bluuebunny/crossref_metadata_2025 huggingface dataset:
The ultrastructure of plexogenic pulmonary arteriopathy
Authors: Smith Paul , Heath Donald , Yacoub Magdi , Madden Brendan , Caslin Anthony , Gosney John
Published: June 2005

The lungs from 16 cases of plexogenic pulmonary arteriopathy obtained at heart‐lung transplantation, half of which had primary pulmonary hypertension, were examined by electron microscopy. From these the probable pathogenesis of pulmonary arterial intimal fibrosis in plexogenic pulmonary arteriopathy was deduced. The earliest detectable change was migration of smooth muscle cells from the media; through the internal elastic lamina into the intima. These cells collected beneath the endothelium and lost many of their myofilaments to become myofibroblasts. They were associated with ground substance but scanty collagen fibrils. As the quantity of interstitial collagen increased, the myofibroblasts reverted to a muscular structure, became elongated, and assumed a regular, circumferential orientation. This later stage coincided with the development of plexiform lesions. At both early and later stages, the muscular pulmonary arteries were contracted but not markedly so, and muscular evaginations were not seen. On the other hand, the cellular intimal proliferations developed early and were occlusive. This suggests that occlusion of small pulmonary arterial vessels by myofibroblasts may be at least as important as vasoconstriction in the early elevation of the pulmonary vascular resistance in primary pulmonary hypertension.
Congenital ECHO virus infection–morphological and virological study of fetal and placental tissue
Authors: Garcia Aparecida Gomes Pinto , Da Silva Basso Nádia Gomes , Fonseca Maria Evangelina Ferreira , Outani Heloisa Novaes
Published: June 2005

A prospective study of 78 pregnant women was undertaken to detect maternal enterovirus infection. Maternal faecal specimens and blood samples, placental and fetal tissue were taken for viral study, electron microscopy, histochemistry, and morphological examination.

We present the post‐mortem findings in three fetuses whose maternal infection was detected before delivery by isolation of ECHO virus type 33 and type 27 from faecal specimens and or placental and fetal tissues. The morphological aspects were similar in all cases and included an acute infection of the placenta and hypoxic/hypotensive injury to fetal organs. In one case, viral particles were detected by electron microscopy of the fetal liver.

This series of cases of intrauterine ECHO virus infection confirms the potential gravity of such infection during pregnancy and the need to prevent enterovtral disease.
Immunohistochemical detection of proliferating lipocytes in regenerating rat liver
Authors: Tanaka Yujiro , Mak Ki M. , Lieber Charles S.
Published: June 2005

For the detection of proliferating lipocytes in regenerating liver, partially hepatectomized rats were injected intrapcritoneally with 5‐bromo‐2′‐deoxyuridine (BrdUrd, 50 mg/kg body weight) and killed 1 h later. Acetone‐fixed frozen liver sections were used for the simultaneous detection of cytoplasmic desmin and BrdUrd‐labelled nuclei of lipocytes using double immunohistochemical procedures. The best results were obtained with the sequences: rabbit anti‐desmin → biotinylated anti‐rabbit IgG → avidin‐biotin‐peroxidase complex → 3,3′‐diaminobenzidine tetrahydrochloride, followed by DNA denaturation → mouse anti‐BrdUrd → anti‐mouse IgG → peroxidase‐anti‐peroxidase complex → 4‐chloro‐l‐naphihol. With this method, cytoplasmic desmin was stained a brown colour, which sharply contrasted with the blue‐stained BrdUrd‐labelled nuclei. Uniabelled nuclei appeared green after counterstaining with methyl green. No cross‐reaction between immunoreagents of desmin and BrdUrd stainings was observed. The labelling index of lipocytes peaked (25.7 percent) 48 h after partial hepatectomy, whereas it was 3.7 per cent in normal rat liver.
Experimental rhabdomyosarcoma with regions like malignant fibrous histiocytoma (MFH)–a true double phenotypic pattern?
Authors: Kosmehl Hartwig , Langbein Lutz , Katenkamp Detlef
Published: June 2005

In murine sarcomas induced by 20‐methylcholanthrene, histological features of malignant fibrous histiocytoma (MFH) as well as rhabdomyosarcoma were found in the same tumour both at light microscopy and at ultrastructural level. The areas showing rhabdomyomatous differentiation expressed vimentin, desmin, muscle‐specific alpha‐actinin, and sometimes myoglobin, but in the MFH areas only vimentin was expressed. A series of allografts in athymic mice, using tumour areas of both histological types, showed in every case a mixed pattern of tumour growth, whether the transplanted tissue was of MFH or rhabdomyosarcomatous type. This suggests that the MFH areas in the original experimental sarcomas were modulated disguised rhabdomyosarcomas. The significance of MFH‐like areas in non‐related soft tissue sarcomas is also discussed.
Use of interactive videodisc for teaching of pathology laboratory cases
Authors: Kumar Kusum , Hodgins Mark
Published: June 2005

Images from a generic videodisc called the 'Slice of Life' were utilized to develop computer assisted interactive videodisc instruction (IVI) units for supplemental independent review of the laboratory cases studied in the general pathology course for year 1 medical students. Four units based on laboratory cases (two on ‘inflammation’ and two on ‘neoplasia’) were developed for a dual‐screen Macintosh system. With this dual‐screen system, specific picture frames from the videodisc could be retrieved on one screen while the other screen displayed the corresponding text. Each unit contained a number of case studies, each of which consisted of a clinical profile, gross picture and description of the tissue removed, sequential photomicrographs at successively higher magnifications from various fields of slides and their descriptions, comparison with the normal histology of the organ involved, interpretation of the observations and comments, final diagnosis, and a quiz at the end. Histopathological features were specifically emphasized. Evaluation of the units obtained from three consecutive classes of students indicated that these units were received as valuable supplemental tools for independent review of the laboratory cases.
Histochemical and immunocytochemical evaluation of copper and metallothionein in the liver and kidney of copper‐loaded rats
Authors: Evering Winston E. , Haywood Susan , Elmes Margaret E. , Jasani Bharat , Trafford James
Published: June 2005

Histochemical methods do not always show a good correlation with analytical measurement of copper content and consequently immunoreactive staining techniques for metaliothionein (MT) have recently been employed for the differential diagnosis of copper‐associated diseases. This study compares histochemical with immunocytochemical methods for the assessment of copper status. Male rats were fed a high copper (1 g/kg) diet for 16 weeks and killed sequentially during this period. The livers and kidneys were analysed for copper and zinc (atomic absorption spectrophotometry), and sections were stained with rubeanic acid and rhodanine for copper and for immunoreactive MT using the DNP localization system. Immunoreactive stains for MT corresponded better with copper content than histochemical stains and were more sensitive, albeit less selective, indicators of copper accumulation. Moreover, major differences in intracellular staining were apparent between the two methods, attributed to differences in copper binding and microcompartmentalization of metal.
Helicobacter pylori is not identified in areas of gastric metaplasia of gall bladder
Authors: Arnaout A. H. , Abbas S. H. , Shousha S.
Published: June 2005

Sections from 200 cholecystectomy specimens were searched microscopically for areas of gastric metaplasia and Helicobacter (Campylobacter) pylori colonization. The presence of gastric metaplasia was confirmed in 31 cases by the presence of tall columnar cells with abundant apical neutral mucin. Examination of haematoxylin and eosin and, in some cases, Warthin–Starry stained sections failed to demonstrate H. pylori in any of these 31 cases or in 29 other cases with ‘bladder neck‐type mucous gland metaplasia’. The failure of H. pylori to colonize areas of gastric metaplasia in the gall bladder confirms their intolerance to high concentrations of bile.
The relation of endotoxaemia to the cause of death and fatal hepatic damage following obstruction of the portal vein
Authors: Shibayama Yuro
Published: June 2005

The present study was undertaken in rats with portosystemic collaterals to clarify whether portal vein occlusion induces fatal hepatic damage and whether endotoxaemla relates to development of lethal hepatic failure following portal vein obstruction. Sudden portal vein occlusion caused severe hepatic necrosis and hepatic dysfunction. However, the mortality rate was only 14 percent, and the hepatic damage recovered 1 week later. Administration of endotoxin immediately after portal vein ligation induced a high mortality rate (86 per cent): the cause of death was thought to be systemic circulatory failure. Endotoxin administration 1 week after portal vein ligation caused marked hepatic dysfunction and hepatic necrosis, but no deaths occurred. These experimental data suggest that hepatic damage following sudden portal vein occlusion is temporary and not fatal provided that portosystemic shunts form adequately. However, it is lethal when complicated by endotoxaemia. Severe hepatic damage and death in chronic portal vein occlusion with portosystemic collaterals cannot be attributed solely to portal vein occlusion, and other factors such as endotoxaemia may play an important role in their development.
Immunohistochemical distribution of c‐erbB‐2 in in situ breast carcinoma—a detailed morphological analysis
Authors: Ramachandra S. , Machin L. , Ashley S. , Monaghan P. , Gusterson B. A.
Published: June 2005

An immunohistochemical study of c‐erbB‐2 expression was carried out on in situ (non‐invasive) breast carcinoma, using antibody 21N, raised to the intracytoplasmic domain of the c‐erbB‐2 oncogene product. Strong membrane staining was observed in 44 out of 74 (59 per cent) cases of ductal carcinoma in situ (DCIS), but none of 48 lobular carcinoma in situ (LCIS) lesions. A detailed comparative morphological evaluation using several different parameters, including histological subtypes, was performed within the DCIS group.

The results showed that there was a significant correlation between c‐erbB‐2 expression and the presence of large cell size, periductal lymphoid cell infiltration, marked nuclear pleomorphism, multinucleation, and a high mitotic rate. Of these, cell size appears to be the most important predictor of c‐erbB‐2 status, followed by the presence of periductal lymphoid cell infiltration.

These results indicate, firstly, that LCIS and DCIS are biologically (as well as histologically) different and, secondly, that a subgroup of DCIS, which is associated with c‐erbB‐2 over‐expression, exists and appears to have distinct histological features. The subgroup of DCIS cases which over‐express c‐erbB‐2 may be a biologically definable category with prognostic importance. These results may therefore have relevance to breast screening programmes, but a larger study incorporating clinical data would be necessary to correlate these findings with clinical outcome.
Ultrastructural study of Helicobacter pylori‐associated gastritis
Authors: Kazi J. L. , Sinniah R. , Zaman V. , Ng M. L. , Jafarey N. A. , Alam S. M. , Zuberi S. J. , Kazi A. M.
Published: June 2005

Endoscopic biopsies of antral mucosa from 26 patients with Helicobacter pylori‐associated gastritis were studied by electron microscopy (EM). Scanning electron microscopy (SEM) showed clustering of H. pylori in the intercellular areas, being entrapped by the microvilli which were decreased at the sites where the bacilli were seen. The observations of SEM were confirmed by transmission electron microscopy (TEM), which showed adherence of the bacilli to the cell surface, producing cup‐shaped depressions in the epithelial cells, and occasionally intracellular infiltration by H. pylori. There were also depletion of mucus granules, degenerative changes, and disruption of intercellular junction complexes of the epithelial cells. Post‐treatment biopsies showed complete disappearance of the bacilli, and ultrastructural changes associated with H. pylori infection were resolved.
Induction of antigen expression of follicular dendritic cells in a monoblastic cell line. A contribution to its cellular origin
Authors: Fliedner A. , Parwaresch M. R. , Feller A. C.
Published: June 2005

Experiments were carried out to elucidate the cellular origin of the dendritic reticulum cell (DRC). The monoblastic cell line THP‐1, the histiocytic cell line U‐937, and the mononuclear cell fraction from peripheral blood (PMC) were stimulated with supernatants from lectin‐stimulated peripheral blood lymphocytes and from stimulated T‐ and B‐cell lines. Differentiation towards DRC was assessed by immunocytochemical demonstration of the DRC‐specific antigen Ki‐M4. Supernatants from isolated peripheral T lymphocytes and from T‐ and B‐cell lines were capable of stimulating THP‐1 to Ki‐M4 antigen expression, whereas U‐937 and the PMC fraction remained negative for this antigen throughout the experiments. These results provide further evidence for a relationship of DRCs with the mononuclear phagocytic cell system and hence for their bone marrow origin. Furthermore, the data suggest that soluble factors of T and/or B cells are involved in mediating the differentiation process of precursor cells to DRC.
Renin gene expression in nephroblastoma
Authors: Lindop G. B. M. , Duncan K. , Millan D. W. M. , Gibson A. A. M. , Patrick W. J. A. , Leckie B. J. , Birnie G. D.
Published: June 2005

Most cases of nephroblastoma have high plasma levels of prorenin which is biologically inactive. Plasma prorenin levels fall to normal following nephrectomy. In order to ascertain whether renin synthesis occurs in nephroblastomas we decided to search for renin‐specific mRNA using a cDNA probe and Northern blot analyses on total RNA purified from snap‐frozen human tumour tissue obtained at nephrectomy. We demonstrated renin‐specific mRNA in 5/11 (45 per cent) nephroblastomas. It was 1.6 Kb in length, similar to the mRNA detected in normal kidney tissue and in kidneys with renal artery stenosis. In one of the cases of nephroblastoma, in which we could detect no normal renin mRNA at 1.6 Kb, the cDNA probe hybridized with a higher molecular weight mRNA 3 Kb in length. We conclude that some nephroblastomas synthesize renin.
Evaluation of nucleolus organizer regions (NORs) by automatic image analysis: A contribution to standardization
Authors: Rüschoff Josef , Plate Karl H. , Contractor Hans , Kern Sigrid , Zimmermann Ralf , Thomas Carlos
Published: June 2005

This study summarizes our experiences with the silver staining of nucleolus organizer regions (AgNORs) in a total of 580 tumours from ten different tissues. In contrast to other investigators, we made use of automatic image analysis for the evaluation of AgNORs. This provided good reproducibility as determined by the standard cumulative means technique and intra‐observer (r1) and inter‐observer (r2) agreement in 30 benign (r1 = 0.83‐0.95, r2= 0.76–0.92) and 50 malignant tissue samples (r1=0.72–0.85, r2 = 0.51–0.78). By using a series of staining times on sections from 30 tissue blocks taken from the ten types of tissue investigated, considerable variation in the argyrophilic staining of NORs in different tissues and in different blocks from one tumour was shown. The mean AgNOR area of resting lymphocytes or connective tissue cells within tissue blocks of the same organ system varied up to four‐fold, even though identical staining times had been used. The most suitable silver reaction time which rendered a good diagnostic difference in the AgNOR content of benign and malignant tissue ranged, for example, in the breast cancer specimens, from 23 to 35 min. We therefore conclude that the staining time has to be adjusted to the individual silver‐binding characteristics of each tissue block or even each section. The use of internal staining standards like lymphocytes or connective tissue cells in the same tissue section is mandatory. This, in turn, is most precisely controlled by morphometry.
Ultrastructural immunogold studies of heparan sulphate proteoglycan in normal human glomeruli and glomerulonephritis
Authors: Moss Jill , Woodrow David F. , Shore Ian , Gower Peter , Phillips Malcolm , Spiro Robert G.
Published: June 2005

The distribution of heparan sulphate proteoglycans (HSPG) has been investigated in normal human glomeruli, membranous glomerulonephritis, mesangial IgA disease, and anti‐glomerular basement membrane disease. HSPG was localized using anti‐bovine HSPG antibody and 10 nm gold‐labelled secondary antibody on paraformaldehyde‐fixed, Lowicryl K4M resin‐embedded kidneys. HSPG was present in all glomeruli and there was a zonation of its distribution in that it was predominantly on the epithelial aspect of the glomerular basement membrane (GBM) and mesangium with little in the central regions of the mesangial matrix. In the cases of immune complex glomerulonephritis, no HSPG was found in the electron‐dense deposits. These findings contrast with our previous studies using the same technique in which type IV collagen and fibronectin were found predominantly on the endothelial aspect of the GBM.
Expression of gastrin‐releasing peptide (human bombesin) gene in large cell undifferentiated carcinoma of the lung
Authors: Hamid Q. A. , Corrin B. , Dewar A. , Hoefler H. , Sheppard M. N.
Published: June 2005

Production of the growth factor gastrin‐releasing peptide (GRP) or human bonbesin has been shown to be a feature of neuroendocrine tumours of the lung, particularly small cell carcinoma, and is possibly responsible for the characteristically rapid growth of this tumour. Large cell undifferentiated carcinoma of the lung (LCC) is also characterized by rapid growth and there is increasing evidence that some LCCs exhibit neuroendocrine differentiation. We therefore investigated GRP/bombesin immunoreactivity and the expression of GRP gene in ten LCCs. Histologically, all were composed of large cells with abundant cytoplasm, open nuclei, and prominent nucleoli, and there was no evidence of squamous, glandular, or neuroendocrine differentiation. At the ultrastructural level, most showed squamous or glandular differentiation but none contained neuroendocrine granules. None of the tumours showed immunoreactivity for GRP/bombesin but seven of the ten showed a focal hybridization signal when treated with 32P‐labelled GRP cRNA probes, indicating the presence of GRP mRNA. This was confirmed by northern‐blot analysis. This study shows for the first time that GRP gene is expressed in LCC. The production of GRP may contribute to the aggressive behaviour of LCC.
Ubiquitin carboxyl‐terminal hydrolase (PGP 9.5) is selectively present in ubiquitinated inclusion bodies characteristic of human neurodegenerative diseases
Authors: Lowe James , McDermott Helen , Landon Michael , Mayer R. John , Wilkinson Keith D.
Published: June 2005

The recent discovery that brain PGP 9.5 is a ubiquitin carboxyl‐terminal hydrolase suggests that the role of this protein should be studied in relation to ubiquitinated cellular inclusions characteristic of several chronic human degenerative diseases.

Formalin‐fixed, paraffin‐processed sections known to contain ubiquitin‐protein conjugate immunoreactivity in cortical Lewy bodies, neurofibrillary tangles, Rosenthal fibres, Pick bodies, spinal inclusions in motor neurone disease, and Mallory's hyaline in alcoholic liver disease were immunostained to localize PGP 9.5. The majority of cortical Lewy bodies in diffuse Lewy body disease showed immunoreactivity for PGP 9.5. In Alzheimer's disease, only a minority of loosely arranged globose‐type neurofibrillary tangles were immunostained together with a minority of neurites surrounding senile plaques. In cerebeilar astrocytomas, the periphery of the majority of Rosenthal fibres was immunostained in addition to strong diffuse cytoplasmic immunostaining in some astrocytes lacking apparent Rosenthal fibres. In Pick's disease, there was no immunostaining of inclusions but there was intense immunostaining of swollen Pick cells. No spinal inclusions in motor neurone disease were stained; however, anterior horn neurones appear to show increased levels of PGP 9.5 compared with those from control cases. No immunostaining of hepatic Mallory's hyaline was demonstrable, which accords with suggestions that PGP 9.5 is a tissue‐specific ubiquitin C‐terminal hydrolase isoenzyme.

The differential detection of a ubiquitin C‐terminal hydrolase in different forms of ubiquitinated inclusion body in the nervous system may form the basis of a method for assessment of the staging of inclusion body biogenesis and give insight into the dynamics of inclusion body formation. In particular, it is possible that the proportion of inclusion‐bearing cells showing high levels of the hydrolase will provide a marker of the level of activity of the degenerative process.
Ultrastructural localization of milk fat globule membrane antigens in human breast carcinomas
Authors: Corcoran David , Walker Rosemary A.
Published: June 2005

The localization of milk fat globule membrane components has been assessed using post‐fixation immunoelectron microscopy with three different antibodies for a group of breast carcinomas of different type and histological differentiation.

For well differentiated carcinomas localization was in relation to the cell membrane, with polarization being evident in a proportion of cases. Moderately differentiated carcinomas showed a combined picture of cell membrane, vesicular, and intracytoplasmic luminal localization. The latter is a feature of infiltrating lobular carcinomas. Poorly differentiated carcinomas exhibited vesicular labelling throughout the cytoplasm, with no cell membrane localization. No labelling was seen over endoplasmic reticulum.

It is proposed that carcinomas exhibit defects in intracellular transport of milk fat globule membrane components resulting in failure of expression at the cell surface and accumulation of vesicles within the cytoplasm, the extent of change relating to tumour differentiation.
High endothelial venules in B‐cell non‐Hodgkin malignant lymphomas
Authors: Pajor Katalin , Pap Tihamér , Csanaky György , Kalász Veronika
Published: June 2005

The number of high endothelial venules (HEVs) per unit area was determined in 55 B‐cell non‐Hodgkin malignant lymphomas (nHMLs) classified according to the Kiel classification. The number of HEVs per unit area was significantly different between low‐ and high‐grade B‐cell nHMLs. In the low‐grade group, a large number of HEVs with preserved structure were present, whereas in high‐grade tumours the HEVs appeared damaged. The present findings support the previous observation attributing prognostic relevance to lymphocyte recirculation in nHMLs.
Early dermato‐pathological signs during bath‐puva therapy
Authors: Piérard‐franchimont Claudine , Nickels‐read Dominique , Mosbah Taha Ben , Estrada Jorge Arrese , Piérard Gérald E.
Published: June 2005

The combination of trioxsalen bath and long‐wave UV‐A is used in dermatology to treat selected patients with psoriasis. Such treatment induces side effects whose clinical aspects have already been described. We report the histological aspects of these lesions. The most striking features occur in the epidermis, where we found vacuolar alterations, disorganization of the stratum malpighi, necrotic cells, and Bowenoid changes. Langerhans cells are reduced in number, and melanocytes may become large and atypical. In addition, factor XIIIa‐positive dendrocytes of the superficial dermis increase in size, and some of them migrate into the epidermis, which is a rare location for these cells.
Chronic serum sickness glomerulonephritis: Heparin enhances the removal of glomerular antigen
Authors: Furness Peter N.
Published: June 2005

Chronic serum sickness glomerulonephritis was induced in rats using 125I‐labelled cationic bovine serum albumin as antigen. During the recovery period the animals were given protamine or polyethyleneimine (PEI), both of which are cationic, or heparin, which is anionic. A control group received saline. The cationic molecules were not shown to influence the rate of removal of antigen from the glomeruli, but heparin increased removal. In subsequent experiments this effect of heparin was confirmed, and a similar effect was demonstrated using low molecular weight heparin. These results indicate that large doses of heparin can enhance the removal of antigen from well‐established glomerular deposits. Fibrin is not detectable in the glomeruli in this model, so this effect of heparin may be independent of its anticoagulant properties.
Clinical, biochemical and immunoelectron microscopical evidence of dual hormone production in a mammosomatotroph cell adenoma
Authors: Cameron Stuart , Allen Ingrid , Ozo Chukwuemiko , Kennedy Laurence , Atkinson Brew , Hadden David
Published: June 2005

This paper reports the occurrence of a rare, yet distinct pituitary adenoma which was surgically removed from a 42‐year‐old male with both clinical and biochemical evidence of acromegaly and mild hyperprolactinaemia. The monomorphic adenoma consisted of mature cells which were ultrastructurally indistinguishable from those of a prolactinoma. Electron immunocytochemistry, including a series of double‐labelling techniques using selected colloidal gold particles as markers, indicated the presence of a pituitary adenoma in which the cells were capable of simultaneously producing growth hormone and prolactin and packaging them within the same secretory granule. This is thought to represent a mammosomatotroph cell adenoma.
Analysis of T‐cell receptor and immunoglobulin gene rearrangements in the diagnosis of Hodgkin's and non‐Hodgkin's lymphoma
Authors: Gledhill Sarah , Krajewski Andrew S. , Dewar A. Edna , Onions David E. , Jarrett Ruth F.
Published: June 2005

The results of genotypic analysis of 29 cases of malignant lymphoma are reported and the application of this technique for differentiating between Hodgkin's disease (HD) and non‐Hodgkin's lymphoma (NHL) is evaluated. Five cases with a differential diagnosis which included HD and NHL were analysed. These results are compared with those obtained for six B‐cell NHLs, nine T‐cell NHLs, and nine cases of HD. This report suggests that gene rearrangement analysis is useful in some cases in which the differential diagnoses includes HD and NHL as the absence of gene rearrangements is more consistent with a diagnosis of HD than of NHL. Two monoclonal antibodies reactive with the variable region of the T‐cell receptor β‐chain and molecular probes to the relevant variable region genes were used to assist in the diagnosis of T‐cell lymphoma. This report confirms that genotypic analysis is useful diagnostically when the results are assessed in the context of the histopathological findings.
Demonstration of Epstein‐Barr viral DNA in formalin‐fixed, paraffin‐embedded samples of Hodgkin's disease
Authors: Libetta Carole M. , Pringle J. H. , Angel Carole A. , Craft A. W. , Malcolm A. J. , Lauder I.
Published: June 2005

The presence of Epstein‐Barr virus (EBV) DNA in formalin‐fixed, paraffin‐embedded samples of Hodgkin's disease (HD) was investigated by Southern blot hybridization using a specific EBV Bam H1W fragment probe. DNA from 16 samples of HD in children and young adults was compared with that from ten samples in adults older than 45 years. A further eight cases of DNA extracted from fresh samples of HD were also studied, in order to compare the results obtained with fresh and fixed tissue samples.

Hybridization was demonstrated in 15 of the 34 cases of HD studied in contrast to 3 or the 34 control specimens of non‐Hodgkin's lymphoma, non‐neoplastic lymph nodes, and carcinomas. No differences between the two age groups compared were found.

The results of this study suggest that there is a definite association between EBV and HD, although the exact nature of this association remains to be established. It has also been shown that DNA of sufficient quality for the detection of EBV DNA can be extracted from formalin‐fixed, paraffin‐embedded material, and that comparable results can be obtained using DNA extracted from fresh and fixed tissue.
QBEnd/10: A new immunostain for the routine diagnosis of Kaposi's sarcoma
Authors: Sankey Elizabeth A. , More Linda , Dhillon Amar P.
Published: June 2005

Kaposi's sarcomas are seen more commonly in routine histopathology laboratories since the advent of the more widespread and aggressive variant of the disease associated with HIV infection. Distinguishing nodular lesions from other spindle cell and vascular tumours can sometimes be difficult. Immunohistochemistry has been disappointing as a diagnostic aid, often requiring special fixation or frozen tissue and even then, staining of spindle cells has been variable. We describe the use of the new IgGl mouse monoclonal antibody raised against human placental endothelial cells, QBEnd/10, on routine formalin‐fixed, paraffin‐embedded tissue. A retrospective study was performed on 22 Kaposi's sarcomas of skin including patch, plaque, and nodular lesions and compared with 38 other vascular and spindle cell tumours from skin. All sections were stained with haematoxylin and eosin, QBEnd/10, Ulex europaeus agglutinin 1 (UEA‐1) and for factor VIII‐related antigen (FVIIIRAg). The results demonstrate that spindle cells in lesions from Kaposi's sarcomas, but not other vascular or spindle cell tumours, immunostain clearly with QBEnd/10. Immunostaining for FVIIIRAg shows only weak and irregular positivity of the spindle cells, whilst staining with UEA‐1 is consistently negative. We find that immunostaining with QBEnd/10 aids the diagnosis of Kaposi's sarcomas and allows their distinction from other spindle cell neoplasms of skin in routinely processed material.
Axonal damage in Crohn's disease is frequent, but non‐specific
Authors: Brewer Douglas B. , Thompson Henry , Haynes Ian G. , Alexander‐williams John
Published: June 2005

We have attempted to confirm the claim by Dvorak and Silen that ‘Crohn’s disease is accompanied by a severe and extensive necrosis of gut axons. ‘which’ may serve to differentiate Crohn's disease from other inflammatory conditions'. In this electron microscope study the diagnoses were withheld until the assessment of axonal damage was completed. We assessed the axonal damage in ileostomy biopsies in 13 cases of Crohn's disease, four cases of ulcerative colitis, and two ‘controls’. In Crohn's disease we found a mean per cent of abnormal axons of 29‐85, in ulcerative colitis of 21.25 per cent, and in the two ‘controls’ of 12.11 and 10.63 percent, respectively. The difference between the 13 cases of Crohn's disease and the six cases of non‐Crohn's disease is not significant. We found considerable numbers of abnormal, very small axons of uncertain nature but probably related to regeneration following surgery. Including or excluding such axons did not significantly alter the incidence of abnormal axons. We conclude that axonal damage is common in chronic inflammatory bowel disease and is not specifically related to Crohn's disease.
Role of endotoxaemia in the development of hepatic failure following hepatic artery occlusion
Authors: Shibayama Yuro
Published: June 2005

The present study was undertaken to examine whether endotoxaemia relates to the development of hepatic failure following surgical ligation or embolization of the hepatic artery. A small amount of endotoxin was given immediately or 1 week after hepatic artery ligation in rats, and liver function tests and morphological examination of the liver were performed at 24 h after the administration. Hepatic artery ligation alone or endotoxin administration alone did not induce functional and morphological alteration of the liver. However, when endotoxin was given immediately after hepatic artery ligation, there was an increase in serum transaminase activity and focal hepatocellular necrosis developed. On the other hand, when endotoxin was given 1 week after hepatic artery ligation, the hepatic injury was not induced because of the development of hepatic arterial collaterals. These data suggest that endotoxaemia may be a cause of hepatic failure following hepatic artery occlusion, and that the risk may persist until the establishment of hepatic arterial collaterals.
Accelerated immunohistochemical staining by microwaves
Authors: Leong Anthony S.‐Y. , Milios James
Published: June 2005

The application of microwave irradiation is extended to include the acceleration of the streptavidin‐biotin peroxidase staining procedure for the detection of a wide range of antigens in paraffin‐embedded sections of both normal and neoplastic tissues. Microwave irradiation is used for all procedural steps requiring incubation so that the entire staining procedure can be completed within 20 min. The technique is simple and convenient, with uniform irradiation times for different tissue antigens. The results obtained equalled that of conventional staining procedures.
Detection of endothelin immunoreactivity and mRNA in pulmonary tumours
Authors: Giaid Adel , Hamid Qutayba A. , Springall David R. , Yanagisawa Masahi , Shinmi Osmu , Sawamura Tatsuya , Masaki Tomoh , Kimura Sadao , Corrin Bryan , Polak Julia M.
Published: June 2005

Paraffin sections of 66 surgically resected lung tumours were immunostained with antisera to human endothelin‐1 and to the C‐terminal peptide of big endothelin. With both antisera, strong immunoreactivity was demonstrated in 11 of 15 squamous cell carcinomas and 11 of 16 adenocarcinomas. Focal immunoreactivity was seen in small cell carcinoma (2/2), large cell carcinoma (2/5), and carcinoid tumours (2s/11). Four lymphomas and three sarcomas did not show endothelin immunoreactivity. Cryostat sections of 22 of the 66 tumours were hybridized with radiolabelled complementary RNA probes prepared from the 3′ non‐coding region of endothelin‐1 cDNA, and the chromosomal genes encoding endothelin‐2 and ‐3. In situ hybridization demonstrated the presence of endothelin mRNAs in 4 of 7 squamous cell carcinomas and in 5 of 8 adenocarcinomas, in a pattern similar to that shown by immunocytochemistry. No hybridization signals were obtained from the other types of tumours. In lung tissue adjacent to the tumours, endothelin‐like immunoreactivity and mRNA were detected in pulmonary endocrine cells and, in some cases, other epithelial cells, and in alveolar capillary endothelial cells. This study demonstrates the expression of endothelin in a number of pulmonary tumours and suggests a possible role for this peptide in the growth and/or differentiation of these tumours.
Bacillus Calmette‐Guéarin‐treated superficial bladder cancer: Correlation of morphology with immunophenotyping
Authors: Leong Anthony S.‐Y. , Wannakrairot P. , Jose J. , Milios J.
Published: June 2005

Thirty‐five biopsies from 19 patients with superficial transitional cell carcinoma of the bladder, treated with intravesical bacillus Calmette‐Guérin (BCG), were assessed histologically and immunohistochemically. Pretreatment biopsies were available for comparison in all cases and five cases of non‐specific cystitis were also examined. The inflammatory infiltrate was assessed with a streptavidin‐biotin‐peroxidase method using UCHL1, MT1, LN3, L26, HAM56, MAC387, Leu7 and anti‐S100 in paraffin sections, and in 18 specimens where frozen tissues were available, Leu 1,2,3,4,14, OKT10, HLA‐DR and anti‐Tac antibodies were applied. Post‐treatment bladder biopsies showed severe oedema and a variable degree of inflammation. A granulomatous inflammation was seen in 11 cases, with granulomas present in six prostatic biopsies and acid‐fast bacilli in two cases. The lymphoid infiltrate in all biopsies were largely T lymphocytes with a predominance of T helper cells present, often as a band‐like infiltrate pressing against the residual epithelium, or the denuded bladder surface, and distributed in the vicinity of the granulomas. Activated lymphocytes were prominent in seven cases, although a moderate infiltrate of such cells was seen in all instances. Tac antigen was only occasionally expressed, and in a few NK cells were present among the infiltrates. In eight cases, HLA‐DR was expressed in epithelial cells following BCG treatment, whereas all pre‐treatment epithelia were negative.
Ubiquitin conjugate immunoreactivity in the brains of scrapie infected mice
Authors: Lowe James , McDermott Helen , Kenward Nigel , Landon Michael , Mayer R. John , Bruce Moira , McBride Patricia , Somerville Robert A. , Hope James
Published: June 2005

Sections of brain from normal mice or clinically‐ill mice infected with either the 87V or the ME7 strains of sheep scrapie were immunostained to show the localization of ubiquitin‐protein conjugates or a specific marker of disease, the scrapie‐associated fibril protein (PrP). In both scrapie models immunoreactive ubiquitin‐protein conjugates were seen in thread‐like structures found throughout the neuropil, in inclusion bodies within vacuolated neurones, and in areas surrounding anti‐PrP positive amyloid plaques. The PrP protein was visualized in diffuse deposits in highly vacuolated parts of the scrapie‐affected brain, and focally in amyloid plaques, microglia and neuronal processes.

The ubiquitin‐protein conjugate staining of scrapie amyloid plaques is very similar to that seen in the plaques of Alzheimer's disease. The ubiquitinated intraneuronal inclusion bodies seen in scrapie resemble the granulovacuolar lesions also seen in Alzheimer's disease, but appear much larger and possibly correspond to material in giant autophagic vacuoles. We suggest that these inclusions may be the result of ubiquitinated abnormal proteins being directed to the lysosomal system, and that scrapie and Alzheimer's disease share at least some common processes of neurodegeneration.
The polymerase chain reaction: Miracle or mirage? A critical review of its uses and limitations in diagnosis and research
Authors: Wright Pat A. , Wynford‐Thomas David
Published: June 2005

Since publication of the polymerase chain reaction (PCR) technique in 1985 (Saiki et al. Science 1985; 230: 1350–1354), there has been an explosion of reports on its use in medicine and science. We critically review its use both as a diagnostic technique and as a research tool, and show the pathologist how to evaluate PCR data and how to avoid the pitfalls of overinterpretation. We discuss the value of PCR in the characterization of genetic defects, prenatal diagnosis, carrier testing, H LA typing, detecting micro‐organisms, identifying activated oncogenes, and in the characterization of leukaemias and lymphomas, and summarize the main applications in biomedical research.
Immunohistochemical identification of mast cells in formaldehyde‐fixed tissue using monoclonal antibodies specific for tryptase
Authors: Walls Andrew F. , Jones David B. , Williams Julie H. , Church Martin K. , Holgate Stephen T.
Published: June 2005

An avidin–biotin enhanced immunoperoxidase procedure using monoclonal antibodies (AA1, AA3, and AA5) prepared against human mast cell tryptase resulted in intense staining of mast cells in paraffin‐embedded tissue. The distribution of mast cells observed was similar to that seen when adjacent serial sections were stained using a standard procedure with toluidine blue, though the immunoperoxidase technique permitted the identification of significantly more mast cells. With monoclonal antibody AA1, immunostaining was entirely specific for mast cell granules, and there was negligible background staining in a range of tissues including lung, tonsil, colon, gastric mucosa, skin, and pituitary. There was no staining of antibody on basophils or on any other normal blood leukocyte. The technique was effective with tissue fixed in either Carnoy's or neutral buffered formalin, though the internal mast cell structure was better preserved with formaldehyde fixation. The immunoperoxidase staining procedure with monoclonal antibody AA1 is a highly specific and sensitive means for the detection of mast cells in routinely processed tissues.
Sequence analysis of amplified t(14;18) chromosomal breakpoints in B‐cell lymphomas
Authors: Eick S. , Krieger G. , Bolz I. , Kneba M.
Published: June 2005

We have explored different strategies for sequencing of major breakpoint (mbr) junctional regions in t(14;18) chromosomal transiocations—the most frequent chromosomal abnormality observed in B‐cell lymphomas. We demonstrate that coupling of the preparation of single‐stranded DNA by asymmetric polymerase chain reaction (PCR) and direct sequencing is the method of choice for the rapid and precise determination of clone‐specific bcl‐2/JH fusion gene sequences. The rapidity, relative ease, and accuracy of the technique, described for the nucleotide sequence analysis of mbr t(14;18) breakpoints, permits the analysis of a relatively large number of samples and should be considered as part of the clinical evaluation of lymphoma patients. Furthermore, by providing sequence information of clone‐specific DNA regions, the procedure should reduce the risk of false‐positive results from PCR.
A genotypic study of low grade B‐cell lymphomas, including lymphomas of mucosa associated lymphoid tissue (MALT)
Authors: Wotherspoon Andrew C. , Pan Langxing , Diss Tim C. , Isaacson Peter G.
Published: June 2005

Genotypic analysis has led to the implication of certain oncogenes in the pathogenesis of specific groups of non‐Hodgkin's lymphoma. Rearrangements of c‐myc are associated with Burkitt's lymphoma and of bcl‐2 with centroblastic/centrocytic lymphoma. Rearrangement of bcl‐1 has yet to be associated with a specific group of lymphoma. In this study DNA from 62 cases of low grade B‐cell lymphoma, classified using the Kiel classification, were analysed by Southern blotting and hybridization with probes to bcl‐1, bcl‐2, and c‐myc. Rearrangements of bcl‐2 were found in a proportion of centroblastic/centrocytic lymphoma comparable to other published studies. Rearrangement of c‐myc was not found in any case studied. Bcl‐1 rearrangement was found in 2/9 cases of B‐CLL, and 3/6 cases of centrocytic lymphoma. This incidence of bcl‐1 rearrangement in centrocytic lymphoma suggests that it is a characteristic change. No rearrangement of bcl‐1, bcl‐2 or c‐myc was found in any case of lymphoma of mucosa associated lymphoid tissue (MALT), providing further evidence that, in spite of having a similar morphology to some other groups of low grade B‐cell lymphoma, lymphomas of MALT comprise a distinct entity.
A sensitive method of screening for dominant T cell clones by amplification of T cell gamma gene rearrangements with the polymerase chain reaction
Authors: Goudie R. B. , Karim Sabeeha N. , Mills K. , Alcorn M. , Lee F. D.
Published: June 2005

A sensitive method of screening for dominant T cell clones in small samples of DNA has been developed using the polymerase chain reaction to amplify and identify T cell gamma receptor gene rearrangements. It can detect such rearrangements in nanogram quantities of DNA from cultured T cell clones, even in the presence of 20–100 parts of polyclonal lymph node DNA, and works with DNA extracted from paraffin sections of cloned T cells which have been fixed in formalin. Presumptive clonal reactions have been obtained in preliminary tests on 8 of 10 unfixed T cell lymphomas but in 0 of 10 reactive lymph nodes.
Zonal distribution of immunoglobulin‐synthesizing cells within the germinal centre: An in situ hybridization and immunohistochemical study
Authors: Close Pauline M. , Pringle James H. , Ruprai Anita K. , West Kevin P. , Lauder Ian
Published: June 2005

Immunohistologic studies have shown that synthesis of cytoplasmic immunoglobulin (cIg) is a normal function of some follicle centre cells (FCCs). The mechanisms regulating this synthesis of immunoglobulin and its function within the germinal centre are still poorly understood. In this study we applied a recently developed in situ hybridization method for the detection of kappa and lambda light chain mRNA to reactive lymph nodes and tonsils in order to investigate further the immunoglobulin‐synthesizing cells of the germinal centre.

FCCs containing detectable levels of light chain mRNA corresponded closely to cells containing cIg. The detection of light chain mRNA rather than its immunoglobulin product was found to be an advantage in that problems associated with the detection of extracellular immunoglobulin were eliminated. This was most apparent in germinal centres where the absence of ‘network’ immunoglobulin led to the observations that immunoglobulin‐synthesizing FCCs are predominantly small centrocytes and that in a proportion of germinal centres they localize in that part of the light zone closest to the dark zone. This zonal distribution of immunoglobulin‐synthesizing FCCs raises the possibility of further functional and micro‐environmental subcompartments within the light zone.
Growth fraction in lung tumours determined by Ki67 immunostaining and comparison with agnor scores
Authors: Soomro Irshad Nabi , Whimster William F.
Published: June 2005

Estimation of the growth fraction with the use of monoclonal antibody Ki67, which recognizes a nuclear antigen in proliferating cells, was compared with the nucleolar organizer region staining in 95 lung tumours. There was nuclear staining in most tumours; 12 tumours were negative. Cytoplasmic staining was observed in another seven tumours. The small cell carcinoma group had the highest mean Ki67 index (23·75); squamous carcinomas had a mean value of 15·71, adenocarcinomas 10·99, and large cell anaplastic 20·76. Carcinoids had few stained cells. Nucleolar organizer regions were demonstrated by the argyrophilic method (AgNOR). No correlation was found between Ki67 indices and AgNOR scores. Kinetic data obtained by the AgNOR technique were less discriminating in view of the overlap between scores of various groups including carcinoids. We conclude that use of the monoclonal antibody Ki67 is a more reliable method of assessing proliferative activity in lung tumours. This antibody may be effective in identifying slowly proliferating tumours which are less sensitive to chemotherapeutic agents.
The glomerular changes in children with reflux nephropathy
Authors: Morita Miyako , Yoshiara Shoichi , White Richard H. R. , Raafat Faro
Published: June 2005

While heavy proteinuria and focal segmental glomerulosclerosis (FSGS) are well‐recognized features of progressive reflux nephropathy in adults, little is known of their early evolution. We have studied the glomerular changes in renal biopsy specimens obtained from 24 patients aged 5·2–18·8 years, in whom urinary protein excretion was measured as early morning urine protein/creatinine ratios, using the Coomassie blue dye‐binding method.

Segmental sclerotic lesions were found in eight biopsies and traced through serial sections to a hilar origin in every instance. There was a strong positive correlation between the extent of glomerular involvement and the amount of proteinuria (P<0·0001). Parahilar hyaline deposits were observed in 16 biopsies, including five of the eight showing FSGS. All unsclerosed glomeruli were enlarged, and the hilar arterioles showed both enlargement and thickening, their walls frequently containing subendothelial hyaline deposits.

Since in most patients renal function was comparatively well preserved, despite extensive loss of renal substances, we believe that these glomerular and vascular changes represent the stages in the evolution of hyperfiltration. Microproteinuria is the earliest clinical manifestation of FSGS, and should be sought routinely in all patients with reflux nephropathy.