bluuebunny/crossref_metadata_2025 huggingface dataset:
Tumours acquire their vasculature by vessel incorporation, not vessel ingrowth
Authors: Thompson W. D. , Shiach K. J. , Fraser R. A. , McIntosh L. C. , Simpson J. G.
Published: June 2005

This study was designed to investigate the early development of tumour circulation using a transplantable mouse mammary adenocarcinoma. Tumour cells (106) were injected subcutaneously into the flank and groups of treated mice were killed at 24 h intervals up to 12 days; the tumours and surrounding tissues being processed by standard histological methods. Sections of tumour were sub‐divided into neoplastic tissue, vessels and connective tissue using computer‐assisted morphometric techniques: the host tissue around the tumour was similarly quantified for vessels and connective tissue. With increasing tumour size, the proportion of vessels within tumours rapidly increased to reach a plateau of approximately 1·5 per cent of tumour volume, a 400 per cent increase on the vascular density of normal subcutaneous tissue. Within tumours, vascular density was always higher at the periphery than the centre. The most pronounced increase in vascular density affected the host tissue around the tumour. It is not clear why vascular development is most prominent outwith the tumour when postulated angiogenic factors, such as tumour angiogenesis factor, are presumably released within. Our results imply, however, that tumours acquire their vasculature by infiltration into, and expansion between, a network of newly formed vessels in the surrounding connective tissue.
Ovarian cancer in women given diethylstilboestrol in pregnancy: Two case reports
Authors: Cohen Ilan , Altaras Marco , Jaffe Richard , Markov S. , Corduba Mario , Aderet Noah Ben
Published: June 2005

Two cases of ovarian cancer that developed many years after exposure to large dozes of diethylstilboestrol during pregnancy are reported. This paper is the first such cases reported.
The immunohistology of Hodgkin's disease—Reed–Sternberg cells and their variants
Authors: Angel Carole A. , Warford Anthony , Campbell Andrew C. , Pringle James H. , Lauder Ian
Published: June 2005

Thirty‐three cases of Hodgkin's disease were analysed by immunoalkaline phosphatase and immunoperoxidase techniques, using a monoclonal antibody panel, including markers of B‐cells, T‐cells, macrophages, granulocytes, and the antibody Ki‐1. Hodgkin's cells were found to express markers generally regarded as T‐cell, B‐cell, myeloid, or monocyte associated. Furthermore, heterogeneity of marker expression was also seen within the Hodgkin's cell population in any single case. Morphological and immunohistological analogies between cells involved in antigen presentation and Hodgkin's cells are described, suggesting possible relationships between these cell types. Anti‐Leu M 1 was not found to be a particularly sensitive marker of Hodgkin's disease under the conditions used in this study.
Leucocyte common antigen is present on osteoclasts
Authors: Athanasou N. A. , Quinn J. , O'D. Mcgee J.
Published: June 2005

Using a large panel of monoclonal antibodies which recognize the leucocyte common antigen (LCA), the presence of LCA on osteoclasts in both fetal and adult human bone specimens has been determined by immunohistochemistry. LCA is evident on the surface of fetal human osteoclasts in bone imprints and cryostat sections. LCA was also found on osteoclasts in specimens of fixed, decalcified osteoarthritic bone. The intensity and pattern of osteoclast reactivity were similar to those of foreign‐body type macrophage polykaryons in inflammatory lesions. These results favour derivation of osteoclasts and their precursors from the multipotential haemopoietic stem cell which produces peripheral blood leucocytes and argues against their origin from a separate stem cell.
Structural distinctions among human breast epithelial cells revealed by the monclonal antikeratin antibodies AE1 and AE3
Authors: Sorenson Stephen C. , Asch Bonnie B. , Connolly James L. , Burstein Nelson A. , Asch Harold L.
Published: June 2005

Two monoclonal antikeratin antibodies, AE1 and AE3, were used in indirect immunocytochemistry to examine keratin expression in normal, benign proliferative, and malignant human breast epithelium. Both antibodies reacted strongly with most luminal cells in ducts and acini of normal gland. While AE1 did not stain myoepithelium, AE3 recognized myoepithelial cells of ducts but not acini, implying a cytoskeletal difference between the myoepithelium of these two components. Moreover, the antibodies reacted differently with the myoepithelium of intracanalicular as compared with pericanalicular types of fibroadenomas. Tumour cells of infiltrating ductal carcinomas with a prominent intraductal component stained more homogeneously with AE1 and AE3 than those without intraductal growth. The results provide evidence for two phenotypes of myoepithelial cells and for the presence of cryptic keratin epitopes in human breast epithelial cells. The finding that neither AE1 nor AE3 is a universal detector of these cells has important clinical and experimental implications.
Pattern of connective tissue development in swine pulmonary vasculature by immunolocalization
Authors: Mills Adrian N. , Haworth Sheila G.
Published: June 2005

Light microscopic immunolocalization studies were carried out on lung tissue from eight newborn and four adult pigs using antibodies to six extracellular matrix components. Antibodies to fibronectin, collagen type IV, and laminin localized on the same structures in adult and newborn lungs. By contrast, antibodies to the interstitial collagens (types I, III, and V) were less extensively localized in the newborn than in the adult, particularly those to type I because the fibres on which they localized in the newborn were thin and sparse. At all ages, antibodies to collagen types III and V co‐localized on type I fibres and also on thin individual fibres which formed networks, more dense in the adult than in the newborn. At all ages, anti‐type III collagen antibodies also localized on smooth muscle cells. In the adult, but not in the newborn, anti‐type I and type V collagen antibodies localized on the connective tissue around the smooth muscle cells in the pulmonary arterial media and vein wall. The dominance of collagen type III suggests greater plasticity in the newborn pulmonary vasculature, which helps explain the recently described rapid changes in arterial wall structure which constitute adaptation to extrauterine life. The postnatal increase in collagen type I helps explain the documented postnatal increase in structural stiffness of the pulmonary arteries.
Immune complex‐induced defects in glomerular basement membrane charge can be “repaired” despite continuing glomerular disease
Authors: Furness Peter N. , Turner David R.
Published: June 2005

There have been conflicting reports over whether or not glomerular immune complex deposits cause proteinuria by neutralizing the glomerular basement membrane anionic charge. We investigated whether the reported discrepancies could arise as a result of repair of the charge layer, in the face of continuing antigen deposition, and how rapidly this repair could be effected. We used a chronic serum sickness model of glomerulonephritis which consistently produces a nephrotic syndrome stable over 8 weeks. Charge distribution was demonstrated by the polyethyleneimine (PEI) method. Subepithelial electron dense deposits were associated with defects in the charge layer of the lamina rara externa at 2 and 4 weeks, but at 8 weeks many of the deposits had become surrounded by a new charge layer. This finding is of relevance to the interpretation of similar studies on human material.
Cellular composition of follicles of follicle centre cell lymphomas in relation to germinal centres of reactive lymph nodes. A morphometrical electromicroscopical study
Authors: Peters J. P. J. , Rademakers L. H. P. M. , De Boer R. J. , Van Unnik J. A. M.
Published: June 2005

The cell spectrum of neoplastic and benign reactive germinal centres was determined ultrastructurally. The degree in which the cell composition found in reactive germinal centres is maintained in analogous structures of follicular lymphomas was investigated by pattern recognition methods and discriminant analysis based on the frequencies of the various lymphoid and non‐lymphoid cell types. The follicular lymphomas included lymphomas with predominantly centrocytes (FCCL, Cb‐cc) and lymphomas with predominantly centroblasts (FCCL, Cb).

Pattern analysis of FCCL Cb, FCCL Cb‐cc and reactive germinal centres indicates that FCCL Cb follicles resemble reactive germinal centres in more aspects than follicles of FCCL Cb‐cc.

Clear statistical differences were encountered between the frequencies of the lymphoid cell types and of the follicular dendritic and histiocytic reticulum cells in follicles of FCCL Cb‐cc and FCCL Cb and reactive germinal centres.

Application of a discriminant analysis using a combination of the frequency of centrocytes and follicular dendritic cells demonstrated that both types of neoplastic follicles and reactive germinal centres were correctly classified on the basis of their cell spectrum. For the three groups the most potent discriminator was the centrocyte, whereas the small and large centroblast were of less value. For discrimination between Cb‐cc follicles and reactive germinal centres again the centrocyte was the most potent discriminator. Discrimination between FCCL Cb follicles and reactive germinal centres of FCCL Cb‐cc follicles can be easily achieved using the frequencies of small centroblasts or centrocytes on their own.

These findings indicate that (1) follicles of both FCCL Cb and FCCl Cb‐cc differ greatly in the cellular composition and not only with respect to their content of centroblasts but also in their content of follicular dendritic cells and (2) they may be considered as neoplasms representing different developmental phases of germinal centres.
Nucleolar organizer regions in small cell tumours of childhood
Authors: Egan M. J. , Raafat F. , Crocker J. , Smith K.
Published: June 2005

A silver colloid technique for nucleolar organizer regions (AgNORs) was applied to paraffin sections of 50 small cell tumours of childhood. These comprised 20 neuroblastomas, 10 Ewing's sarcoma, and 20 rhabdomyosarcomas, including spindle cell, myxoid, and round cell variants. On the basis of the number of AgNORs, one could differentiate between the three groups. The differences observed were statistically significant (P = 0·01−0·001). The subtypes of rhabdomyosarcoma could not be distinguished from one another on the basis of AgNOR counts. We suggest that this simple histochemical technique may be a useful adjunct to the range of special techniques now available to assist the pathologist in the diagnosis of small round cell tumours.
Prognostic factors in breast cancer: Immunohistochemical staining for SP1 and NCRC 11 related to survival, tumour epidermal growth factor receptor and oestrogen receptor status
Authors: Wright C. , Angus B. , Napier J. , Wetherall M. , Udagawa Y , Sainsbury J. R. C. , Johnston S. , Carpenter F. , Horne C. H. W.
Published: June 2005

It has been proposed that intense immunohistochemical staining using the monoclonal antibody NCRC 11 is indicative of a better prognosis in breast cancer, and that expression of pregnancy specific glycoprotein (SP1) and epidermal growth factor receptors (EGFR) are poor prognosis indicators. In order to evaluate the significance of staining for NCRC 1 1 to prognosis and to these other variables, we studied two series of breast cancer patients. In one series (n = 93), staining with NCRC 11 and for SP1 was correlated with prognosis: in neither case was there any overall relationship between staining and survival. However, when the patients were stratified by tumour histological (Bloom's) grade, SP1 expression was associated with a poorer prognosis in grade II tumours (p <0·025). In a further series (n = 94), NCRC 11 staining was carried out and fresh tumour samples were assayed biochemically for oestrogen receptor (OER) and EGFR. NCRC 11 showed a negative correlation with EGFR status (p = < 0·05) but no significant association with OER status. Thus despite our demonstration of a negative correlation with an indicator of poorer prognosis, we were unable to demonstrate any direct relationship between staining with NCRC 11 and survival.
Appendicitis revisited: A comparative study of Malawian and English appendices
Authors: Rode J. , Dhillon A. P. , Hutt M. S. R.
Published: June 2005

The incidence of appendicitis shows a marked variation between populations which has been attributed to dietary differences. Neural mechanisms and serotonin discharge from subepithelial neurosecretory cells have been previously implicated in pain referable to the appendix and appendicitis. Forty consecutive appendicectomy specimens from Malawi were studied by staining with haematoxylin and eosin, an alcian blue – PAS diastase sequence coupled with lead haematoxylin (PbH) and immunohistology for serotonin and NSE. The findings were compared with those in appendices removed at the Middlesex Hospital, London, to see if there were any differences between a population with a low risk of appendicitis (Malawi) and a high risk population (England). Acute transmural appendicitis was seen in fewer appendices from Malawi (27·5 per cent) than in English appendices (58 per cent). Subepithelial neurosecretory cells identified with PbH were present in 20 per cent of appendices from Malawi and 69 per cent of English appendices. These cells in both series showed immunohistochemical staining for serotonin. Nerve hyperplasia identified by staining for NSE in the appendix was present in 17·5 per cent and 81 per cent of non‐inflamed appendices from Malawi and England respectively.

Appearance of subepithelial neurosecretory cells and hyperplasia appear to be concomitants of an increased risk of appendicitis. Neural mechanisms may participate in adapting to a low residue diet and in some cases may generate appendicitis.
Immunohistologically definable light chain restriction in autoimmune disease
Authors: Jasani Bharat
Published: June 2005

Analysis of serum immunoglobulins from patients suffering from autoimmune disease has shown that pathogenically relevant autoantibodies directed at organ specific antigens are light chain restricted, i.e., they are entirely lambda or kappa type in a given patient.1–3 Furthermore, plasma cells involved at tissue level in the production of such antibodies, for example in Graves' disease, have also been shown to express a marked light chain bias as judged immunohistologically.4 On the basis of these findings, a study was conducted to determine the light chain status of tissue plasma cell infiltrates associated with Sjögren's disease, a known autoimmune disease. Of the six cases examined, all six showed a marked lambda light chain bias, with two patients exhibiting a monotypic plasma cell infiltrate of IgA, lambda isotype. The significance of the overall observations is discussed in the context of other examples of light chain restricted B‐cell responses and the generally increased incidence of benign and malignant B‐cell neoplasia in autoimmune disease.
An immunocytochemical study of lymphocyte and macrophage populations in the bone marrow of patients with non‐Hodgkin's lymphoma
Authors: Nash John R. G. , Smith Steven R. , Mackie Michael J.
Published: June 2005

Pairs of bone marrow trephine samples from 67 patients with known or suspected non‐Hodgkin's lymphoma (NHL) were collected. One sample was processed for morphological examination, the other for frozen section immunohistology, using a panel of monoclonal antibodies (MAB) reacting with lymphocyte and macrophage determinants, detected by the immuno‐alkaline‐phosphatase (APAAP) method. Forty‐one cases showed definite (36) or suspected (5) involvement of the marrow by NHL. Most were examples of lymphocytic or centroblastic/centrocytic NHL. The pattern of immunostaining confirmed the presence of NHL in these cases: the phenotype of the neoplastic cells was broadly consistent with nodal histology, where available. In ten cases, the marrow showed no evidence of involvement by NHL, and in 15, the eventual diagnosis was an abnormality of the myeloid series. These two groups of marrows not involved by NHL both showed a ‘reactive’ pattern of immunostaining, comprising polyclonal B‐cells, T‐helper and suppressor/cytotoxic cells and macrophages. We conclude that immunohistological examination of the bone marrow is useful in cases where the specimen shows morphological evidence of NHL, including those that are only ‘suspect’, but does not detect lymphoma where there is no morphological evidence of involvement.
Characterization of lipid accumulation in the gallbladder mucosa of the ground squirrel fed a lithogenic diet
Authors: Pemsingh Ramjeet S. , Macpherson Brian R. , Scott Gerald W.
Published: June 2005

Richardson's ground squirrels (Spermophilus richardsonii) of both sexes were fed a 2 per cent cholesterol‐enriched diet for intervals of 12, 18, and 24 h; 3, 5, and 7 days; and 2, 3, 10, and 20 weeks. It was shown that free (unesterified) cholesterol, phospholipid, and cholesterol ester accumulated in specific regions of the gallbladder mucosa during cholelithiasis. Electron microscopy revealed the presence of lipids inter‐ and intracellularly as early as 12 h after ingestion. By 7 days, lipids were seen in dilated endoplasmic reticulum, as well as in supranuclear and basal regions of epithelial cells. Histochemical localization revealed free cholesterol in dilated endoplasmic reticulum and residual bodies at the ultrastructural level. Neutral lipid was observed by light microscopy in the supranuclear and basal regions of the cells. In 10‐ and 20‐week treated animals, lipid droplets were also seen in the lamina propria and macrophages. The lesion induced by cholesterol ingestion persisted throughout the experimental period, and while different from that in human tissue, it was similar to those observed in experimental canine cholesterosis.
Lectin expression in carcinoid tumours of the gastrointestinal tract
Authors: Raju Gangaraju C. , Lee Yoke‐Sun
Published: June 2005

The binding of peroxidase‐conjugated Dolichos biflorus (DBA), Triticum vulgaris (WGA), Canavalia ensiformis (Con A), Arachis hypogaea (PNA), Lotus tetragonolobus, and Bandeiraea simplicifolia I (BSAI) to gastrointestinal carcinoid tumours was studied. The results indicate that carcinoid tumour cells express certain carbohydrates similar to those present in the adjacent surface epithelium. The differences in the lectin‐binding properties of carcinoid tumours of different sites of the gastrointestinal tract are closely related to the regional differences in the lectin binding of adjacent surface epithelium. These observations therefore form a useful basis for further studies in the application of lectin histochemistry to elucidate the histogenesis of carcinoid tumours.
The lectin‐binding characteristics of aflatoxin B1 induced lesions in the rat liver
Authors: Pritchard Deborah J. , Butler William H.
Published: June 2005

Hepatocellular carcinoma was induced in rats by administering aflatoxin B1 (AFB1) for 6 weeks. Malignant tumours were preceded by foci and nodules of altered hepatocytes of three histological types, composed of basophilic, eosinophilic, and vacuolated cells. In addition, there were areas of altered hepatocytes that were considered as hyperplastic.

Lectins were used as histochemical markers to compare the expression of membrane glycoproteins in hepatocellular carcinomas and hepatic nodules with non‐nodular or control hepatocytes. There were marked changes in the lectin‐binding patterns of the hepatocellular carcinoma cells and the eosinophilic nodules. The lectin‐binding patterns of basophilic nodules, vacuolated nodules, and hyperplastic areas were similar to non‐nodular or untreated hepatocytes. The similarity in the lectin‐binding changes of the eosinophilic nodules and hepatocellular carcinomas suggests that the eosinophilic nodules may be an early stage in the development of carcinoma.
The Royal Free Hospital school of medicine
Authors: Scheuer P. J. , Chappell M. E.
Published: June 2005

The teaching of pathology at the Royal Free Hospital has evolved over the past few decades in a stepwise manner, moving away from a didactic lecture‐ and classroom‐based approach to one in which staff time is mainly spent in small‐group teaching. We present an outline of our current teaching programme in the belief that it differs substantially from that in many other medical schools.
Immunohistochemical definition of antigenic determinants of pregnancy‐associated α2‐glycoprotein (α2‐PAG) using monoclonal antibodies
Authors: Smith Nicholas M. , Horne Charles H. W. , Carpenter Frederick H. , Stigbrand Torgny , Carlsson‐Bostedt Lena
Published: June 2005

Human pregnancy‐associated α2‐glycoprotein (α2‐PAG) is a high molecular weight glycoprotein present in normal sera. The protein is present in high concentration in the sera of pregnant females and in abnormally low concentration in association with conditions connected with abnormalities of mucosal immunity.

Indirect immunoperoxidase techniques using poly‐ and monoclonal antibodies were employed to identify pregnancy‐associated α2‐PAG in different tissues. Four monoclonal antibodies were selected from a battery of antibodies with defined specificities in order to ascertain reactivity with various epitopes of the antigen. The antibodies were applied to paraffin sections of breast, colon, salivary gland, and tonsil, and different fixation regimes were used in the preparation of the tissues. The polyclonal antibodies were found to stain plasma cells and epithelial lumina evenly in all the tissues included whereas the monoclonal antibodies were shown to stain certain components selectively. In breast and salivary glands, all four monoclonal antibodies could identify α2‐PAG, but in tonsil and colon, only two were reactive. This difference in epitope expression might reflect the internal processing of α2‐PAG, and lack of availability of certain epitopes may be indicative of functional blocking of certain domains.
The fine structure and histochemistry of human bile duct in obstruction and choledocholithiasis
Authors: Hopwood D. , Wood Rab , Milne G.
Published: June 2005

Biopsies from the common bile ducts from seven patients undergoing surgery for biliary obstruction due to stones or malignancy were studied histochemically and electron microscopically. The surface of the bile duct is lined by a tall epithelium which extends into diverticula. Apically, they contain some neutral and sialated mucosubstances. Fucosyl residues were found in the Golgi apparatus and along the apical cell membrane. The latter is lined by microvilli. There was a well‐developed rough endoplasmic reticulum and Golgi apparatus and a small number of apical secretory droplets. Large numbers of lipid droplets were present basally in some cells. Lipid‐containing macrophages were also seen intra‐epithelially and in the lamina propria. This suggests a possible pathway for lipid transport. The glands were lined by cuboidal cells, some containing much mucus—sulphated, sialated, and neutral with a basal nucleus. A well‐developed endoplasmic reticulum and Golgi apparatus were found with abundant secretory droplets. The glandular epithelium contained lysozyme, α‐1‐antitrypsin, and α‐1‐antichymotrypsin. These may play a protective role. The lamina propria contained scattered smooth muscle cells amongst the fibroblasts and inflammatory cells.
Xenografting of normal colonic mucosa in athymic mice
Authors: Verstijnen Cees P. H. J. , Kate Joop Ten , Arends Jan W. , Schutte Bert , Bosman Fred T.
Published: June 2005

Colonic mucosa was implanted subcutaneously into nude mice in order to investigate the potential use of xenografts as an in vivo model system for the study of colonic epithelial proliferation and differentiation. The xenografts were followed for 5 weeks. Proliferation was studied by bromodeoxyuridine (BrdU) incorporation followed by visualization of the DNA‐synthesizing cells with an anti BrdU monoclonal antibody. Differentiated cells were visualized by histo‐chemical staining of goblet‐ and enterochromaffin cells and of columnar cells by immunoperoxidase staining for carcinoembryonic antigen (CEA), secretory component, and serotonin.

In different strains of mice different observations were made. Less immunocompetent strains (Balb c nu/nu, NMRI nu/nu) developed abscesses at the site of the xenograft as well as wasting disease. These phenomena almost never occurred in CD‐1 nu/nu mice. The success rate was highest in CD‐1 nu/nu mice. After about 1 week, only crypt base cells remained vital and started to repopulate crypts with epithelial cells showing normal colonic differentiation features such as CEA, secretory component, and serotonin immunoreactivity and mainly sulphomucin production. After longer periods of time, crypts started to form cyst‐like structures due to accumulation of secretion products and dead cells. DNA‐synthesizing cells were seen in the basal areas of recognizable crypts and in the cyst‐like structures in a random distribution. These results indicate that normal colon mucosa xenografted into nude mice maintains its proliferative and differentiating capacity for at least 5 weeks.
Monoclonal S‐100 immunocytochemistry of pigmented naevi with tactile‐like corpuscles
Authors: Suzuki Toshimitsu , Ajioka Yoichi , Kogure Shoji
Published: June 2005

We examined 12 cases of pigmented naevi with tactile‐like corpuscles by immunohistochemistry using polyclonal and monoclonal antibodies against S‐100 protein. Naevus cells, the tactile‐like corpuscles, and the attached lamellae and fibroblastic cells gave a strong reaction with S‐100 protein and the β‐subunit antibodies. The same cells gave only a weak reaction with the a‐subunit antibody with the exception of about 4 per cent of corpuscles and 1–1.5 per cent of naevus cells, whether pigmented or not, which gave a strong reaction with this antibody. These results suggest that the non‐pigmented naevus cells and the constituent cells of the corpuscles are closely related and the latter might derive from the former by organoid differentiation.
Cellular composition of the spleen after human allogeneic bone marrow transplantation
Authors: Dilly Susan A. , Sloane John P.
Published: June 2005

The cellular composition of the spleen has been assessed in 18 patients who died 15–326 days after receiving allogeneic marrow for leukaemia. The white pulp showed marked lymphocyte depletion with no germinal centres, very few B cells, and rare plasma cells. The marginal zone was unrecognizable but there were moderate numbers of T cells in the periarteriolar lymphatic sheaths (PALS), showing great variation in CD4/CD8 ratio. The percentage of CD4+ cells decreased with time post transplant. CD8+ cells were reduced in patients with graft‐versus‐host disease (GvHD) who also showed no increase in cells staining for activation markers. No T cells were detected expressing immature phenotypes and no differences were detected between patients who received marrow purged or unpurged of T cells. Macrophage numbers appeared normal. Extramedullary haemopoiesis (EMH) was predominantly in the red pulp greater than 30 days after transplantation but more commonly in the white pulp before then. Pyknotic cells were common in seven cases and appeared to be associated with EMH rather than GvHD. Chimaeric studies demonstrated small numbers of donor cells in the PALS at 26 days and larger numbers at 56 days.
Studies on cell proliferation and tracer localization in the kidneys of guinea pigs with experimental autoimmune anti‐tubular basement membrane nephritis
Authors: Gorfien Stephen F. , Kitazawa Kozo , Brodkin Marc A. , Noble Bernice , Andres Giuseppe A. , Brentjens Jan R.
Published: June 2005

The mitotic activity in kidneys of guinea pigs with experimental autoimmune anti‐tubular basement membrane (TBM) nephritis was investigated using autoradiographic techniques to determine the uptake of [3H]thymidine by actively dividing cells. It was observed in these animals that cells of proximal tubules, distal tubules, cortical and medullary interstitium, medullary collecting ducts, and loops of Henle took up significantly greater amounts of [3H]thymidine when compared with normal animals.

In addition, the behaviour of horseradish peroxidase (HRP) and goat anti‐HRP IgG in extraglomerular sites in the kidneys of these animals was studied. Contrary to what was expected, these tracers appeared to be less concentrated in the tubules and interstitium of animals with anti‐TBM disease, with tracer movement restricted in areas of disrupted TBM. The significance of these observations is discussed.
Optimal preservation of p21 ras immunoreactivity and morphology in paraffin‐embedded tissue
Authors: Going J. J. , Williams A. R. W. , Wyllie A. H. , Anderson T. J. , Piris J.
Published: June 2005

Specific immunostaining of p21 ras protein by the well‐characterized pan‐ros antibody Y13‐259 is achieved in paraffin sections of human and animal tissues fixed in periodate‐lysine‐paraformaldehyde‐dichromate (PLPD). Intensity of staining is as good as in cryostat sections, with superior histological detail. Localization to plasma membrane is demonstrated in rodent cells genetically manipulated to express abundant p21 ras (the FHO5T1 cell line), both in preparations suspended in agar after culture in vitro and in those growing as tumour in vivo. Strong positive staining is observed in neoplasms of human breast and colon, tissues in which there is independent evidence of elevated ras gene expression. The superior morphology afforded by this technique allows clear characterization of p21 ras expression in small premalignant lesions for which other methods of detection of oncogene expression are not appropriate.
Axillary lymph node status in carcinoma of the breast
Authors: Lewis Vanessa , Rice Paul
Published: June 2005

In this prospective study, axillary clearance specimens from consecutive routine mastectomies and wide excisions for carcinoma were divided into three segments by sutures in the theatre: apical (furthest from the tumour), mid and basal (nearest to the tumour) segments. Each specimen was then randomized to either formalin or Carnoy's fixation. Following fixation, lymph nodes were dissected from each of the three segments, sectioned once through the hilum, and examined microscopically.

There was a significant (P < 0·05) difference in the total number and number of reactive lymph nodes harvested from Carnoy's compared with formalin‐fixed material. The number of nodes containing metastatic carcinoma was not significantly different (P = 0·64).

In several cases, involved apical segment lymph nodes were found with no involvement of the proximal groups; this observation may have serious implications for the procedure of proximal lymph node sampling in the surgical management of breast cancer. It is suggested that clearing of axillary contents with Carnoy's fixative does not provide any additional information of clinical significance, although it may allow the pathologist to identify lymph nodes rapidly.
Assessment of the effects of age and smoking on the media of muscular pulmonary arteries
Authors: Fernie June M. , Lamb David
Published: June 2005

The effects of age and smoking on the media of muscular pulmonary arteries were studied in 23 subjects without cardio‐pulmonary disease. The group ranged in age from 19 to 81 years and included 12 non‐smokers and 11 smokers. Twelve representative tissue blocks were taken from each subject and all cross‐sectionally cut arteries were measured from histological sections using a digitizer. Medial area was measured and artery size defined as total length of internal elastic lamina. Only arteries measuring less than 1500 μm length of internal elastic lamina were studied.

For all subjects, the relationship between medial area and length of internal elastic lamina was a curve, which was linearized by plotting square root of medial area against artery size. These individual linear regression equations (y=A+bx) were used to ‘predict’ for each subject the amount of muscle in the walls of arteries of a specific size: 500, 1000, and 1500 μm length of internal elastic lamina.

Using this type of summary data it was found that neither age nor smoking affected the media. Individual (intersubject) variation was evident in the amount of muscle present in any size of artery, particularly in the smallest arteries. In arteries measuring 500 μm length of internal elastic lamina, the amount of muscle was positively and significantly correlated with the absolute weight of the right ventricle.
Increased intracellular levels of calcitonin gene‐related peptide‐like immunoreactivity in pulmonary endocrine cells of hypoxic rats
Authors: Springall D. R. , Collina G. , Barer G. , Suggett A. J. , Bee D. , Polak J. M.
Published: June 2005

The mammalian respiratory tract contains innervated groups of endocrine cells which are believed to respond to hypoxia. We have demonstrated the involvement of a specific regulatory peptide produced by the cells, calcitonin generelated peptide (CGRP), in this response. Cells immunoreactive for CGRP or for protein gene product 9·5 (PGP 9·5), a general marker of nerves and endocrine cells, were quantified in sections of lungs from hypoxic (21 days, 10 percent 02) and normoxic rats. An immunostaining method employing supra‐optimal dilutions of primary antiserum was used. This detects variations in antigen concentration which may be masked if the routine, optimal dilution is used. The number of CGRP‐immunoreactive endocrine cells was significantly (P < 0·001) greater in the lungs of hypoxic rats (76·9 ± 10·1 cells/cm2, mean ± SEM) compared with controls (19·7 ± 2·4). However, the numbers of PGP 9·5‐immunoreactive cells were the same in both groups (81·3 ± 12·2, hypoxic; 79·5 ± 9·8 control), suggesting that the total number of endocrine cells did not change. It is concluded therefore that the apparent increase in CGRP‐immunoreactive endocrine cells in hypoxic rat lungs is due to increased intracellular levels of the peptide. Since CGRP is a vasodilator, this could have important implications in the vasoconstrictor response to hypoxia.
Chromophobe cell renal carcinoma and its variants—a report on 32 cases
Authors: Thoenes W. , Störkel ST. , Rumpelt H.‐J. , Moll R. , Baum H. P. , Werner S.
Published: June 2005

This paper reports on 32 chromophobe cell renal carcinomas observed in 697 renal cell cancers (RCC) of adults (peak in the sixth decade of life). The chromophobe cell‐type differs from other types of RCC macroscopically, the cut‐surface being predominantly of grey‐beige colour. Histologically, there are two variants: one is the typical (light) variant (n=22) and the other is eosinophilic (n=10). Both variants have in common (a) reaction of the cytoplasm with Hale's acid iron colloid; (b) electron microscopic detection of cytoplasmic microvesicles (150–300 nm), frequently with ‘inner vesicles’, and (c) low glycogen content in comparison with the clear cell carcinoma. Immunocytochemical investigations on the intermediate filaments show a positive reaction for cytokeratins No. 18 (uniformly) and Nos. 7 and 19 (to varying extents) for both variants, whereas vimentin was not found in any of these carcinomas, in contrast to the clear‐cell type. The cytomorphological grading revealed predominantly G II tumours. A lymph node metastasis was found in one patient.

On the basis of the mortality curves determined, the prognosis for patients with chromophobe cell carcinomas is more favourable than that of the clear‐cell type.

In terms of differential diagnosis, on the one hand, the typical (light) variant of the chromophobe cell RCC must be delimited from the clear‐cell RCC, and on the other hand, the eosinophilic variant must be distinguished from the chromophilic or ‘granular’ RCC. Microscopic, histological, histochemical, electron microscopic, and intermediate filament analysis results document that the chromophobe cell type of RCC is a distinct entity. The implications for the nomenclature of RCC, especially with regard to the ‘granular’ type, are discussed.
Transcutaneous measurement of PO2 and PCO2 in the dermis at the site of the tuberculin reaction in healthy human subjects
Authors: Spence V. A. , Beck J. Swanson
Published: June 2005

The respiration of the skin at the site of a delayed hypersensitivity reaction in tuberculin skin tests was studied by transcutaneous measurement of dermal oxygen and carbon dioxide tensions in normal individuals who had been immunized with BCG: six reactions were strong positives, four were weak positives, and four without induration were regarded as negative. The tcPO2 fell over the first 2 days of the reaction and remained low for the next 2 days: the severity of the changes was greater in the ‘strong’ reactions than in the ‘weak’ reactions. The tcPCO2 showed a reciprocal rise over the first 2 days and, although still high, tended to recover over the fourth day. These results indicate that local hypoxia and hypercapnia are prominent features of the positive tuberculin test, probably as a consequence of the respiration of the infiltrating lymphocytes and monocytes. It is likely that similar respiratory changes occur in those chronic inflammatory diseases where delayed hypersensitivity reactions make a contribution to the pathogenesis of the lesion.
Ultrastructural characteristics of the contact zones between langerhans cells and lymphocytes
Authors: Concha Miguel , Figueroa Carlos D. , Caorsi Italo
Published: June 2005

The present work was carried out to look for the ultrastructural substrate of the contact zones between Langerhans cells and lymphocytes. A high resolution electron microscopic analysis of the contact zones between Langerhans cells and lymphocytes was performed. The material used for this study was obtained from experimental contact dermatitis in mice, and human cervical squamous carcinoma and mycosis fungoides. Three types of cell—cell binding were found. Type I is a junction‐like structure characterized by the presence of intercellular bridges. It is suggested that this contact might represent a fixation mechanism between the two cells. Type II is characterized by a glycocalyx—glycocalyx continuity. An immunological function—recognition and antigen presentation—is proposed for this type of contact. Type III is a septilaminar tight contact area which seems to be a gap junction.

It is suggested that all these types of physical contact might be the morphological expression of interaction between antigen presenting cells and lymphocytes.
Extraskeletal myxoid chondrosarcoma characterized by microtubular aggregates in the rough endoplasmic reticulum and tubulin immunoreactivity
Authors: Suzuki Toshimitsu , Kaneko Hiroshi , Kojima Kuniji , Takatoh Masanaga , Hasebe Kei‐Ichi
Published: June 2005

A case is reported of a 66‐year‐old female with an extraskeletal myxoid chondrosarcoma which had originated in the lateral region of the right knee. The tumour tissue of the primary, recurrent, and metastatic deposits in the lungs was examined by electron microscopy and immunohistochemistry. Almost all the sarcoma cells in every tumour specimen harboured immunoreactivity to both a‐ and β‐subunits of S‐100 protein. A large population of cells in the subcutaneous tumour at autopsy had numerous parallel arrays of microtubules within the rough endoplasmic reticulum in addition to the well‐described ultrastructural features indicative of chondroblastic origin. These structures were present in round to polygonal, but not in fibroblastic, tumour cells. Tubulin immunoreactivity in the tumour cells showed the same tendency, being frequently positive in the large cells of the subcutaneous tumour but weakly positive in the fibroblastic and medium‐sized cells of the recurrent and metastatic tumours. The parallel arrays of intracisternal microtubules therefore may be composed of tubulin protein, as in ordinary cytoplasmic microtubules.
Immunocytochemical demonstration of p21ras in normal and transitional cell carcinoma urothelium
Authors: Dunn Timothy L. , Seymour Gregory J. , Gardiner Robert A. , Strutton Geoffery M. , Lavin Martin F.
Published: June 2005

Activation and/or overexpression of the protein product of the ras gene family (p21ras) has been implicated in the development of various cancers, including bladder carcinoma. We have used the anti‐p21ras monoclonal antibody, RAP‐5, to assess the level and pattern of expression in formalin‐fixed, paraffin‐embedded tissue sections of both normal and malignant urothelium. All 14 random normal bladder biopsies and 67 of 68 transitional cell carcinomas of the urinary bladder were positively stained with the RAP‐5 antibody. In normal urothelium, p21ras staining tended to be localized to the superficial cell layer. With increasing histological grade and/or depth of invasion of the tumour, a greater proportion of tissue sections demonstrated a staining pattern which was more uniform with respect to the different epithelial cell types. Serially diluting the primary antibody did not reveal any significant differences in the staining patterns observed. Despite the change in staining pattern with increasing grade, these results suggest that p21ras expression by itself is not a useful indicator of the malignant phenotype.
Immunohistochemical differences between intracranial germinomas and their gonadal equivalents. An immunoperoxidase study of germ cell tumours with epithelial membrane antigen, cytokeratin, and vimentin
Authors: Nakagawa Yoshio , Perentes Elias , Ross Gary W. , Ross Alan N. , Rubinstein Lucien J.
Published: June 2005

Twenty‐six intracranial germ cell tumours (11 germinomas, 10 teratomas, 2 endodermal sinus tumours, 1 teratocarcinoma, and 2 undifferentiated embryonal carcinomas) and 26 gonadal germ cell tumours (13 testicular seminomas, 2 ovarian dysgerminomas, 9 ovarian teratomas, and 2 myometrial choriocarcinomas) were studied by immunoperoxidase with monoclonal antibodies (MAbs) against epithelial membrane antigen (EMA), cytokeratin, and vimentin. Typical tumour cells in three of the 11 germinomas (two of the latter being situated in the posterior fossa) expressed both EMA and cytokeratin, whereas those in the seminomas and dysgerminomas did not. In one seminoma, a few multinucleated giant cells expressed cytokeratin. In three of seven germinomas, vimentin‐positive tumour cells were found, but all seminomas and dysgerminomas were negative. In the other forms of intracranial and gonadal germ cell tumours, epithelial and mesenchymal elements displayed the expected patterns of immunoreactivity to the respective determinants.

The immunoperoxidase differences between the intracranial germinomas and their gonadal equivalents indicate that, in the former, early epithelial or mesenchymal differentiation of the primordial germ cells may be present. The findings draw attention to the heterogeneous cellular composition of these otherwise morphologically homogeneous‐appearing tumours and, especially in the posterior fossa, to their transitional links to the immature teratomas.
An immunocytochemical study of the germinal layer vasculature in the developing fetal brain using Ulex europaeus 1 lectin
Authors: Gould Stephen J. , Howard Susan
Published: June 2005

The characteristics of the germinal matrix vasculature were studied in the developing fetal brain using immunocytochemical methods. A preliminary comparative immunocytochemical study was made on six fetal brains to compare endothelial staining by Ulex europaeus I lectin with that of antibody to Factor VIII related antigen. Ulex was found to stain germinal layer vessels better than Factor VIII related antigen. Subsequently, the germinal layers of a further 15 fetal and preterm infant brains ranging from 13 to 35 weeks' gestation were stained with Ulex europaeus I to demonstrate the vasculature. With increasing gestation, there was a gradual increase in vessel density, particularly of capillaries. This was not a uniform process. A plexus of capillaries was prominent immediately beneath the ependyma while the more central parts of the germinal matrix contained fewer, but often larger diameter, vessels. The variation in vessel density which was a feature of the later gestation brains may have implications for local blood flow and may be a factor in haemorrhage at this site.
Chronic serum sickness glomerulonephritis: Modification of the immune response influences the rate of removal of mesangial electron‐dense deposits
Authors: Furness Peter N. , Turner David R.
Published: June 2005

We have used a chronic serum sickness model of glomerulonephritis to investigate whether gross interference with the immune system can influence the rate of removal of antigen and established electron‐dense deposits from the glomerulus. Radio‐labelled cationized bovine serum albumin (BSA) was used as antigen. During the 2 weeks after the cessation of injections, the rate of removal of antigen from isolated glomeruli and from renal cortex, liver, spleen, and lung was measured. The rate of removal of mesangial and subepithelial deposits was assessed by point‐counting. Urinary excretion of free and protein‐bound isotope was also measured.

Having quantified the rate of removal of antigen and deposits from the glomerulus, we attempted to influence the rate of removal by interfering with the immune response in the course of recovery. Contrary to our expectations, stimulation of the immune system with antigen in Freund's complete adjuvant, 4 days after the last injection of antigen, inhibited the removal of antigen and mesangial electron‐dense deposits. Prednisolone had no detectable effect, but large doses of a non‐nephritogenic form of the antigen (native BSA) enhanced removal. Removal of antigen and mesangial deposits was inversely correlated with the levels of circulating anti‐BSA antibody suggesting that specific antibody, circulating through the mesangium, inhibits the removal of antigen which is already trapped at that site. None of the forms of intervention applied during recovery produced a detectable change in the rate of removal of subepithelial deposits.
p21 ras protein expression in benign and malignant
Authors: Walker Rosemary A. , Wilkinson Nafisa
Published: June 2005

The ras oncogenes encode for GTP binding and GTPase active proteins of relative molecular mass 21 000 (p21ras) which are involved in the transduction of stimuli for cell proliferation. There have been conflicting reports about the detection and significance of expression of p21ras protein in human breast disease as determined by immunohistochemistry.

The antibody Y13‐259, which detects a single protein of Mr 21 000, has been applied immunohistochemically to frozen sections of normal, benign proliferative breast, fibroadenomas, and carcinomas. Uniform staining of normal breast epithelium and myoepithelium was found, with occasional stronger staining in areas of epithelial hyperplasia in benign breast disease. Contrary to previous reports, decreased expression, usually heterogeneous, was found in half of the carcinomas examined. Thirty per cent of the carcinomas exhibited heterogeneous staining stronger than that of normal breast, interpreted as increased expression of p21ras protein. This did not relate to tumour grade or node status but showed a significant correlation with proliferation rate as determined by the monoclonal antibody Ki‐67.

This study confirms previous reports that p21ras protein expression is a feature of normal cells, and has identified increased expression in 30 per cent of tumours associated with higher proliferation rates, which is a lower incidence than previously claimed when a different antibody was employed.