bluuebunny/crossref_metadata_2025 huggingface dataset:
Angiogenic activity of fibrin degradation products is located in fibrin fragment E
Authors: Thompson W. D. , Smith E. B. , Stirk C. M. , Marshall F. I. , Stout A. J. , Kocchar A
Published: June 2005

The source of angiogenic activity of fibrin degradation products has been sought in a series of experiments, applying degradation products from different types of fibrin and fibrinogen to the chick chorioallantoic membrane. The presence of platelets or fibronectin during clotting was not essential for activity, and neither was crosslinking. Fibrinogen degradation products were non‐stimulatory, as was serum. Molecular sieve column chromatography indicated a range of active fragments. Admixture of active fibrin degradation products with antifibrin fragment E, but not D, antiserum neutralized activity. Preparations containing only fibrin fragment E retained activity. A commercial preparation of fibrinogen fragment E was inactive until treated with thrombin. These experiments point to fibrin fragment E being the source of angiogenic activity, with thrombin cleavage being the essential step in generating activity.
Evaluation of PGP9.5 in the diagnosis of Hirschsprung's disease
Authors: Sams Virginia R. , Bobrow Lynda G. , Happerfield Lisa , Keeling Jean
Published: June 2005

The ability of an acetylcholinesterase‐stained frozen section to detect an increase in large cholinergic nerve fibres within the muscularis mucosae and extending into the lamina propria was a significant step forward in the diagnosis of Hirschsprung's disease (HD). However, such frozen section diagnosis is not always possible. The purpose of this study was to assess the ability of PGP9.5 to detect this pattern of mucosalnen/e fibre staining immunohistochemically. Sixtyfour specimens were included in the study. Twenty‐six of these had been diagnosed as HD by conventional means. All cases were stained immunohistochemically with PGP9.5, S100, and anti‐neurofilaments (NF). Twenty‐four cases of HD were also stained with neurone‐specific enolase (NSE). PGP9.5 reliably stained fibres in the mucosal and submucosal plexuses, and ganglion cells, when the latter were present. This positive staining of ganglion cells was more intense than that seen with NSE, and the positive fibre staining was more intense than that seen with NF. Increased lamina propria fibres were detected with PGP9.5 in only 37 per cent of HD cases compared with S100 positive staining in 60 per cent of cases. However, when S100 staining was assessed alone, it gave a higher false‐negative rate in diagnosing HD than PGP9.5 used alone. Therefore we would recommend the use of PGP9.5 and S100 together for the immunohistochemical diagnosis of HD in formalin‐fixed biopsies.
Adhesion receptors of intimal and subintimal cells of the normal synovial membrane
Authors: Demaziere A. , Athanasou N. A.
Published: June 2005

The distribution of cell adhesion molecules (CAMs) and matrix proteins in the normal synovium of four subjects was studied by immunohistology in order to determine the factors governing the cellular and tissue organization of the intimal and subintimal compartments. Basement membrane proteins, laminin, and collagen type IV, as well as vitronectin and fibronectin, were identified in the intima and there was corresponding expression of integrin and non‐integrin receptors (e.g., CD29, CD49b, CD49d, CD49e, CD49f, CD51, CD61, CD44) for these matrix proteins. There were notable differences in CAM expression between intimal, subintimal, and vascular compartments of the synovial membrane. Phenotypic heterogeneity for CAMs involved in cell–cell interactions, particularly CDlla, CD 11 b, ICAM‐1, and HLA‐DR, was also present. The range of CAMs expressed by synovial and endothelial cells not only indicates a structural role for these antigens, but also suggests that they may control leucocyie traffic into the membrane, including recruitment of cells into the synovial lining.
Asbestos‐stimulated tumour necrosis factor release from alveolar macrophages depends on fibre length and opsonization
Authors: Donaldson Kenneth , Li Xiao Yang , Dogra Shashi , Miller Brian G. , Brown Geraldine M.
Published: June 2005

Fibre length has been shown to be an important factor in the ability of respirable fibres to cause lung fibrosis and cancer. We have reported that a long sample of amosite asbestos is more carcinogenic and fibrogenic than a short sample of similar diameter. These amosite asbestos samples were studied with regard to their ability to stimulate the release of the pro‐inflammatory cytokine tumour necrosis factor (TNF) from rat alveolar macrophages in vitro. The long fibre sample was found to stimulate substantially greater release of the cytokine than the short sample. Furthermore, on treatment of the fibres with rat immunoglobulin G (IgG), there was an increase in the ability of both the long and the short sample to stimulate TNF secretion, although the long sample retained by far the greatest activity. Coating of the fibres with a range of other proteins had no substantial effect on their ability to stimulate TNF secretion. Quartz and titanium dioxide (TiO2) were included as control particles and the TNF‐stimulating activity of quartz was notably increased by opsonization with IgG. TiO2 showed a similar low activity to that of the short fibre sample of amosite but this again could be modestly increased by opsonization with IgG. The simulation of TNF release caused by treatment with immunoglobulin‐opsonized long fibre amosite could be inhibited by treatment of the macrophages with the protein kinase C‐inhibitor staurosporine. The study demonstrates a fibre length‐related ability to stimulate cytokine secretion by alveolar macrophages, and its enhancement by opsonization with IgG. This is likely to be relevant to the relationship between fibre length and pathogenic potential to the lung.
Expression of the c‐fos oncogene in chemically‐induced mouse tumours and in human skin tumours
Authors: Urabe Atsumichi , Nakayama Juichiro , Taniguchi Shun'Ichiro , Terao Hiroshi , Hori Yoshiaki
Published: June 2005

Using a polyclonal antibody to fos oncoprctein and an immunofluorescent technique, we investigated expression of the fos oncogene in chemically‐induced mouse tumours and human premalignant and malignant skin lesion. In the chemically induced tumours, the nuclei of almost all carcinoma cells stained uniformly with this antibody, while positive cells were observed in the outermost layers in the benign papillomas. In human tumours, a greater degree of nuclear staining was observed in cases of squamous cell carcinoma than in tissues from patients with Bowen's disease. Basal cell carcinoma and malignant melanoma with histological evidence of invasiveness of the tumour cells showed a higher expression of the fos gene product than that seen in histologically circumscribed tumour nests. Thus, a higher expression of the fos oncogene is closely related to the malignant progression of tumour cells, in particular, the extent of invasiveness.
Effect of cholesterol on the position of segmental lesions in unilaterally nephrectomized rats
Authors: Green Nicholas J. , Howie Alexander J. , Rayner Hugh C. , Walls John
Published: June 2005

Different positions of segmental lesions within glomeruli may correspond to different pathogenetic mechanisms. The effect of a high cholesterol diet on the position of lesions had not previously been investigated. This was studied in rats following unilateral nephrectomy, as a change in position would suggest a different mechanism of damage. Thirty‐two female WAG/ola rats had unilateral nephrectomy. Half the rats were given a diet supplemented with 4 per cent cholesterol and 1 per cent cholic acid. At death, six at 10 weeks after nephrectomy and the rest at 24 weeks, kidney sections were examined microscopically. There were significantly more segmental lesions in the cholesterol‐fed rats than in the controls, and these lesions were almost entirely at the glomemerular hilum in both groups. Significantly more glomeruli contained foamy cells in the cholesterol‐fed group, both within lesions and away from them. These findings confirmed that in reduced renal mass, segmental lesions are mainly hilar. The diet increases the number of glomeruli affected by lesions, but these are still mainly hilar. Therefore one possibility is that hypercholesterolaemia worsens the hyperfiltration effect on glomeruli. The diet also produces foamy cells scattered throughout the glomeruli but these do not appear to develop into segmental lesions.
Epithelial (E‐) and placentae (P‐) cadherin cell adhesion molecule expression in breast carcinoma
Authors: Rasbridge S. A. , Gillett C. E. , Sampson S. A. , Walsh F. S. , Millis R. R.
Published: June 2005

Two members of the cadherin family of intercellular adhesion molecules are found in normal breast tissue: E‐ (epithelial) cadherin is present in both luminal and myoepithelial cells, whereas P‐ (placental) cadherin is confined to myoepithelium. There is experimental evidence that loss of E‐cadherin is associated with increased invasiveness of malignant cells in vitro, which stimulated us to examine the presence and distribution of E‐ and P‐cadherin in breast carcinomas by means of immunohistochemical staining.

E‐Cadherin was present in all in situ and invasive ductal carcinomas examined, although it had a patchy distribution and the staining was of variable intensity. However, in 83 per cent of invasive lobular carcinomas and all lobular carcinomas in situ there was complete loss of E‐cauherin expression. In the remaining 17 per cent of invasive lobular tumours, E‐cadherin appeared to have an abnormal distribution within the cytoplasm with variable expression on the cell membrane. P‐Cadherin, by contrast, was absent from all benign breast luminal epithelium and 25 carcinomas of ductal and lobular type. It was found in only one carcinoma of lobular type. We suggest that loss of cell‐cell adhesion mediated by E‐cadherin plays a part in the characteristic morphology of lobular carcinomas.
Expression of xenobiotic metabolizing enzymes in breast cancer
Authors: Murray Graeme I. , Weaver Richard J. , Paterson Pamela J. , Ewen Stanley W. B. , Melvin William T. , Danny M. Burke
Published: June 2005

We have studied the expression of different xenobiotic metabolizing enzymes in primary operable breast cancer of no special type. The expression of two forms of cytochrome P450, microsomal epoxide hydrolase, and three classes of glutathione S‐transferase was investigated using immunohistochemistry. The tumours were characterized by consistent expression of microsomal epoxide hydrolase and by variable expression of the two forms of cytochrome P450 and the three types of glutathione S‐transferase. Cytochrome P450 1A and cytochrome P450 3 A were identified in 39 and 22 per cent of tumours, respectively. In each case, immunostaining was present only in areas of invasive carcinoma. Epoxide hydrolase was identified in 89 per cent of tumours and glutathione S‐transferases pi, mu, and alpha were identified in 56, 65, and 44 per cent of tumours, respectively. Immunoreactivity for epoxide hydrolase and glutathione S‐transferases was identified in both tumours and non‐neoplastic breast tissue. The presence of different xenobiotic metabolizing enzymes may have a role in determining the intrinsic drug resistance of breast cancer to a variety of anti‐cancer drugs, and the expression of these enzymes can readily be assessed using immunohistochemistry.
Retinoblastoma (rb) gene product expression in lymphomas. Correlation with Ki67 growth fraction
Authors: Martínez Juan C. , Piris Miguel A. , Sánchez‐Beato Margarita , Villuendas Raquel , Orradre Juan L , Algara Patricia , Sánchez‐Verde Lidia , Martínez Pedro
Published: June 2005

The retinoblastoma susceptibility gene (Rb) has been characterized as a tumour suppressor gene. Rb protein is involved in cell‐cycle control, regulating gene transcription. The absence of Rb protein in inherited retinoblastoma has been proved to be the result of inactivation of both Rb alleles through mutation or deletion, according to the general model for suppressor genes. The frequent detection of Rb gene alterations in human tumours (retinoblastoma, osteosarcoma, bladder carcinoma, small‐cell lung carcinoma) and the correlation with clinical outcome found in some tumours prompted us to study Rb gene expression in lymphoid tumours in an attempt to determine whether Rb gene expression is related to histological type and degree of aggrcssivity in human lymphomas.

To establish normal levels of Rb protein, its expression was analysed in vitro on cytospin preparations from normal and pokeweed mitogen (PWM) or phytohaemagglutinin (PHA)‐stimulated peripheral blood lymphocytes (PBLs), using a monoclonal antibody (PMG3‐245). Rb protein expression in vivo was quantified using a computer analysis system (CAS) on frozen sections from reactive and neoplastic lymphoid tissue. As a control of tissue preservation, and to compare Rb expression and growth fraction, the tumours and cells were labelled simultaneously with the Ki67 monoclonal antibody.

Normal and stimulated lymphocytes showed a gradual increase of Rb protein during progression of the cell cycle, with a peak in the M phase. G0–G1 cells had no detectable levels of Rb protein, suggesting that the Rb gene may act as a ‘status quo’ cellular growth fraction control mechanism. In reactive lymphoid tissue, Rb protein was mainly expressed in germinal centres (lymph nodes, tonsils) and cortical thymocytes.

There was a statistically significant correlation between Ki67 and Rb expression in non‐Hodgkin's lymphomas (NHLs), most low‐grade lymphomas having very weak or undetectable levels of Rb protein. The majority of high‐grade lymphomas have higher Rb protein levels. However, a group of ten lymphomas with no Rb protein was found, in spite of a Ki67 range similar to that found in the Rb‐positive lymphomas. This lack of Rb expression in a subset of high‐grade NHLs suggests that Rb deregulation, through deletion, mutation, or lack of transcription, may play a role in this group of tumours. Further studies should focus on Rb genetic alterations.
Malignant potential of aneuploid pancreatic endocrine tumours
Authors: Lee C. S. , Charlton I. G. , Williams R. A. , Dhillon A. P. , Rode J.
Published: June 2005

Evaluation of the malignant potential of pancreatic endocrine tumours is difficult by histological criteria alone. Nuclear deoxyribonucleic acid (DNA) cytometric analysis can provide significant prognostic and biological information in a number of solid human tumours. Thus, the DNA profiles and nuclear parameters of 39 patients with pancreatic endocrine tumours diagnosed from 1969 to 1989 were studied, using computerized video image analysis. Of the 39 patients with tumours, 27 cases did not have evidence of metastatic disease at the time of investigation. The majority of these (N=21) showed a diploid profile. Five were aneuploid and one was tetraploid. In the 12 tumours that proved malignant, three had tissue of both the primary tumour and the metastases analysed; six had material only from the primary tumour, although metastatic disease was established clinically; and another three only had tissue from the secondary tumour studied. The majority of the malignant cases were aneupioid (N =7); three were dipioid; one showed a diploid primary but an aneuploid secondary; and one showed a hyperdiploid pattern. The percentage of nuclei with a DNA content greater than 5N (5CER) did not seem to contribute further to the assessment nor did the means of the nuclear areas and shape factors. It appears that ploidy studies may be useful in predicting malignant potential of pancreatic endocrine tumours. There is a high rate of immunostaining for insulin by benign pancreatic endocrine tumours and this may be a useful adjunct to distinguish them from their malignant counterparts. However, there is no statistically significant correlation between the expression of various hormones and the DNA ploidy of tumour cells.
Immunolocalization of heparinbinding growth factors (HBGF) types 1 and 2 in rat liver. Selective hyperexpression of HBGF‐2 in carbon tetrachloride‐induced fibrosis
Authors: Charlotte Frederic , Win Khin Maung , Preaux Anne‐Marie , Mavier Philippe , Dhumeaux Daniel , Zafrani Elie Serge , Rosenbaum Jean
Published: June 2005

Ito cells play a major role in liver fibrosis but the mechanisms controlling their activation in vivo are poorly understood. Heparin‐binding growth factors (H BGF) types 1 and 2 are mitogenic for cultured Ito cells. They have been found in liver extracts but their cellular localization is unknown. We have studied by immunohistochemistry KBGF‐1 and ‐2 expression in normal rat liver and in carbon tetrachloride (CCl4)‐induced fibrosis. In normal liver, HBGF‐1 was present only in sinusoidal cells whereas HBGF‐2 was also detected in endothelial cells lining major vessels. At the acute stage of CC14 intoxication, HBGF‐2 was expressed in centrilobular clusters of mononuclear phagocytes that were surrounded by many HBGF‐2‐negative Ito cells. In the later stages, HBGF‐2 was expressed by Ito cells within the fibrous bands. No modulation of HBGF‐1 expression was noted at any stage. These results suggest that (1) at the acute stage of CC14 intoxication, HBGF‐2 produced by mononuclear phagocytes could participate in the recruitment of Ito cells; and (2) during the CCl4‐induced fibrotic process, HBGF‐2 could contribute to Ito cell proliferation and the synthesis of fibrosis components. In this in vivo model of hepatic fibrosis, the hyperexpression of HBGF‐2 is a relatively specific event since the expression of a structurally related molecule, HBGF‐1 was not modulated.
Epstein‐Barr virus (EBV) in enteropathy‐associated T‐cell lymphoma (EATL)
Authors: Pan Langxing , Diss Tim C. , Peng Huaizheng , Lu Qilong , Wotherspoon Andrew C. , Thomas J. Alero , Isaacson Peter G.
Published: June 2005

Using Southern hybridization analysis, we have detected the Epstein‐Barr virus (EBV) genome in 36 per cent (4/11) of enteropathy‐associated T‐cell lymphoma (EATL), a frequency much higher than that seen in nodal T‐cell lymphomas in which we were able to show EBV DNA in only 3 per cent (1/30) of the cases examined. Using a terminal fragment probe, monoclonal proliferation of EBV in infected cells was demonstrated in three of the four EBV‐positive EATL cases (in one case, insufficient signal prevented the determination of EBV clonality). The EBV genome and an early transcript, EBER1, were identified in tumour cells by in situ hybridization. Expression of latent membrane protein (LMP) was detected in two EBV DNA/RNA‐positive EATL cases. In view of the known oncogenic properties of EBV and the putative central role of LMP in EBV‐induced cell immortalization, the results of this study suggest that the virus may play an aetiological role in the pathogenesis of EATL.
Participation of macrophages in segmental endocapillary proliferation preceding focal glomerular sclerosis
Authors: Saito Takao , Ootaka Tetsuya , Sato Hiroshi , Furuta Takashi , Sato Toshinobu , Soma Jun , Abe Keishi , Yoshinaga Kaoru
Published: June 2005

We studied infiltrating cells in the glomeruli of eight cases with focal segmental endocapillary proliferation (FSEP) using monoclonal antibodies to leukocyte common antigen, T cells, B cells, and monocytes/macrophages (Mo/Mφ). It was demonstrated by sequential biopsies performed in five cases that FSEP preceded focal glomerular sclerosis (FGS). Cell types in FSEP were compared with those in FGS from 17 patients with persistent nephrotic syndrome, ten non‐nephrotic patients, and eight patients with nephrotic syndrome which was initially responsive to steroid therapy but relapsed, as well as minimal change specimens from nine nephrotic patients. In the glomeruli, the mean total leukocyte counts increased significantly in the FSEP group (P < 0.01). The serial sections in FSEP revealed that Mo/Mφ were the predominant cells and were localized in areas of endocapillary proliferation. T‐cell or B‐cell infiltration was less marked. The extensive intracapillary distribution of p150,95 antigen belonging to the integrin family and acting as a C3bi receptor suggested that FSEP may be mediated by adhesion molecules expressed on Mo/Mφ. These findings indicate that Mo/Mφ may play a key role in FGS which shows endocapillary proliferation in the initial stage.
Study of renal tubular glycoconjugates in tubulointerstitial damage using conjugated lectins
Authors: Khan Tahir Naeem , Sinniah Raja
Published: June 2005

Changes in the glycoconjugates of renal tubules associated with tubular damage were studied by lectin histo‐chemistry, and their possible significance was determined. The excretion of various saccharides in tubular casts may also serve as markers of renal tubular damage. Renal tissues from 55 cases with various glomerular diseases including ten controls were studied. The patients were divided into two groups: one with tubulointerstitial lesions (TILs) (30 cases), and the other without (15 cases). Our results showed a wide spectrum of changes, predominantly in the disease group with TILs. The brush border of the proximal tubules showed significantly increased staining with Triticum vulgaris (WGA) and decreased staining with Canavalia ensiformis (Con A) lectins in both disease groups. The distal tubules revealed a significant increase in the apical and a decrease in the basal staining with Arachis hypogaea (PNA) and WGA lectins, respectively, in cases with TILs. The significant decrease in the basal domain staining was also seen with WGA lectin in cortical ducts. The vulnerability of various segments of the tubules in the process of TILs was clearly demonstrable. It appeared that the distal tubules were the most vulnerable anatomical segments around which the TILs began and later spread to involve other segments of tubules.
Expression of the C‐erbB‐3 protein in gastrointestinal tract tumours determined by monoclonal antibody RTJ1
Authors: Rajkumar Thangarajan , Goden Calvin S. R. , Lemoine Nicholas R. , Gullick William J.
Published: June 2005

We have examined the expression of the c‐erbB‐3 protein in a wide range of tumours of the gastrointestinal (GI) tract using immunocytochemical staining. Two antibodies were employed, a polyclonal antibody, 49.3, and a new monoclonal antibody, RTJ1, both raised to a synthetic peptide from the cytoplasmic domain of the human c‐erbB‐3 protein. The antibodies recognize c‐erbB‐3 by immunoprecipitation and Western blotting of lysate prepared from a cell line engineered to overexpress the protein. Both antibodies also detect expression of the protein in formalin‐fixed, paraffin‐embedded human tissues. The RTJ1 monoclonal antibody gave superior staining, showing lower background and more pronounced cell surface immunoreactivity. The c‐erbB‐3 protein was found in normal epithelial cells throughout the GI tract, in squamous epithelium of the oropharanyx and oesophagus, in the parietal cells of the stomach, and in the surface enterocytes of the small and large bowel. Seventy‐six tumours arising at these sites were examined for c‐erbB‐3 protein expression. Widely varying levels of expression were seen, from absent to intense staining with cell membrane accentuation.
Trabecular bone does not have a fractal structure on light microscopic examination
Authors: Cross Simon S. , Rogers Suzanne , Silcocks Paul B. , Cotton Dennis W. K.
Published: June 2005

The fractal dimension of the boundary of trabecular bone in 62 biopsies was measured on histological sections using a box‐counting method implemented on a microcomputer image analysis system. The calculated fractal dimension had a mean value of 0.99 with a normal distribution. Since this value is not greater than the topological dimension (1), trabecular bone, when examined by light microscopy, does not have a fractal structure. Conventional Euclidean dimensions will continue to be the most useful measurements in bone histomorphometry.
Tumour progression and metastatic behaviour in vivo correlates with integrin expression on melanocytic tumours
Authors: Schadendorf Dirk , Gawlik Christian , Haney Uwe , Ostmeier Hermann , Suter Ludwig , Czarnetzki Beate M.
Published: June 2005

In order to evaluate the significance of adhesion molecules expressed on melanocytic tumours for progression and prognosis in vivo, we studied integrin expression (VLA‐1 to VLA‐6, CD18, CD51, CD61) on 10 naevi, 40 primary malignant melanomas, and 11 metastases by immunohistology using the APAAP technique. Evaluation was done by grouping the percentage of positive tumour cells in six categories. Statistical analysis (Wilcoxon rank test, Scheffe test) revealed significant differences in the expression of VLA‐1 (P <0.0001), VLA‐2 (P=0.0001), VLA‐5 (P=0.0093), VLA‐6 (P=0.0232), and CD61 (P0.0002) between naevi and primary melanomas. Comparing primary melanomas with metastases, a statistically significant decrease in the expression of VLA‐1, VLA‐2, and VLA‐6 was detectable, as well as a significant increase in VLA‐4 and VLA‐5. There was no correlation between integrin expression and tumour type (superficial spreading melanoma, nodular melanoma, lentigo maligna melanoma), regression and ulceration. Changes of VLA‐1, VLA‐4, and VLA‐6 expression correlated with the tumour thickness of the primary melanoma, but only VLA‐4 and VLA‐6 expression on primary melanomas correlated significantly with the development of metastases (P=0.024 and P=0.001). These changes of integrin expression during tumour progression particularly, the data showing an increase of VLA‐4, and a decrease of VLA‐6 expression support the concept that integrins are a new additional set of prognostic markers which indicate predisposition to the development of metastases.
Patterns of integrin common chain β1 and collagen IV immunoreactivity in hepatocellular carcinoma. Correlations with tumour growth rate, grade and size
Authors: Patriarca Carlo , Roncalli Massimo , Gambacorta Marcello , Cominotti Marzia , Coggi Guido , Viale Giuseppe
Published: June 2005

Thirty cases of hepatocellular carcinoma (HCC) were investigated immunocytochemically for expression of β1 integrin molecule and of collagen IV. Immunoreactivity was related to the tumour proliferation index, as detected by PCNA immunostaining, and to tumour size and grade. Membrane β1 integrin immunoreactivity was deteccted in the neoplasticc cells of all cases, though two different staining patterns were clearly recognized. In 14 cases, β1 integrin immunoreactivity was confined to the cell‐stroma interface, showing the same polarized pattern as the non‐neoplastic cell counterpart. This staining pattern was associated significantly (P 0.0001) with low PCNA labelling (i.e. less than 20 per cent of neoplastic cells showing nuclear immunostaining. Conversely, 16 cases showed non‐polarized pericellular β1 integrin immunostaining. This staining pattern was significantly associated (P < 0.0001)) with high PCNA labelling (more than 20 per cent of immunoreactive cells) and with tumour size greater than 4 cm in diameter (P < 0.0001). β1 Integrin, collagen IV, and PCNA immunoreactivities, however, did not correlate with the histological grade. The data emphasize that neoplastic progression of HCCs may be correlated with an aberrant expression of adhesion molecules and with a disruption of the collagen IV complement of basal membranes.
β1 and β4 integrin expression in methacarn and formalin‐fixed material from in situ ductal carcinoma of the breast
Authors: Hanby Andrew M. , Gillett Cheryl E. , Pignatelli Massimo , Stamp Gordon W. H.
Published: June 2005

The integrins are αβ heterodimeric transmembrane proteins mediating cell‐substratum as well as cell‐cell interactions. Previous distribution studies on integrin expression have been limited by the requirement for cryostat sectioned tissues, and consequent poor resolution. We have examined 40 examples of ductal carcinoma in situ (DCIS) for the expression of both β1 and β4 integrin chains. These showed strong polarized membrane staining of residual myoepithelial cells (correlating with expression of smooth muscle specific actin) and of the basement membrane region with β1 and β4 antibodies respectively. In 12 out of 40 cases, the DCIS was negative for the β1 chain and a variable pattern of reactivity was seen in the remaining cases. The β4 chain was detected focally and weakly in the tumour cells of 7/40 DCIS and strongly in one; all of these cases were also positive for the β1 chain. Of the 22 cases where co‐existent invasion was present, the infiltrating component showed either a similar degree or a diminution of the extent of immunostaining when compared with the in situ component; only one showed enhanced staining (β1 only). This study demonstrates that two of the main β chains, β1 and β4, can be effectively demonstrated on methacarn and cold (4°C) formalin‐fixed tissues by avidin‐biotin indirect immunoperoxidase staining and that the results are similar to those achieved using frozen tissue.
Induction of nitric oxide synthase in the neo‐vasculature of experimental tumours in mice
Authors: Buttery L. D. K. , Springall D. R. , Andrade S. P. , Riveros‐Moreno V. , Hart I. , Piper P. J. , Polak J. M.
Published: June 2005

Maintenance of blood flow is an important factor in sustaining tumour growth. Functional studies have previously demonstrated a reduction in tumour blood flow with selective inhibitors of nitric oxide (NO) synthesis, L‐NAME (NG‐nitro‐L‐arginine‐methylester) and L‐NMMA (NG‐monomethyl‐L‐arginine), when administered locally to tumours derived from murine colon 26 adenocarcinoma and B16 melanoma cells. The type of NO synthase which might be responsible for this locally‐derived NO and the site of synthesis was not described. Here we have investigated the distribution of immunoreactivity and the biochemical characteristics of the enzymes synthesizing NO in the same murine model. Adenocarcinoma (colon 26) or melanoma (B16) cells were introduced into a sponge matrix implanted subcutaneously in mice. After 7, 12, and 14 days, the implants were removed and frozen sections were immunostained with rabbit antisera to constitutive and inducible isoforms of NO synthase. Immunoreactivity with antisera to inducible NO synthase was detected in the vasculature of neoplastic implants, with and without the sponge, at 12 and 14 days. The enzyme was not evident in 7‐day‐old tumours, in non‐neoplastic implants, in areas of tissue outside the tumour, or in adenocarcinoma or melanoma cells. Enzyme activity was measurable in homogenates of neoplastic implants removed at day 7 and was found to be Ca2+/calmodulin‐independent. Immunoreactivity with antisera to inducible NO synthase was seen principally in the endothelium of newly‐formed capillaries, identified by immunostaining for von Willebrand factor in serial sections. Immunoreactivity with antiserum to constitutive NO synthase was not evident in either neoplastic or non‐neoplastic implants. These results suggest that neoplasms can modulate the synthesis of NO in their vasculature by inducing the generation of NO synthase and hence the production of NO which could be very important in maintaining the tumour blood flow.
Distribution of the extracellular matrix components in human glomerular lesions
Authors: Büyükbabani Nesimi , Droz Dominique
Published: June 2005

Most glomerular pathologies are associated with alterations of the matrix compartment. Using reagents directed against the α/α2 and α3 chains of type IV collagen [α1/α2(IV), α3(IV)], laminin, heparan sulphate proteoglycan (HPG), fibronectin, collagen I, and collagen III, we investigated the modifications of the glomerular matrix components in several human glomerular lesions compared with normal kidney. In type I membranous glomerulo‐nephritis (MGN) (nine cases), we did not observe alterations in the matrix component distribution. In MGN types II and III (five cases), the spikes and chainettes were made of the α3(IV) chain, laminin, and HPG, while the α1/α2(IV) chains were localized along the subendothelial side of the glomerular basement membrane (GBM). In focal and segmental glomerulosclerosis (six cases), fibronectin, α1/α2(IV) chains, laminin, and small amounts of interstitial collagens were detected within the collapsed capillary loops; the newly formed matrix material between the podocytes and the GBM contained the α1/α2(IV) chains, laminin, and HPG but not the α3(IV) chain. In crescentic glomerulo‐nephritis (six cases), fibronectin was the most abundant and, in purely cellular crescents, the unique component. A basement membrane‐like network containing laminin, HPG, α1/α2(IV) chains, and interstitial collagens developed in a second step between the crescent cells. Interstitial collagens were present in the crescent framework, even when the integrity of Bowman's capsule was preserved. In membranoproliferative glomerulonephritis (five cases), we observed strong accumulation of fibronectin in the thickened mesangial spaces together with accumulation of laminin, α1/α2(IV) chains, and HPG; type I collagen was also present in the central part of the mesangial areas. This study shows that each glomerular lesion is characterized by particular alterations of the matrix components.
Proliferation in malignant mesothelioma as determined by mitosis counts and immunoreactivity for proliferating cell nuclear antigen (PCNA)
Authors: Ramael Marc , Jacobs Werner , Weyler Joost , van Meerbeeck Jan , Białasiewicz Pjotr , van den Bossche Jan , Buysse Corinne , Vermeire Paul , Van Marck Eric
Published: June 2005

In order to assess its discriminating and prognostic value, we studied immunoreactivity for proliferating nuclear cell antigen (PCNA) in human malignant mesothelioma (31 cases) and in human non‐neoplastic mesothelium (33 cases with reactive mesothelium and 20 cases of normal mesothellum) using the murine monoclonal antibody PC 10. We also compared it with mitosis counts expressed as the mitotic volume index (MV index). There were differences between malignant mesothelioma, reactive mesothelium, and normal mesothelium for percentage of PCNA immunoreactive cells (mean ± SD; 27 ± 9, 9·5 ± 5·1, and 3·6 ± 1·6, respectively) and for their MV index (20·3 ± 4·5, 9·4 ± 2·1, and 3·6 ± 0·6, respectively). The median actuarial survival was 10·1 months for patients with less than 25 per cent PCNA immunoreactive cells, 9·4 months for patients with less than 20 mitoses per mm2 of tumoural tissue, 5·9 months for patients with more than 25 per cent PCNA immunoreactive cells, and 5·3 months for patients with more than 20 mitoses per mm2 of tumoural tissue. Our results suggest that PCNA immunoreactivity is useful in discriminating between neoplastic and non‐neoplastic mesothelium and that it may have prognostic value in malignant mesothelioma.
Expression of the ret proto‐oncogene product in human normal and neoplastic tissues of neural crest origin
Authors: Nakamura Takuro , Ishizaka Yukihito , Nagao Minako , Hara Mitsuru , Ishikawa Takatoshi
Published: June 2005

The histological localization of the ret proto‐oncogene (proto‐ret) product was examined in neural crest‐derived and neuronal tissues together with their neoplastic counterparts by immunohistochemistry using a polyclonal antibody. Schwann cells, neurons, sympathetic ganglia, and cells of the adrenal medulla were positive for the proto‐ret product, whereas melanocytes were negative. Positive results were obtained from neural crest‐derived tumours such as schwannoma (69 per cent, 11/16), neurofibroma (59 per cent, 13/22), neuroblastoma (80 per cent, 4/5), phaeochromocytoma (100 per cent, 3/3) and medullary thyroid carcinoma (100 per cent, 3/3). The antibody reacted with all of the 22 astrocytomas examined. With negative proto‐ret expression in melanocytic tumours, proto‐ret expression was considered to correlate with the differentiation of some lineages of neural crest‐derived cells.
Microvascular lesions in reflux oesophagitis and Barrett's oesophagus
Authors: Sbarbati Andrea , Ricci Francesco , Inaspettato Giorgio , Lazzara Giuseppe , Cordiano Claudio , Osculati Francesco
Published: June 2005

In 30 patients with reflux oesophagitis, the mucosal microvessels have been studied by transmission electron microscopy. The aim of the study was to test the hypothesis that a microvascular injury is involved in reflux oesophagitis, and to clarify if the epithelial metaplasia is correlated with a contemporaneous modification of the microvasculature in Barrett's oesophagus. In squamous epithelium‐lined mucosa, signs of microangiopathy were found in all patients and capillaries regularly showed an interrupted, duplicated, or thickened basal lamina. In areas of columnar metaplasia, capillaries showed an ectatic lumen and a thin basal lamina without duplications or interruptions; endothelial cells had a thin rim of cytoplasm with many fenestrations. These findings demonstrate that microangiopathy is associated with epithelial damage in reflux oesophagitis.
Integrin expression in squamous neoplasia of the cervix
Authors: Hughes D. E. , Rebello G. , Al‐Nafussi A.
Published: June 2005

Epithelial cell‐basement membrane interactions are important in maintaining tissue architecture and function, and the anatomical and functional relationships between epithelial cells and their basement membranes are clearly altered in malignancy. These interactions are thought to be largely mediated by the integrins, a family of heterodimeric transmembrane glycoproteins, each consisting of an α and a β chain. Epithelial integrins mainly belong to the β1 (VLA) subfamily, which forms receptors for matrix macromolecules such as fibronectin, laminin, and collagen. There is evidence that integrin expression changes in some epithelial malignancies, possibly in relation to invasive potential. Integrin expression in cervical neoplasia was studied by immunohistochemical examination of prospectively collected colposcopic biopsies. Well‐characterized monoclonal antibodies against β1–4, αl–6, and αV integrins were used to examine normal, koilocytic, and dysplastic cervical squamous epithelium, and invasive squamous carcinoma. β1, β4, α2, α3, α6, and αV were expressed by the basal layer of normal cervical squamous epithelium and by dysplastic cells in CIN (cervical intraepithelial neoplasia) 1 and 2, with none being lost and no new chains acquired. In CIN3, these integrins were either expressed throughout the ectocervical epithelium or restricted to the basal layer. In the latter cases, integrin expression was retained to a greater degree by dysplastic squamous epithelium within endocervical glands. These patterns could not be correlated with age or smear history in the cases examined. Patterns of integrin expression in neoplastic cervical epithelium therefore differ from those of normal cervical epithelium. It is possible that these changes may be related to changes in cellular function occurring during neoplastic progression.
Nuclear quantitative grading by discriminant analysis of renal cell carcinoma samples. A patient survival evaluation
Authors: Artacho‐Pérula Emilio , Roldán‐Villalobos Rafael , Martínez‐Cuevas Juan F. , López‐Rubio Fernando
Published: June 2005

Specimens from 60 cases of renal cell carcinoma (RCC) were graded employing quantitative nuclear data combined with multivariate discriminant analysis. Evaluation of patient survival was analysed with respect to quantitative microscopic and qualitative features. Both morphometric and stereological estimators were used to establish the nuclear size and form pattern of the RCC specimens. Tumoural dedifferentiation paralleled progressive increases in nuclear elongation and in two‐ and, especially, three‐dimensional—mean nuclear volume (MNV)—size parameters. Using stepwise discriminant analysis, 85‐0 per cent of the specimens were correctly classified when differentiating grade 2 and 3 tumours. It is concluded that simple and realistic estimates of MNV are the best discriminator for objective grading in patients with RCC. Univariate survival analysis demonstrated the important significance of several features such as MNV, clinical stage, and nuclear discriminant and histopathological tumour grades. Nuclear form factor PE, area, and perimeter were also significant. A prognosis study based on the Cox model using a stepwise selection of parameters showed that only MNV has an independent prognostic role when examining all investigated quantitative parameters. The clinical stage was the best prognostic feature when all quantitative and qualitative characteristics were included in the analysis.
Transformation from SAA2‐fibrils to AA‐fibrils in amyloid fibrillogenesis: In vivo observations in murine spleen using anti‐SAA and anti‐AA antibodies
Authors: Arai Kazumori , Miura Katsutoshi , Baba Satoshi , Shirasawa Haruyuki
Published: June 2005

Early amyloid fibrillogenesis from serum amyloid A protein (SAA) has been observed in the murine spleen after an injection of casein‐Freund's complete adjuvant in the presence of amyloid enhancing factor, using anti‐SAA C‐terminal (anti‐SAA) and anti‐amyloid A (AA) antibodies. In Western immunoblotting of sera, both SAA1 and SAA2 reached a maximum after 24 h and began to decrease after 48 h. In spleen extracts, SAA2, but not SAA1 or AA, was found from 48 h, when amyloid was first deposited in the marginal zone. Electron microscopic immunohistochemistry of this stage showed reaction products from SAA in the marginal zone as fine granules along the cell membrane of mononuclear cells and focal intercellular aggregates, which contained fine fibrils originating from the cell membrane. Amyloid nodules, surrounded by mononuclear cells, developed from this stage. In the nodules, fibrils were positive for anti‐SAA only in the vicinity of the cell membrane, while anti‐AA stained fibrils throughout. Our hypothesis for fibrillogenesis is thus as follows: Serum SAA2 is specifically deposited on mononuclear cells in the marginal zone and polymerized extracellularly into fibrils, retaining its antigenicity (SAA2 amyloid fibrils); these fibrils are then processed to AA amyloid fibrils in situ by cleavage of the C‐terminal portion of SAA2.
Apoptosis in bladder cancer as related to standard prognostic factors and prognosis
Authors: Lipponen P. K. , Aaltomaa S.
Published: June 2005

Four hundred cases of transitional cell bladder cancer were reviewed by light microscopy for the presence of apoptotic cells in the primary tumour biopsy specimens. The number of apoptotic cells/mm2of neoplastic epithelium (apoptotic index, AI) was related to various histological features and prognosis. AI was related significantly to high T‐category, high grade, DNA aneuploidy, large nuclear morphometric variable values, and high proliferation rate of cancer cells. Tumours showing overexpression of p53 oncoprotein in over 10 per cent of the nuclei had significantly higher AI values than p53‐negative tumours. Multivariate regression analysis showed that AI was independently predicted by mitotic index, mean nuclear area, and papillary status. Progression, recurrence‐free survival, and survival of superficial tumours were all related significantly to AI. In multivariate survival analysis, T‐category, papillary status, grade, and mitotic index had independent prognostic value, while the recurrence‐free survival of Ta‐T1 tumours was related independently to AI. The results show that AI is related particularly to mitotic activity in transitional cell bladder tumours, while AI as assessed by light microscopy hardly has any independent prognostic significance.
Immunolocalization of serum amyloid A and AA amyloid in lysosomes in murine monocytoid cells: Confocal and immunogold electron microscopic studies
Authors: Chronopoulos S. , Laird D. W. , Ali‐Khan Z.
Published: June 2005

Murine AA amyloid (AA) protein represents the amino‐terminal two‐third portion of SAA2, one of the isoforms of serum amyloid A. Whether plasma membrane‐bound or lysosomal enzymes in activated murine monocytoid cells degrade SAA2 to generate amyloidogenic AA‐like peptides is not clearly understood, although AA has been localized in the lysosomes. Here we show, using confocal and immunogold microscopy (IEM), that both SAA and AA localize in lysosomes of activated monocytoid cells from amyloidotic mice. Rabbit anti‐mouse AA IgG (RAA) and two monoclonal antibodies against murine lysosome‐associated membrane proteins (LAMP‐1 and LAMP‐2) were used to immunolocalize SAA/AA and lysosomes, respectively. Confocal analysis co‐localized both anti‐RAA and anti‐LAMP‐1/LAMP‐2 reactivities in the perikaryal organelles which by IEM proved to be electron‐dense lysosomes. LAMP‐1/LAMP‐2‐specific gold particles were also localized on lysosomal and perikaryal AA. The results suggest sequestration of SAA into the lysosomes. Since monocytoid cells are not known to phagocytose native amyloid fibrils, our results implicate lysosomes in AA formation.
CD4‐positive cells from patients with IgA nephropathy demonstrate increased mRNA of cytokines that induce the IgA switch and differentiation
Authors: Lai Kar Neng , Lui Siu Fai , Li Philip K. T. , Ho Rainbow T. H. , Leung Joseph C. K. , Chui Y. L. , Lim P. L.
Published: June 2005

IgA nephropathy (IgAN) is characterized by raised serum IgA1 deposits. We have previously shown increased T‐cell activation in IgAN. Recently, transforming growth factor‐β (TGF‐β) has been shown to induce IgA isotype switch at a clonal level and interleukin 5 (IL5) promotes differentiation into IgA‐bearing B cells. In the present study we have examined the TGF‐β and IL5 mRNA expression by mitogen‐activated CD4‐positive T cells from patients with IgAN (n=25), patients with other primary nephritides (CGN) (n=24), and healthy control subjects (n=25). The cytokine genes were analysed by reverse transcription (RT)‐polymerase chain reaction (PCR) and were semi‐quantitated by normalizing the differences occurring during RT and PCR using a housekeeping gene, β‐actin. CD4‐positive T cells from IgA nephritic patients expressed a higher level of IL5 mRNA than healthy controls (P<0.01) and patients with CGN (P<0.005). CD4‐positive T cells from IgA nephritic patients expressed a higher level of TGF‐β mRNA than healthy controls (P<0.01) but no difference was demonstrated on comparison with CGN patients. Elevated TGF‐β mRNA expression in patients with CGN probably reflects its other important function as a (sclerogenic) factor involved in the glomerulosclerosis found in these nephritides. Our data suggest that there is increased expression of cytokine genes which induce the IgA isotype switch and differentiation; these immunological abnormalities may be important in the pathogenesis of IgAN.
The expression of alpha B‐crystallin in epithelial tumours: A useful tumour marker?
Authors: Pinder S. E. , Balsitis M. , Ellis I. O. , Landon M. , Mayer R. J. , Lowe J.
Published: June 2005

Alpha B‐crystallin is a lens protein showing homology with small heat‐shock proteins. We have previously demonstrated its expression in non‐lenticular normal and diseased human tissues by immunostaining with a polyclonal antibody. In view of the expression seen in normal renal tubular epithelium, we have assessed the immunoreactivity of a variety of epithelial tumours, to determine the usefulness of alpha B‐crystallin as a specific renal tumour marker. Carcinomas arising from tissues which normally express alpha B‐crystallin, such as colo‐rectal and thyroid carcinomas, showed a varying pattern and degree of immunoreactivity. The most consistently positive tumours, however, with typically strong cytoplasmic and cell membrane staining, were renal cell carcinomas, 90 per cent of which showed positive immunoreactivity. This pattern of staining, while not absolutely specific, is a useful aid to the diagnosis of renal carcinoma. When a metastic deposit or a small biopsy is being assessed, anti‐alpha B‐crystallin may be included in a panel of antibodies, the pattern of staining of which may direct the search for the primary site of the tumour.