bluuebunny/crossref_metadata_2025 huggingface dataset:
Abnormal expression of MHC class II and ICAM‐1 by melanocytes in vitiligo
Authors: Al Badri Adnan M. T. , Foulis Alan K. , Todd Pamela M. , Garioch Jennifer J. , Gudgeon Janet E. , Stewart D. Graeme , Gracie J. Alastair , Goudie Robert B.
Published: June 2005

The relationship between damage to cutaneous melanocytes and antimelanocyte autoimmunity in vitiligo is unclear. We have demonstrated abnormal expression of MHC class II molecules by perilesional melanocytes in 13/21 patients with vitiligo and a six‐fold increase in the number expressing the intercellular adhesion molecule ICAM‐1. These molecules have important roles in normal antigen presentation and activation of helper T lymphocytes, and their expression by melanocytes may contribute to the abnormal immune response in vitiligo. MHC class II is not expressed by melanocytes in psoriasis and is unlikely to be induced in vitiligo by cytokines released from activated non‐melanocyte‐specific T lymphocytes.
Ultrastructural features of fetal erythroid precursors infected with parvovirus B19 in vitro: Evidence of cell death by apoptosis
Authors: Morey Adrienne L. , Ferguson David J. P. , Fleming Kenneth A.
Published: June 2005

Human parvovirus B19 cannot be cultured in standard cell lines and relatively little is known about the intracellular life‐cycle of the virus. In this study, ultrastructural features of B19 infection were examined using haemopoietic cell suspension cultures derived from human fetal liver. Erythroblasts from infected cultures frequently contained crystalline arrays of both full and empty virus‐like particles. The number and size of these arrays increased with the duration of culture, and their location changed from exclusively nuclear at 24 h post‐infection to both nuclear and cytoplasmic at 3 days post‐infection. Arrays were occasionally found in cytoplasmic protuberances which appeared to be pinching off from the cell. The location of the arrays corresponded to the distribution of viral capsid protein determined by immunolabelling at the light microscope level. Cells containing viral crystalline arrays also exhibited nucleolar degeneration, extreme margination of the nuclear heterochromatin, and cytoplasmic vacuolation. These features are typical of cells undergoing individual programmed cell death or ‘apoptosis’. The triggering of apoptosis in erythroid precursors by parvovirus B19 may help to explain the apparent lack of a strong inflammatory response to fetal B19 infection and may have implications for understanding the mechanisms of viral spread throughout the host.
AgNOR counts have no prognostic value in breast cancer
Authors: Toikkanen Sakari , Joensuuf Heikki
Published: June 2005

The prognostic significance of silver‐stained nucleolar organizer regions (AgNORs) was retrospectively studied among 230 patients with operable invasive breast cancer from a defined urban population. The number of AgNORs was evaluated from routinely processed paraffin sections by light microscopy at a total magnificiation of × 630. The number of AgNORs had no predictive value on the 8‐year survival rate corrected for intercurrent deaths (P= 0.2). A high number of AgNORs (<2.7, the median value) was not related to any other prognostic variable studied except a low S‐phase fraction measured by flow cytometry. AgNOR counting does not appear to be a useful prognostic variable in breast cancer.
Immunoglobulin heavy chain rearrangements in primary brain lymphomas. A study using PCR to amplify CDR‐III
Authors: Albrecht Steffen , Bruner Janet M. , Segall Gary K.
Published: June 2005

Primary brain lymphomas (PBLs) have only rarely been analysed for immunoglobulin heavy chain (IgH) rearrangements. In this study, DNA was extracted from paraffin blocks in 23 cases of PBL and examined for IgH rearrangements using the polymerase chain reaction (PCR) to amplify the complementarity‐determining region III (CDR‐III) of rearranged IgH genes. Fifteen of the cases were phenotyped on paraffin‐embedded tissue using a pan‐B and pan‐T antibody (L26 and UCHL‐1, respectively). The remaining eight cases were not phenotyped for lack of tissue. For comparison, we used DNA extracted from paraffin blocks of normal brain, lymph nodes with lymphoid hyperplasia, and non‐lymphoid malignancies. PCR products were examined by polyacrylamide gel electrophoresis. Among the ten B‐cell PBL; four had a pattern indicative of IgH rearrangement, one had a germline pattern, and five had no detectable PCR products. Among the five T‐cell PBLs, one had a germline pattern and four had no detectable products. Among the eight untyped PBLs, two had IgH rearrangement, four had a germline pattern, and two gave no detectable products. DNA from non‐lymphoid tissues gave a consistent germline pattern, while DNA from polyclonal lymphoid populations (lymph node) had a pattern of polyclonal IgH rearrangement. In a dilution study, a clonal rearrangement could be detected as long as the clone's DNA constituted at least 10 per cent of the total DNA. PCR to amplify CDR‐III can be successfully applied to DNA extracted from paraffin blocks, and it detected a clonal rearrangement in 50 per cent of cases that gave a detectable pattern. This allows clonality analysis of tissue unsuitable for conventional Southern blot analysis. Furthermore, B‐cell PBLs have IgH rearrangements similar to those of extracranial B‐cell neoplasms.
A novel monoclonal antibody (FUN‐1) identifies an activation antigen in cells of the B‐cell lineage and Reed—Sternberg cells
Authors: Nozawa Yoshihiro , Wachi Eiko , Tominaga Kunihiko , Abe Masafumi , Wakasa Haruki
Published: June 2005

The characterization of a new monoclonal antibody (MoAb) recognizing a human B‐cell activation antigen, designated FUN‐1, is described in this paper. Immunoprecipitation revealed that FUN‐1 recognizes an antigen with a molecular weight (MW) of 75 kD. FUN‐1 reacts with pokeweed mitogen‐activated B lymphocytes and monocytes of peripheral blood, but not with unstimulated lymphocytes or granulocytes. It also reacts with large lymphoid cells in germinal centres, Epstein–Barr virus‐transformed B cell lines, large B‐cell lymphomas, Ki‐1‐positive anaplastic largecell lymphomas, and Reed–Sternberg cells of Hodgkin's disease, but not with Concanavalin A‐activated T cells, acute lymphoblastic leukaemias, T‐cell lymphomas, or low‐grade B‐cell leukaemias. These findings indicate that FUN‐1 recognizes a previously unreported B‐cell activation antigen. This MoAb appears to be useful for the study of maturation and differentiation in the B‐cell lineage as well as for the immunohistochemical diagnosis of B‐cell lymphomas and Hodgkin's disease.
Immunoreactive transforming growth factor alpha and epidermal growth factor in oral squamous cell carcinomas
Authors: Christensen Maria E. , Therkildsen Marianne H. , Poulsen Steen S. , Bretlau Poul
Published: June 2005

Forty oral squamous cell carcinomas have been investigated immunohistochemically for the presence of transforming growth factor alpha (TGF‐α) and epidermal growth factor (EGF). The same cases were recently characterized for the expression of EGF‐receptors. TGF‐α was detected with a monoclonal mouse antibody and EGF with polyclonal rabbit antiserum. Thirty‐five of the tumours were positive for TGF‐α and 26 of the tumours for EGF. None of the poorly differentiated tumours was positive for EGF, but they all were for TGF‐α. In sections including normal differentiated oral mucosa, the cells above the basal cell layer were positive for both TGF‐α and EGF. The same staining pattern was observed in oral mucosa obtained from healthy persons. In moderately to well differentiated carcinomas, the immunoreactivity was mainly confined to the cytologically more differentiated cells, thus paralleling the situation observed in the normal differentiated oral mucosa. In four cases, material was available from both a primary tumour and a metastasis. Three of these were positive for TGF‐α and EGF with the same staining pattern as that of the primary tumours. This investigation together with our previous results confirms the existence of TGF‐α, EGF, and EGF‐receptors in the majority of oral squamous cell carcinomas and their metastases.
The expression of the trefoil peptides pS2 and human spasmolytic polypeptide (hSP) in ‘gastric metaplasia’ of the proximal duodenum: Implications for the nature of ‘gastric metaplasia’
Authors: Hanby Andrew M. , Poulsom Richard , Elia George , Singh Sukhdev , Longcroft Janet M. , Wright Nicholas A.
Published: June 2005

Expression of pS2 protein (an oestrogen‐induced gene discovered in the MCF‐7 breast carcinoma cell line) and its homologue human spasmolytic polypeptide (hSP) was analysed, using immunohistochemistry and in situ hybridization to their mRNAs, in the proximal duodenum of 17 partial gastrectomy specimens removed from individuals with chronic peptic ulceration. Eight were found to have gastric‐type metaplasia. In gastric metaplasia, mRNAs for pS2 and hSP, and pS2 peptide antibody were co‐localized in the cells covering the duodenal villi, pS2 immunostaining was diffusely cytoplasmic in nature. A similar pattern was seen in Brunner's gland ducts. The trefoil peptide localization in gastric metaplasia closely resembles that seen in superficial gastric epithelium and the distal Brunner's gland duct, which in turn shares morphological similarities with gastric epithelium. We therefore conclude that gastric metaplasia may be the result of an expansion of the surface component of the Brunner's gland duct. The function of these trefoil peptides is at present unknown, but their distribution elsewhere suggests an involvement in reparative mechanisms. The similarities between gastric foveolar and Brunner's gland duct epithelium may derive from common restitution‐enhancing features pertinent to a locally harsh environment.
Cartilage and bone formation in repairing achilles tendons within diffusion chambers: Evidence for tendon–cartilage and cartilage–bone conversionin vivo
Authors: Rooney Paul , Walker Duncan , Grant Michael E. , McClure John
Published: June 2005

Rodent Achilles tendons were subjected to midpoint tenotomy and allowed to recover for various times in situ before the operated tissue was removed, placed into a Millipore diffusion chamber, and inserted intraperitoneally into syngeneic hosts. Diffusion chambers were then removed at weekly intervals, such that the total time after the operation (i.e., time allowed to recover in situ plus time within the diffusion chamber) was up to 8 weeks, and examined histologically. Ectopic cartilage was produced within the diffusion chamber after a total of 4 weeks but only if the first 2 weeks of recovery were in situ. With increasing time, calcified cartilage, osteoid and bone were also observed. Overall, the evidence suggests that the cartilage forms via a direct conversion from tendon tissue and that the bone may form as a result of differentiative changes of hypertrophic chondrocytes.
Comparison between the monoclonal antibodies Ki‐67 and PC 10 in 125 malignant lymphomas
Authors: Sabattini Elena , Gerdes Johannes , Gherlinzoni Filippo , Poggi Simonetta , Zucchini Luca , Origioni Giuseppe Meliell Francesco , Del Vecchio Maria T. , Leoncini Lorenzo , Falini Brunangelo , Pileri Stefano A.
Published: June 2005

The monoclonal antibody (MAb) Ki‐67 detects a nuclear proliferation‐associated antigen which corresponds to a non‐histone protein with a molecular weight of 395 and 345 kD. Its prognostic relevance has been assessed in both lymphoid and non‐lymphoid tumours. The MAb PC10 picks up the proliferating cell nuclear antigen (PCNA), which is a 36 kD nuclear protein associated with the cell cycle Whereas Ki‐67 works only in fresh material. PC10 detects a fixation‐resistant epitope of PCNA. Preliminary data have revealed a linear relationship between Ki‐67 and PC 10 reactivity in normal lymphoid tissue and in non‐Hodgkin's lymphomas (NHLs). We applied Ki‐67 and PC 10 to frozen and routine sections, respectively, from 25 examples of Hodgkin's disease (HD) (14 nodular sclerosis, 6 lymphocyte predominance, 5 mixed cellularity) and 100 NHLs (corresponding to the main varieties of the updated Kiel classification). The results obtained can be summarized as follows: (1) both MAbs gave rise to extremely variable results within the same category of NHLs; (2) most Hodgkin and Reed‐Stern berg cells (50‐98 per cent) were labelled by the reagents; (3) Ki‐67 and PC10 stained a similar ratio of neoplastic cells in 65 and 76 per cent of NHL and HD cases, respectively; in the remaining instances, no correspondence was observed, the PC10‐positive elements usually outnumbering the Ki‐67‐positive ones significantly. These discrepancies, which might be due to low PCNA catabolism and/or PCNA expression by quiescent cells, underline the need for further kinetic and clinico‐pathologic studies in order to define the specific relevance of PC 10.
Molecular mechanisms involved in intraepithelial lymphocyte migration: A comparative study in skin and tonsil
Authors: Uccini Stefania , Ruco Luigi P. , Monardo Francesca , La Parola Ilaria Lesnoni , Cerimele Decio , Baroni Carlo D.
Published: June 2005

Intraepithelial lymphocyte migration is a biological process frequently observed in skin and tonsil. Using immuno‐histochemistry, we have studied the molecular bases of this process in seven skin biopsies involved by mycosis fungoides (MF) and in 12 tonsils, four involved by B‐chronic lymphocytic leukaemia (B‐CLL) and eight by lymphoid follicular hyperplasia (LH). In the skin, intraepidermal T‐lymphocyte infiltration was associated with narrowing and fragmentation of the basement membrane, as shown by an anti‐collagen type IV antibody. Immunostaining of serial sections with an anti‐collagenase type IV antibody revealed that collagenase type IV was localized in the upper dermis and Strictly co‐distributed with collagen type IV, suggesting that enzymatic digestion played a role in the alterations of the basement membrane. Further migration through the epidermis was mediated by expression on keratinocytes of intercellular adhesion molecule‐1 (ICAM‐1) and of leukocyte‐function associated antigen‐1 (LFA‐1) on infiltrating lymphocytes. In the tonsil, intraepithelial infiltration was mediated by the expression of vascular cell adhesion molecule‐1 (VCAM‐1) by epithelial cells and of very late antigen‐4 (VLA‐4) by infiltrating lymphocytes. Further intraepithelial lymphocyte migration was then established, as already shown in the skin, by ICAM‐1/LFA‐1 interaction. Lymphocyte recruitment from the systemic circulation was studied using antibodies directed against endothelial leukocyte adhesion molecule‐1 (ELAM‐1), ICAM‐1, and VCAM‐1. These adhesion molecules were highly expressed by blood vessels in the upper dermis of MF and the percentage of ELAM‐1 +/VCAM‐1 + vessels was significantly higher than that observed in tonsils. Our data suggest that distinct molecular mechanisms are used by lymphocytes in intraepithelial migration in the skin and in tonsils.
Immunohistochemical staining of ras oncogene product in neoplastic and non‐neoplastic mesothelial tissues: Immunoreactivity for N‐ras and lack of immunohistochemical staining for Ha‐ras and K‐ras
Authors: Ramael Marc , Deblier Ivo , Eerdekens Christa , Lemmens Greet , Jacobs Werner , Van Marck Eric
Published: June 2005

Immunohistochemical staining of 36 malignant mesothcliomas and 45 cases of non‐neoplastsic mesothelium including 20 specimens with signs of hyperplasia were investigated using murine monoclonal antibodies directed against p21 ras protein, Ha‐ras protein, K‐ras protein, and N‐ras protein. All cases of non‐neoplastic mesothelium and the majority of the malignant mesotheliomas (78 per cent) showed cytoplasmic and often submembranous immunoreactivity in more than 50 per cent of the cells with both the pan‐ras and N‐ras antibody. No immunoreactivity was observed for Ha‐ and K‐ras. There was no statistically significant difference with respect to immunoreactivity between neoplastic and non‐neoplastic mesothelium or between the various mesothelioma subtypes.
Metabolic requirements for phorbol ester‐induced cytoskeletal changes in renal epithelium
Authors: Rahilly Mave A. , Fleming Stewart
Published: June 2005

Alteration of the cytoskeleton and cell–substratum adhesion are important in the progression of renal carcinoma. We have previously shown that treatment of normal human renal epithelium with phorbol esters mimics the changes seen in renal carcinoma cells. In this study we have demonstrated that the phorbol ester‐induced cytoskeletal reorganization is inhibited in the presence of deoxyglucose but not by cycloheximide. We have also shown that treatment of cells with cytochalasin B results in the formation of immature stress fibres restricted to the cell–substratum contact regions. These results suggest that the actin filaments elongate from the focal contacts and that structural rearrangement caused by phorbol esters is an energy‐dependent phenomenon but is independent of de novo protein synthesis.
Induction of drug‐metabolizing enzymes in human pancreatic cancer and chronic pancreatitis
Authors: Foster John R. , Idle Jeffrey R. , Hardwick James P. , Bars Remi , Scott Patrick , Braganza Joan M.
Published: June 2005

Chronic pancreatitis and pancreatic cancer have both been linked with occupational exposure to organic chemicals. These chemicals are known to be metabolized within the liver by the cytochrome P‐450 family of enzymes, and indeed are able to induce levels of these enzymes as evidence of their interaction. The purpose of this study was therefore to see if these enzyme systems were altered in chronic pancreatitis and pancreatic cancer. Immunocytochemistry of four phase I drug‐metabolizing enzymes (cytoxhromes P‐450 IIIA1, P‐450IIE, P‐450 IA2, and NADPH cytochrome P‐450 oxido‐reductase) and one phase II enzyme [glutathione S‐transferase (GST) 5‐5] was therefore performed on pancreas and/or liver biopsy samples from organ donors and compared with patients with chronic pancreatitis or pancreatic cancer. In samples from donor subjects, the types and levels of drug‐metabolizing enzymes in hepatocytes were similar to those seen in pancreatic acinar cells. In material from patients with chronic pancreatitis or pancreatic cancer, cytochrome P‐450 enzyme levels were greater in both the liver and the pancreas than those seen in the donor group, while GST levels were unchanged. Islets of Langerhans showed high levels of P‐450 IA2 in the donor group, with clear induction of P‐450 IIIA1 and NADPH cytochrome P‐450 oxidoreductase in patients with chronic pancreatitis but not in the pancreatic cancer group. Levels of GST 5‐5 were also induced in the islets. The present findings raise the possibility of an aetiological relationship between elevated levels of drug‐metabolizing enzymes and the subsequent development of disease.
Ultimobranchial body nests in human fetal thyroid: An autopsy, histological, and immunohistochemical study in relation to solid cell nests and mucoepidermoid carcinoma of the thyroid
Authors: Harach H. R. , Vujanić G. M. , Jasani B.
Published: June 2005

The mid portion of the lateral thyroid lobes from 64 fetuses was systematically analysed for the presence of ultimobranchial body nests. The nests showed a prevalence of 1 /24 (4.2 per cent) or 13/40 (32.5 per cent) depending on whether a single‐ or a multi‐step sectioning method was employed, respectively, and showed anatomical, morphological, and histochemical features similar to those of ultimobranchial postnatal thyroid solid cell nests. Histochemical studies revealed the presence of mucosubstances in 73 per cent of the cases, calcitonin‐immunoreactive cells in 36 per cent, and both carcinoembryonic antigen and high‐molecular‐weight cytokeratin‐immunoreactive epidermoid cells in 85.7 per cent, respectively. These findings indicate that these markers, which are also expressed by solid cell nests of the thyroid, are of value for the detection and tracing of ultimobranchial tissue to earlier stages of development. The findings of this study support the hypothesis that mucoepidermoid carcinomas of the thyroid are of ultimobranchial tissue origin.
In situ end‐labelling detects DNA strand breaks in apoptosis and other physiological and pathological states
Authors: Ansari B. , Coates P. J. , Greenstein B. D. , Hall P. A.
Published: June 2005

We have investigated the use of a novel technique, in situ end‐labelling, as a means of the specific identification of apoptotic cells in formalin‐fixed, paraffin‐processed tissue sections. The technique relies on the presence of DNA strand breaks in apoptotic cells, caused by activation of endogenous nuclease activity during the process of cell death. These strands are labelled with a non‐isotopic reporter molecule in the presence of a DNA polymerase, and labelled DNA is identified immunohistochemically. We show that in situ end‐labelling stains cells with the morphological characteristics of apoptosis, and greatly simplifies their identification. Furthermore, in two model systems, the number of labelled cells parallels the number of cells undergoing apoptosis as measured by alternative techniques. The ability of the Klenow fragment of DNA polymerase to label apoptotic nuclei suggests that the characteristic DNA fragmentation seen during this process involves the formation of DNA breaks with a 5′ overhang. in situ end‐labelling will be valuable for the identification and quantitation of apoptosis in a range of normal tissues and in a variety of pathological states. However, the technique is not specific for programmed cell death, and results must be interpreted with caution and correlated with morphological criteria of apoptosis.
Sequencing and quantitative assessment of mutant and wild‐type mitochondrial DNA in paraffin sections from cases of MELAS
Authors: Love S. , Nicoll J. A. R. , Kinrade E.
Published: June 2005

MELAS (mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke‐like episodes) is a clinically devastating disease of children and young adults. The cause of the stroke‐like episodes is not known. We have sequenced the mitochondrial DNA (mtDNA) in archival paraffin‐embedded material from two cases. In only one of these did the mitochondrial tRNALeu(UUR) gene contain the nucleotide 3243 A‐to‐G mutation that is most commonly responsible for MELAS. In this case, we determined the relative proportion of mutant:wild‐type mtDNA in sections of the central nervous system and other tissues by PCR amplification, PalI digestion, DNA electrophoresis, and scanning densitometry of the ethidium bromide‐stained gels. The technique allowed the proportion of mitochondria that contain the mutant genome to be compared with the histological findings in immediately adjacent sections of tissue. The mutant mtDNA was detectable in most tissues, the percentage of mtDNA ranging from barely detectable levels to 78 percent. The relative amount of mutant mtDNA correlated poorly with the distribution of histological lesions, both within the central nervous system and in other tissues examined. The proportion was high in tissues such as liver, kidney, adrenal, and pancreas that appeared histologically normal. Relatively low levels were present in some regions of the central nervous system, such as the occipital lobe, which contained many of the characteristic infarct‐like lesions. These observations do not support previous speculation that the distribution of these lesions reflects that of the defective mitochondria. The results emphasize the usefulness of the polymerase chain reaction in correlative histogenetic studies.
An immunohistochemical study of the long‐term effects of androgen administration on female‐to‐male transsexual breast: A comparison with normal female breast and male breast showing gynaecomastia
Authors: Burgess Helen E. , Shousha Sami
Published: June 2005

This study was aimed at assessing the effects of therapeutic long‐term administration of androgens on normal human female breast. Sections from mastectomy specimens of 29 female‐to‐male transsexuals who had prolonged androgen administration prior to surgery were examined using routine light microscopy and immunohistochemical techniques. For comparison, sections from ten ‘normal’ female breast reduction mammoplasty specimens and ten cases of gynaeco‐mastia were similarly examined. Haematoxylin and eosin‐stained sections were assessed for the prevalence of elements of the normal breast and benign breast lesions. Immunoperoxidase techniques were performed to study the distribution of a variety of breast‐associated antigens and receptors. The results were assessed semi‐quantitatively.

The prevalence of normal acini and ducts, fibrosis, cysts, and apocrine metaplasia in transsexual specimens was not statistically different from that seen in normal controls. However, transsexual specimens had a significantly higher prevalence of microcalcification than normals. The majority of transsexual specimens were positive for gross cystic disease fluid protein‐15, lactoferrin, and progesterone and oestrogen receptors, and negative for B72.3 and pS2. These findings were not significantly different from those in normal controls.

All ten gynaecomastia specimens were positive for oestrogen and progesterone receptors. The prevalence of oestrogen receptors was significantly higher than that seen in transsexuals and normal controls, but the prevalence of progesterone receptors was only significantly higher than that seen in transsexuals.

It is concluded that long‐term androgen administration does not appear to have any significant lasting effect on the normal human female breast, as demonstrated by a wide range of histological and immunohistological criteria.
Increased expression of cytokeratins 8, 18 and vimentin in the invasion front of mucosal squamous cell carcinoma
Authors: Schaafsma H. E. , Velden L.‐A. Van Der , Manni J. J. , Peters H. , Link M. , Ruiter D. J. , Ramaekers F. C. S.
Published: June 2005

The immunohistochemical expression patterns of cytokeratins 8 and 18 and vimentin were examined in frozen sections of 120 human mucosal squamous cell carcinomas with special emphasis on the topological distribution in the tumour. This was done in order to evaluate in squamous cell carcinoma a particular expression pattern observed recently by us in transitional cell carcinoma of the urinary tract and designated as an ‘interface phenomenon’;. This phenomenon implying maximum expression of cytokeratins 8 and 18 at the tumour front, and to a lesser extent also in areas of intratumorous stroma contact, was also found in about 50 per cent of the squamous cell carcinomas examined. It was even found for vimentin, which contrasted with transitional cell carcinoma. The percentages of occurrence of the phenomenon varied for the different sites of origin of the tumour. Tumour grade did not influence the results. These findings further support the idea that invasive carcinoma cells interacting with the stromal micro‐environment display a characteristic intermediate filament phenotype that deviates from the pattern expected on the basis of their direction of differentiation. These changes might eflect phenotype involved in invasive, migrating, and proliferating activities.
Intrahepatic distribution of transforming growth factor‐alpha (TGFα) during liver regeneration following carbon tetrachlorideinduced necrosis
Authors: Burr Andrew W. , Carpenter Martyn R. , Hines Julie E. , Gullick William J. , Burt Alastair D.
Published: June 2005

The distribution of transforming growth factor‐alpha (TGFα) in rat liver during regeneration was studied immunohistochemically using two antibodies, one a polyclonal (26T) raised against a synthetic peptide corresponding to the 17 C‐terminal amino acids of the mature rat protein, and the other a monoclonal (Ab‐2) raised against recombinant human protein. In normal liver, immunoreactive TGFα was detected in perivenular hepatocytes using both antibodies. No sinusoidal cells were found to contain the peptide. In response to carbon tetrachloride (CCI4)‐induced necrosis, an initial increase in the intensity of immunoreactivity was noted at 24 h following exposure to the toxin. This coincided with the period immediately preceding the peak of hepatocyte proliferation; Ab‐2 immunoreactive cells outnumbered 26T‐positive cells. Thereafter there was a reduction in the number of TGFα‐positive cells, but by day 4 the level of immunoreactivity had returned to that of normal liver. Using bromodeoxyuridine labelling, spatial and temporal relationships between TGFα expression and cell proliferation were identified, supporting the concept that this peptide plays an important role in the in vivo regenerative response to hepatic injury via an autocrine mechanism.
Cellular and molecular aspects of hepatic fibrosis
Authors: Burt Alastair D.
Published: June 2005

Hepatic fibrosis, a consequence of most forms of chronic liver disease, is a dynamic process involving complex interactions between several cell types, the net result of which is accumulation of several distinct extracellular matrix (ECM) proteins. The resultant disruption of intrahepatic blood flow contributes to the development of portal hypertension. The effects, however, are not merely a space‐occupying phenomenon; by changing the composition of the ECM, fibrosis may also alter hepatocyte function via cellular integrins. The principal source of ECM proteins in normal and fibrotic liver is the perisinusoidal cells which lie in the space of Disse. The response of this cell population to acute and chronic liver injury has been studied in detail. Perisinusoidal cells proliferate and become activated following hepatocyte necrosis. This phenomenon is transient in acute injuries, but in chronic liver disease, continued activation is associated with phenotypic modulation of perisinusoidal cells to myofibroblasts. This process is mediated by various cytokines including TGF‐β and PDGF. Some of the growth factors involved are derived from activated Kupffer cells and there is evidence of a complex interplay between mediators; injured sinusoidal endothelial cells and platelets are possible additional sources. Accumulation of ECM proteins in fibrosis can be explained not only by increased synthesis, but also by decreased degradation. There is growing evidence that in fibrotic liver there is decreased interstitial collagenase activity. This is, at least in part, due to expression of a tissue inhibitor of metalloproteinase, TIMP‐1, by activated perisinusoidal cells.
Monoclonal antibody 4C7 recognizes an endothelial basement membrane component that is selectively expressed in capillaries of lymphoid follicles
Authors: Jaspars Lies H. , Van Linden Hans C. Der , Scheffer George L. , Scheper Rik J. , Meijer Chris J. L. M.
Published: June 2005

In order to define compartment‐related structures within the extracellular matrix of human lymphoid organs, monoclonal antibodies (MAbs) were generated by immunizing mice with stromal fragments of human tonsils. One MAb (4C7) was selected which recognized an endothelial basal membrane component that is selectively expressed in capillaries of lymphoid follicles. The epitope was also present in follicles within chronically inflamed synovial membrane and in a hyperplastic thymus of a patient with myasthenia gravis. B‐cell non‐Hodgkin's lymphomas with a follicular growth pattern expressed the antigen in neoplastic follicles, whereas diffuse growing lymphomas lacked the antigen. The restricted distribution pattern suggests involvement of the 4C7‐defined antigen in the organization of the follicular compartment within human lymphoid tissue.
Presence of Epstein‐Barr virus harbouring small and intermediate‐sized cells in Hodgkin'S disease. Is there a relationship with Reed‐Sternberg cells?
Authors: Jiwa N. M. , Kanavaros P. , De Bruin P. C. , Valk P. Van Der , Horstman A. , Vos W. , Mullink H. , Walboomers J. M. M. , Meijer C. J. L. M.
Published: June 2005

Forty‐four cases of Hodgkin's disease (HD), mostly of the nodular sclerosing type, were investigated for the presence of Epstein‐Barr virus (EBV) by polymerase chain reaction (PCR) and DNA and RNA in situ hybridization (DISH, RISH), as well as by immunohistochemistry for the detection of latent membrane protein‐1 (LMP‐1) of EBV. in situ hybridization (ISH) was combined with immunohistochemistry to correlate the presence and activity of the virus at the cellular level. In 18/34 (53 per cent) cases, EBV‐DNA sequences could be detected with the PCR method. In 12/18 positive cases, DISH and RISH were also positive. In the remaining six EBV‐PCR positive cases, two were also positive with RISH and LMP‐1, whereas no positive signal with DISH could be obtained. All DISH and/or RISH positive cases were also positive for LMP‐1. With RISH, not only the Reed‐Sternberg cells and their mononuclear variants (RS cells) stained positive, but also small and intermediate cells frequently reacted with the EBV‐specific probes (EBER‐1 and ‐2). Double staining with cellular markers (CD3, CD20, CD45, CD45RO, CD68, and the lectin PNA) revealed that most of the smaller EBER‐positive cells frequently did not express T, B, or histiocytic markers, but that they, as well as the RS cells, showed cytoplasmic and membranous staining with PNA. These smaller EBER‐positive cells were not found in EBV‐PCR negative HD. EBER‐positive RS cells were almost always LMP‐1 positive, as well as a substantial proportion of the intermediate‐sized cells, whereas the majority of the small EBER‐positive cells remained LMP‐1 negative. In EBV‐PCR positive non‐malignant lymph nodes, only a few EBER‐1 and ‐2 positive cells could be observed. As in infectious mononucleosis, these cells frequently expressed the B‐cell marker CD20. Although we cannot exclude the fact that the majority of the smaller EBV‐positive cells in HD belong to reactive EBV‐infected lymphocytes, our data favour the hypothesis that at least some of these smaller cells may belong to the reservoir of neoplastic cells in HD.
Reciprocal expression of CD34 and cell adhesion molecule ELAM‐1 on vascular endothelium in acute cutaneous graft‐versus‐host disease
Authors: Norton J. , Sloane J. P. , Delia D. , Greaves M. F.
Published: June 2005

The expression of CD34 and endothelial leucocyte adhesion molecule‐1 (ELAM‐1) on endothelial cells was studied in skin biopsies from normal human donors and recipients of allogeneic bone marrow. Both molecules were demonstrable on a variable number of cells in normals with no significant change after marrow transplantation in patients with no clinical evidence of skin disease. In patients with graft‐versus‐host disease (GvHD), however, there was a striking decrease in CD34‐positive cells with a corresponding increase in ELAM‐1 positivity. These changes were similar to those occurring in cultured endothelial cells after the addition of certain cytokines. They were seen only in the presence of a lymphocytic infiltrate and were not observed in the early stages of GvHD before lymphocytic infiltration was discernible. The reciprocal expression of these molecules may thus be important in the adhesion of lymphocytes to endothelium and their entry into the skin in GvHD and may be modulated by local cytokine release.
Proliferation of reactive and neoplastic human tissue mast cells. An immunohistochemical study using the antibody PC10 (anti‐PCNA)
Authors: Horny Hans‐Peter , Schumacher Udo , McCullagh Paul , Wehrmann Manfred , Rochf William R. , Kaiserling Edwin
Published: June 2005

Studies on the proliferative compartment of human tissue mast cells (MCs) and their tumours (mastocytosis) have not been performed. We have used the monoclonal antibody PC 10 to study MCs in reactive or hyperplastic states (chronic non‐specific lymphadenitis, n = 10; benign and malignant solid tumours, n = 5) and in the various subtypes of mastocytosis (urticaria pigmentosa, n = 22; solitary mastocytoma of the skin, n = 7; systemic mastocytosis; n = 8; malignant mastocytosis, n = 4). The identification of PC10‐positive MC nuclei was achieved by double staining. We found no PC10‐positive MCs in reactive or hyperplastic states, or in 14 of 22 cases of urticaria pigmentosa. PC 10‐positive MCs could be identified in all other mastocytoses but mostly in very low numbers. The mean percentages of PC10‐positive MCs amounted to 0.5 in eight positive cases of urticaria pigmentosa, 1.2 in mastocytoma, 0.7 in sytemic mastocytosis, and 4.0 in malignant mastocytosis. The difference between the latter form of mastocytosis and each of the other subtypes proved to be significant (P<0.05). The very smali proliferative compartment in the cutaneous and sytemic variants of mastocytosis is in accord with their favourable prognosis Most of the patients with systemic mastocytosis in the present study are all alive and well up to 12 years after diagnosis. In contrast, most of the patients with malignant mastocytosis died within 1 year of diagnosis.
Patterns and composition of basement membranes in colon adenomas and adenocarcinomas
Authors: Visser Robbert , Arends Jan W. , Leigh Irene M. , Bosman Fred T.
Published: June 2005

We studied the distribution of type IV collagen and type VII collagen in the basement membranes of normal mucosa of the colon, adenomas, and adenocarcinomas using immunoperoxidase and immunofluorescence techniques. In normal mucosa, we found regular type IV collagen‐positive basement membranes, lining vascular structures and mucosal epithelia. These basement membranes, however, lacked type VII collagen. In adenomas of the colon, intact basement membranes were observed through type IV collagen staining. Type VII collagen staining was also detected, but only in connection with dysplastic epithelium. Adjacent to the dysplastic epithelium in adenomas, histologically normal epithelium also showed type VII collagen staining along the basement membrane, but this was restricted to the epithelium of the luminal surface. These areas were also investigated for expression of keratins 8, 18, and 19, and keratins 5 and 8 (monoclonal antibodies NCL‐5D3 and RCK 102, respectively), but altered differentiation was not detected using this technique. In adenocarcinomas of the colon, type IV collagen was irregularly deposited in the basement membrane of neoplastic tubules. Type VII collagen staining was detected only in well or moderately differentiated carcinomas and in higher amounts. Our findings therefore reveal a transient expression of type VII collagen in the transition of dysplastic epithelium into carcinoma, suggesting the involvement of type VII collagen in the process of early invasion.
Synthesis and degradation of basement membranes in benign and malignant salivary gland tumours. A study by in situ hybridization
Authors: Soini Ylermi , Autio‐Harmainen Helena
Published: June 2005

In this study we investigated the mRNA expressions of 72 kD and 92 kD type IV collagenases, al (IV) chain of type IV collagen, and laminin Bl chain mRNAs in a set of malignant and benign salivary gland tumours and compared the results with non‐neoplastic salivary gland tissue. While only a few cases expressed 72 kD type IV collagenase mRNA or a1 (IV) chain of type IV collagen mRNA in tumour cells, 92 kD type IV collagenase and laminin mRNA synthesis could be seen in the neoplastic cells of many tumours. Stromal fibroblasts or endothelial cells demonstrated mRNA synthesis for all these proteins to a variable degree except for Warthin's tumours, in which no synthetic activity for any of the proteins could be seen. Since signals for 92 kD type IV collagenase mRNA could be seen in non‐neoplastic epithelial cells of the salivary gland, the synthesis of 92 kD type IV collagenase by tumour cells can be regarded as an intrinsic property of salivary gland epithelial cells. The pattern of mRNA synthesis for 72 kD and 92 kD type IV collagenases follows that observed in other tumours, in which the stromal cells also mainly synthesize 72 kD type IV collagenase while epithelial tumour cells more readily express 92 kD type IV collagenase mRNA. The synthesis of type IV collagenases by malignant tumours has been suggested to be of crucial importance for invasion and metastasis. The expression of both collagenases in benign tumours, such as pleomorphic adenomas, indicates that more complex mechanisms, such as disturbances in the regulation of enzyme activity, are involved in the spread and invasion of salivary gland tumours than merely the synthesis of 72 kD and 92 kD collagenases.
The specificity of integrin‐ligand interactions in cultured human renal epithelium
Authors: Rahilly Maeve A. , Fleming Stewart
Published: June 2005

Members of the β1 integrin family are present at the basolateral membrane of human renal tubuiar epithelium in vivo and at the ventral surfaces of cultured renal epithelial cells, at‐the sites appropriate for cell substratum adhesion. In this study we have proven that these molecules are indeed functional in mediating cell substratum attachment in normal human renal epithelium by using monoclonal antibodies to integrin α subunits to block initial cell attachment. The importance of arginine‐glycine‐aspartic acid (RGD) recognition by cell surface receptors in various extracellular ligands was also examined using synthetic peptides. RGDS peptide strongly inhibited attachment to plain plastic or fibronectin‐coated substrata but had no effect on cell adhesion to laminin‐coated coverslips.
Transforming growth factor alpha and CD68 immunoreactivity in giant cell tumours of bone: A study on the nature of stromal and giant cells, and their interrelations
Authors: Hasegawa Tadashi , Hirose Takanori , Seki Kunihiko , Sano Toshiaki , Hizawa Kazuo
Published: June 2005

To clarify the nature of neoplastic cells, 17 giant cell tumours of bone were studied histologically and immunohisto‐chemically. L1 antigen and S‐100 protein were not detected in the tumour giant cells and stromal cells, although present in non‐neoplastic macrophages. The giant cells in all the lesions, some stromal cells, and osteoclasts in the normal bone showed CD68 and transforming growth factor alpha (TGFα) immunoreactivity. Fibrohistiocytic antigen, factor XIIIa, was expressed in large numbers of stromal cells in all lesions. Some stromal cells expressed α‐smooth muscle actin and osteocalcin. These immunohistochemical results suggested that the stromal cells of giant cell tumours of bone showed histiocytic and occasional myofibroblastic and osteoblastic differentiation. Proliferating cell nuclear antigen was demonstrated in the nuclei of the stromal cells only, indicating that these were the sole proliferating elements. TGFα produced by the giant cells and some stromal cells may play a role as a mediator for the attraction and/or proliferation of the precursor cells, and may suppress the activity of osteoblastic stromal cells, resulting in restricted bone formation in giant cell tumours.
Immunohistochemical comparison of cutaneous histiocytoses and related skin disorders: Diagnostic and histogenetic relevance of MS‐1 high molecular weight protein expression
Authors: Goerdt Sergij , Kolde Gerhard , Bonsmann Gisela , Hamann Kathrin , Czarnetzki Beate , Andreesen Reinhard , Luger Thomas , Sorg Clemens
Published: June 2005

Twenty‐nine cases of Langerhans cell histiocytosis (LCH), non‐Langerhans cell histiocytoses (N‐LCH), noninfectious granulomas, and fibroblast‐related lesions were examined with a panel of monoclonal and polyclonal antibodies on freshly frozen tissue sections to characterize the macrophage phenotype of N‐LCH syndromes. MS‐1 high molecular weight extracellular protein, specific for sinusoidal endothelial cells and dendritic perivascular macrophages in normal human organs, was expressed by N‐LCH cells but was not found in LCH cells, epithelioid cells in sarcoidosis, or palisading histiocytes in granuloma annulare. The subcellular location of MS‐1 protein, i.e., cytoplasmic vs. peripheral/extracellular, allowed discrimination of small and large (foamy or multinucleated) N‐LCH cells. MS‐1‐positive cells, which were found intermingled in cellular dermatofibromas but not in fibrous dermatofibromas, differed from MS‐1‐positive N‐LCH cells by their dendritic morphology, and thus rather resembled their normal dermal counterparts. A preserved functional relationship of these two MS‐1‐positive cell types was indicated by the fact that N‐LCH and cellular dermatofibromas were the only lesions found to be highly vascularized. As expected, CD1a showed high specificity for LCH, while CD34 was predominantly expressed by fibroblast‐related lesions; in cellular dermatofibromas, CD34 and MS‐1 expression partially overlapped. The other antigens tested showed non‐specific or overlapping patterns of expression. In conclusion, assessment of MS‐1 protein expression (in addition to assessment of CD1a and CD34) promises to be of diagnostic value in the discrimination of N‐LCH from related skin disorders, and it may indicate a common differentiative pathway for most N‐LCH disease entities.
The behaviour of liposarcoma in tissue culture. A specific growth pattern
Authors: Tardio Juan , Escalona‐zapata Julio , Diez‐nau D. Dolores
Published: June 2005

There have been few studies of liposarcoma in tissue culture. We report a study of 27 cases of liposarcoma including examples of all subtypes in tissue culture. All the cases showed a uniform growth pattern of glass adherence by polygonal cells with short processes distributed in a random fashion. The cytoplasm of these cells became progressively loaded with glycogen followed by lipid droplets. These lipid droplets tended to fuse progressively to form a single or dominant vacuole. At the same time, the cells tended to adopt a rounded shape. This pattern of growth, which was seen in all subtypes of liposarcoma with minimal variation, is different from that shown by other soft tissue tumours and is similar to the growth of embryonal fat in tissue culture.
Cathepsin D immunocytochemical assays in breast carcinomas: Image analysis and correlation to prognostic factors
Authors: Charpin Colette , Devictor Bénédicte , Bonnier Pascal , Andrac Lucile , Lavaut Marie Noëlle , Allasia Claude , Piana Lucien
Published: June 2005

Immunocytochemical assays of cathepsin D were assessed in a series of breast carcinomas (n=257) using monoclonal M1G8 anti‐total cathepsin D and the avidin‐biotin‐peroxidase complex. Cathepsin immunoreactivity was compared in frozen and paraffin sections. All tumours were anti‐cathepsin‐positive. Positive staining was observed in carcinoma and stromal cells and in the extracellular matrix. The amount of immunodetectable cathepsin in tissue was measured by computer‐assisted image analysis (SAMBA 2005). Both the percentage of immunostained tumour surface and the mean optical densities were processed as continuous variables for statistical analysis and correlated with prognostic factors. It was shown that cathepsin D was independent of the tumour size, the lymph node status, hormone receptors, and pHER‐2/neu overexpression. Cathepsin was significantly correlated with anti‐EGFR (P=0.012) and Ki67 (P=0.002) immunoreactivity, tumour grade (P=0.032), vascular invasion (P=0.0081), proliferation index (P=0.0045), and, to a lesser extent with AgNORs (P=0.0504) and the degree of hyperploidy (P=0.057). Tissue fixation and paraffin embedding significantly decreased cathepsin immunoreactivity. These results show that cathepsin D is not a totally independent prognostic factor in breast carcinomas.