bluuebunny/crossref_metadata_2025 huggingface dataset:
AgNOR area in interphase nuclei of human tumours correlates with the proliferative activity evaluated by bromodeoxyuridine labelling and Ki‐67 immunostaining
Authors: Trerè Davide , Farabegoli Fulvia , Cancellieri Alessandra , Ceccarelli Claudio , Eusebi Vincenzo , Derenzini Massimo
Published: June 2005

The area of silver‐stained proteins associated with interphase nucleolar organizer regions (AgNORs) was compared with labelling data obtained by bromodeoxyuridine (BrdU) incorporation and Ki‐67 immunostaining in 25 tumours of different origins and two non‐neoplastic lesions of the thyroid. Our data demonstrate a highly significant correlation between the mean area occupied by the AgNOR proteins masured by an image processing system and the proliferative indices evaluated by BrdU labelling (r = 0·89, P < 0·001) and Ki‐67 immunostaining (r = 0·86, P < 0·001). AgNOR protein area measurement is therefore proposed as a simple, inexpensive, and reliable method of evaluating the proliferative activity in routinely processed tumour samples.
Insulitis in type 1 (insulin‐dependent) diabetes mellitus in man—macrophages, lymphocytes, and interferon‐γ containing cells
Authors: Foulis A. K. , McGill M. , Farquharson M. A.
Published: June 2005

This study sought to determine, firstly, the relative frequency of lymphocytes and macrophages and, secondly, the percentage of lymphocytes containing interferon‐γ in inflamed islets (insulitis) of patients with type 1 (insulindependent) diabetes. Autopsy pancreases of 12 patients who had died of recent‐onset type 1 diabetes and one prediabetic patient who had died of cardiomyopathy were examined immunohistochemically. In the 87 islets that were studied, the lymphocyte macrophage ratio was 9·7:1 and approximately 40 per cent of the lymphocytes contained interferon‐γ. Interferon‐γ release in the insulitis process may be involved in the pathogensis of type 1 diabetes.
Demonstration of insulin and glucagon mRNA in routinely fixed and processed pancreatic tissue by in‐situ hybridization
Authors: Shorrock Kenneth , Roberts Phillipa , Pringle James H. , Lauder Ian
Published: June 2005

Human insulin and glucagon mRNA were identified in routinely processed pancreatic tissue by non‐radioactive in‐situ hybridization using digoxigenin‐labelled oligoncleotide probes. Cocktails of synthetic oligonucleotides complementary to human insulin and glucagon mRNA were labelled with digoxigenin using terminal deoxynucleotidyl transferase (Tdt). Specific hybrids were detected with alkaline phosphatase‐labelled anti‐digoxigenin antibody and visualized by BCIP‐nitroblue tetrazolium indicator substrate. The results showed highly sensitive and specific staining of islet cells on a range of routinely formalin‐fixed and paraffin‐embedded tissues. Post‐mortem pancreatic tissue from adults and stillborn neonates yielded acceptable signals as long as tissue morphology was well preserved. Preliminary investigations using pancreatic endocrine cell tumours gave clear easily interpretable signals which were comparable to conventional immunostaining. The application of this technique promises to be of value in the investigation of pancreatic disease.
The expression of parathyroid hormone messenger RNA in normal and abnormal parathyroid tissue
Authors: Kendall C. H. , Roberts Phillipa A. , Pringle J. H. , Lauder I.
Published: June 2005

The distribution and expression of preproparathyroid hormone (PTH) mRNA were investigated in parathyroid tissue from 57 parathyroidectomy specimens. PTH mRNA was detected by in situ hybridization using digoxigeninlabelled oligonucleotide probes. Cell morphology was seen to correlate with PTH mRNA expression. Strong expression of PTH mRNA was confined to cells which on haematoxylin and eosin staining had large vesicular nuclei. These included both vaculated and non‐vacuolated cells. Chief cells with small dark nuclei and scanty cytoplasm had little or no expression. In both adenoma and chief cell hyperplasia, the striking difference from normal was the greatly increased proportion of cells expressing PTH mRNA. In adenomas, the rim of uninvolved parathyroid tissue showed PTH mRNA expression similar to that of normal parathyroid. In hyperplasia, there was frequently concordance of staining within individual nodules. The findings establish morphological criteria for activity of parathyroid tissue and support current concepts of the different pathogenesis of hyperplasia and adenoma. The expression of PTH mRNA in oxyphil change and parathyroid carcinomas was also investigated.
Leakiness of gastric superficial and foveolar cells. A quantitative electron microscopic study using tannic acid
Authors: Hopwood D. , Milne G. , Penston J.
Published: June 2005

Two adjacent antral biopsies were taken endoscopically from 41 patients attending a clinic for the investigation of upper gastrointestinal tract symptoms. One was processed for histopathological grading and assessment of the Helicobacter pylori by light microscopy. The other was fixed in tannic acid, which demonstrates cell leakiness, and processed for electron microscopy. On each grid, the light (electron lucent) and dark (electron dense—leaky) gastric superficial and foveolar cells away from mechanical trauma of biopsy were counted, along with the number of H. pylori which were in contact with the epithelial cells.

There was a significant relationship between histological grade of gastritis and the extent of electron microscopic damage, i.e., the degradation of normal to leaky cells and the number of H. pylori per leaky cell.

If the patients were divided into two groups with minimal and maximal gastritis and compared against each other, then there was a significant difference between the two groups with respect to surface cell damage, polymorphs per unit area, the number of glands invaded by polymorphs, light/dark cells, Helicobacter grades assessed light microscopically or totals counted in contact with epithelial cells by electron microscopy, the number of Helicobacter per cell, and the age of the patients. There was no difference between the two groups on their endoscopic appearances or treatment with H2 blockers or NSAIDS.
Immunohistochemical demonstration of keratin 7 in routinely fixed paraffin‐embedded human tissues
Authors: Van Niekerk Catharina C. , Jap Paul H. K. , Ramaekers Frans C. S. , Van De Molengraft Fred , Poels Lambert G.
Published: June 2005

The immunoreactivity of OV‐TL 12/30, a monoclonal anti‐keratin 7 antibody (Mab), was investigated on frozen as well as paraffin‐embedded human tissues. Its reactivity patterns were compared with another well‐characterized monoclonal antibody to keratin 7 (RCK 105), and with broadly cross‐reacting monclonal (OV‐TL 12/5) as well as polyclonal (pKer) keratin antisera. In frozen sections of normal and malignant human tissues both keratin 7 Mabs gave similar staining patterns. The immunoreactivity for OV‐TL 12/30 and the polyclonal antibody (pKer) in tissue sections fixed in 4 per cent formalin or Bouin solution, was completely restored when pretreated with 0·1 per cent pronase, 0·1 per cent trypsin in phosphate‐buffered saline (PBS) or with 0·5 per cent pepsin in 0·01 N HCl. Except for loss of immunoreactivity on human normal stomach surface epithelium and glandular mucous cells, Mab OV‐TL 12/30 reacted strongly positive with essentially all those formalin‐ or Bouin‐fixed paraffin‐embedded tissues that had been shown to stain in non‐fixed, frozen sections. In addition to the good correlation in normal human tissues, a complete correlation between the reactivity on frozen and paraffin‐embedded human carcinomas (n = 86) was found as well. While both RCK 105 (anti‐keratin 7) and OV‐TL 12/5 (anti‐keratin 5, 7, 14, 19) did not stain on paraffin‐embedded sections, the polyclonal control antiserum (pKer) lost immunoreactivity in some cell types (e.g. mucous cells in compound glands, hepatocytes, pancreatic acinar cells, and proximal and distal convoluted tubules of the kidney). Our study shows that the keratin 7 Mab OV‐TL 12/30 is an excellent marker for tumour histopathology since it is reactive in paraffin‐embedded formalin‐fixed human tissues.
Composition and organization of cell–substratum contacts in normal and neoplastic renal epithelium
Authors: Rahilly Maeve A. , Salter Donald M. , Fleming Stewart
Published: June 2005

We have investigated the molecular basis of the organization of cell–substratum contact in normal and neoplastic renal epithelium. The several components of focal contacts and non‐collagenous basement membrane glycoproteins have been identified by antibodies, detected by immunofluorescene and viewed by laser scanning microscopy. Tubular epithelial cells grown on glass coverslips expressed laminin at the cell periphery. Co‐localized with laminin were receptors of the β1 integrin class, and the cytoplasmic plaque proteins vinculin and talin. When basement membrane glycoproteisn laminin or fibronectin were exogenously supplied by growing cells on coated coverslips, the vinculintalin‐ and integrin‐containing contact became organized into linear arrays with intervening free cell membranes. Under these conditions the actin cytoskeleton became highly organized. By contrast some tumour cells showed a reduction in focal contact molecules and a failure to organize these structures in response to laminin or fibronectin. This loss of cell matrix interaction may be important during the progression of renal tumours.
Human papillomavirus 6, 11, and 16 in laryngeal papillomas
Authors: Dickens Paul , Srivastava Gopesh , Loke Shee Loong , Larkin Siepie
Published: June 2005

Twenty‐seven cases of benign laryngeal papillomas, both single and multiple variants, were analysed for human papilomavirus (HPV) by DNA slot‐blot hybridization chiefly to determine the pattern of infection in Hong Kong Chinese. DNA was extracted from paraffin blocks of formalin‐fixed tissue and probed separately for HPV 6, 11, 16, and 18. Sixteen cases (59 per cent) showed the presence of at least one of these four HPV genomes. Thirteen cases (48 per cent) were positive for HPV 11 only. Three other cases (11 per cent) showed triple positivity for HPV 6, 11, and 16. None were positive for HPV 18. The predominance of HPV 11 infection contrasts with other series which have shown either an almost equal distribution of HPV 6 and 11 or a predominance of HPV 6. The finding of HPV 16 in three cases was unexpected. Using the polymerase chain reaction (PCR) with primers complementary to the upstream regulatory region of the HPV 16 viral DNA, the presence of HPV 16 genome was confirmed in all three cases. As the number of HPV 16‐positive cases in this study is small, analysis of more cases using fresh biopsy material and a wider range of HPV type‐specific PCR primers is warranted to determine the relative incidence of HPV subtypes in these benign laryngeal papillomas.
Apoptosis in cultured rat hepatocytes: The effects of tumour necrosis factor α and interferon γ
Authors: Shinagawa Tadashi , Yoshioka Kentaro , Kakumu Shinichi , Wakita Takaji , Ishikawa Tetsuya , Itoh Yuji , Takayanagi Masahiro
Published: June 2005

We investigated the cytotoxic effects of tumour necrosis factor α (TNF α) and interferon γ(IFN γ) on rat hepatocytes in culture. Under phase contrast microscopy, we found a small number of dying hepatocytes in control cultures, each having been transformed into a cluster of small spheres. Under transmission electron microscopy, these cells showed the characteristics of apoptosis. TNF α and a combination of TNF α and IFN γ exerted a cytotoxic effect, whereas IFN γ showed no significant cytotoxicity when assessed by neutral red assay and by measuring LDH activity in culture medium. Under phase contrast microscopy, the number of apoptotic cells increased with the addition of either TNF α or IFN γ, and markedly with the addition of both. DNA extracted from apoptotic cells cultured with TNF α and IFN γ was fragmented, and a set of bands of the ‘200 bp ladder’, which is characteristic of the DNA of apoptotic cells, was observed in agarose gel electrophoresis. These findings indicate that cultured hepatocytes die from apoptosis. TNF α killed cultured rat hepatocytes by increasing apoptosis, and this effect was potentiated by the addition of INF γ, which by itself was also weakly cytotoxic.
Cell proliferation
Authors: Hall Peter A.
Published: June 2005

The composition of a series of critical summaries is necessarily idiosyncratic, but in this compliation I have been guided by the recent publication of reviews and monographs on the subject.1–3 consequently, it seems reasonable to focus on more fundamental areas of research which may lie outside the normal reading of most pathologists. Nevertheless, I regard these burgeoning areas as being likely to impinge on our research, if not our practice, in the near future. I will concentrate on four areas that I consider of particular interest. First, regulation of the cell cycle; second, arrest of proliferation; third, programmed cell death; and finally, the generation of novel markets of prolkiferation.
The epidermal growth factor receptor in human pancreatic cancer
Authors: Lemoine Nicholas R. , Hughes Christine M. , Barton Claire M. , Poulsom Richard , Jeffery Rosemary E. , Klöppel Günter , Hall Peter A. , Gullick William J.
Published: June 2005

The epidermal growth factor receptor (EGFR) and its ligands are thought to be important in the control of proliferation of many epithelial systems, including the exocrine pancreas. Abnormalities in expression of two of the known ligands of the EGFR, transforming growth factor alpha and epidermal growth factor, occur frequently in ductal adenocarcinoma of the human pancreas. We have examined an archival series of cases of pancreatic pathology for expression of the EGFR using the anti‐EGFR antiserum 12E and found that there is almost ubiquitous overexpression of EGFR in pancreatic cancer and in chronic pancreatitis. Southern blot analysis showed no evidence of amplification or rearrangement of the EGFR gene. We conclude that an autocrine loop involving the EGFR system may be involved in the genesis of both neoplasia and reactive hyperplasia of pancreatic ductal epithelium.
Oestrogen receptor and epidermal growth factor receptor expression in male breast carcinoma
Authors: Fox Stephen B. , Rogers Suzanne , Day Christopher A. , Underwood James C. E.
Published: June 2005

Male breast carcinomas are probably hormone‐dependent, but receptor studies are few because this is a relatively rare tumour, We have studied 21 cases of male breast carcinoma immunohistochemically for oestrogen receptor (ER) and epidermal growth factor receptor (EGFR) expression employing the antibodies ER‐ICA and 12E on formalinfixed, paraffin‐embedded material. In our series, 86 per cent of male breast cancers were ER‐positive and 76 per cent were EGFR‐positive. Male breast carcinomas do not exhibit the inverse correlation between ER and EGFR expression that characterizes female breast carcinomas. Owing to the limitations of a small series, we were unable to comment on the relationship between ER and EGFR expression and patient survival. However, the relatively high incidence of ER expression may provide a growth advantage for this tumour in a male environment characterized by low levels of oestrogen. In addition, high EGFR expression may also contribute to a poor prognosis independent of ER status.
Peroxisomal enzymes in normal and tumoral human breast
Authors: Bouhtoury Fatima El , Keller Jean‐Marie , Colin Suzanne , Parache Robert Michel , Dauça Michel
Published: June 2005

The presence of peroxisomes and their enzymatic content were investigated and compared in healthy and neoplastic human breast epithelial cells using cytochemical studies at the ultrastructural level as well as Western blot and biochemical analyses. Ultrastructural cytochemistry revealed the presence of these organelles in both normal and neoplastic breast tissues. Their mean diameter was 0.27 ± 0.11 μm. No significant difference was noted between numbers of peroxisomes in normal and neoplastic breast epithelia. Catalase, D‐amino acid oxidase, and urate oxidase were found to be expressed in mammary carcinoma and in surrounding non‐malignant tissue when the postnuclear supernatant fractions prepared from homogenates were assessed by Western blot techniques. Their specific activities and that of fatty acyl CoA oxidase as determined spectrophotometrically were found to be diminished in the tumour when compared with the control tissue. On the other hand, no significant difference was found in the specific activity of the L‐a‐hydroxy acid oxidase of normal and neoplastic human breast tissues. Investigations of the relationship between peroxisomal enzymes and tumour grade revealed that catalase, urate oxidase, and fatty acyl CoA oxidase activites in breast neoplastic tissues belonging to grade III were significantly lower than in the adjacent normal tissues.
A study of cell proliferation in formalin‐fixed, wax‐embedded bone marrow trephine biopsies using the monoclonal antibody PC10, reactive with proliferating cell nuclear antigen (PCNA)
Authors: Wilkins B. S. , Harris S. , Waseem N. H. , Lane D. P. , Jones D. B.
Published: June 2005

We have investigated proliferation in bone marrow trephine biopsies from 32 patients with normal or abnormal haemopoiesis, using the monoclonal antibody PC10, which detects proliferating cell nuclear antigen (PCNA), together with immunohistochemical markers of haemopoietic cell lineage.

PCNA immunostaining revealed the pattern of proliferation within individual haemopoietic lineages in normal marrow. Two unexpected observations were made: of erythroid cells, only pro‐erythroblasts and occasional early normoblasts reacted, and positivity of megakaryocytes was unrelated to nuclear lobulation or CD61 expression.

The pathological cases represented conditions in which haemopoiesis is increased (reactive hyperplasia, chronic granulocytic leukaemia, myeloproliferative and myelodysplastic syndromes, megaloblastic anaemia). Increases in the number, and disturbances of the spatial organization, of PCNA‐expressing cells were present to a variable extent in all cases.

Sheets of PCNA‐positive megaloblastoid erythrocytes were frequently found in myelodysplastic and myeloproliferative tissue, associated with marked disturbances in the spatial organization of all haemopoietic lineages. Cases of megaloblastic anaemia due to vitamin B12/folate deficiency also demonstrated greatly increased erythroid PCNA expression, with positivity in some giant metamyelocytes.

In addition to reflecting increased proliferation, elevated PCNA expression in some bone marrow pathologies may be due to altered kinetics of the protein induced by disturbances in growth factor production.
Morphometric analysis of nucleolar organizer regions in benign and malignant peritoneal effusions using backscattered electron microscopy
Authors: Lim S. Megan , Duggan Máire A. , Ruff Monica , Rahim Shameem , McGregor S. Elizabeth , Green Francis H. Y.
Published: June 2005

Nucleolar organizer regions are loops of DNA associated with silver‐stainable proteins (AgNORs). In general, malignant cells have more and larger AgNORs than benign cells. An inconsistent argyrophilic method and difficulties a objectively evaluating AgNORs account for some of the reluctance to utilize AgNOR staining as a diagnostic tool to differentiate benign and malignant lesions. Section from paraffin‐embedded cell blocks of 10 cases of benign and nalignatnt peritoneal effusions were stained with a modified AgNOR method. Backscattered electron imaging in the Scanning electron microscope, together with image analysis, was used to evaluate more objectively a number of AgNOR parameters and to determine which measurement was the most reliable discriminant of the two types of fluids. One hundred nuclei per case were identified and imaged. In contrast to benign nuclei, AgNORs in malignant nuclei were more numerous (P 0.0001). and larger (P0.0001) A cut‐off mean AgNOR area of 1.1 μm2 (P 0.0001) correctly categorized all malignant (1.1 μm2) and benign (>1.1 μm2) cases. This system's objectivity and specificity could be and to enhance the cytological interpretation of effusions, where the separation of reactive mesothelial cells and malignant cells is extremely difficult.
αB crystallin expression in nonlenticular tissues and selective presence in ubiquitinated inclusion bodies in human disease
Authors: Lowe J. , McDermott H. , Pike I. , Spendlove I. , Landon M. , Mayer R. J.
Published: June 2005

αB crystallin is a lens protein which has homology with the small heat‐shock proteins and is also expressed in nonenticular tissues. Polyclonal antibodies have been raised to a synthetic peptide corresponding to residues 1–10 of αB crystallin. The antiserum detects a 20 kDa polypeptide on nitrocellulose replicas after polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate of extracts of heart muscle known to be rich in αB crystallin. Staining of normal human tissues reveals immunoreactivity of lens capsular epithelium, skeletal muscle, cardiac muscle, smooth muscle. renal tubular epithelium, Schwann cells, and glial cells, as has been described by other workers. In addition, positive staining of normal thyroid epithelium, colonic epithelium, and stratified squamous epithelium was seen.

Tissues known to contain ubiquitinated inclusion bodies were immunostained with the anti‐αB‐crystallin antiserum. Staining of cortical Lewy bodies, astrocytic Rosenthal fibres, and hepatic Mallory bodies was seen, but only a propertion of inclusions were positive. Neurones containing the ubiquitinated inclusions of Alzheimer's disease were only very rarely stained and the ubiquitinated inclusons of motor neurone disease were not detected by the antiserum. Reactive astrocytes in cerebral tissues were strongly immunostained.

The results suggest that αB crystallin is involved in the formation of ubiquitinated inclusion bodies that have issociated intermediate filaments and support previous observations on the localization of a brain‐specific ubiquitin carboxy‐terminal hydrolase which similarly divides ubiquitinated filamentous inclusions in the central nervous system into two main groups.
Replication of endothelial cells and smooth muscle cells induced in vivo by hypercholesterolaemia and materials released from platelet‐rich white thrombus
Authors: Asada Yujiro , Hayashi Tohru , Tanimura Nobuhiko , Kisanuki Atsushi , Sumiyoshi Akinobu
Published: June 2005

Endothelial cell injury is considered to be the primary event in atherogenesis. In this study, we investigated the effects of hypercholesterolamemia, and substances released from platelet‐rich thrombi, individually and together on endothelial cells and the wall of the rabbit in vivo. We divided 24 rabbits into four groups: I was a control group on a normal diet; II was a tubing group on a normal diet, in which polyethylene tubing was inserted into the ascending aorta; III was a group being fed a cholesterol diet; and IV was a combined group being fed a cholesterol diet with polyethylene tubing in the ascending aorta. Segments from the descending thoracic and abdominal aortas which were not injured directly by tubing were examined morphologically and for [3H] thymidine incorporation into endothelial cells and smooth muscle cells. The descending aortas of groups II, III, and IV showed various degrees of endothelial cell damage. [3H]Thymidine incorporation into endothelial cells and aortic wall was increased in groups II, III, and IV (most in group IV) as compared with group I.

These data indicate that hypercholesterolaemia and substances released from activated platelets and/or white mural thrombi can cause endothelial damage which may result in endothelial and smooth muscle cell proliferation. In addition, a combination of these two factors showed an additive effect on the endothelial injury and regeneration in vivo.
Immunohistological detection of human parvovirus B19 in formalinfixed, paraffin‐embedded tissues
Authors: Morey Adrienne L. , O'neill Hugh J. , Coyle Peter V. , Fleming Kenneth A.
Published: June 2005

Human parvovirus B19 is a cause of aplastic crises in patients with haemolytic anaemias, prolonged bone marrow failure in the immunosuppressed, and fetal death secondary to non‐immune hydrops. The immunohistological detection of parvovirus B19 in formalin‐fixed, paraffin‐embedded tissues has not previously been reported, and definitive diagnosis of infection in such specimens has relied on the use of specialized DNA hybridization and amplification techniques. A new monoclonal antibody to B19 capsid proteins, R92F6, was found to be capable of labelling infected cells in paraffin‐embedded tissues from all 19 cases of parvovirus‐related fetal hydrops tested, and in bone marrow from a child with congenital immunodeficiency and chronic parvovirus infection. Viral antigen was detected both in cytoplasmic and in nuclear distributions using the alkaline phosphatase anti‐alkaline phosphatase (APAAP) technique without preceding proteolytic digestion. The viral epitope recognized appears to be highly conserved, as speicmens were obtained over a 13‐year period from widely spaced locations in the U.K. Antibody R92F6 should facilitate rapid diagnosis of parvovirus B19 infection in routinely processed and archival specimens.
Detection of Epstein‐Barr virus in lymphoid tissues of patients with infectious mononucleosis by in situ hybridization
Authors: Prange Eberhard , Trautmann Johann C. , Kreipe Hans , Radzun Heinz J. , Parwaresch Mohammad R.
Published: June 2005

Although the immunological response during infectious mononucleosis (IMN) has been studied in detail, little is known about the spread of Epstein–Barr virus (EBV) in lymphoid organs or the topographical distribution of the infected cells. In this study, EBV was detected in 11 lymph nodes, 4 tonsils, and 1 spleen of 16 patients with IMN. The predominant cell type positive for the EBV genome was identified as small lymphocytes localized chiefly within typical T areas, preferentially in perifollicular and interfollicular regions of the lymph node. A few endothelia of epithelioid venules were also found to be positive. Furthermore, a small number of sinus lining cells of lymph nodes exhibited labelling. Altogether, only a small number of cells, not exceeding 1 per cent of all cells, were infected with EBV. Our results show that only a small number of lymphocytes carry the EBV and that besides B lymphocytes, other cell constituents of lymphatic tissues are infected by EBV during IMN.
Oestrogen and progesterone receptor immunocytochemistry in human hyperplastic and neoplastic endometrium
Authors: Snijders Marc P. , De Goeij Antony F. , Koudstaal Johan , Thunnissen Erik B. , De Haan Jelte , Bosman Fred T.
Published: June 2005

Proliferative disorders of the endometrium may be associated with autocrine and paracrine actions between stromal and epithelial cells. To determine whether the stromal–epithelial relation with respect to oestrogen receptor (OR) and progesterone receptor (PR) is disturbed in (pre)malignant endometrium immunocytochemical OR and PR expression was quantitated by computerized image analysis. This was studied in the stromal and epithelial cells of endometrial specimens diagnosed as hyperplasia (n = 14), atypical hyperplasia (n = 16), and adenocarcinoma (n = 33). Paraffin sections were used for optimal preservation of histomorphology. A progressive loss of OR and PR content occurred with increasing malignant transformation. Stromal cells in atypical hyperplasia (P = 0.0007) and well‐differentiated adenocarcinoma (P = 0.0008) exhibited a relative loss of PR content as compared with epithelial cells (P = 0.036 and P = 0.17, respectively). In atypical hyperplasia, the decrease in stromal PR content was not in parallel with persistent stromal OR immunostaining. Furthermore, stromal PR expression in atypical hyperplasia was significantly (P = 0.004) lower than in the surrounding hyperplasia, whereas the stromal OR staining as well as the epithelial OR and PR staining did not differ significantly. These observations may reflect a disturbance in hormonal interrelationships between endometrial cells in the development of endometrial neoplasia, indicating that stroma may modulate epithelial growth by paracrine mechanisms.
Contact between abluminal microvilli and collagen fibrils in metastatic adenocarcinoma and mesothelioma
Authors: Carstens Per H. B.
Published: June 2005

It can be difficult to differentiate between epithelial and mixed type mesothelioma and metastatic adenocarcinoma. To distinguish between the two, it has recently been suggested that the presence of abluminal microvilli making contact with collagen fibrils through discontinuities in the basal lamina was characteristic of mesothelioma but not of metastatic adenocarcinoma. This study presents three patients out of 39 who had adenocarcinoma metastatic to the pleural and abdominal cavities, in which direct contact was observed between abluminal microvilli and collagen fibrils. All 39 examples of mesothelioma which were also examined demonstrated this feature. This phenomenon should therefore not be used as a discriminator between mesothelioma and metastatic adenocarcinoma. An additional patient with metastatic adenocarcinoma in an axillary lymph node demonstrates that this phenomenon is not limited to the pleural or abdominal cavities.
Onset and evolution of pulmonary vascular disease in young children: Abnormal postnatal remodelling studied in lung biopsies
Authors: Hall Susan M. , Haworth Sheila G.
Published: June 2005

Pulmonary arterial structure was examined in lung biopsies from 26 children with severe pulmonary hypertensive congenital heart disease, aged 2 months–18 years, in whom the mean pulmonary arterial pressure was 55 (range 35–105) mmHg, using light, transmission, and scanning electron microscopy. Qualitative and quantitative techniques were applied and findings compared with those in age‐matched controls.

At 2 months, the smooth muscle cells showed hyperplasia, hypertrophy (mean cell diameter increased, P<0.001), and accelerated differentiation. In all pulmonary hypertensive cases aged less than 6 months, the contractile myofilament concentration was similar to the normal concentration at 6 months. Medial connective tissue was excessive for age. Smooth muscle cells within the intima (intimal proliferation) were first seen at 7 months, lying beneath a new internal elastic lamina. They showed a reduction in myofilament concentration in the more fibrotic lesions.

In all cases, endothelial cells were abnormally thick (P<0.001) and elongated. Cytoskeletal remodelling was indicated by an increase in micro‐ and intermediate filament volume density (P<0.05 for both). The severity of endothelial damage was related to vessel size and position in the arterial pathway.

These findings indicate that pulmonary vascular disease begins at or soon after birth with abnormal pulmonary vascular remodelling which leads to obliterative pulmonary vascular disease.
1,1‐dichloroethylene‐induced alterations in glutathione and covalent binding in murine lung: Morphological, histochemical, and biochemical studies
Authors: Moussa Madeleine , Forkert Poh‐Gek
Published: June 2005

Non‐ciliated Clara cells of the pulmonary bronchiolar epithelium are preferentially damaged by administration of 1,1‐dichloroethylene (1,1‐DCE) to mice. In this study, an in vivo system was utilized to investigate the dose‐dependent effects of 1,1‐DCE (75, 125, 175, and 225 mg/kg) on covalent binding and on reduced glutathione (GSH) in murine lung. Treatment of mice with each dose level of 1,1‐DCE elicited significant decreases in GSH content and resulted in covalent binding of [14C]1,1‐DCE in a dose‐dependent manner. Histochemical staining for GSH in lungs of control mice revealed positive cellular sites in alveolar septa and bronchiolar epithelium, with the highest staining intensities in Clara cells. Staining was reduced after exposure to 75 and 125 mg/kg 1,1‐DCE, and at higher doses it was abolished in alveolar septa and retained in bronchiolar epithelium, albeit at considerably reduced intensities. Heterogeneity with respect to staining intensities was consistently observed in the Clara cell population in both control and 1,1‐DCE‐treated mice. Progressive increases in covalent binding and decreases in GSH content correlated with increasing severities of Clara cell injury. These results show a dose dependence in regard to the magnitudes of [14C]1,1‐DCE binding, the alterations in cellular GSH, and the severities of Clara cell necrosis.
Cellular localisation of C‐myc product in human colorectal epithelial neoplasia
Authors: Royds Janice A. , Sharrard R. Michael , Wagner Bart , Polacarz Steven V.
Published: June 2005

Aberrant expression of c‐myc has been implicated in the development of colorectal carcinomas. We have used monoclonal antibodies 6E10 and 9E10, raised against mid‐sequence and C‐terminal peptides of the c‐myc protein, to study the distribution of myc protein in normal and diseased bowel at the light microscope and ultrastructural levels. Normal mucosa showed staining only of some nuclei in the proliferative zones of crypts. In adenomas, staining varied from predominantly nuclear to pancellular to focal or pancytoplasmic. Moderately well differentiated areas of carcinomas gave strong focal cytoplasmic staining, while in poorly differentiated tumours staining was pancytoplasmic. Electron microscopy with these antibodies detected myc protein associated with dense chromatin and, where cytoplasmic staining occurred, with polyribosomes. Tumours showed a reduced staining of nuclear pores compared with normal tissue. Comparison of staining patterns with 6E10 and 9E10 in normal tissue, adenomas, and tumours suggests that tumour progression is associated with an accumulation of cytoplasmic c‐myc protein, perhaps resulting from alterations to the C‐terminus which reduce the efficiency of nuclear targeting of the protein and thus disrupt the regulation of the cell cycle.
P53 protein expression in lymphomas and reactive lymphoid tissue
Authors: Villuendas Raquel , Piris Miguel A. , Orradre Juan L. , Mollejo Manuela , Algara Patrocinio , Sanchez Lydia , Martinez Juan C. , Martinez Pedro
Published: June 2005

P53 is a tumour suppressor gene, located in the short arm of chromosome 17, which encodes for a nuclear protein involved in the control of cellular growth, regulating the entry of the cell into the S‐phase. P53 mutations have been identified in a progressively increasing number of human malignancies. Nuclear p53 protein is usually present in non‐tumour cells in minute concentrations, due to its short half‐life. In contrast, tumours with p53 mRNA mutations show a higher nuclear protein concentration, detectable by immunohistological techniques, due to stabilization by complexing with other proteins such as heat‐shock protein or wild‐type p53 protein.

Leveis of nuclear p53 protein detected by immunohistochemistry with the monoclonal antibody PAb 1801 were measured with the aid of an image analysis system in 83 non‐Hodgkin's lymphomas (NHLs) and 13 cases of Hodgkin's disease, as well as in 14 cases of normal thymus, reactive tonsils, and lymphadenitis. High levels of p53 protein (>5 per cent of the cells) were present only in high‐grade lymphomas (in the proportion 13/55), with a peak incidence in Burkitt's lymphoma (5/8 cases). Lower levels (<5 per cent) of p53 protein were detected in low‐grade B‐ and T‐cell lymphomas, as well as in most of the cases of Hodgkin's disease, where p53 protein was selectively present in Hodgkin and Reed‐Sternberg cells. In 5/14 reactive tonsils or lymph nodes, occasional p53‐positive cells were identified.

These results suggest a relationship between levels of p53 protein and the aggressiveness of NHL. The sporadic presence of p53‐positive cells in human reactive tissues suggests that its expression could depend not only on mutation, but also on stabilization of the protein through complexing with other nuclear proteins. It is not known if this mechanism is also responsible for the p53 nuclear accumulation in high‐grade lymphomas.
A tumour promoter induces alterations in vinculin and actin distribution in human renal epithelium
Authors: Rahilly Maeve A. , Fleming Stewart
Published: June 2005

Loss of cell–substratum adhesion is an important factor during tumour progression. We have previously described reduced focal contact components and poorly organized cytoskeletal actin in renal cell carcinomas. In this study, we have used the potent tumour promoter TPA on cultured human renal epithelium to mimic neoplastic transformation. The morphological changes induced by TPA were examined by phase contrast and fluorescence microscopy. TPA treatment caused rounding up of cells and loss of adhesion to either fibronectin or laminin substrata. Cytoskeletal actin was redistributed from orientated stress fibre bundles to a perinuclear circumferential arrangement. This was accompanied by a progressive reduction in the number of vinculin‐containing contacts with accumulation of vinculin in punctate spots in the perinuclear region. These altered membrane‐cytoskeletal interactions induced by TPA are entirely reversible and mimic epigenetic changes which occur during tumour progression.
Apposition of fibroblasts to mast cells and lymphocytes in normal human lung and in cryptogenic fibrosing alveolitis. Ultrastructure and cell perimeter measurements
Authors: Heard Brian E. , Dewar Ann , Corrin Bryan
Published: June 2005

The perimeters of mast cells and lymphocytes in human lungs were measured in electron micrographs by digitizer to determine the percentage of perimeter apposed to fibroblast (PPAF). Fibroblasts were apposed to the majority of mast cells. The median PPAF for mast cells in normal lung was 50.3 per cent, and in cryptogenic fibrosing alveolitis (CFA), 35.7 per cent. Although the value in CFA was lower, the difference was not statistically significant (median difference 11.8; 95 per cent confidence interval (–19.6, 25.1); P = 0.65). The PPAF range overall was 3.8–94.1 per cent. There was similar apposition of fibroblasts to lymphocytes, and no statistical differences were found when median PPAF results for mast cells and lymphocytes were compared for normal and CFA lung.

The high degree of percentage apposition, accurately quantified in this study, shows that fibroblasts, mast cells, and lymphocytes are ideally arranged structurally in normal alveolar walls, to facilitate the many physiological interactions which are currently being uncovered. The present study also shows that apposition persists in pathological states, e.g., CFA lung, but because all these cells are increased in number in CFA lung, apposition is easier to identify here than in normal lung.
X‐ray microanalysis of bulk hydrated specimens of neoplastic and non‐neoplastic human urothelium
Authors: Hopkins Diane M. , Morris James A. , Oates Kenneth , Huddart Henry , Staff William G.
Published: June 2005

Specimens of neoplastic and non‐neoplastic human urothelium were rapidly frozen against a copper block immersed in liquid nitrogen, planed in a cryoultramicrotome and then examined by low‐temperature scanning electron microscopy (LTSEM) following coating by aluminium. Energy dispersive X‐ray microanalysis (XRMA) was performed on the superficial urothelial cells and on gelatin blocks mounted in parallel which had known concentrations of sodium, potassium, chloride, and phosphorus. The neoplastic urothelial cells had significantly less phosphorus (P <0.05) than non‐neoplastic cells and the ratios K+/P,K+/Na+, and K+/Cl− were significantly higher in neoplastic cells than in non‐neoplastic cells (P <0.00001). These results are consistent with those expected in cells with a sustained increase in intracellular pH caused by stimulation of Na+/H+ ion membrane exchange.
Macrophage and perisinusoidal cell kinetics in acute liver injury
Authors: Johnson Sarah J. , Hines Julie E. , Burt Alastair D.
Published: June 2005

Perisinusoidal cells (PSCs) are currently regarded as the major source of extracellular matrix proteins during hepatic fibrogenesis in response to liver injury. However, the cellular mechanisms underlying their response to injury are not fully understood. One hypothesis is that the PSCs are stimulated by peptide growth factors produced by hepatic macrophages (Kupffer cells) in response to parenchymal cell damage. In this study we have investigated the kinetics of the PSC and macrophage populations in acute carbon tetrachloride‐induced hepatic injury in rats. PSCs were identified immunohistochemically by detection of cytoplasmic desmin; monocytes and macrophages were detected using the monoclonal antibodies ED1 and ED2; cells in S phase were identified by immunohistochemical detection of nuclear‐incorporated bromodeoxyuridine. The results showed an expansion of the desmin‐positive PSC population, predominantly within the damaged perivenular zones, which reached a peak on days 3 and 4 following administration of carbon tetrachloride; this was contributed to by local PSC proliferation. The PSC response was preceded by an expansion of the macrophage population resulting from both local macrophage proliferation and influx of blood monocytes. These results are in keeping with the hypothesis that the PSC response to acute liver injury is mediated, at least in part, by hepatic macrophages.
The distribution of LH39 basement membrane epitope in the tumour stroma of oral squmaous cell carcinomas
Authors: Almeida Bernice M. , Challacombe Stephen J. , Eveson John W. , Morgan Peter R. , Purkis Patricia E. , Leigh Irene M.
Published: June 2005

LH39 monoclonal antibody detects a novel component of epithelial and endothelial basement membranes. The expression of LH39 antigen was investigated by immunohistochemistry in 55 oral squamous cell carcinomas and compared with 15 pyogenic granulomas of skin and oral mucosa, 20 non‐specific oral ulcers, and 20 specimens of normal oral mucosa. The distribution of this basement membrane epitope was compared with that of other basement membrane components, type IV collagen, and laminin. LH39 monoclonal antibody labelled basement membrane surrounding small blood vessels in normal human organs. In oral squamous cell carcinomas, in contrast to the other basement membrane antigens, the LH39 epitope was not detectable in vessels within histologically recognizable tumour stroma. Neovascularization is known to attend malignant neoplasms and this finding was interpreted as absence of LH39 antigen within newly formed vessels. In support of this hypothesis, LH39 immunoreactivity was absent in newly formed blood vessels within pyogenic granulomas and the granulation tissue within ulcers. As increasing neovascularization is reported to correlate with a rising rate of metastasis, assessment of tumour angiogenesis may be of value in selecting patients for initial aggressive therapy.
Optimization of non‐isotopic in situ hybridization on formalin‐fixed, paraffin‐embedded material using digoxigenin‐labelled probes and transgenic tissues
Authors: Fleming Kenneth A. , Evans Mark , Ryley K. Claire , Franklin David , Lovell‐badge Robin H. , Morey Adrienne L.
Published: June 2005

The sensitivity of non‐isotopic in situ hybridization (NISH), particularly on formalin‐fixed, paraffin‐embedded (FFPE) clinical tissues, has been the subject of controversy. Generally, NISH has been regarded as being less sensitive than radiolabelled procedures, although some reports have contradicted this. Accordingly, tissues from mice which were transgenic for variable amounts of the human α‐1‐antitrypsin gene were used to optimize the NISH procedure and to estimate the sensitivity. This approach showed that prolonged incubation of slides in final substrate resulted in high sensitivity—about 13 kb of target DNA. However, this prolonged incubation crucially depended on achieving minimal non‐specific background staining. Many factors affected the degree of background staining, but five were particularly important. First, the method of mounting cut sections onto slides. Second, the length of the probe (ideally less than 400 bp). Third, the procedure for proteolytic digestion. Fourth, the denaturation technique, and fifth, the quality of the dextran sulphate used in the hybridization mix. The optimized protocol showed variable patterns of mRNA distribution in the transgenic mouse livers, while DNA distribution appeared uniform.
Reduction in apoptosis relative to mitosis in histologically normal epithelium accompanies fibrocystic change and carcinoma of the premenopausal human breast
Authors: Allan David J. , Howell Anthony , Roberts Stephen A. , Williams Gerrard T. , Watson Robert J. , Coyne John D. , Clarke Robert B. , Laidlaw Ian J. , Potten Christopher S.
Published: June 2005

The aims of this study in 227 premenopausal women were (a) to determine the mitotic index (MI), the thymidine labelling index (LI), and the apoptotic index (AI) within the epithelial cells of histologically ‘normal’ human breast biopsy material removed away from the site of either a fibroadenoma or a carcinoma; and (b) to relate differences in the kinetic indices of the ‘normal’ epithelium to the pathology in the same breast diagnosed as fibroadenoma alone (125 patients), fibroadenoma with accompanying mild fibrocystic change (79 patients), or carcinoma (23 patients). Ratios of the average indices (AI/MI, AI/LI, MI/LI) were also calculated to minimize uncertainties related to the total cell population counted, the denominator in the LI, MI, and AI All indices and ratios of indices were corrected for age, averaged over the cycle, and expressed as log‐transformed values for analysis. Significant (P<0.001) reductions in AI and in apoptosis relative to mitosis (reduced AI/MI) were found in ‘normal’ epithelium from breasts having fibrocystic change (AI = 0.17 ± 0.02; AI/MI = 1.01 ± 0.18) and carcinoma (AI = 0.19 ± 0.04; AI/MI=0.8 ± 0.29), compared with breast with fibroadenoma alone (AI = 0.27 ± 0.03; AI/MI = 1.29 ± 0.39). In the absence of significant differences in MI and LI between the ‘normal’ tissue groups, this finding raises the possibility that reduced epithelial cell apoptosis might be causally associated with the development of fibrocystic change and with an increased risk of development of carcinoma. Reduced apoptosis may lead to a shift in tissue kinetics towards expansion of cell numbers, and also to the preservation of genetically aberrant cells, favouring neoplastic development. The indices, however, are complex and must be interpreted with caution.