bluuebunny/crossref_metadata_2025_split huggingface dataset:
Ultrastructure of rat pulmonary arterioles after neonatal exposure to hypoxia and subsequent relief and treatment with monocrotaline
Authors: King Andrew P. , Smith Paul , Heath Donald 
Published: June 2005

AbstractA group of rats was born in and spent the first 4 weeks of life at a simulated altitude of 3550 m. Two animals were killed immediately afterwards and the remaining 16 were allowed to recover for various times up to a maximum of 12 weeks at sea‐level atmospheric pressure. On ultrastructural examination, the pulmonary arterioles of hypoxic rats showed muscularization, the new layer of mature smooth muscle cells containing abundant organelles and myofilaments. These cells were bounded by prominent elastic laminae. During the recovery period, the medial layer became progressively thinned, but the cells still retained some characteristics of smooth muscle by 12 weeks' recovery. When a similar group of ten hypoxic rats was allowed to recover for 12 weeks before being given monocrotaline, there was early enlargement of the residual smooth muscle cells in the media of pulmonary arterioles and within 5 weeks there was again a thick layer of medial smooth muscle. This was in contrast to the sparse, weakly muscularized arterioles seen in eight similarly treated rats born under normoxic conditions. The relevance is discussed of these findings to the rare occurrence of primary pulmonary hypertension in people who were born at high altitude but returned to sea‐level during childhood.
Fractal patterns for cells in culture
Authors: Vilela Marcelo J. , Martins Marcelo L. , Boschetti Sandro R. 
Published: June 2005

AbstractThe present study aims to understand the growth of human malignant tumours in vitro, using the geometry of fractals as a method of analysis. The fractal dimensions of HN‐5 and MDCK cell growth patterns have been measured. The first results may suggest the possibility of distinct growth processes characterized by different (time‐dependent) effective fractal dimensions for MDCK and HN‐5 cells. If this is true, the fractal dimension may yet prove to be a useful discriminant for comparing different diagnostic categories.
Non‐Hodgkin's lymphoma, coeliac disease, and Epstein–Barr virus: A study of 13 cases of enteropathy‐associated T‐ and B‐cell lymphoma
Authors: Ilyas M. , Niedobitek G. , Agathanggelou A. , Barry R. E. , Read A. E. , Tierney R. , Young L. S. , Rooney N. 
Published: June 2005

AbstractA group of 166 patients with coeliac disease was followed for a period of up to 25 years. During this time, 17 patients developed intestinal tumours that were diagnosed as lymphoma, of which 15 cases were available for review. Eleven of the lymphomas were of T‐cell type (enteropathy‐associated T‐cell lymphoma, EATL) and two were of B‐cell type. Two cases were reclassified as undifferentiated carcinoma. The interval between the diagnosis of enteropathy and the onset of lymphoma varied from less than 2 months in four patients to more than 5 years in seven. Seven of the T‐cell and both B‐cell lymphomas were investigated for the presence of Epstein–Barr virus (EBV) by in situ hybridization (ISH) using probes against Epstein–Barr virus‐encoded RNAs (EBERs) and by immuno‐histochemistry with EBV‐specific monoclonal antibodies. All EATL cases were negative, suggesting that EBV is not an important factor in these cases. In one of the B‐cell cases, EBV was detectable by ISH and immunohistochemistry in most tumour cells in the mesenteric lymph nodes, but not in any of the tumour cells in the primary ileal tumour, indicating that in this case EBV infection was a late event in the neoplastic process. These results show that lymphoma may develop any time after the onset of coeliac disease and that in our cases of EATL, EBV was not an important factor. In some cases of EBV‐related neoplasia, virus infection may be a late event.
Determination of transforming growth factor beta1 mRNA expression in breast carcinomas by in situ hybridization
Authors: Walker Rosemary A. , Gallacher Barbara 
Published: June 2005

AbstractThe expression of transforming growth factor beta1 (TGF‐β1) mRNA has been determined in 16 breast carcinomas using in situ hybridization and compared with TGF‐β protein as detected by antibodies against TGF‐βT1 and TGF‐β1 plus TGF‐β2. Digoxigenin‐labelled riboprobes were used, with alkaline phosphatase and immunogold silver detection systems. TGF‐β1 mRNA was only detected in carcinomas in which TGF‐β1 protein was found (9 of 16 cases) and not in those with prominent reactivity for TGF‐β2. RNA preservation was poor in two other cases in which TGF‐β1 protein had been detected. In general, those tumours with greater numbers of cells labelled for TGF‐β1 mRNA had prominent reactivity for TGF‐β1 protein. The mRNA was localized to cancer cells with no labelling of stromal cells, although in a small number of cases scanty staining for TGF‐β1 protein had been observed in stromal cells. The incidence of detection of TGF‐β1 mRNA is lower than the published data from Northern analysis studies of breast carcinomas, suggesting that only higher levels of TGF‐β1 mRNA expression are being detected by in situ hybridization. However, this approach has provided useful information about the cellular sites of expression of TGF‐β1 in breast carcinomas.
The intracellular polymerase chain reaction for small CMV genomic sequences within heavily infected cellular sections
Authors: Ray R. A. , Smith M. , Sim R. , Nystrom M. , Pounder R. E. , Wakefield A. J. 
Published: June 2005

AbstractThe indirect intracellular polymerase chain reaction (in situ PCR) combines the potential sensitivity of the polymerase chain reaction (PCR) with the high specificity and morphological preservation of in situ hybridization (ISH). This study describes a method for the amplification of small, specific cytomegalovirus (CMV) genomic sequences [100 base pairs (bp)] within large formalin‐fixed, paraffin‐embedded tissue sections. A heat‐resistant glue surrounds the section, creating a well which contains a relatively large volume of isotonic reaction solution without a covering mineral oil layer; this optimizes morphological preservation, permits the evaluation of large sections, and allows both denaturation steps and up to 40 cycles of in situ PCR to be performed, whilst progressively concentrating the reaction solution by evaporation during thermal cycling. ISH was performed using a non‐isotopic DNA probe with specificity for CMV, with or without prior in situ PCR amplification, both for samples on slides (fibroblasts and lung) and in suspension (fibroblasts). Samples on slides were evaluated by both blinded studies and image analysis, comparing the intensity of signal (P<0·003) and the numbers of positive cells detected (P<0·007), with or without intracellular amplification. Cells in suspension were analysed by blinded studies on cytospins and by gel electrophoresis of cell lysates. Successful intracellular amplification was achieved in this high copy model.
A megakaryocyte analysis of the bone marrow in patients with myelodysplastic syndrome, myeloproliferative disorder and allied disorders
Authors: Ohshima Koichi , Kikuchi Masahiro , Takeshita Morishige 
Published: June 2005

AbstractA morphometric analysis was performed on aspirate clots of bone marrow to identify the presence of atypical megakaryocytes after immunohistological staining with a monoclonal antibody against Factor VIII. This study included cases of myelodysplastic syndrome (MDS), myeloproliferative disorder (MPD), aplastic anaemia (AA), idiopathic thrombocytopenic purpura (ITP), chronic myelogenous leukaemia (CML), and control cases free from any haematological disease. Quantitative and qualitative abnormalities of megakaryocytes were assessed using an image analyser and a personal computer to perform a morphometric analysis of the number (per mm2), arrangement (μm), nuclear size (μm2), cell size (μm2), nuclear size/cell size (N/C) ratio, and nuclear contour index (NCI). Micromononuclear megakaryocytes were detected in MDS, while in MPD, large over‐mature magakaryocytes were observed to increase in number. In AA, the megakaryocytes decreased dramatically in number without showing any morphological abnormality. In CML, the megakaryocytes also increased without any remarkable morphological abnormality. In ITP, only the NCI increased. The above results show that MDS is characterized by the presence of atypical micromegakaryocytes, while MPD is characterized by atypical large over‐mature megakaryocytes.
Pathogenesis of segmental glomerular changes at the tubular origin, as in the glomerular tip lesion
Authors: Howie A. J. , Lee S. J. , Sparke J. 
Published: June 2005

AbstractSegmental abnormalities at the glomerulo‐tubular junction, or tip changes, are a common and early feature in several experimental and human renal disorders. Scanning and transmission electron microscopic study of an experimental model, Lewis rats given anti‐glomerular basement membrane antibodies, showed that at 5 days there were monocytes/macrophages in glomerular capillary loops next to the tubular origin, with abnormalities of visceral epithelium, including prolapse of groups of these cells into the tubular origin. At 12 days, there was contact and adhesion between glomerular capillary loops and Bowman's capsule. Transmission electron microscopic study of human renal biopsies showed that the human tip changes resembled the later lesions seen in rats, with intracapillary foam cells confirmed by immunohistological study to be monocytes/macrophages. These findings show that abnormalities of the glomerular tuft precede adhesion to Bowman's capsule and that the earliest changes recognized by light microscopy in human renal biopsies are relatively late in their natural history, meaning at least several days old. Prolapse of visceral epithelial cells into the tubular origin may explain the characteristic site of tip changes.
Expression of p53 protein has no independent prognostic value in breast cancer
Authors: Pietiläinen Timo , Lipponen Pertti , Aaltomaa Sirpa , Eskelinen Matti , Kosma Veli‐Matti , Syrjänen Kari 
Published: June 2005

AbstractA series of 392 female breast carcinomas was analysed immunohistochemically for expression of p53 protein with special emphasis on the role of p53 as an independent prognostic factor. Altogether, 54·8 per cent of the carcinomas expressed p53 protein, with the mean [standard error (SE)] fraction of positive nuclei being 17·1 per cent (1·2 per cent). Expression of p53 protein was independent of tumour metastasis at diagnosis, axillary lymph node status, tumour diameter, histological type, tubule formation, proportion of intraductal growth, margin formation, necrosis, DNA ploidy, and S‐phase fraction. A high fraction of p53‐positive nuclei was significantly related to patient age under 70 years, high grade, severe nuclear pleomorphism, dense infiltration of tumour by lymphocytes, high mitotic index, and high apoptotic index (for all,P<0·05). Impaired survival probability in the entire cohort (P=0·05) and in the axillary lymph node‐positive (ANP) tumours (P=0·015) was associated with a fraction of p53‐positive nuclei less than 25 per cent, while in the axillary lymph node‐negative (ANN) tumours, expression of p53 had no prognostic value. In multivariate analysis, independent prognostic predictors included axillary lymph node status, tumour diameter, and mitotic index. In the ANN tumours, tumour diameter, fraction of p53‐positive nuclei, and tumour grade were independent prognostic factors, whereas in the ANP tumours, diameter and mitotic index were the two independent prognostic factors. The results suggest that abnormal expression of p53 protein is only a weak independent prognostic factor in female breast cancer.
In situ detection of Epstein–Barr virus in non‐small cell lung carcinomas
Authors: Wong Maria P. , Chung L. P. , Yuen S. T. , Leung S. Y. , Chan S. Y. , Wang Elaine , Fu K. H. 
Published: June 2005

AbstractEpstein–Barr virus (EBV) is strongly associated with nasopharyngeal carcinoma and lymphoepithelioma‐like carcinomas (LELC) of foregut‐derived organs. Recently this group of EBV‐associated carcinomas has been expanded by the identification of the virus in conventional adenocarcinomas of the stomach. In situ hybridization (ISH) using a sensitive digoxigenin‐labelled EBER RNA probe was performed on 167 consecutive unselected primary non‐small cell lung carcinomas, to determine the frequency of EBV association in these tumours. Nine cases (5·4 per cent) showed strong EBER signals in the tumour cell nuclei. By immunohistochemistry, four of the EBER‐positive tumours showed patchy expression of the viral latent membrane protein (LMP‐1) and none showed any expression of the EBV nuclear antigen 2 (EBNA2). Morphologically, all the positive tumours were LELC, whereas no conventional type of non‐small cell lung carcinoma showed EBV association. The LELC presented a morphological spectrum from undifferentiated to squamoid or glandular differentiation. The patients showed a male to female ratio of 8:1. The mean age at presentation was 48 years. Smoking was not a risk factor. All patients were alive at follow‐up periods of 23–52 months. Southern blot analysis performed on eight of the nine positive tumours showed a clonal episomal form of EBV, suggesting the clonal expansion of an infected tumour cell early in oncogenesis. These characteristics of the EBV‐associated lung tumours justify their consideration as a distinct clinicopathological entity.
Expression of bcl‐2 oncoprotein in renal cell tumours
Authors: Paraf François , Gogusev Jean , Chrétien Yves , Droz Dominique 
Published: June 2005

AbstractExpression of bcl‐2 is associated with inhibition of apoptosis and extension of cell survival. The importance of apoptosis in relation to the development and progression of renal cell neoplasia remains undefined so far. In order to determine the expression of bcl‐2 oncoprotein in normal and neoplastic renal cells, 37 renal tumours were investigated by immunolabelling, including 13 clear cell carcinomas, ten tubulopapillary carcinomas, four chromophobic renal cell carcinomas, and ten oncocytomas. Twenty‐six samples of adjacent normal renal tissue served as controls. bcl‐2 expression was correlated with cell proliferation activity as estimated by Ki67 antigen expression, and p53 protein expression in the tumour samples. The results demonstrate that in the normal kidney, positive bcl‐2 immunostaining was present in glomerular parietal epithelial cells, in distal tubular cells, and in sparse proximal tubule cells. Renal cell tumours showed heterogeneous bcl‐2 expression according to the tumour cell type. While the majority of carcinomas of clear cell type were usually negative or contained sparsely distributed positive cells, all tubulopapillary carcinomas were consistently positive for bcl‐2. In oncocytomas and chromophobic carcinomas, there was a low percentage of bcl‐2 immunoreactive tumour cells; some nuclear bcl‐2 positivity was detected in one chromophobic tumour. These findings indicate variable bcl‐2 oncoprotein expression in different types of renal cell tumours, with the highest level of expression in tubulopapillary carcinomas. No clear relationship was found between nuclear grade, cell proliferation activity, and level of bcl‐2 expression. p53 protein was detected in only one tubulopapillary carcinoma.
Expression of vascular endothelial growth factor in renal vascular disease and renal allografts
Authors: Gröne Hermann‐Josef , Simon Mathias , Gröne Elisabeth F. 
Published: June 2005

AbstractVascular endothelial growth factor (VEGF) is a dimeric glycoprotein that exerts a proliferative effect specifically on endothelial cells. VEGF can increase vascular permeability and collagenase activity, is chemotactic for monocytes, and may dilate blood vessels. It can be induced by phorbol ester and cAMP in both mesenchymal and epithelial cells. In vitro cell culture experiments suggest that VEGF is upregulated by oxygen deprivation. In this study we tested whether in vivo acute and/or chronic reduction of renal blood flow by vascular obstruction would result in increased expression of VEGF mRNA and protein. Three normal kidneys, five human kidneys with narrowing of preglomerular vessels by vascular rejection or by vasculitis, and eight kidneys with nephrosclerosis and/or diabetic nephropathy were examined. In situ hybridization with 35S‐labelled riboprobes showed a pronounced expression of VEGF mRNA in acutely hypoxic proximal and distal tubules of both the cortex and medulla; VEGF protein was demonstrated in the epithelia of these tubules by immunohistochemistry. In kidneys with chronically reduced blood flow, the majority of atrophic tubules were negative for VEGF mRNA and protein, although interstitial cells expressed VEGF mRNA. In arcuate arteries showing intimal and adventitial fibrosis, some medial smooth muscle cells were positive for VEGF mRNA. In glomeruli with segmental sclerosis, viable podocytes showed a prominent signal for VEGF mRNA. Mesangial cells did not express VEGF in the cases studied. It is possible that hypoxia itself led to the upregulation of VEGF in tubular epithelia and vascular smooth muscle cells. The vasodilatory and permeability‐promoting effects of the endothelial growth factor produced by damaged tubular epithelia may constitute a mechanism to alleviate a decrease in blood flow and substrate availability and to re‐establish vascular integrity.
Thymic epithelial neoplasms in transgenic mice expressing SV40 T antigen under the control of an erythroid‐specific enhancer
Authors: Teitz Tal , Chang Judy C. , Kan Yuet Wai , Yen T. S. Benedict 
Published: June 2005

AbstractThe β‐globin locus control region is known to be a powerful erythroid‐specific enhancer. In an attempt to produce immortalized erythroid percursor cells, transgenic mice have been generated with the simian virus (SV) 40 T antigen gene under the control of the locus control region. As previously reported, transgenic mice did not develop erythroleukaemia, but rather succumbed to insulinomas and poorly differentiated rhabdomyosarcomas. This paper describes additional mice containing this transgene that developed thymomas of the mixed epithelial/lymphocytic type, in which only the epithelial component expressed the T antigen. Epithelial cell lines have been established from these tumours. This system may be useful in future studies on the pathogenesis of thymomas and the function of thymic epithelial cells.
Inhibition of gastric cell proliferation by acetaldehyde
Authors: Matysiak‐Budnik T. , Kärkkäinen P. , Methuen T. , Roine R. P. , Salaspuro M. 
Published: June 2005

AbstractHelicobacter pylori possesses alcohol dehydrogenase activity and is capable of producing acetaldehyde from ethanol in vitro. Acetaldehyde is a toxic and reactive compound and has been shown to inhibit the proliferation of many different cell lines in vitro. To study the effects of acetaldehyde on the proliferation of gastric epithelial cells in vivo, we employed an immunohistochemical method after labelling proliferating cells with 5′‐bromo‐2′‐deoxyuridine in rats receiving acetaldehyde intragastrically. Chronic (16 weeks) exposure of gastric mucosa to acetaldehyde given to rats in their drinking water in concentrations of 10 or 20 mM resulted in significant (P<0·05) inhibition of gastric epithelial cell proliferation, expressed as 332 ± 36, 348 ± 8, and 695 ± 15 proliferating cells per ten high‐power ( × 400) fields in the groups drinking 10 mM acetaldehyde, 20 mM acetaldehyde, and in controls respectively. In an acute study, significant inhibition of proliferation was observed after as few as 4 days of exposure to acetaldehyde, but only when a higher dose (50 mM) of acetaldehyde was given (438 ± 44 versus 615 ± 19 in controls, P<0·05). The inhibition of gastric cell renewal by acetaldehyde may play a role in the pathogenesis of ethanol‐ and/or H. pylori‐associated gastric diseases by inhibiting normal gastric mucosal protection and repair.
Protein extraction and western blotting from methacarn‐fixed tissue
Authors: Lalani El‐Nasir , Golding Matthew , Hudson Mark , Chieffi Gabriella , Stamp Gordon , Anilkumar T. V. , Sarraf Catherine , Alison Malcolm R. 
Published: June 2005

AbstractPolyacrylamide gel electrophoresis (PAGE) of proteins with subsequent western blotting has become a routine technique for the analysis of proteins from both cultured cells and fresh whole tissue. We have developed a method to extract proteins from methacarn‐fixed tissue which renders them suitable for SDS‐PAGE and western blotting. With a panel of antibodies to specific intermediate filaments, transforming growth factor‐α (TGF‐α), and albumin, immunohistochemistry was performed in parallel with western blotting on sections cut from methacarn‐fixed samples of normal rat liver and liver from rats treated under a regime which induces oval cell proliferation. Immunohistochemistry enabled the determination of changes in tissue distribution and abundance of the target proteins, which was mirrored by the corresponding western blot data. This technique can be especially effective when used in conjunction with immunohistochemistry. Tissue samples are easy to prepare, avoiding the precautions which need to be taken when handling fresh tissue (Abstract:J Pathol1994;173S: No. 41).
Long‐term follow‐up in early gastric cancer: Evaluation of prognostic factors
Authors: Ranaldi Renzo , Santinelli Alfredo , Verdolini Roberto , Rezai Banafsheh , Mannello Bruno , Bearzi Italo 
Published: June 2005

AbstractFour hundred and fourteen cases of early gastric cancer (EGC), diagnosed between 1977 and 1993, were studied. The percentage of EGC increased from 1977 to 1984, but thereafter remained more or less stable, despite a continuous increase in the number of endoscopic examinations. Three hundred and ninety‐six patients were followed up. Twenty‐nine patients died from the tumour, giving a 5‐year survival rate of 82·8 per cent. The ‘large’ size type of EGC, the presence of submucosal penetration, and lymph‐node metastasis showed a highly significant association with a lower survival rate. A small number of patients died despite the presence of ‘favourable’ prognostic factors. Other still unknown factors may therefore be important in determining the aggressive behaviour of certain EGCs.
Expression of basic fibroblast growth factor and fibroblast growth factor receptor in advanced gastric carcinoma
Authors: Ueki Takashi , Koji Takehiko , Tamiya Sadafumi , Nakane Paul K. , Tsuneyoshi Masazumi 
Published: June 2005

AbstractThe expression of basic fibroblast growth factor (bFGF) and fibroblast growth factor receptor (FGFR) mRNA was examined in gastric carcinomas by immunohistochemistry and in situ hybridization, respectively. In the 20 advanced carcinomas examined, bFGF was found in 14 (70·0 per cent) and was confined to the tumour cells, whereas FGFR mRNA was demonstrated in 12 (60·0 per cent) and seen in both tumour cells and endothelial cells. The bFGF and FGFR mRNA‐positive carcinomas were larger, were more frequently classified as undifferentiated adenocarcinoma, more frequently invaded the serosal layer, and had a higher rate of lymph node metastases than the bFGF and FGFR mRNA‐negative carcinomas. Patients with bFGF and FGFR mRNA‐positive carcinomas appear to die earlier than those with bFGF and FGFR mRNA‐negative tumours. The values for the carcinomas that were positive for either bFGF or FGFR mRNA fell between these two groups. The findings suggest that the autocrine/paracrine bFGF/FGFR channel is associated with undifferentiated gastric carcinomas and may lead to a poorer prognosis.