bluuebunny/crossref_metadata_2025 huggingface dataset:
The effect of cyclosporin a on cationized bovine serum albumin‐induced nephropathy in NZW rabbits
Authors: Bass Paul S. , Wang Yiming , Nawab Mashal Al , Evans Beryl , Thomas Hywel , Davies David R.
Published: June 2005

The effect of cyclosporin A (CyA) was studied on the morphology and protein excretion of a rabbit chronic serum sickness nephritis using cationized bovine serum albumin (cBSA). One group of rabbits was given intravenous (i.v.) immunizing doses of cBSA and Escherichia coli endotoxin. One week later, these animals began a 6‐week i.v. injection schedule of cBSA only. A second group followed the same injection protocol, but was given intramuscular (i.m.) CyA for 3 days prior to the immunizing dose of cBSA/endotoxin and throughout the subsequent cBSA schedule. A third group was given i.m. CyA only. Regular blood samples for CyA levels were taken from animals given the drug. Two 24‐h urine samples were obtained from all animals in the study. Analysis of the blood samples showed that immunosuppressive levels of CyA were achieved after two i.m. doses of CyA. These levels were maintained during the course of the schedule. Morphologically, all rabbits completing the cBSA only injection schedule showed evidence of an immunemediated glomerulopathy with variably severe membranous and endocapillary proliferative change. Less than half the rabbits in the cBSA/CyA group showed any evidence of membranous change. The glomeruli of animals given CyA only were normal. No morphological evidence of CyA toxicity was seen in any animal given the drug. The proteinuria profiles, however, suggested that as well as reducing protein excretion in rabbits given cBSA, CyA may interact with the immunizing dose of cBSA to produce an early, reversible, nephrotoxic effect. These results show that (a) immunosuppressive levels of CyA can be efficiently obtained and maintained by the i.m. route, (b) commencing CyA treatment before initiating cBSA injections ameliorates cBSA glomerulopathy in the majority of rabbits tested, and (c) using this protocol, no significant CyA‐induced morphological effects are seen.
Diabetic glomerulosclerosis—immunogold ultrastructural studies on the glomerular distribution of type IV collagen and heparan sulphate proteoglycan
Authors: Woodrow David , Moss Jill , Shore Ian , Spiro Robert G.
Published: June 2005

We have undertaken an ultrastructural immunogold investigation of the distribution of type IV collagen and heparan sulphate proteoglycan (HSPG) in glomeruli from the kidneys of one normal control and three patients with diabetes mellitus and proteinuria. The sample included both diffuse and nodular diabetic glomerulosclerosis. In the control and diabetic kidneys, the type IV collagen was present predominantly on the endothelial aspect of the glomerular basement membrane (GBM), and by contrast the HSPG was found mainly on the epithelial side. In the mesangium in both control and diabetic glomeruli, type IV collagen was found predominantly in the central regions, while HSPG was mostly restricted to the region beneath the epithelial cells. Consequently, where there is a marked increase in mesangial matrix with nodule formation in diabetics there is a corresponding increase in the amount of type IV collagen but not of HSPG. Although the three diabetic patients were proteinuric, the HSPG was not decreased in the thickened GBMs.
The ultrastructure of pulmonary arteries and arterioles in emphysema
Authors: Smith Paul , Rodgers Brian , Heath Donald , Yacoub Magdi
Published: June 2005

The lungs from three cases of pulmonary emphysema obtained at heart‐lung transplantation were examined by electron microscopy to determine the origin of intimal longitudinal muscle and the formation of muscular tubes in small pulmonary arteries and arterioles. The earliest change consisted of migration of mature smooth muscle cells from the media of small pulmonary arteries, through gaps in the internal elastic lamina, into the subendothelial space. Most of these cells then adopted a longitudinal orientation, maintained a muscular phenotype, and became enmeshed in a web of elastic fibres. A small minority, immediately subjacent to the endothelium, were orientated circularly and, in some vessels, were enclosed by rudimentary internal and external elastic laminae to form early muscular tubes. Pulmonary arterioles, which are normally devoid of a media, contained several layers of circularly orientated smooth muscle cells, some of which also formed muscular tubes. It is postulated that the limited migration of mature smooth muscle cells seen in states of chronic hypoxia is mediated by a different stimulus from that causing the florid invasion of the intima by immature smooth muscle, with subsequent transformation into myofibroblasts, which characterize plexogenic pulmonary arteriopathy.
In situ detection of oxidative stress in rat hepatocytes
Authors: Mochida Satoshi , Masaki Naohiko , Ohta Yasuhiko , Matsui Atsushi , Ogata Itsuro , Fujiwara Kenji
Published: June 2005

In rat hepatocytes in primary culture incubated with nitro blue tetrazolium, formazan content was increased by addition of t‐butyl hydroperoxide, a potent oxidant, in a dose‐related manner, but not by addition of valinomycin, which kills hepatocytes through mitochondrial damage. This increment after t‐butyl hydroperoxide addition was not seen in hepatotyctes preincubated with deferoxamine mesylate, a ferric iron chelator which inhibits radical formation. Liver perfusion with nitro blue tetrazolium and t‐butyl hydroperoxide in rats produced formazan deposition faintly on the surface of hepatocytes throughout the liver and prominently in the cytoplasm of some hepatocytes, which was attenuated when performed following deferoxamine mesylate perfusion. When liver perfusion with nitro blue tetrazolium was performed in carbon tetrachloride‐intoxicated rats, formazan deposition appeared diffusely in hepatocytes in the centrilobular areas. Similar deposition was also observed on the surface and in the cytoplasm of hepatocytes in the periportal and mid‐zonal areas in rats undergoing post‐ischaemic reperfusion. Liver perfusion with nitro blue tetrazolium can detect in situ oxidative stress in hepatocytes and may be a useful tool for studying the role of lipid peroxidation in rat liver injury.
A study of adhesion molecules as markers of progression in malignant melanoma
Authors: Denton Karin J. , Stretch Jonathan R. , Gatter Kevin C. , Harris Adrian L.
Published: June 2005

Adhesion molecules are substances which are involved in the interactions between cells, and between cells and the extracellular matrix in both benign and malignant tissues. Two members of this group—intercellular adhesion molecule‐1 (ICAM‐l) and MUC18—have previously been found to be expressed on melanoma; however, studies seeking a correlation between expression and metaslatic behaviour have yielded conflicting results. In this study we investigated the expression of these two antigens and that of a number of other adhesion molecules [VCAM‐1, ELAM, and the neural cell adhesion molecule (NCAM)] on a range of benign and malignant melanocytic lesions. Both ICAM‐1 and MUC18 were found on a high percentage of all melanocytic lesions including benign naevi. VCAM‐1 was found to be expressed on 79 per cent of benign naevi, 62 per cent of primary melanomas less than 1‐5 mm in depth, and 6 per cent of thick primaries. The antigen was present on 14 per cent of lymph node metastases and on no extranodal deposits. This suggests that loss of melanoma cell adhesion mediated by VCAM‐1 may be important in the development of metastatic melanoma.
Immunohistochemical studies on the innervation of human transplanted liver
Authors: Dhillon Amar P. , Sankey Elizabeth A. , Wang Jiahua H. , Wightman A. Kay , Mathur Sandip , Burroughs Andrew K. , Scheuer Peter J.
Published: June 2005

The changing pattern of innervation in the human transplanted liver was studied from the day of transplantation to 5 years later. Seven liver biopsies from non‐transplant controls, 37 liver biopsies from 22 transplant patients, and one of these biopsied livers removed at retransplantation, were available for the study. Sections were immunostained for protein gene product 9.5 (PGP 9.5), neurone‐specific enolase (NSE), S‐100 protein, and vasoactive intestinal poly‐peptide. NSE and PGP 9.5 demonstrated nerves most successfully in our tissues. Staining for most small nerves was reduced by day 5 post‐transplantation. Scanty fine nerves could be detected from day 13 to day 241 in occasional biopsies. Consistently identifiable immunostaining of PGP 9.5 and NSE nerve fibres was again apparent in portal areas after this time in all but one case. The findings indicate that in transplanted liver limited reinnervation can eventually take place. This could be due to either proliferation of intrinsic nerves, or regrowth of extrinsic nerves, or both.
Dermatopathological aspects of restrictive dermopathy
Authors: Piérard‐franchimont Claudine , Piérard GéRald E. , Estrada Trinh Hermanns‐Lě Jorge Arrese , Verloes Alain , Mulliez N.
Published: June 2005

We present an immunopathological and electronmicroscopic study of the skin of two newborns affected by restrictive dermopathy. Evidence of abnormal maturation was found in the epidermis, cutaneous appendages, dermis, and hypodermis. Our observations confirm two previous descriptions. We emphasize some unreported data concerning the L1 antigen and Factor XIIIa in the skin. The L1 antigen is expressed in the interadnexal epidermis, but not in hair follicles. This is the reverse pattern compared with normal skin. Factor XIIIa is poorly expressed in dermal dendrocytes, which appear rare compared with controls. The multiple defects in maturation found in all cutaneous tissues suggest a qualitative or quantitative aberration in control mechanisms of tissue interactions.
Interstitial pneumonia in simian immunodeficiency virus infection
Authors: Baskerville A. , Ramsay A. D. , Addis B. J. , Dennis M. J. , Cook R. W. , Cranage M. P. , Greenaway P. J.
Published: June 2005

Interstitial pneumonia unrelated to Pneumocystis carinii or other infections was observed histopathologically in 5 of 25 rhesus monkeys infected with simian immunodeficiency virus (SIV). The predominant lesion was lymphocytic infiltration of interalveolar septa and hyperplasia of peribronchial and perivascular lymphoid tissue. Immunohistochemical staining using a panel of antibodies against human T and B lymphocytes, macrophages, and immunoglobulins showed that peribronchial aggregates and interstitial infiltrates were predominantly B cells, whereas perivascular masses consisted mainly of T cells. One animal with a primary B‐cell lymphoma of the spinal cord had secondary plasmacytoid lymphomatous nodules throughout the lung which were accompanied locally by reactive B‐cell lymphoid follicles. Another animal also had large areas of diffuse alveolar fibrosis and epithelial metaplasia to a bronchiolar type. In two monkeys, branches of the pulmonary arteries showed intimal proliferation and organizing occlusive thrombi, some of Which were mineralized.
Parathyroid hormone‐related peptide: Expression in fetal and neonatal development
Authors: Burton P. B. J. , Moniz C. , Quirke P. , Malik A. , Bui T. D. , Juppner H. , Segre G. V. , Knight D. E.
Published: June 2005

Hypercalcaemia frequently complicates the clinical management of cancer. Many factors have been implicated in the pathogenesis of this humoral hypercalcaemia of malignancy, the most recent candidate being parathyroid hormonerelated peptide (PTHrP). Until now, this peptide has been detected only in some normal and transformed adult tissues. In recent years, it has become apparent that tumours are capable of expressing and secreting factors previously elaborated only during fetal life. Many of these factors act to stimulate the growth of both tumour and fetal cells in an autocrine manner. The data presented here demonstrate that PTHrP is expressed in the human and rat fetus throughout gestation. Immunocytochemistry reveals a gestationally related, changing pattern of expression which is paralleled by changes in mRNA transcription. These data sunport the hypothesis that PTHrP may function as a fetal growth factor.
Differential expression of integrin alpha chains by renal epithelial cells
Authors: Rahilly Maeve A. , Fleming Stewart
Published: June 2005

We have investigated the expression of integrin chains by human renal epithelial cells during in vivo renal differentiation and in vitro cell culture on different extracellular matrices. Using the immunoperoxidase technique to visualize the binding of monoclonal antibodies to different integrin chains in fetal and adult kidneys, we found a change during development from α1β1, α3β1, and α4β1 ‐positive blastemal cells to α2β1, α3β1, and α6β1 ‐positive epithelial cells. The pattern of integrin expression correlates with the presence in the extracellular matrix of the appropriate ligands. In in vitro cell culture experiments, renal epithelium expressed α3 and α5 integrins on all extracellular matrices. Integrins α2 and α6 were found only in cells grown on a laminin‐containing substratum. Fibronectin and α5 integrin co‐localized on the ventral surface of cells grown on a laminin substratum and at the periphery of cells on glass coverslips. These results suggest that there is a close relationship between integrin α chain usage and the presence of appropriate ligands in the extracellular matrix.
Successive infection of coxsackievirus B3 and encephalomyocarditis virus: An animal model of chronic myocarditis
Authors: Okada Ikutaro , Matsumori Akira , Tomioka Nobuyoshi , Kawai Chuichi
Published: June 2005

Successive infection of coxsackievirus B3 and encephalomyocarditis virus was investigated as a disease model of chronic myocarditis. Four‐week‐old C3H/He mice were inoculated with coxsackievirus B3 and then inoculated with encephalomyocarditis virus at 8 weeks old. The hearts were evaluated on histopathological changes compared with those of non‐infected mice and mice infected with either virus alone. At 10 weeks old, the hearts of the mice infected successively with both viruses showed co‐existence of fibrosis surrounding calcified lesions and marked cellular infiltration with myocardial necrosis. These findings resembled chronic active myocarditis in humans, unlike the lesions due to either virus alone. At 12 weeks old, the hearts of all the infected mice showed fibrosis with scarce cellular infiltration. The successively infected hearts also showed a significantly higher heart weight to body weight ratio than that of the non‐infected control mice, and localized wall thinning in the damaged regions. Thus, we conclude that successive infection additively causes myocardial damage that resembles chronic myocarditis and may produce a heart condition similar to dilated cardiomyopathy.
Herpes simplex virus type 1 DNA is present in specific regions of brain from aged people with and without senile dementia of the Alzheimer type
Authors: Jamieson Gordon A. , Maitland Norman J. , Wilcock Gordon K. , Yates Celia M. , Itzhaki Ruth F.
Published: June 2005

We have investigated the possible involvement of viruses, specifically Herpes simplex virus type 1, in senile dementia of the Alzheimer type (SDAT). Using the highly sensitive polymerase chain reaction, we have detected the viral thymidine kinase gene in post‐mortem brain from 14/21 cases of senile dementia of the Alzheimer type and 9/15 elderly normals. The temporal cortex and hippocampus were usually virus‐positive; in contrast, the occipital cortex was virus‐negative in 9/9 SDAT cases and 5/5 elderly normals. Temporal and frontal cortex from younger normals (five infants and five middle‐aged) were negative. Thus, the presence of Herpes simplex virus type 1 DNA is a region‐dependent feature of the aged brain.
A sensitive method for the detection of poly‐A tails of mRNA using a biotin‐labelled heteropolymer OF dT:rA
Authors: Markovic Boban , Kwan Yiu‐Lam , Nicholls E. Max , Walsh Claire , Crouch Roger L.
Published: June 2005

We have developed a highly sensitive non‐radioactive in situ hybridization technique that enables us to study the production of mRNAs in tissues. As part of the validation procedure of our methods, we examined various methods of detecting poly‐A RNA tails of mRNA. We have used three types of biotin‐labelled probes complementary to poly‐A sequences: a 25‐mer poly‐dT oligonucleotide, a polymer of dT, and a heteropolymer of dT:rA. All the probes had the same specificity of reactivity but the heteropolymer of dT:rA gave the strongest signals as visualized histochemically by the use of alkaline phosphatase as the detection enzyme. All the probes tested for poly‐A detection showed reactivity. The poly‐dT oligonucleotide showed a strength of signal comparable to published results. The biotinylated polymer of dT gave a stronger signal than that of the oligonucleotide, and the heteropolymer was the strongest of all. The strong signal seen with the heteropolymer probe is due to probe complexing during hybridization, in which additional binding between sense and antisense strands of the probe (i.e. poly‐rA and poly‐dT) amplifies the number of biotin molecules at the hybridization site; this strategy has been exploited by us as a means of visualizing low copy numbers of specific mRNAs.
The demonstration of a subset of carcinoid tumours of the appendix by in situ hybridization using synthetic probes to proglucagon mRNA
Authors: Shaw Paul A. V. , Pringle James H.
Published: June 2005

Previous studies using immunohistochemistry have shown variable hormone production by carcinoid tumours of the appendix. In order to confirm the existence of a specific subset of these tumours, in situ hybridization using synthetic oligonucleotide probes to detect prc‐proglucagon and pre‐proinsulin mRNA was performed in formalin‐fixed, paraffin‐ embedded material from eight tubular carcinoids, 12 insulin carcinoids, and two mucinous carcinoids. The results were correlated with standard silver and mucin stains. All tubular carcinoids but none of the insular or mucinouscarcinoids contained proglucagon mRNA. Proinsulin mRNA was not detected in any of the tumours. Tubular carcinoids of the appendix constitute a definable subset of appendiceal carcinoids which have a similar distribution and prognosis to typical insular carcinoids and can be diagnosed on hamaloxylin and eosin‐stained sections confirmed by routine special stains. The main need for recognition is to avoid confusion with mucinous carcinoids. which have a worse prognosis and may require more aggressive treatment.
Frequent expression of Epstein‐Barr virus latent membrane protein‐1 in tumour cells of Hodgkin's disease in HIV‐positive patients
Authors: Audouin Josée , Diebold Jacques , Pallesen Gorm
Published: June 2005

Epstein‐Barr virus (EBV) is believed to be implicated in the aetiology of non‐Hodgkin's lymphomas developing in immunodeficient individuals including AIDS patients. EBV has also been associaled with Hodgkin's disease (HD), where the genomes have been demonstrated in the Hodgkin and Reed‐Sternberg cells in some of the cases. Recent evidence has shown that EBV genomes are transcribed in these cells, because the EBV‐encoded latent membrane protein‐1 (LMP‐1) can be demonstrated in the tumour cells in about half of the HD cases in HIV‐negative patients using immunohistochemistry. LMP‐1 is of special interest as a possible oncogenic agent because of its strong transforming capacity in vitro. In this study we have examined the expression of LMP‐1 in HD of HIV‐positive patients compared with HD in HIV‐negative patients. We investigated 18 lymph nodes from 16 HIV‐positive patients with HD (eight mixed cellularity, nine nodular sclerosis, one unclassified) using the CS. 1–4 anti‐LMP‐1 monoclonal antibodies, which can usually be applied successfully to archival biopsy material. In each case, 50–90 per cent of the tumour cells were labelled. Staining was excellent for both fixatives used (4 per cent buffered formalin, Bouin's fluid).

It is concluded that EBV‐encoded LMP‐1 is firmly associated with HD of HIV‐positive patients. This is most conspicuous in the nodular sclerosing subtype HD in HIV‐positive patients, in which 100 per cent were LMP‐1 positive as compared with 32 per cent of nodular sclerosis HD in HIV‐negative cases in a previously published series. This difference is statistically significant (P< 0.001). The possible biological and clinical significance of this difference should therefore be studied in larger series.
Assessment of rejection in orthotopic human heart transplantation using proliferating cell nuclear antigen (PCNA) as an index of cell proliferation
Authors: Mann Jessica M. , Jennison Stephen H. , Moss Eileen , Davies Michael J.
Published: June 2005

Myocardial biopsies taken during the management of cardiac transplantation were stained for proliferating cell nuclear antigen (PCNA). Counts of PCNA‐positive interstitial cells were compared, in retrospect, with the reported histological grade of rejection. Biopsies without rejection had negligible numbers of PCNA‐positive cells. Ascending grades of rejection were paralleled by an increase in the number of PCNA‐positive cells [grade 1,13±35 (mean ± SD); grade 2a, 38±40; grade 2b, 91±75; grade 3, 170±78]. While highly significant, in statistical terms, the overlap in the counts between different grades means that prediction of rejection from the PCNA count alone is not feasible. Biopsies graded as 0 or 1 and which immediately preceded more severe rejection episodes showed no increase in PCNA‐positive cells. The majority of PCNA‐positive cells are fibroblasts, although in grade 2b and 3 rejection a small population of PCNA‐positive T lymphocytes occurs. PCNA staining is also seen in cardiac myocytes immediately after transplantation, during rejection episodes, and late after transplantation in the absence of rejection. The positive PCNA staining of cardiac myocytes probably reflects DNA synthesis that occurs with the shift toward polyploidy in hypertrophy.
TDM35—a new monoclonal antibody to the XH1 cervical carcinoma cell line. Characterization and immunoperoxidase localization in benign and malignant tissues
Authors: Chana Treena , Han Xin , Dagg Belinda , Lawrie Andrew S. , Gooding Roger P. , Eustace Damian L. S. , Fletcher Christopher D. M. , Heyderman Eadie
Published: June 2005

The murine monoclonal IgG1 kappa antibody TDM35 was raised against the cervical carcinoma cell line XH1. The antibody recognizes 18.5–66 kDa NCA‐like glycoproteins and immunostains a variety of formalin‐fixed, paraffin‐embedded normal, benign, and malignant tissues. It is of value in the diagnosis of carcinoma of the exocrine pancreas and it identifies foci of squamous and glandular differentiation in other tumours. TDM35 should form a useful addition to a panel of antibodies for the evaluation of epithelial lesions.
Flow cytometric DNA‐histogram analysis: Non‐stoichiometric fluorochrome binding and pseudo‐aneuploidy
Authors: Kubbies Manfred
Published: June 2005

Detection of aneuploid subpopulations using flow cytometry requires stoichiometric binding of nucleic acid‐specific fluorochromes onto DNA. It is shown that parameters like cell type specificity and differentiation stage, cell cycle stage, loss of DNA‐integrity, cell preparation, and cytochemistry affect fluorochrome binding to DNA and give rise to the appearance of pseudo‐aneuploid cell populations. Intercalating as well as non‐intercalating fluorochromes show non‐stoichiometric DNA‐labelling in cell populations with identical DNA content, and pseudo‐aneuplpidy was found in flow cytometers equipped with either arc lamps or argon lasers. Pseudo‐aneuploidy was never observed with intercalating and non‐intercalating fluorochromes within identical specimens, consisting of cells of various differentiation states (e.g., bone marrow) or containing large numbers of dead cells. Therefore, fluorochromes exhibiting different base‐pair specificities or steric binding modes should be applied to be sure of the correct interpretation of small levels of hypo‐ or hyper‐diploidy (±xs 20 per cent).
Collagen fibril structure of normal, aging, and osteoarthritic cartilage
Authors: Hwang W. S. , Li B. , Jin L. H. , Ngo K. , Schachar N. S. , Hughes G. N. F.
Published: June 2005

The collagen architecture in normal, aging, and osteoarthritic articular cartilage was studied optically using a new silver staining technique based on specimens from 50 autopsy cases, four amputated limbs, and six osteoarthritic knees. In the normal articular cartilage, the collagen fibrils in the superficial zone were compactly arranged into layers of decussating flat ribbons mostly parallel to the artificial split lines. The fibrils showed a tendency to condense into vertical arcade columns undergirded by tangential bundles in the intermediate zone. In the deep zone, the fibrils formed a random meshwork with a slight preponderance of vertical fibrils in the perilacunar region.

Three types of early degradative lesions involving the collagen network were identified. Type I lesions consisted of focal superficial disruptions related to age and friction. Type II lesions consisted of focal disruptions of tangential fibrils in the intermediate zone leading to cyst formation, probably representing a form of local stress failure. Type III lesions were found in the patella and consisted of marked swelling of the superficial zone, the cause of which was unknown. Lesions of varying severity were seen within each of the three types; the morphological changes of the more severe lesions overlapped with those of clinically overt osteoarthritis.
Proliferative activity determined by DNA flow cytometry and proliferating cell nuclear antigen (PCNA) immunohistochemistry as a prognostic factor in prostatic carcinoma
Authors: Visakorpi Tapio
Published: June 2005

Proliferative activity was measured in 165 paraffin‐embedded prostatic carcinomas using DNA flow cytometric analysis of the S‐phase (SPF) and G2/M‐phase fractions and CAS 200 image analysis of the proliferating cell nuclear antigen (PCNA) expression defined immunohistochemically by PC10 and 19A2 monoclonal antibodies. No significant associations were found between the flow cytometric and the two immunohistochemical measures of cell proliferation. Of the four indices, only SPF, S + G2/M, and immunostaining with 19A2 antibody were associated with the poor histological grade of the tumour. High SPF and S + G2/M were significantly associated with poor 10‐year overall survival (P<0·001) and prostatic carcinoma‐specific survival (P< 0·01). Multivariate analyses of prostatic carcinomaspecific survival in patients with non‐metastatic disease (M0‐stage) indicated that only S + G2/M, T‐stage, and histological grade (only if re‐evaluated by a single pathologist) had independent prognostic significance. High‐level PCNA staining (> 16 per cent of cells stained) with 19A2 antibody was associated with poor prognosis only in univariate analysis, and PC10 immunostaining had no prognostic value. In conclusion, a high proliferative activity as defined by flow cytometric S + G2/M is an independent predictor of poor survival in patients with non‐metastatic prostatic carcinoma. PCNA immunostaining from formalin‐fixed, paraffin‐embedded prostatic carcinomas has little, if any, prognostic value.
The peritubular myofibroblasts in the testes from normal men and men with Klinefelter's syndrome. A quantitative, ultrastructural, and immunohistochemical study
Authors: Martin R. , Santamaría L. , Nistal M. , Fraile B. , Paniagua R.
Published: June 2005

The ultrastructure and immunostaining with antibodies against actin, desmin, and vimentin were studied in the peritubular myofibroblasts of testes from normal men and men with Klinefelter' syndrome (KS). The seminiferous tubules were classified into five types (a–e), related to the progressive degree of sclerosis measured as thickening of the lamina propria. In control testes, only types a and b tubules were present, whereas the testes from men with KS showed types b, c, d, and e tubules. The ultrastructural study revealed abundant microfilament bundles with electron‐dense bodies in the cell periphery of the myofibroblasts in a and b tubules. In c tubules, the microfilament bundles of the myofibroblasts were lacking in electron‐dense bodies. Myofibroblasts in tubules d and e showed scanty microfilament bundles. Immunostaining of peritubular myofibroblasts with anti‐actin antibodies was intense in tubule types a–c and scanty in types d and e. Immunostaining of myofibroblasts with anti‐desmin antibodies was intense in tubule types a and b, and negative in types c–e. Immunostaining with anti‐vimentin antibodies was weak in tubule types a–c and intense in types d and e. Quantitative study revealed that with the progression of sclerosis, the number and volume per cross‐sectioned tubule of actin‐containing cells and, mainly, desmin‐containing cells decrease while the number and volume of vimentin‐containing cells increase.
Immunoreactivity of proliferating cell nuclear antigen compared with bromodeoxyuridine incorporation in normal and neoplastic rat tissue
Authors: Wijsman Jan H. , Van Dierendonck Jan‐Hein , Keijzer Rob , Van De Velde Cornelis J. H. , Cornelisse Cees J.
Published: June 2005

Monoclonal antibodies (MoAbs) against proliferating cell nuclear antigen (PCNA) represent a potentially useful tool for cell kinetic analysis of tumours. Because in paraffin‐embedded tissue the relationship between PCNA immu‐noreactivity and tumour cell proliferation is not well characterized, we have compared PCNA positivity as detected by the PC10 MoAb with the bromodeoxyuridine labelling index (BrdUrd‐LI) in two different transplantable hormone‐dependent rat mammary tumours. Together, these two tumour models (MCR‐83 and EMR‐86) cover a wide range of S‐phase fractions. Evaluating 31 methacarn‐fixed tumours, a strong but non‐linear relationship (r=0·98) was obtained. PCNA‐positive fractions were invariably higher than corresponding BrdUrd‐LIs and also higher than the estimated growth faction: growth fractions as determined by continuous BrdUrd labelling of the tumour and stromal cell population in EMR‐86 carcinomas were 12 and 26 percent lower than PCNA‐positive fractions, implying that a certain fraction of non‐cycling cells can also express PCNA. A dramatic disturbance in the relationship of PCNA positivity and the BrdUrd‐LI was observed in the EMR‐86 model after growth arrest induced by hormonal ablation: PCNA immu‐noreactivity remained detectable for at least 3 days, whereas the BrdUrd‐LI decreased almost immediately. In comparison, PCNA immunoreactivity persisted for a much shorter period in small intestinal cells that had stopped DNA replication when moving from the crypt towards the villus. It is concluded that although differences in PCNA expression exist between various tissues, PCNA as detected by the PC10 MoAb may be used in tumours as an operational marker for the growth fraction. However, this seems to be valid only under conditions of unperturbed growth.
Delay in fixation does not affect the immunoreactivity of proliferating cell nuclear antigen (PCNA)
Authors: Start R. D. , Cross S. S. , Clelland C. , Silcocks P. B. , Rogers K. , Smith J. H. F.
Published: June 2005

The effect of delayed fixation on the immunoreactivity of proliferating cell nuclear antigen (PCNA) was investigated using eight breast carcinomas. Topologically shuffled samples of each tumour were immersed in fixative at times of 0·5. 1,2,4,6,18, and 24 h after surgical removal. In addition to a PCNA index (percentage of positive cells per 1200 tumour cells), a semi‐quantitive PCNA grading system was used, based on estimates of more than or less than 50 per cent of positive tumour cells at each time interval. The PCNA index of six tumours increased by a mean of 10 per cent with a fixation delay of 24 h. The PCNA grade of all eight tumours showed no change with delayed fixation.
Morphometry of gastric carcinoma: Its association with patient survival, tumour stage, and DNA ploidy
Authors: Hamilton P. W. , Wyatt J. I. , Quirke P. , Watt P. C. H. , Arthur K. , Ward D. C. , Johnston D.
Published: June 2005

Morphometric image analysis of nuclear features was performed on tissue from 46 patients who had had curative resections for gastric cancer. Clinical, pathological, flow cytometric, and follow‐up data were available for these patients, which were drawn from a larger, previously reported series.1 The morphometric data were compared with patient survival, clinico‐pathological status, and DNA ploidy. Univariate survival analysis revealed that morphometric parameters were not significantly related to survival, but examination of clinico‐pathological data showed lymph node involvement, involvement of the resection margin, and lymphatic invasion to be significantly associated (P<0·01) with patient prognosis. Multivariate survival analysis using the Cox model found only lymph node and resection margin involvement to be independently related to survival. Comparison of morphometric results with the clinico‐pathological parameters showed various features, relating to nuclear size, and its variation to be significantly associated (P<0·01) with the presence of lymphatic invasion, resection margin involvement, and tumour pattern (intestinal/diffuse). A comparison of morphometry with flow cytometric analysis in these cases showed that nuclear size was not significantly related to either DNA aneuploidy or the DNA proliferative index.
Heparin causes partial removal of glomerular antigen deposits by a mechanism independent of its anticoagulant properties
Authors: Furness Peter N. , Drakeley Simon
Published: June 2005

We have previously reported that heparin can enhance the removal of antigen from the glomeruli of rats with chronic serum sickness glomerulonephritis. We have performed two studies to investigate the relevance of anticoagulation to this effect. The first experiment was essentially a repeat of those described before, but instead of using heparin, anticoagulation was achieved using Ancrod, which causes depletion of fibrinogen. The amount of antigen in the glomeruli of these rats at the end of the experiment did not differ from controls. In the second experiment, both kidneys were excised from rats with glomerulonephritis induced by a radiolabelled cationic antigen. Of each pair of kidneys, one was perfused for 30 min at 5 ml/min with Hank's balanced salt solution (HBSS) at 37°C. The other was perfused with HBSS to which a total of 4500 units of heparin had been added. The antigen content of isolated glomeruli was subsequently measured. In every case, the kidney perfused with heparin had less antigen per glomerulus than the control contralateral kidney. The ability of heparin to enhance removal of glomerular immune complex deposits is therefore not mediated by anticoagulation.
Epidermotropism of T cells correlates with intercellular adhesion molecule (ICAMI) expression in human papillomavirus (HPV)‐induced lesions
Authors: Viac J. , Chardonnet Y. , Euvrard S. , Schmitt D.
Published: June 2005

Adhesion molecules play an important role in inflammatory reactions. Among them, ICAM1, a ligand for the lymphocyte function‐associated antigen (FLA1) of leucocytes, may be expressed by antigen‐presenting cells and keratinocytes in various inflammatory disorders. As cell‐mediated immune responses play a great role in HPV infections, we investigated the expression of ICAM1 and correlated it with the presence of LFA1‐positive cells by immunohistochemistry on serial frozen sections of a series of non‐regressing cutaneous and mucosal HPV‐induced lesions. ICAM1 expression by keratinocytes was observed only in intensely infiltrated lesions of condylomas and laryngeal papillomas. Its induction was usually correlated with the presence of LFA1‐positive cells (mainly CD8‐positive cells) which were in close apposition to ICAM1‐positive proliferative epithelial cells expressing also, in some cases, HLA‐DR antigen. ICAM1 was not correlated with the presence of HPV DNA or viral antigen. In moderately infiltrated lesions, keratinocytes did not express ICAM1, and LFA1 ‐positive cells were not observed in the epidermis. In all lesions, ICAM1 was more intense on endothelial cells than in normal skin; infiltrating cells (lymphocytes and dendritic cells) may also express this antigen but intraepithelial Langerhans cells were devoid of anylabelling. These studies provide further evidence that T‐lymphocyte mechanisms are important in the host response to HPV‐induced lesions. ICAM1 expression correlates with a lesional infiltrate but not with HPV infection and probably results in a more efficient initiation of the immune reaction.
Histological preservation after in situ hybridization to archival solid tumour sections allows discrimination of cells bearing numerical chromosome changes
Authors: Van Dekken H. , Kerstens H. M. J. , Tersteeg T. A. , Verhofstad A. A. J. , Vooijs G. P.
Published: June 2005

In this study, non‐isotopic in situ hybridization (ISH) was used for the cytogenetic and histological examination of urological (prostatic adenocarcinoma) and endocrine (phaeochromocytoma) tumour cell nuclei in 4μm paraffinembedded tissue sections. In order to investigate preservation of tissue morphology, standard heat denaturation was compared with a mild enzymatic treatment for the production of singletranded (ss)‐DNA for ISH. Numerical analysis by ISH with chromosome‐specific repetitive DNA probes for chromosomes 1, 7, and 11 revealed overrepresentation of chromosome 7 in the phaeochromocytoma (P < 0·01). The constitutional underrepresentation of the Y chromosome was easily detected in the prostate tumour (P < 0·01) when probed for chromosomes 7, 16, and Y. The enzymatic treatment appeared superior to heat denaturation with respect to tissue architecture in the phaeochromocytoma, while no clear difference was observed in the prostatic cancer. ISH probe patterns were similar for the two types of denaturation in both tumours (P≥ 0·20). We conclude that (1) ISH can be used for the identification of numerical cytogenetic changes in solid tumour cell nuclei within archival tissue sections; and (2) mild ‘denaturation’ protocols, replacing heat, are preference in retaining tissue architecture in fragile tumour specimens.
Polycystic kidney disease in the CBA/N immunodeficient mouse
Authors: Rahilly Maeve A. , Samuel Kay , Ansell J. D. , Micklem H. S. , Fleming S.
Published: June 2005

We describe a polycystic lesion of the kidney in the CBA/N mouse with an X‐linked recessive immunodeficient syndrome. There is progressive cystic dilatation affecting all parts of the nephron. The cyst lining is composed of a single layered epithelium with focal nuclear crowding and the formation of micropapillary structures. The cystic epithelial ceils show subnuciear vacuolation. Focal basement membrane thickening is also a feature. There is no significant inflammatory infiltrate present within these kidneys. Electron microscopic examination reveals that the subnuciear vacuolation is due to loss of the membrane infoldings at the basal pole of the epithelial cell with fluid accumulation within the extracellular space. The basement membrane thickening is due to expansion of the lamina densa. These changes are not present at birth but develop progressively with age. The finding of a polycystic kidney lesion in these mice offers an opportunity to investigate the relationship between the immune system and renal cyst formation.
PrP protein is associated with follicular dendritic cells of spleens and lymph nodes in uninfected and scrapie‐infected mice
Authors: McBride Patricia A. , Eikelenboom Piet , Kraal Georg , Fraser Hugh , Bruce Moira E.
Published: June 2005

Abnormal forms of a host protein, PrP, accumulate in the central nervous system in scrapie‐affected animals. Here, PrP protein was detected immunocytochemically in tissue sections of spleen, lymph node, Peyer's patches, thymus, and pancreas from uninfected mice and from mice infected with a range of mouse‐passaged scrapie strains and bovine spongiform encephalopathy (BSE). In the spleen, lymph node and Peyer's patches, PrP‐positive cells were identified as follicular dendritic cells (FDC) by their location, appearance, and immune complex trapping function, whereas in the thymus they appeared to be two types of stromal cells: interdigitating cells (IDC) and cortical epithelial cells. In pancreas, PrP‐containing cells were confined to the islets of Langerhans. Although the distribution of PrP immunola belling was the same in tissues from scrapie‐affected and uninfected mice, there was evidence that PrP accumulated in abnormal forms in FDC of infected mice. If, as is likely, PrP is essential for agent replication, our results suggest that FDC are the site of scrapie and BSE replication in the spleen and lymph node.
The relationship between wall tension, lamellar thickness, and intercellular junctions in the fetal and adult aorta: Its relevance to the pathology of dissecting aneurysm
Authors: Berry Colin L. , Sosa‐Melgarejo Jorge A. , Greenwald Stephen E.
Published: June 2005

It is known that the distribution of stress and strain in the vessel wall is not uniform. We believe that this explains the location of the plane of dissection in dissecting aneurysms of large elastic arteries. We have investigated the effects of non‐uniformity of stress and strain on the thickness of each elastic lamella and on the distribution of intercellular junctions in the media of developing and adult rats, to seek evidence to support this hypothesis. Intercellular junctions were identified by transmission electron microscopy of whole wail sections. A morphomeiric study of elastic tissue distribution was made on an image analysis computer. Differences were analysed using one‐way analysis of variance.

There are between six and eight elastic lamellae in the aorta of rats. In the fetus, only the internal elastic lamella is complete; the others were not fully formed by term. In the adult, the inner five elastic lamellae were thicker than the remaining two or three, and smooth muscle cells in the thicker lamellar units had more cell‐cell contacts of all types examined. These data support the concept of a difference in stress‐resisting properties of the aortic wall on the junctions between the inner two‐thirds and the outer third of the media.

The findings indicate that, as proposed in theoretical models the innermost lamellae support the high tension. In the adult aorta, the structure is modified to enhance the capacity to resist stress in the internal two‐thirds of the media. This leaves a clear change in form at the junction with the external third of media, where tearing might be expected with exposure to high levels of stress. We suggest that this is critical in the location of aortic dissections.
Immunohtstochemical analysis of the p53 oncoprotein on paraffin sections using a series of novel monoclonal antibodies
Authors: Bártek Jiřía , Bártková jiřina , Lukáš Jiří , Stašková Zdenka , Vojtěšek Bořivoj , Lane David P.
Published: June 2005

Alterations of the p53 tumour suppressor gene are considered critical events in multistage carcinogenesis of a wide range of human cancers. In an attempt to elucidate the role of various p53 mutations in tumorigenesis and to investigate their relationship to the p53 protein accumulation and subcellular localization, we have raised a new series of 21 mouse monoclonal antibodies (MAbs) to human recombinant p53. The new MAbs (designated the Bp53 series) appear to recognize mainly denaturation‐resistant epitopes in immunoblotting and the majority of them are suitable for immunostaining of p53 in cultured cells and frozen sections. Furthermore, at least three MAbs (Bp53‐11, Bp53‐12, and Bp53‐28) proved to be reliable reagents for immunohistochemistry on paraffin‐embedded specimens. The immunohistochemical analysis of paraffin sections from 118 human tumours of various histogeneses with Bp53‐11 and Bp53‐12 showed nuclear accumulation of the p53 protein in variable proportion of tumour cells in 76 cases (64 per cent). The influence of three parameters of tissue processing (type of fixative, period of fixation, and duration of autolysis) on p53 protein detection was also investigated. The results of this study provide the necessary basis for wider application of these novel MAbs as tools in both routine hisiopathology and functional analyses of the p53 oncoprotein.
Electron microscopy of rectal biopsies in HIV‐positive individuals
Authors: Blanshard C. , Ellis D. S. , Tovey G. , Gazzard B. G.
Published: June 2005

A series of 83 rectal biopsies was examined by electron microscopy (EM), 78 from HIV‐seropositive patients (64 with diarrhoea, 9 with weight loss, and 5 asymptomatic) and 5 from controls without risk factors for HIV infection. Although EM identified the likely cause of the diarrhoea in only nine cases, the majority of symptomatic HIV‐seropositive patients had abnormal biopsies including the presence of tubuloreticular structures, loss of microvilli, and oedema and disruption of autonomic nerves. Neuronal abnormalities were particularly common in patients with cryptosporidiosis, but no ultrastructural marker was specific to ‘pathogen‐negative’ diarrhoea.
Ultrastructural aspects of tumour invasion in the central nervous system
Authors: McGrady B. J. , McCormick D. F. , Toner P. G.
Published: June 2005

Tumour invasion in the central nervous system (CNS) was studied using a mouse melanoma cell model. Attention was concentrated on the third ventricle as this site is distant from any implantation artefact. Transmission electron microscopy (TEM) revealed the formation of tumour cell aggregates within the ventricle. Underlying ependymal cells showed loss of cilia and microvilli. Progressive thinning of the ependymal cells took place until continuity of the ependyma was lost through ependymal retraction. Tumour cells were then in direct contact with sub‐ependymal neuroglial processes. Further invasion was then accomplished by penetration between neuropil processes, followed by tumour cell infiltration along the intact basement membrane of cerebral capillaries with displacement of astrocyte foot processes from the capillary basement membrane.
Experimental adriamycin nephropathy. Fine structure, morphometry, glomerular polyanion, and cell membrane antigens
Authors: Rossmann P. , Matoušovic K. , Bohdaneck M.
Published: June 2005

Two doses of adriamycin (2 mg/kg i.v.) were administered to young adult rats with a 22‐day interval and the kidneys were examined 51, 79, and 107 days after the first injection. Light microscopy showed early prominent intraglomerular microcystic spaces and progressive, though not severe, segmental glomerulosclerosis with focal interstitial scarring and round‐cell infiltration. The main features of the ultrastructure were widespread fusion of foot processes, focal cytoplasmic rarefaction of podocytes, and non‐specific sclerosis of the mesangium without signs of severe degenerative changes or mesangiolysis. The Segmental microcystic structures corresponded to agglomerted intrapodocytic vacuoles. The scatter of glomerular polyanion sites visualized by polyethylenimine–phosphotungstic acid resembled that of control animals but the quantitative assessment revealed a significant reduction. CD4‐positive cells were the predominant element of the interstitial infiltrates. Most of the infiltrating cells expressed Ia antigens, whereas la‐positive intraglomerular resident mononuclear cells were depleted. The adriamycin lesion resembles the post‐five‐sixths nephrectomy ablation nephropathy in the subpopulations of the interstitial infiltrate but differs from it in the reduction of intraglomerular resident macrophages, the absence of destructive mesangial damage, and reduced segmental glomerulosclerosis. The main feature is the prolonged damage of podocytes, especially of their cytoskeletal system.