five hundred and fifty‐nine lymph nodes from 77 cases of Dukes' B carcinoma were examined for lymph node micrometastases by immunocytochemical staining for cytokeratin AE1:AE3. Micrometastases were detected in 19 cases (25 per cent). Cell clusters were present in ten cases, the remaining nine cases displaying only single cells. The presence of micrometastases was unrelated to age (P = 0·06), sex (P = 0·32), tumour site (P = 0·37), tumour size (P = 0·67), or tumour differentiation (P = 0·66). Ten‐year survival estimates by the Kaplan–Meier lifetable method was 47 per cent in patients with and without micrometastases (χ2 = 0·35 and 1 df, P = ns). The presence of nodal micrometastases detectable only by immunocytochemistry in patients with Dukes' B colorectal cancer does not justify reassignment to a more advanced disease stage.
Glycosylphosphatidylinositol (GPI)‐anchored membrane proteins are constitutively down‐regulated in psoriatic skin
Authors: Venneker Gerard T. , Das Pranab K. , Meinardi Marcus M. H. M. , v Marle Jan , v Veen Henk A. , Bos Jan D. , Asghar Syed S.
Published: June 2005

Hyperproliferation of keratinocytes (KCs) in psoriasis has been found to be associated with excessive activation of a phospholipase C (PLC)/protein kinase C (PKC) signal transduction system. The molecular species of PLCs which are activated in psoriasis have not been thoroughly investigated. It was envisaged that if glycosylphosphatidyl‐inositol (GPI)‐specific PLC was activated in the membrane of psoriatic epidermal cells, it would render these cells devoid of those proteins which are anchored to the cell membrane through their GPI moiety. In order to test this possibility, four GPI proteins (CD 16, CD55, CD58, and CD59) were determined immunohistochemically in normal and psoriatic skin. In normal skin, CD55 and CD59 were strongly expressed on epithelium and vascular structures, whereas CD16 and CD58 were strongly expressed only on epithelium. The expression of all four GPI proteins was decreased in non‐lesional psoriatic skin and virtually abolished in lesional psoriatic skin. A control transmembrane protein, CD46, was strongly expressed in normal and non‐lesional psoriatic skin, and its expression was not significantly decreased in psoriatic lesions. The absence or reduction of GPI proteins was not seen in the lesions of several other inflammatory and proliferative diseases studied.
Cell cycle‐related variation and tissue‐restricted expression of human cyclin D1 protein
Authors: Bartkova Jirina , Lukas Jiri , Strauss Michael , Bartek Jiri
Published: June 2005

Recent evidence from genetic studies suggests that abnormalities of some of the members of the cyclin superfamily may be intimately associated with tumourigenesis, most likely through deregulation of the cell cycle control. In an attempt to elucidate the potential role of cyclin D1 (a gene located within the 11q13 amplicon and a candidate BCL‐1, PRAD‐1 oncogene) in the pathogenesis of human neoplasias, we have developed and characterized a novel monoclonal antibody specifically recognizing cyclin D1 protein in various assays including immunohistochemistry on frozen and paraffin sections. Using the DCS‐6 antibody as a tool, we now show a characteristic cell cycle‐dependent variation of the cyclin D1 protein in human cultured cells and report on the first immunohistochemical study of this G1 cyclin in a range of normal human tissues and breast carcinomas. Analysis of normal tissues revealed generally low levels of cyclin D1 protein, mainly restricted to the proliferative zones of some epithelial tissues, and the lack of its expression in several human tissues including lymph nodes, spleen, and tonsils. In contrast, pronounced overexpression/nuclear accumulation of cyclin D1 was found in 37 per cent of cases in a series of 35 primary ductal carcinomas of the breast. We conclude that the DCS‐6 antibody provides a potentially useful tool for the establishment of simple methods suitable for verifying any diagnostic and/or prognostic value of this novel marker on large series of histological specimens and opens the way for biochemical, immunocytochemical, and immunohistochemical studies of the role played by cyclin D1 aberrations in human oncogenesis.
Nm23 ‘anti‐metastatic’ gene product expression in colorectal carcinoma
Authors: Royds Janice A. , Cross Simon S. , Silcocks Paul B. , Scholefield John H. , Rees Robert C. , Stephenson Timothy J.
Published: June 2005

Nm23 is a putative metastasis suppressor gene and alterations in this gene have been reported in colorectal carcinomas suggestive of a role for nm23 in the dissemination of these tumours. In this study we used an affinity purified polyclonal antibody, ab‐11, on formalin‐fixed, paraffin‐embedded sections of colorectai carcinomas from 46 patients in a three‐stage avidin–biotin complex immunoperoxidase technique. Follow‐up of these patients was until time of death or for 5 years, with a mean time of 31·2 months. Two observers scored the staining results from 1 to 3 according to the proportion of tumour cells positive. The association of nm23 staining with survival, sex, age, vascular invasion, and Dukes' stage was determined using Cox's regression model. The association of death from colorectal cancer and nm23 status reached marginal significance in this study (P= 0.0417). Moreover, there is some suggestion of a protective effect from nm23 as the relative risk of dying from colorectal cancer for each increment of nm23 positivity is 0·573 (95 per cent confidence limits 0·34–0·98).
Immunohistochemical analysis of extracellular matrix components in synovial sarcoma
Authors: Guarino Marcello , Christensen Lise
Published: June 2005

Little attention has been paid to the composition of the extracellular matrix in synovial sarcoma, a tumour showing both epithelial and mesenchymal phenotypes. As extracellular matrix participates actively in interactions between epithelial and mesenchymal tissues, further knowledge of the pathogenesis of this tumour may be provided by the study of extracellular matrix components. Therefore, we have analysed the immunohistochemical distribution of type I, III, and IV collagen, fibronectin, laminin and tenascin in four cases of synovial sarcoma. The pattern of immunoreactivity for these molecules varied according to the tissue phenotype of the tumour. Mesenchymal tissue labelled mainly for type I and III interstitial collagen and fibronectin. The epithelial component was surrounded by a laminin and type IV collagen‐positive basement membrane, but punctate pericellular reactivity for laminin and type IV collagen was also detected among some mesenchymal cells. Tenascin was strongly expressed in the mesenchymal tissue immediately around epithelial structures and weakly or not at all expressed in the monophasic tumours and in mesenchymal tissue distant from epithelial elements in the biphasic tumours. These results suggest some resemblances between synovial sarcoma and the embryonic development of epithelia from mesenchymal cells, providing further support for the concept of an epitheliogenesis from the mesenchyme in these tumours.
Measurement of nuclear dna content in histological and cytological specimens: Principles and applications
Authors: Carey Francis A.
Published: June 2005

Nuclear DNA content can be measured in histological and cytological material by the techniques of image and flow cytometry. The measurement depends on the availability of stoichiometric markers for DNA. Results are expressed in DNA histograms from which tumour DNA ploidy and cell proliferation status can be estimated. Flow cytometry analyses large numbers of cells and gives the most meaningful cell cycle data. Image cytometry is more specifically targeted to the population of interest and can accurately measure rare events. A large body of literature exists relating DNA cytometry to clinical outcome, but there are considerable inconsistencies between clinical series which may be due, in many instances, to problems of histogram interpretation and tumour sampling. Rapid ‘user‐friendly’ systems currently under development may increase the clinical acceptance of DNA cytometry.
Numerical abnormalities of chromosome 7 in human prostate cancer detected by fluorescence in situ hybridization (FISH) on paraffin‐embedded tissue sections with centromere‐specific dna probes
Authors: Zitzelsberger Horst , Szücs Sandor , Weier Heinz‐Ulrich , Lehmann Lars , Braselmann Herbert , Enders Susanne , Schilling Albrecht , Breul Jürgen , Höfler Heinz , Bauchinger Manfred
Published: June 2005

Fluorescence in situ hybridization (FISH) using chromosome‐specific α‐satellite DNA probes for chromosomes 7, 8, and 12 was performed on paraffin‐embedded tissue sections and touch imprint preparations of 53 cases of human prostate cancer. Subsequent haematoxylin and eosin (H & E) staining of the hybridized tissue sections allowed unambiguous assignment of hybridization signals either to tumour or to non‐tumorous parenchyma. Fifty‐three cases of human prostate cancer were evaluated for numerical aberrations of chromosome 7. Scoring 200 cells of tumour and non‐tumorous parenchyma in each case revealed abnormalities exclusively in tumour parenchyma in 41 cases (77 per cent). Ten of 41 cases (24 per cent) showed trisomy 7, and 15 cases (37 per cent) monosomy 7 or trisomy 7 in combination with monosomy 7, respectively. Sixteen cases (39 per cent) exhibited polysemy 7 in cells of the tumour parenchyma. In the tumour tissue in one case, different polyploid clones (triploid, tetraploid) and polysomy 7 could be identified by double hybridization with chromosome‐specific DNA probes for chromosome 7, plus 8 or 12. The indicated numerical aberrations of chromosome 7 were correlated with 78 per cent of advanced pathological stages or poorly differentiated tumours (pT3/4 or G3) of prostate carcinomas. A statistical analysis of the data revealed significant relationships of particular numerical abnormalities of chromosome 7 to different pathological categories (pT, G, pN) of tumour classification. For the T‐classification, the frequency of cells carrying polysomy 7 and polysomy 7/+7 increases significantly from pT1 to pT3/4 (P = 0·022). A significant increase from G1 to G3 also became apparent for the total frequencies of numerical abnormal cells (P = 0·05).
Immunohistochemical localization of syndecan‐1 in normal and pathological human uterine cervix
Authors: Inki Pirjo , Stenbäck Frej , Grenman Seija , Jalkanen Markku
Published: June 2005

Expression of syndecan‐1, a cell surface proteoglycan that binds growth factors and extracellular matrix components, was studied in normal and pathological human uterine cervix using immunohistochemical methods. Normal cervical squamous epithelium showed positive staining for syndecan‐1 in all cell layers, except the basal cell layer, whereas endocervical columnar epithelium stained weakly. In non‐neoplastic reactive lesions, metaplastic squamous cells were positive for syndecan‐1, whereas columnar cells showed weak or negative staining. In cervical condylomas, cells showing koilocytotic atypia were positive for syndecan‐1. The progression of cervical intraepithelial neoplasia (CIN) grade I to grade III was associated with reduced syndecan‐1 expression and localization of syndecan‐1 to more superficial cell layers. In squamous cell carcinomas (SCCs), syndecan‐1 expression correlated with histological differentiation, being absent from most poorly differentiated tumours. The results suggest that loss of syndecan‐1 from atypical cells is an early event during cervical carcinogenesis and show a close association of syndecan‐1 expression with preserved epithelial morphology and differentiation.
Co‐expression of endothelial cell and macrophage antigens in Kaposi's sarcoma cells
Authors: Uccini Stefania , Ruco Luigi P. , Monardo Francesca , Stoppacciaro Antonella , Dejana Elisabetta , La Parola Ilaria Lesnoni , Cerimele Decio , Baroni Carlo D.
Published: June 2005

The histopathogenesis of Kaposi's sarcoma (KS) was investigated using immunocytochemistry in 12 skin biopsies obtained from two AIDS patients, nine patients with the classic form, and one organ‐transplant patient. KS cells revealed a peculiar antigenic profile, being characterized by co‐expression of endothelial and macrophage markers. KS cells were stained for von Willebrand factor (vWF); for vascular endothelial (VE) cadherin, an endothelial specific adhesion molecule; and for PECAM/CD31. In addition, they expressed the macrophage antigens PAM‐1, CD68, and CD14, and were positive for vitronectin receptor and alpha‐1,5,6/beta‐1 integrins. KS cells were weakly stained for ICAM‐1 in 6 of 12 cases and were negative for VCAM‐1 and E‐selectin.
Immunohistochemical investigation of epidermal growth factor receptor expression in ameloblastomas
Authors: Ueno Shigeru , Miyagawa Tomoko , Kaji Ryuichi , Mushimoto Kozo , Shirasu Rikiya
Published: June 2005

Immunohistochemical analysis was performed to determine the localization of epidermal growth factor receptor (EGFR) in ameloblastomas. Ameloblastoma samples were classified into follicular, plexiform, and basal cell types. The number of cases in each category was 17, 19 and 3, respectively. Ameloblastomas, disregarding their histological type, consist of two cell forms: peripheral columnar cells and central stellate cells. The frequency of EGFR expression was much higher in the latter than in the former (P<0.005). On analysis with respect to histological types, the frequency of EGFR expression in columnar cells was not significantly different between the follicular and the plexiform types, but was observed more frequently in the stellate cells in the follicular than in the plexiform ameloblastomas (P<0.05). This pattern of EGFR expression was not consistent with the PCNA staining pattern, but was similar to that of keratin expression which we have reported previously. The present study suggests that EGFR expression in ameloblastomas is closely associated with tumour differentiation, and squamous differentiation in particular.
Development of papillary renal cell tumours is associated with loss of Y‐chromosome‐specific DNA sequences
Authors: Kovacs Gyula , Tory Kalman , Kovacs Angela
Published: June 2005

Twenty‐two papillary renal cell tumours were analysed by Southern hybridization using eight DNA probes from homologous regions of the X and Y chromosomes and two Y‐chromosome‐specific DNA probes. Sixteen of the 19 papillary renal cell tumours of male patients showed the loss of Y‐chromosome‐specific sequences. No loss of heterozygosity was detected in three tumours that developed in females. The frequency of loss of the Y chromosome was established in 50 non‐papillary renal cell carcinomas as well. Only seven of the 31 non‐papillary renal cell carcinomas obtained from male patients had lost the Y‐chromosome‐specific sequences, whereas no allelic loss was found in 19 non‐papillary tumours obtained from female patients. Papillary renal cell tumours show a strong male preponderance (6:1) and loss of Y chromosome in 84 per cent of the cases, whereas non‐papillary renal cell carcinomas show only a slight male preponderance (1:5:1) and the Y chromosome is lost in only 22 per cent of the cases. These data suggest that a tumour suppressor gene is localized at one of the homologous regions of the X and Y chromosomes, the homozygous inactivation of which is associated with the development of papillary renal cell tumours.
Alterations in the glomerular charge barrier in human lupus nephritis
Authors: Al‐Nawab M. D. , Davies D. R.
Published: June 2005

Renal tissue taken from ten patients with proteinuria and diagnosed as suffering from human lupus nephritis was examined by a post‐embedding charge labelling method using cationic gold (CG). The method allows localization of the anionic charge in the glomerular epithelial cell coat and/or glomerular basement membrane (GBM). Our results demonstrate that in human lupus nephritis there are gaps in the charge barrier associated with immune deposits found in the lamina rara externa of the GBM. These data suggest that the loss of glomerular fixed anionic charges in the GBM may play an important role in the development of proteinuria in patients suffering from lupus nephritis.
Immune complex deposition in Bowman's capsule is associated with parietal podocytes
Authors: Gibson Ian W. , Downie Thomas T. , More Ian A. R. , Lindop George B. M.
Published: June 2005

We have recently documented the presence of podocytes lining part of Bowman's capsule at the vascular pole, in adult human kidney. In this study, we describe the deposition of immune complexes in Bowman's capsule in association with these parietal podocytes. We examined 1 year's consecutive human renal biopsies (n=170). Transmission electron microscopy (TEM) revealed 18 cases in which parietal podocytes were present. Of these 18, there were 11 cases of glomerulonephritis, in which immune complexes were demonstrated in the capillary tuft by both TEM and direct immunofluorescence microscopy. In seven of these 11 cases, TEM showed immune complex‐type deposits in Bowman's capsule, always associated with parietal podocytes. These deposits were similar in size, appearance, and distribution to the deposits in the capillary tuft. By contrast, non‐specific electron densities within Bowman's capsule were found beneath both squamous parietal cells and parietal podocytes. In four cases, Bowman's capsule also showed focal positive immunostaining for complement components and/or fibrinogen. Both parietal and visceral podocytes showed similar fusion of pedicels. We suggest that filtration through parietal podocytes may be responsible for immune complex deposition and subsequent damage to the vascular pole of the glomerulus in human renal disease.
Pathogenesis of IgA nephropathy: In vitro activation of human mesangial cells by IgA immune complex leads to cytokine secretion
Authors: Chen Ann , Chen Wei‐Perng , Sheu Lai‐Fa , Lin Ching‐Yuang
Published: June 2005

IgA immune complex (IC) plays a crucial role in the pathogenesis of IgA nephropathy (IgAN). As IgA‐IC is not itself cytotoxic, other mediators may be involved in the pathogenesis. In order to elucidate the mechanisms by which IgA‐IC mediates renal injury in IgAN, the ability of IgA‐IC to ‘activate’ cultured human mesangial cells (HMC) was studied. HMC were incubated with nephritogenic IgA‐IC, containing a MOPC‐315 plasmacytoma‐derived IgA anti‐dinitrophenyl (DNP) and DNP‐conjugated bovine serum albumin. The cells showed morphological changes, an accelerated rate of proliferation, and increased production of interleukin‐1 (IL‐1), interleukin‐6 (IL‐6), platelet activating factor (PAF) and generation of superoxide anion. The enhancement of IL‐1 and IL‐6 mRNA expression in HMC incubated with IgA‐IC was identified by dot blot analysis. Northern blot hybridization also demonstrated an augmented IL‐6 mRNA expression in HMC treated with IgA‐IC. These results suggest that nephritogenic IgA‐IC may amplify the proliferation of HMC and the production of immune/chemical mediators and superoxide anion thereby resulting in the renal lesions of IgAN.