bluuebunny/crossref_metadata_2025 huggingface dataset:
Expression of a nuclear envelope protein recognized by the monoclonal antibody BU31 in lung tumours: Relationship to Ki‐67 antigen expression
Authors: Rowlands David C. , Bunce Christopher M. , Crocker John , Ayres Jon G. , Johnson Gerald D. , Ling Noel , Brown Geoffrey
Published: June 2005

The production of the murine monoclonal antibody BU31 is described. This antibody identifies a nuclear envelope protein which is expressed in some but not all cells, and which resembles statin, a protein reported to be expressed by non‐proliferating cells. BU31 was applied onto frozen sections of a series of 78 lung tumours and the staining patterns were compared with those obtained with Ki‐67. There was an inverse correlation between the proportion of tumour nuclei labelled with the two reagents (r= −0.61, 95 per cent confidence intervals −0.73 to −0.45). However, the four neuroendocrine neoplasms were BU31‐negative. Squamous cell carcinomas often showed a peripheral distribution of the cells stained positively with Ki‐67, whereas BU31 tended to label centrally situated cells. These observations are consistent with the concept that the antigen recognized by BU31 is expressed by non‐proliferating cells in these tumours.
Fractal geometric analysis of material from molar and non‐molar pregnancies
Authors: Cross Simon S. , Howat Alec J. , Stephenson Timothy J. , Cotton Dennis W. K. , Underwood James C. E.
Published: June 2005

Histological sections from 25 non‐molar pregnancies, nine partial hydatidiform moles, and 16 complete hydatidiform moles were examined (diagnosis was taken as the consensus of seven experienced histopathologists) and the fractal dimension was measured using a box‐counting method implemented on a microcomputer‐based image analysis system. The fractal dimensions of the different diagnostic categories were normally distributed with a mean of 1.50 for non‐molar pregnancies, 1.44 for partial moles, and 1.42 for complete moles. All the measured fractal dimensions were greater than the topological dimension (1), demonstrating that the specimens had a fractal element to their structure. There was a significant difference between the fractal dimensions of non‐molar pregnancies and complete moles (P=0.0005), but not between partial moles and non‐molar pregnancies (P=0.0823) or complete and partial moles (P=0.4400). Using the fractal dimension to predict the histopathological diagnosis assigned 56 per cent of the cases to the correct category with a kappa statistic of 0.26, so the fractal dimension, used alone, is not a useful morphometric discriminant in the diagnosis of molar and non‐molar pregnancy.
Detection of specimen contamination in routine histopathology by HLA class II typing using the polymerase chain reaction and sequence specific oligonucleotide probing
Authors: Bateman Adrian C. , Leung Shu T. , Howell William M. , Roche William R. , Jones David B. , Theaker Jeffrey M.
Published: June 2005

This study evaluates the use of human leukocyte antigen (HLA) class II PCR‐SSO typing for the investigation of suspected specimen contamination in four routine surgical histopathology cases. Two cases were of patients undergoing transurethral resection of the prostate gland. The third case was a patient undergoing excision of a subcutaneous small cell carcinoma. The fourth case was a patient undergoing reversal of a Hartman's procedure for adenocarcinoma of the colon. Tissue was extracted from routinely processed formalin‐fixed, paraffin‐embedded material and HLA class II typed, using a non‐radioactive polymerase chain reaction‐sequence specific oligonucleotide probe technique (PCR‐SSO), using probes specific for the DRB, DQA, and DQB loci. The accuracy of PCR‐SSO typing of DNA extracted from paraffin biopsy material was confirmed by concordant results for DRB, DQA, and DQB typing in five separate control individuals from whom frozen and paraffin lymph node biopsies were available. PCR‐SSO typing was also successful in the four study cases and revealed that contamination had occurred in two cases, eliminating this possibility in the remaining cases. This study demonstrates that DNA can be reliably amplified and accurately typed from routinely processed material, and reveals a useful new application for PCR‐SSO tissue typing.
Co‐expression of M‐CSF transcripts and protein, FMS (M‐CSF receptor) transcripts and protein, and steroid receptor content in adenocarcinomas of the ovary
Authors: Kommoss F. , Wölfle J. , Bauknecht T. , Pfisterer J. , Kiechle‐Schwarz M. , Pfleiderer A. , Sauerbrei W. , Kiehl R. , Kacinski B. M.
Published: June 2005

The co‐expression of M‐CSF (CSF‐1) and its receptor in specimens of ovarian cancer has recently been reported. Preliminary results have already suggested a possible influence of steroids on FMS (M‐CST receptor) expression. Fifty‐five non‐pretreated FIGO stage III/IV ovarian adenocarcinomas were studied for M‐CSF transcripts and protein, as well as FMS transcripts and protein, using standard molecular biological techniques (Northern blot, slot blot analysis) and immunocytochemistry (ICC). Steroid receptor content was measured by DCC analysis in 44/55 specimens; in addition, ER/PR (oestrogen/progesterone) ICC was performed in 32/55 specimens. All tumours were shown to contain M‐CSF‐specific mRNA. Likewise, M‐CSF protein was detected by ICC in the stroma and over the epithelium in all specimens. However, while most tumours were shown to contain FMS‐specific mRNA, only 64 per cent of cases showed significant expression of FMS protein by tumour epithelium as shown by ICC. A statistically significant positive correlation was found between M‐CSF and FMS mRNA expression levels. A week non‐significant positive correlation was noted between FMS mRNA expression levels and tumour grade. Carcinomas were ER‐positive in 66 per cent (DCC) or 34 per cent (ICC), and PR‐positive in 73 per cent (DCC) or 34 per cent (ICC). A statistically significant positive correlation between ER (DCC) and M‐CSF mRNA expression levels was found. Weak non‐significant correlations were present between ER (ICC) and FMS (ICC), as well as between PR (DCC) and FMS mRNA expression. Our study supports earlier propositions that co‐expression of M‐CSF and its receptor could regulate growth in ovarian carcinomas via an autocrine loop.
Hitherto unrecognized arterioles within hepatocellular carcinoma
Authors: Himeno Hiromi , Enzan Hideaki , Saibara Toshiji , Onishi Saburo , Yamamoto Yasutake
Published: June 2005

The distribution of blood vessels in human hepatocellular carcinoma was studied with the anit‐α‐smooth muscle actin monoclonal antibody by light and electron microscopy, and with morphometric analysis. α‐Smooth muscle actin‐positive arterioles were never observed in lobules or pseudolobules of non‐cancerous areas, but were frequently seen within hepatocellular carcinomas. Morphometric analysis revealed that most of these arterioles measured between 10 and 25 μm in diameter. The morphology of intratumoural arterioles differed considerably from that of conventional arteries in the portal tracts of the non‐cancerous area. The presence of abundant intratumoural arterioles can explain the angiographic hypervascularity of hepatocellular carcinoma and provides a pathological basis for its susceptibility to hypoxia and for arterial embolization as a therapeutic strategy.
p53 Protein, PCNA staining, and DNA content in follicular neoplasms of the thyroid gland
Authors: Czyz Wojciech , Joensuu Heikki , Pylkkänen Liisa , Klemi Pekka J.
Published: June 2005

Forty‐nine follicular adenomas and 11 follicular carcinomas of the thyroid were investigated by immuno‐histochemistry for the expression of p53 protein and proliferating cell nuclear antigen (PCNA). The DNA ploidy and the S‐phase fraction (SPF) of the neoplasms were analysed by flow cytometry. Twelve adenomas (24 per cent) and six carcinomas (55 per cent) were DNA non‐diploid (P=0·07). The carcinomas had a higher proliferation rate than the adenomas when assessed either by SPF size (median 9·9 per cent vs. 2·9 per cent, P=0·0003) or by PCNA staining intensity (P<0·0001). Some scattered nuclei in two (4 per cent) adenomas and in three (27 per cent) carcinomas stained positively for p53 (P=0·04). The two adenomas with positive staining for p53 were subserially sectioned, but no signs of invasion were found; both patients are alive and well 6 and 7 years after surgery. One of the two adenomas showing positive p53 nuclear staining was DNA aneuploid, and both were positive in PCNA staining, but their SPFs were low (2·1 and 3·3 per cent). We conclude that p53 protein expression is not confined to follicular carcinomas; scattered p53‐positive cells may also be present in histologically and clinically benign follicular adenomas. Because both follicular adenomas and carcinomas may be DNA aneuploid and their SPF and PCNA staining distributions overlap, the distinction between follicular adenoma and carcinoma should still be based on histological criteria.
Early increase precedes a depletion of endothelin‐1 but not of von Willebrand factor in cutaneous microvessels of diabetic patients. A quantitative immunohistochemical study
Authors: Properzi Giuliana , Terenghi Giorgio , Gu Xu‐Hong , Poccia Gianfranco , Pasqua Raffaele , Francavilla Sandro , Polak Julia M.
Published: June 2005

Endothelin‐1 (ET‐1) is a vasoconstrictor peptide which is produced by endothelial cells. The subcellular distribution of ET‐1 in human skin and the variation of immunostaining for ET‐1 by light microscopy in skin biopsies of diabetic patients have been analysed using immunohistochemistry and image analysis quantification. Skin biopsies were collected from 17 patients with type 1 diabetes of different durations and with presence or absence of microangiopathy in the retina; skin biopsies of healthy subjects were utilized as controls. The distribution of ET‐1 immunoreactivity (IR) at both light and electron microscopy was compared to that of von Willebrand factor (vWf), a general marker of total cutaneous microvessels. Immunohistochemistry revealed that in controls the distribution of immunostaining was similar for ET‐1 and vWf, being localized to microvessels in all areas of the skin. However, at the electron microscopical level ET‐1‐IR was localized in the endothelial cytoplasm rather than in specific organelles, while vWf immunostaining was associated with Weibel‐Palade bodies. ET‐1‐IR was observed in 4/8 (50 per cent) biopsies from healthy subjects; this increased to 81.8 per cent in biopsies of patients affected by diabetes for less than 10 years and decreased to 16.6 per cent in patients with diabetes for more than 10 years. Quantification of ET‐1 staining showed a significant decrease of ET‐1‐IR in patients affected by diabetes for more than 10 years compared with those affected by diabetes for less than 10 years (P<0.05). Also, the percentage of biopsies showing positive ET‐1 staining was lower in patients with retinopathy than in patients without retinopathy. On the contrary, vWf‐IR was observed in all skin specimens and its quantification showed no differences between diabetic patients and controls. These changes are not related to variations in the number of blood vessels, and it is suggested that they reflect a possible functional alteration of the endothelial cells during diabetes.
High prevalence of pancreatic islet amyloid in patients with end‐stage renal failure on dialysis treatment
Authors: de Koning E. J. P. , Fleming K. A. , Gray D. W. R. , Clark A.
Published: June 2005

Islet amyloid polypeptide (IAPP) is the main proteinaceous component of pancreatic islet amyloid, which is a characteristic feature of type 2 diabetes. The factors responsible for amyloid deposition are unclear. Patients with end‐stage renal failure (ESRF) on dialysis treatment have increased insulin resistance which is associated with hypersecretion of beta‐cell products. Furthermore, elevated concentrations of circulating IAPP are found in these patients due to reduced renal clearance of IAPP. To determine the prevalence of islet amyloid in this group of patients, pancreas was examined from 23 non‐diabetic [aged 62 (29‐79) years, median and range] and four type 2 diabetic [aged 67 (56‐72) years] patients with ESRF on dialysis treatment. Pancreatic specimens from 30 non‐diabetic control subjects [aged 67.5 (56‐86) years] and 14 type 2 diabetic subjects without renal disease [aged 69 (48‐86) years] were used as control groups. Islet amyloid was present in all type 2 diabetic patients with ESRF and in 12 out of 14 type 2 diabetic control subjects (86 per cent). Amyloid deposits were found in 8 out of 23 non‐diabetic patients with ESRF (35 per cent), which was a higher prevalence than that found in non‐diabetic control subjects (3 per cent) (P<0.01). This may be related to undiagnosed (pre)diabetes. Elevated secretion rates of IAPP due to insulin resistance and high circulation IAPP concentrations as a result of severely reduced renal clearance of IAPP will cause high pericellular concentrations of IAPP. This condition is likely to enhance amyloid fibril formation in pancreatic islets similar to that observed in type 2 diabetes.
Interphase cytogenetics: Analysis of numerical chromosome aberrations in isolated cells
Authors: Herrington C. S. , Cooper K. , McGee J. O'D.
Published: June 2005

Probes which recognize pericentromeric repetitive sequences can be used to determine numerical chromosome aberrations in interphase nuclei. Under appropriate stringency conditions, these probes decorate only the chromosome from which they were derived. It has been assumed that abnormalities of signal number in interphase nuclei reflect aneuplodiy rather than proliferation, but this has not been clearly demonstrated. In this paper, three alphoid probes (D3Z1, D11Z1, and DXZ1) were localized to the appropriate chromosomes and the factors governing the production of reproducible signal distributions from three aneuploid cervical carcinoma‐derived epithelial cell lines were investigated. Abnormalities of signal number represent numerical chromosome aberrations rather than changes associated with proliferation. Using four simple fules of interpretation, reproducible results can be obtained with minimal technical variation and selection bias. These results demonstrate that pericentromeric repetitive probes can be used reproducibly to determine numerical chromosome aberrations independent of cell proliferation in interphase nuclei, a necessary prerequisite for the application of this approach to the analysis of human tumours.
Combined enzyme histochemical and ultrastructural study on cryostat sections of pig heart to detect early reperfusion damage after ischaemia
Authors: Frederiks Wilma M. , Tukkie Raymond , Gründeman Paul F. , Hoebe Christine , Schellens Jacques P. M.
Published: June 2005

In cardiac surgery, recognition of peroperative myocardial infarction may improve patient selection for prolonged circulatory support. The value of enzyme histochemistry to discriminate between reversible and irreversible myocardial damage at short periods of reperfusion was studied in an in vivo model of regional ischaemia in pig hearts. The left anterior descending coronary artery (LADCA) was clamped for 45 min followed by 2 h reperfusion (group 1, n=3). Post‐mortem heart tissue showed markedly decreased activities of lactate dehydrogenase (LDH) and beta‐hydroxybutyrate dehydrogenase (BDH) as demonstrated in cryostat sections, accompanied by massive leakage of LDH in the venous effluent. The depleted areas showed irreversible cell damage based on the presence of flocculent densities in mitochondria. In group 2 (n=6), LADCA flow was reduced to 40 per cent of the base‐line value followed by 2 h reperfusion. Heart tissue showed normal LDH and BDH activities, except for some cells surrounding blood vessels, which activity was minimally decreased. Flocculent densities in mitochondria were never observed. We conclude that enzyme histochemistry of LDH and BDH activity on cryostat sections is a useful tool for detecting irreversible myocardial cell damage as early as 2 h reperfusion after ischaemia of the pig heart. The technique has potential applications in the detection of peroperative infarction in human biopsies.
Developing diagnostic techniques: The role of magnetic resonance imaging in tumour staging
Authors: Argent Julie , Beer Trevor
Published: June 2005

Magnetic resonance (MR) is a state‐of‐the art imaging modality which does not use ionizing radiation. It is now widely available as an imaging technique in the U.K. and is no longer confined to specialist centres. MR has now become part of the clinician's diagnostic armamentarium and is not merely a research utility.

A magnetic field and radio‐frequency waves are used to excite protons and produce an image. Protons exist in many different environments but for imaging purposes the hydrogen protons in fat and water are used. The rate at which excited protons relax is described by two characteristic times, T1 and T2, which vary in different tissues. T1‐weighted images show normal anatomy, whilst T2‐weighted images generally highlight abnormal tissue. Injection of a paramagnetic gadolinium‐based contrast agent enhances T1‐weighted images by reducing relaxation times. Initially it was felt that MR would be tissue‐specific,1 enabling correlation with histopathological findings. Unfortunately, research and clinical use has shown that MR does not fulfil these expectations at present.2 Future developments such as spectroscopy may improve tissue specificity and hence diagnostic accuracy.

The value of MR for imaging the neuro‐axis and musculo‐skeletal system is well established. New developments are increasing its applications in breast, thorax, and liver imaging. There has been much research into the role of MR in staging malignant tumours, particularly in the areas of bladder and lung carcinomas. The present article concentrates on this specific application.
Overexpression of the tumour suppressor gene p53 is not implicated in neuroendocrine tumour carcinogenesis
Authors: Wang Da‐Gong , Johnston Colin F. , Anderson Neil , Sloan James M. , Buchanan Keith D.
Published: June 2005

The tumorigenesis of neuroendocrine tumours remains poorly understood, although a minority, the familial multiple endocrine neoplasia (MEN 1 and MEN 2), are known to be of uncommon genetic origin. Mutation of the tumour suppressor gene, p53, is now known to be a common genetic alteration in about half of all types of non‐endocrine cancers. In the present study, immunocytochemistry using the monoclonal anti‐p53 antibody, DO‐7, has been employed to investigate the accumulation of p53 immunoreactivity in a wide range of primary neuroendocrine tumours. Tumours (n=109) were fixed and processed to paraffin wax according to a constant protocol. Sections were subjected to microwave antigen retrieval prior to immunostaining for p53. Positive nuclear immunostaining was obsered in one medullary carcinoma of the thyroid (MCT), one lung carcinoid, and five small cell carcinomas of the lung (SCCL). All other tumour samples were consistently negative. As the neoplasia investigated in this study comprised a wide spectrum of neuroendocrine tumour types and ranged from minute, relatively benign lesions to malignant metastasizing disease and as there was no relationship between the presence of p53 overexpression and clinico‐pathological features, the present study suggests that p53 gene mutation may be relatively unimportant in the genesis of neuroendocrine tumours.
A sensitive staining method for NORs
Authors: Yekeler H. , Erel Ö. , Yumbul A. Z. , Doymaz M. Z. , Doǧan Ö. , Özercan M. R. , Iplikçi A.
Published: June 2005

A sensitive staining method for nucleolar organizer regions (NORs) is described, using blue toning of AgNORs. NORs are loops of DNA which are transcribed into ribosomal RNA. NORs can be demonstrated by staining with silver nitrate, since NOR‐associated proteins are argyrophilic, producing structures termed AgNORs. Normal blood lymphocytes were stained with both methods. The number and resolution of NORs increased 2–3 times by blue toning (30 mmol/l FeCl3, 11 mmol/l potassium hexacyanoferrate(III), and 33 mmol/l oxalic acid) compared with silver staining. A significant difference in the number of NORs was noticed between silver‐stained and blue‐toned cells (P<0.001). The blue toning technique thus appears to be more sensitive in detecting NORs than the AgNOR method and may prove a useful alternative for applications in histopathology.
Mutation at chromosome 11q23 in human non‐familial breast cancer: A microdissection microsatellite analysis
Authors: Koreth John , Bethwaite Peter B. , McGee James O' D.
Published: June 2005

Allelotypic evaluation of loss of heterozygosity (LOH) has been instrumental in the identification of tumour suppressor genes. Here we report a high incidence of LOH at chromosome 11q23 in non‐familial breast cancers within situ, invasive, and metastatic tumour cells microdissected from archival haematoxylin and eosin (H & E) sections for polymerase chain reaction (PCR)‐LOH analysis at polymorphic microsatellite loci. Ninety‐four cases of non‐familial breast cancer were examined at the D11S29 microsatellite locus on chromosome 11q23. Eighty‐three cases (88 per cent) were informative and 35 cases overall (42 per cent) had LOH at this locus, comprising 23 per cent ofin situ, 36 per cent of invasive, and 28 per cent of metastatic cancers. The DNA from those cancer cells with LOH was amplified at microsatellite loci D11S554 (11p12–p11.2) and D11S534 (11q13). In 19 of 67 cases overall (28 per cent), LOH occurred solely at 11q23. There was an association between LOH at 11q23 and tumour size ≥2 cm (P<0.01) in the overall results and the invasive cancers. The data revealed heterogeneity for LOH at D11S29 inin situ, invasive, and metastatic cells from the same case. In general, however, there was concordance between LOH (or its absence) inin situand invasive disease. We conclude that the distal part of the long arm of chromosome 11 contains a region involved in breast carcinogenesis and that there is molecular heterogeneity at this chromosomal region in individual breast cancer cells.
Expression of the ret proto‐oncogene in phaeochromocytoma. An in situ hybridization and northern blot study
Authors: Matias‐Guiu Xavier , Colomer Anna , Mato Eugenia , Cuatrecasas Miriam , Komminoth Paul , Prat Jaime , Wolfe Hubert
Published: June 2005

The expression and distribution of ret proto‐oncogene mRNA were investigated in five phaeochromocytomas of both familial and sporadic types by in situ hybridization (ISH) using digoxigenin‐labelled cRNA probes and Northern blot (NB) analysis with random priming labelled cDNA probes. The probes corresponded to the tyrosine kinase domain of the gene. An excellent correlation was found between the ISH and NB results. By both techniques, the expression of the ret proto‐oncogene was detected in three of the five cases, two of four familial tumours, and the only sporadic tumour that was studied. The results confirm that ret is frequently expressed in phaeochromocytomas and suggest that it might be an important event in their development and/or progression.
A murine osteoblast cell line (MC3T3) produces A soluble elastogenic compound
Authors: Grosso Leonard E. , Lacey David L.
Published: June 2005

Elastofibromas are localized proliferations of mesenchymal cells that produce an exuberant amount of elastin‐rich extracellular matrix. Recently periosteal fibroblasts have been proposed to be the proliferating cell. The hypothesis has been tested that osteocytes or osteoblasts contribute to the formation of elastofibromas by secreting a compound(s) that enhances elastin production. Media conditioned by murine calvarial osteoblasts (MC3T3) increased tropoelastin synthesis in bovine ligamentum nuchae fibroblasts. Addition of MC3T3‐conditioned medium to bovine ligamentum nuchae fibroblast cultures produced a two‐fold increase in tropoelastin RNA. The maximal increase in tropoelastin RNA was between 16 and 24 h; tropoelastin mRNA had returned to control values by 40 h. A similar increase in tropoelastin protein production was detected. The soluble elastogenic compound was neither inteleukin‐1 (IL‐1) nor IL‐6. These results support the hypothesis that an interaction between bone and perosteum may be involved in the formation of elastofibromas.
Absence of p53 gene mutation and infrequent overexpression of p53 protein in hepatoblastoma
Authors: Chen Tse‐Ching , Hsieh Ling‐Ling , Kuo Tseng‐Tong
Published: June 2005

Ten cases of hepatoblastoma were studies for overexpression of p53 protein by immunohistochemistry and for possible p53 gene mutation by single strand conformation polymorphism (SSCP) analysis and direct DNA sequencing of the polymerase chain reaction products. Only one case of the macrotrabecular type at stage IV showed overexpression of p53 protein. No DNA mobility shift was found in any of the cases studies by SSCP analysis. DNA sequencing performed on the case showing overexpression of p53 protein revealed no mutation within exons 5 to 8. The associated adrenal cortical carcinoma of the same case also showed overexpression of p53 protein, but no mutation of the p53 gene. These results indicate that mutation of the p53 gene is infrequent in hepatoblastoma. This observation supports the view that mutation of the p53 gene is not as important in the oncogenesis of childhood neoplasms as in adult cancers.
Human malignant mesothelial cells: Variability of ultrastructural features in established and nude mice transplanted cell lines
Authors: Fleury‐Feith Jocelyne , Kheuang Laurence , Zeng Lin , Bignon Jean , Boutin Christian , Monnet Isabelle , Jaurand Marie‐Claude
Published: June 2005

The aim of this study was to determine, by transmission electron microscopy, the differentiation features of 21 human malignant mesothelioma cell lines (HMCLs) established from 13 specimens of 12 confirmed human malignant mesotheliomas, and of tumours induced in nude mice injected with 16 HMCLs. Fifty per cent of HMCLs showed typical mesothelial differentiation (long and slender microvilli, desmosomes, perinuclear intermediate filaments); 29 per cent did not show differentiation; and the remainder were poorly differentiated. Three human tumour specimens gave several different HMCLs; the cell lines obtained from a given tumour exhibited variable mesothelial differentiation. Eleven HMCLs were compared with the native tumour. Four were similar to the tumour and seven were less well differentiated, in most cases in relation to their microvilli. With six HMCLs, tumours induced in nude mice were less well differentiated than the corresponding cell lines, whereas with four HMCLs, tumours were equally or better differentiated. However, in most nude mice tumours, typical mesothelial microvilli were present. These results show that cell lines established from malignant mesothelioma may exhibit dedifferentiated features. However, while the variability in ultrastructural differentiation may result from the culture microenvironment, it could also be related to the state of differentiation, of the native tumour sample and to tumour cell heterogeneity.
Nuclear and cytoplasmic bcl‐2 expression in endometrial hyperplasia and adenocarcinoma
Authors: Chan W. Kong , Mole Margaret M. , Levison David A. , Ball Richard Y , Lu Qi‐Long , Patel Kirtika , Hanby Andrew M.
Published: June 2005

The bcl‐2 proto‐oncogene, which inhibits programmed cell death (apoptosis), has recently been found to be cyclically expressed in human endometrium. In order to investigate its role in endometrial hyperlasia and neoplasia, bcl‐2 expression was studied in 25 cases of endometrial carcinoma and 20 cases of endometrial hyperplasia (eight simple, two complex, and ten atypical hyperplasias). Uniform intense cytoplasmic bcl‐2 expression was found in all cases of non‐atypical hyperplasia, and less strong positivity in eight out of ten cases of atypical hyperplasia. In well‐differentiated carcinomas, nine out of ten showed weak to moderate bcl‐2 expression, whereas six out of seven poorly differentiated carcinomas were bcl‐2‐negative. Moderately differentiated tumours were an intermediate group, with six out of eight being positive. Widespread localization of bcl‐2 protein to the chromosomes of dividing cells was also demonstrated, regardless of cytoplasmic bcl‐2 expression, with rare staining of interphase nuclei. Our findings suggest a role for bcl‐2 in the natural history of endometrial neoplasia and studies are needed to determine its usefulness as a prognostic marker. The finding of bcl‐2 localization to chromosomes has important implications for its mode and site of action.
Differential expression of the c‐kit proto‐oncogene in germ cell tumours
Authors: Izquierdo Miguel A. , Van Der Valk Paul , Van Ark‐otte Jannette , Rubio Gonzalo , Germa‐Lluch Jose R. , Ueda Ryuzo , Scheper Rik J. , Takahashi Takashi , Giaccone Giuseppe
Published: June 2005

The c‐kit proto‐oncogene product and its ligand stem cell factor play an important role in haematopoiesis, spermatogenesis, and melanogenesis. Using an anti‐c‐kit antiserum raised against a synthetic peptide, we studied the immunohistochemical expression of the c‐kit gene product in 60 germ cell tumours (GCTs) (53 testicular, 7 extragonadal), derived from primary GCTs in 45 cases and metastatic tumours in 15 cases. Twenty‐eight out of 28 seminomas showed c‐kit membranous staining in the majority of cells. A similar pattern of expression was seen in intratubular germ cell neoplasia. Nine out of 29 (32 per cent) non‐seminomas displayed cytoplasmic, but not membranous, c‐kit immunoreactivity in occasional cells. In three mixed GCTs, c‐kit expression was limited to the seminoma component. In normal testis, c‐kit expression was observed in some basal tubular cells, corresponding to undifferentiated spermatogonia. These results suggest a role for c‐kit in the oncogenesis of GCT, where down‐regulation of c‐kit might be a critical step during progression from seminomas to non‐seminomas. Immunohistochemical analysis of c‐kit should be considered as a diagnostic aid for GCT and in particular may be helpful in the identification of certain extragonadal seminomas.
Role of endothelial cells in the development of glomerular lesions of mesangiocapillary glomerulonephritis
Authors: Muda Andrea Onetti , Feriozzi Sandro , Crescenzi Anna , Faraggiana Tullio
Published: June 2005

The histological and ultrastructural changes of mesangiocapillary glomerulonephritis (MCGN) are not unique to this entity: splitting of basement membrane (BM) is seen in a number of conditions with an altered coagulation pattern. The distribution of endothelial cells in the glomerular capillaries in five cases of MCGN was studied by light and electron microscopy and immunocytochemistry; endothelial cells were stained with peroxidase or FITC‐conjugated antibodies against Factor VIII‐related antigen or CD34 antigen and observed with conventional light/immunofluorescence microscopy and confocal laser scanning microscopy for three‐dimensional reconstruction. Electron microscopy was performed with colloidal gold labelling. Endothelial cells were shown to be present within the duplicated basement membranes of capillaries and continuity was demonstrated with endothelial cells lining the capillary lumina. These results suggest that endothelial cells as well as mesangial cells can participate in the formation of the characteristic double contour of capillary walls in MCGN, especially in the early stages. There are some similarities to the changes seen in larger arteries as a consequence of thrombotic disease.
Quantitation and prognostic value of breast cancer angiogenesis: Comparison of microvessel density, Chalkley count, and computer image analysis
Authors: Fox Stephen B. , Leek Russel D. , Weekes Michael P. , Whitehouse Ruth M. , Gatter Kevin C. , Harris Adrian L.
Published: June 2005

In some studies of breast cancer, quantitation of immunohistochemically highlighted microvessel ‘hot spots’ has been shown to be a powerful prognostic tool. However, the antibody used, the number and size of the ‘hot spots’ assessed, and the stratification of patients into high and low vascular groups vary between studies. Furthermore, little is known about the relationship between microvessel density and other vascular parameters. These uncertainties and the laborious nature of the technique make it unsuitable for diagnostic practice. Both manual and computerized image analysis techniques were used in this study to examine the relationship between microvessel density and the vascular parameters in different sized microscopic fields in a pilot series of 30 invasive breast carcinomas. Automated pixel analysis of immunohistochemical staining, Chalkley point counting, and observer subjective vascular grading were also assessed as more rapid methods of measuring tumour vascularity. A Chalkley count was also performed on a further 211 invasive breast carcinomas. Significant correlations were observed between manual microvessel density and luminal perimeter (r=0·6, P=0·0004), luminal area (r=0·56, P=0·002), and microvessel number (r=0·57, P=0·0009) by computerized analysis. There were also significant correlations between the microscopic hot spots of 0·155 mm2 and 0.848 mm2 for microvessel number (r=0·81, P>0·00005), luminal perimeter (r=0·78, P<0·00005), and luminal area (r=0·65, P=0·0001). In addition, a significant correlation was observed between microvessel density and both subjective vascular grade (P=0·002) and Chalkley count (P=0·0001). A significant reduction in overall survival was observed between patients stratified by Chalkley count in both a univariate (P=0·02) and a multivariate (P=0·05) analysis in the 211 invasive breast carcinomas. These findings show that Chalkley counting is a rapid method of quantifying tumour angiogenesis and gives independent prognostic information which might be useful in diagnostic practice.
Similarities and differences among antisocial and psychopathic self‐report inventories from the perspective of general personality functioning
Authors: Hicklin Jabbar , Widiger Thomas A.
Published: June 2005

There are currently many alternative self‐report inventories for the assessment of antisocial and psychopathic personality traits. The hypothesis of the current study was that similarities and differences among them can be meaningfully understood with respect to their representation of common personality traits. The current study explored this hypothesis using as a point of comparison the Five Factor Model (FFM) of general personality functioning. Six self‐report measures currently being used in antisocial and psychopathy research were administered, along with a self‐report measure of the FFM. Differences were obtained across the six inventories with respect to how they related to the domains and facets of the FFM that were consistent with FFM models of psychopathy and antisocial personality disorder. Implications for the theoretical and clinical understanding of findings obtained with the respective inventories, and the personality disorder constructs they assess, are discussed. Copyright © 2005 John Wiley & Sons, Ltd.
Palladium‐catalyzed three‐component coupling polycondensation of aromatic diiodide, aromatic bis(boronic acid propanediol ester), and norbornadiene: A new route for poly(p‐phenylene vinylene)
Authors: Ishibe Satoko , Tomita Ikuyoshi
Published: June 2005

A novel synthetic method for soluble precursor polymers of poly(p‐phenylene vinylene) (PPV) derivatives by the palladium‐catalyzed three‐component coupling polycondensation of aromatic diiodides, aromatic bis(boronic acid) derivatives, and norbornadiene is described. For example, the polymerization of 1,4‐diiodo‐2,5‐dioctyloxybenzene, benzene‐1,4‐bis(boronic acid propanediol ester), and norbornadiene at 100 °C for 3 days provided a polymer consisting of the three monomer units in a 97% yield (number‐average molecular weight = 3100, weight‐average molecular weight/number‐average molecular weight = 1.37). A derivative of PPV was produced smoothly by the retro Diels–Alder reaction of the polymer both in a dodecyloxybenzene solution and in a film at 200 °C in vacuo. © 2005 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 43: 3403–3410, 2005
Modeling of the linear viscoelastic behavior of low‐density polyethylene
Authors: Stanescu P. , Majesté J. C. , Carrot C.
Published: June 2005

We predict the linear viscoelastic behavior of low‐density polyethylene from both the molecular‐weight distribution and the individual structure of each species in the sample. The “structure map” of the samples was derived from SEC measurements. This map is a three‐dimensional representation of the seniority distribution, and represents the probability of existence of a segment with seniority i in a molecule of molecular weight M. Moreover, results from the kinetics of the free radical polymerization of polyethylene show that the molecular weight of the segments increases according to their seniority. Finally, tube dilatation was generalized to the case of polydisperse samples. The solvent behavior of the relaxed segments was included through a continuous function of time that describes the instantaneous state of the entanglement network in the sample. The comparison between the theoretical predictions and the experimental data shows a good agreement over the whole experimental frequency range. © 2005 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 43:1973–1985, 2005
Formation of new banded spherulites in polyimides
Authors: Liu Xikui , Tang Jialing , Zheng Yuanyuan , Gu Yi
Published: June 2005

The crystalline morphology and structural development of aromatic polyimides during an optimum continuous thermal imidization procedure were examined by means of polarized optical microscopy and X‐ray diffraction. During thermal imidization, 3,3′,4,4′‐benzophenonetetracarboxylic dianhydride/1,3‐diaminobenzene polyimide samples formed complicated spherulites, which, in addition to zigzag Maltese crosses, also showed concentric extinction rings, which are characteristic of banded spherulites. The factors affecting the formation of banded spherulites were studied. The initial imidization conditions dramatically affected the formation of the banded spherulite morphology: slow heating (0.5 °C/min) or fast heating (20 °C/min) led to relatively small polyimide spherulites and less identifiable extinction rings. The morphological features were also affected by the molecular weight of the polyimide: higher molecular weight samples showed typical banded spherulites, whereas low‐molecular‐weight samples formed degenerated banded spherulites. In all the spherulites formed in 3,3′,4,4′‐benzophenonetetracarboxylic dianhydride/1,3‐diaminobenzene polyimides, special zigzag Maltese crosses, instead of normal Maltese crosses, were observed. The relationship between the imidization procedure and the spherulite morphology formation was also studied. X‐ray and Fourier transform infrared together revealed that after several minutes of thermal treatment, the crystallization was nearly complete, with a 42.5% degree of crystallinity; meanwhile, only some poly(amic acid) converted to the corresponding polyimide, with a 27% degree of imidization. The crystalline morphology and structure formed in the initial stage of the imidization process were maintained during the following imidization processing at an elevated temperature. © 2005 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 43: 1997–2004, 2005
Factors that influence Spanish‐ and English‐speaking participants' decision to enroll in cancer randomized clinical trials
Authors: Ellington Lee , Wahab Stephanie , Sahami Martin Shadi , Field Rosemary , Mooney Kathleen H.
Published: June 2005

Cancer randomized clinical trial (RCT) participation is low, particularly among ethnic and racial minorities. Hispanic enrollment is far below their representation in the US population, yet their cancer burden is higher. Little is known from the patient perspective about factors which influence the decision to enroll in RCTs. We asked Spanish‐ and English‐speaking individuals what factors influence decisions about cancer RCT participation. Eight focus groups were conducted with 55 participants (25 Spanish and 30 English‐speaking). The groups were taped, transcribed, and analyzed for themes. Six major themes emerged: patient–provider communication, personal relationship with provider, involvement of significant others in decision making, role of faith, need for information, and impact of discrimination on decision making. Both similarities (e.g. need for comprehensive information) and differences (e.g. need for provider acknowledgement of emotional and spiritual concerns) were found between Spanish‐ and English‐speaking participants. Among Spanish‐speaking participants, level of education was differentially related to decision‐making themes. Implications for providers are discussed. Copyright © 2005 John Wiley & Sons, Ltd.
Disorders of pyruvate carboxylase and the pyruvate dehydrogenase complex
Authors: Robinson B. H. , MacKay N. , Chun K. , Ling M.
Published: June 2005

The most common defect associated with deficiency of the pyruvate dehydrogenase (PDH) complex occurs in the E1 component, specifically due to mutations in the X‐linked E1α gene. Clinical sequelae of these mutations, which range from severe neonatal lactic acidosis to carbohydrate‐sensitive ataxia, can be different in males and females depending on the nature of the mutation and, in the case of females, on the X‐inactivation pattern in different tissues. Males have a high representation of missense mutations among the patient cohort, while females are much more likely to have DNA rearrangements, particularly toward the 3′ end of the coding sequence of the gene. Missplicing mutations involving exon 6 deletion have been reported, as has a missense mutation conferring true thiamin‐responsiveness of the enzyme and the patient's clinical symptoms.

Pyruvate carboxylase deficiency, on the other hand, is a true autosomal recessive disease, though it has high occurrences in particular ethnic groups, especially in Algonkian‐speaking Amerindians and in Arabs. In the former group the defect is a simple type in which material cross‐reactive to pyruvate carboxylase antibody is present in cultured cells (CRM+ve). In the latter group, cross‐reacting material is rarely present (CRM−ve). The CRM+ve patients can survive into teenage years with careful supervision, while the CRM−ve patients have complications due to hyperammonaemia and dysfunction of the urea cycle and rarely survive beyond 3 months of life.
Disorders of gluconeogenesis
Authors: van den Berghe G.
Published: June 2005

Gluconeogenesis, or the formation of glucose from mainly lactate/pyruvate, glycerol and alanine, plays an essential role in the maintenance of normoglycaemia during fasting. Inborn deficiencies are known of each of the four enzymes of the glycolytic — gluconeogenic pathway that ensure a unidirectional flux from pyruvate to glucose: pyruvate carboxylase, phosphoenolpyruvate carboxykinase, fructose‐1,6‐bisphosphatase, and glucose‐6‐phosphatase. In this paper, the clinical picture, pathophysiology, diagnostic tests, genetics, treatment and prognosis of the deficiencies of fructose‐1,6‐bisphosphatase and phosphoenolpyruvate carboxykinase are reviewed.
Clinical presentation of mitochondrial disorders in childhood
Authors: Munnich A. , Rötig A. , Chretien D. , Cormier V. , Bourgeron T. , Bonnefont J. ‐P. , Saudubray J. ‐M. , Rustin P.
Published: June 2005

Respiratory‐chain deficiencies have long been regarded as neuromuscular diseases. In fact, oxidative phosphorylation, i.e. adenosine triphosphate (ATP) synthesis by the respiratory chain, does not occur only in the neuromuscular system. Indeed, a number of non‐neuromuscular organs and tissues are dependent upon mitochondrial energy supply. For this reason, a respiratory‐chain deficiency can theoretically give rise to any symptom, in any organ or tissue, at any age and with any mode of inheritance, owing to the twofold genetic origin of respiratory enzymes (nuclear DNA and mitochondrial DNA, mtDNA). In recent years, it has become increasingly clear that genetic defects of oxidative phosphorylation account for a large variety of clinical symptoms in childhood. Among 100 patients with respiratory‐chain deficiencies identified in our centre, 56% presented with a non‐neuromuscular symptom and 44% were referred for a neuromuscular problem. It appears that the diagnosis of a respiratory‐chain deficiency is difficult initially when only one symptom is present. In contrast, this diagnosis is easier to consider when two seemingly unrelated symptoms are observed
5,10‐Methylenetetrahydrofolate reductase deficiency. Clinical and biochemical features of a further case
Authors: Haan E. A. , Rogers J. G. , Lewis G. P. , Rowe P. B.
Published: June 2005

We report the case of a boy with 5,10‐methylenetetrahydrofolate reductase deficiency. The clinical features consisted of severe mental retardation, spasticity and seizures remaining static to 7 years of age followed by a phase of rapid deterioration and death at 7 1/2 years of age. The main biochemical findings were homocystinaemia, homocystinuria, a normal methionine level in plasma and cerebrospinal fluid, an increased excretion of methionine in urine and a very low level of folate in the cerebrospinal fluid. The activity of 5,10‐methylenetetrahydrofolate reductase was greatly reduced in the patient's lymphocytes and liver.
Excessive excretion of β‐alanine and of 3‐hydroxypropionic,R‐ andS‐3‐aminoisobutyric,R‐ andS‐3‐hydroxyisobutyric andS‐2‐(hydroxymethyl)butyric acids probably due to a defect in the metabolism of the corresponding malonic semialdehydes
Authors: Pollitt R. J. , Green A. , Smith R.
Published: June 2005

A new metabolic disorder characterised by the excessive excretion of β‐alanine, 3‐hydroxypropionic acid,R‐andS‐3‐amino‐ and 3‐hydroxyisobutyric acids andS‐2‐(hydroxymethyl)butyric acid is probably due to deficient activities of malonic, methylmalonic and ethylmalonic semialdehyde dehydrogenases. These dehydrogenation reactions could be mediated by one enzyme, or by enzymes with a common subunit, and bothR‐ andS‐methylmalonic semialdehydes seem to be equally affected. The patient is now aged 4 years and has developed normally. He has a persistent gross hypermethioninaemia which is probably unrelated to the other biochemical abnormalities.
Dolichol metabolism in cultured skin fibroblasts from patients with “neuronal” ceroid lipofuscinosis (Batten's disease)
Authors: Paton B. C. , Poulos A.
Published: June 2005

Dolichol metabolism was investigated in skin fibroblast cultures from normal individuals and patients with Batten's disease. Incorporation of [3H]mevalonolactone and [14C]acetate into the dolichol fraction of total lipid extracts was similar in cells from normal individuals and patients with Batten's disease. [14C]Acetate incorporation into dolichol in non‐saponifiable lipid extracts was compared with incorporation into dolichol in total lipid extracts, and no difference in the proportion of dolichol esterified to fatty acids was found in Batten's cells as compared to normal cells. The rate of loss of radioactivity from the dolichol pool after prelabelling with [14C]acetate was also similar in cells from Batten's and normal individuals. Thus, in the fibroblast system used, no evidence was found to support the hypothesis that Batten's disease is due to a defect in dolichol metabolism.
Immunochemical study of ornithine transcarbamylase deficiency
Authors: Kodama H. , Nagayama H. , Shimoizumi H. , Okabe I. , Kamoshita S.
Published: June 2005

Deficiency of ornithine transcarbamylase (OTC, EC 2.1.3.3) (McKusick 31125), a mitochondrial enzyme of the urea cycle, appears to be one of the most frequent causes of inherited ammonia intoxication, and evidence indicates that the enzyme is X‐linked. Recently, different kinds of mutation have been reported (Cathelineauet al., 1972; Briandet al., 1982). We carried out detailed studies on the molecular properties of the enzyme from two heterozygous females, using an antibody to bovine OTC which cross‐reacted with the human enzyme.