bluuebunny/crossref_metadata_2025 huggingface dataset:
Biliary epithelial cells from the liver of patients with primary biliary cirrhosis: Isolation, characterization, and short‐term culture
Authors: Joplin Ruth , Strain A. J. , Neuberger J. M.
Published: June 2005

Since biliary epithelial cells of the middle‐sized interlobular bile ducts are targets for lymphocyte‐mediated damage in patients with primary biliary cirrhosis (PBC), we have developed a method for isolating and maintaining these cells in short‐term tissue culture. Intrahepatic biliary epithelial cells were isolated from small segments of liver removed at the time of transplantation. Cells were separated from a collagenase digest by immunomagnetic separation using Dynabeads coupled to a monoclonal antibody (HEA125) specific for a biliary epithelial cell surface antigen. The yield was approximately 3 x 105 cells/g of liver. The isolated cells were characterized morphologically and ultrastructurally using light and electron microscopy, and immunocytochemically using HEA125 and anti‐cytokeratin, anti‐vimentin and anti‐asialoglycoprotein receptor antibodies. By these criteria cells were judged to be identical to biliary epithelial cells from normal liver. The cells could be maintained in short‐term tissue culture for up to 4 weeks without loss of biliary epithelial cell markers. Availability of interlobular biliary epithelial cells will be of value in future investigations of the pathogenetic mechanisms of PBC.
The osteoclast, which derives from a haemopoietic stem cell, is not depleted in aplastic anaemia
Authors: Flanagan Adrienne M.
Published: June 2005

The osteoclast derives from the haemopoietic stem cell but its relationship with the other progeny of the haemopoietic system is unknown. Osteoclast numbers were assessed in patients with aplastic anaemia and were found not to be depleted compared with a control population. This suggests that the osteoclast may develop along a separate lineage which is independent of the colony forming unit granulocyte, erythroid cell, monocyte, and megakaryocyte (CFU GEMM).
Epidermal growth factor (EGF/URO) induces expression of regulatory peptides in damaged human gastrointestinal tissues
Authors: Wright Nicholas A. , Poulsom Richard , Stamp Gordon W. H. , Hall Peter A. , Jeffery Rosemary E. , Longcroft Janet M. , Rio Marie‐Christine , Tomasetto Catherine , Chambon Pierre
Published: June 2005

The pS2 gene encodes for a small cysteine‐rich protein, and was originally found by differential screening of a cDNA library from the human breast carcinoma cell line, MCF‐7. The presence of pS2 is closely correlated with oestrogen dependence in breast carcinomas. While the function of pS2 is unknown, pS2 protein has been shown to be homologous with the gastrointestinal peptide hormone pancreatic spasmolytic polypeptide (PSP) and its human counterpart hSP, in which a 5‐cysteine domain is tandemly repeated. The 5′ flanking region of the pS2 gene contains an enhancer region responsive to oestrogens and to epidermal growth factor (EGF/URO). We now report that pS2 and hSP expression occurs in a wide range of endodermally‐derived tissues, including the duodenum, the pancreas, and in a recently defined cell lineage associated with chronic gastrointestinal ulceration. In each case, this expression was associated with secretion of immunoreactive EGF/URO. We further show that the co‐expression of pS2 and hSP in gastric surface epithelial cells is also associated with the secretion of EGF/URO in the subjacent mucous neck cells. Our results indicate that local EGF/URO secretion induces pS2 and hSP in adjacent cells, and that these molecules are then available to participate in pathophysiological responses. The finding of similar patterns of EGF/URO, hSP and pS2 expression in association with chronic damage suggests that this is a fundamental response in the healing of these tissues.
Steroid receptors and Ki‐67 reactivity in ovarian cancer and in normal ovary: Correlation with dna flow cytometry, biochemical receptor assay, and patient survival
Authors: Isola J. , Kallioniemi O.‐P. , Korte J.‐M. , Wahlström T. , Aine R. , Helle M. , Helin H.
Published: June 2005

Steroid hormone receptors and reactivity for Ki‐67 proliferation antigen were studied immunohistochemically in non‐neoplastic post‐menopausal human ovary and in 29 ovarian cancers. In the normal ovary, oestrogen (OR) and progesterone receptors (PR) were found in the surface epithelium and PR also in the ovarian stroma. Of the ovarian carcinomas 38 per cent (11/29) contained OR and 69 per cent (20/29) PR. Oestrogen receptor expression was confined to malignant cells, whereas PR was present occasionally also in the tumour stroma. In most cases, ORs and PRs were found only in a small population of cancer cells. The growth fractions assessed by the percentage of Ki‐67‐positive cells ranged from 1 to 59 per cent (mean 19·7 per cent) with a significant correlation (r = 0·74, P < 0·0001) to S‐phase values (mean 12·9 per cent, range 1·2–25·9 per cent) determined by DNA flow cytometry. High Ki‐67 (≥ 15 per cent) and S‐phase levels (≥ 7·5 per cent) correlated with advanced disease stage and patient survival but not with OR or PR status, suggesting that hormone‐receptor pathways and proliferative activity are not related in ovarian cancer. Positive OR status, however, identified patients with a better prognosis (P = 0·02), suggesting a correlation with tumour differentiation. The independent prognostic value of oestrogen receptor status and Ki‐67 remains to be determined, but the prognostic impact of Ki‐67 was comparable to that of S‐phase values.
Loss of alkaline phosphatase activity in duodenal mucosa: A marker for precursors of gastric metaplasia?
Authors: Kühl Philippa , Baczako Karl , Stanescu Adrian , Malfertheiner Peter
Published: June 2005

Biopsy specimens from duodenal mucosa in 34 patients with upper gastrointestinal symptoms and endoscopically abnormal mucosa (including duodenitis, active duodenal ulcer, and healed duodenal ulcer) and in 9 patients with histologically normal mucosa were examined histologically for gastric metaplasia and endogenous alkaline phosphatase (AP) activity.

Using haematoxylin and eosin (H&E) and periodic acid‐Schiff (PAS), we found gastric metaplasia in 91‐2 per cent (31 out of 34) of patients with altered duodenal mucosa and in 33·3 per cent (3 out of 9) of patients with histologically normal, non‐inflamed duodenal mucosa (P < 0·001). To characterize gastric metaplasia further, histochemical methods for AP activity were applied to duodenal mucosa specimens. No AP activity was detected in complete metaplastic cells, but focal or diffuse loss of AP activity was frequently shown in otherwise normal appearing enterocytes next to metaplastic cell groups. Focal loss of AP activity was also detected in seven out of nine healthy controls ( = 77·8 per cent) which appeared normal when stained with H&E.

Our results suggest that the loss of AP activity in enterocytes may be an early marker of developing gastric metaplasia or at least a morphological manifestation of cell damage.
CD 1a and S100 antigen expression in skin Langerhans cells in patients with breast cancer
Authors: Wilson Alan J. , Maddox Peter H. , Jenkins David
Published: June 2005

Langerhans cells (LCs) are dendritic, antigen‐presenting cells found in the epidermis. This study investigates the effect of early breast cancer on the expression of CD1a and S100 antigens by these cells. LCs were counted and expressed as cells/mm of epithelial basement membrane on biopsies from the skin overlying the tumour and from biopsies distant from the tumour. A control study was performed on normal breast skin, not adjacent to a lesion, from women with benign breast disease.

The LC count of 18 patients undergoing biopsy for benign breast disease indicated a mean of 26 cells mm [95 percent confidence interval (CI) 23–29] and a S100/CD1a ratio of 70 percent.

In 35 cases of early breast cancer, the CD1a‐positive LC count in the epidermis overlying the carcinoma (mean 26/mm; 95 per cent CI 23–29) was similar, but the count made on biopsies distant from the tumour (mean 21/mm; 95 per cent CI 19–23) was significantly smaller. The percentage ratio of S100/CD1a LCs was 71 per cent over the carcinoma and 84 per cent in the distant biopsies.

The changes were not associated with the presence of nodal metastases or the oestrogen and progesterone status of the primary tumour.

The reduction in LC numbers provides a link between decreased monocyte function and the decreased skin hypersensitivity responses found in patients with breast cancer.
The histopathology of the testis and epididymis in AIDS—a post‐mortem study
Authors: Dalton A. D. A. , Harcourt‐Webster J. N.
Published: June 2005

The right testis and epididymis were removed at necropsy from 31 patients who had died with AIDS and examined microscopically. The testes showed interstitial fibrosis in 22 cases, thickening of seminiferous tubule basement membranes in an overlapping 21, and varying degrees of maturation impairment in all. Arrest of spermatogenesis usually occurred at the spermatocyte or spermatid stage. Five epididymides showed an obstructive picture, two with an associated epididymal cytomegalovirus infection. We discuss the possible pathogenesis of the changes in the HIV‐infected male genital system and briefly comment on the clinical implications of our findings.
NB84: A new monoclonal antibody for the recognition of neuroblastoma in routinely processed material
Authors: Thomas Jaiye O. , Nijjar Jaspal , Turley Helen , Micklem Kingsley , Gatter Kevin C.
Published: June 2005

A new monoclonal antibody NB84 recognizing an uncharacterized molecule of 57 kD was produced using human neuroblastoma tumour tissue as a source of antigen. The antibody was selected for its ability to give reliable staining on conventional formalin‐fixed, paraffin‐embedded material. In the present study, NB84 was assessed for its usefulness in the differential diagnosis of paediatric small cell tumours. It gave reliable staining of 17/19 (90 per cent) neuroblastomas and was negative on all other tumours apart from Ewing's sarcomas, of which 9/14 (64 per cent) showed mainly focal positivity. It is concluded that in association with a panel of markers against leucocyte, muscle, epithelial, and neural markers NB84 provides considerable assistance in the recognition of neuroblastoma and thus is important in enabling the most appropriate therapy to be given.
Expression of c‐erbB‐2 oncoprotein in salivary gland tumours: An immunohistochemical study
Authors: Kernohan N. M. , Blessing K. , King G. , Corbett I. P. , Miller I. D.
Published: June 2005

Sections of formalin‐fixed, paraffin‐embedded primary salivary gland tumours were stained with a monoclonal antibody to the c‐erbB‐2 oncoprotein to determine the incidence and significance of expression of this protein. The series of 131 tumours comprised 33 cases of pleomorphic adenoma, 2 of malignant mixed tumour, 1 oxyphil adenoma, 31 Warthin's tumours, 4 basal cell adenomas, 6 mucoepidermoid carcinomas, 14 acinic cell carcinomas, 19 adenoid cystic carcinomas, 3 squamous carcinomas, and 18 poorly differentiated adenocarcinomas. Positive staining, as defined in previous studies, was present in five tumours (three cases of poorly differentiated adenocarcinoma, one mucoepidermoid carcinoma, and one adenoid cystic carcinoma). A review of the medical records of all patients did not disclose any clear difference between the clinical behaviour of positive and negative cases over a period of follow‐up that ranged from 18 to 120 months. The findings of this study indicate that the protein product of the c‐erbB‐2 proto‐oncogene is infrequently expressed in salivary gland tumours, and when it is localized on the tumour cell surface membrane, there is no clear evidence that this determines the biological behaviour of the tumour.
The AMeX method: A multipurpose tissue‐processing and paraffin‐embedding method. III. Extraction and purification of RNA and application to slot‐blot hybridization analysis
Authors: Sato Yuichi , Mukai Kiyoshi , Furuya Shuichiroh , Shimosato Yukio
Published: June 2005

RNA was extracted from tissues processed by a new fixation and paraffin‐embedding method (the AMeX method) and examined by Northern blot analysis and slot‐blot analysis. The RNA extraction method for AMeX‐processed tissue sections after the deparaffinization step was the same as that for fresh materials. The total amount of cellular RNA extracted from AMeX‐processed mouse liver tissue was slightly less than that extracted from fresh tissue. In tissues of malignant lymphoma, the total amount of cellular RNA extracted from 25 sections each 20μm thick was about 1·6–1·8 μg/mm2, regardless of the histological subtype and period of storage. The extracted RNA was moderately degraded, and usually could not be used for Northern blot hybridization analysis. The intensity of ethidium bromide staining and the hybridization signals of RNA extracted from AMeX‐processed tissues were usually reduced in comparison with RNA from fresh material, but specific signals could be detected by slot‐blot hybridization analysis. We have demonstrated previously that the AMeX method preserves high‐molecular‐weight DNA and various antigens. Since the present study showed that information on mRNA can be obtained from AMeX‐processed tissue, the versatility and usefulness of this method were further proven.
Long‐term survival in breast cancer related to overexpression of the c‐erbB‐2 oncoprotein: An immunohistochemical study using monoclonal antibody NCL‐CB11
Authors: Dykins Roger , Corbett Ian P. , Henry James A. , Wright Christopher , Yuan Jun , Hennessy Colm , Lennard Thomas J. W. , Angus Brian , Horne Charles H. W.
Published: June 2005

In a previous series we have shown poor short‐term (3–5 years) survival for patients with tumours overexpressing the c‐erbB‐2 oncoprotein. In this study we employed archival paraffin‐embedded tissue from patients who underwent mastectomy 10–12 years prior to assessment (n = 187). Immunohistochemical staining was carried out by an indirect immunoperoxidase technique using the novel monoclonal antibody NCL‐CBll. Tumours were scored according to intensity of membrane staining. Patient and tumour information was obtained by scrutiny of clinical records. Survival analysis was carried out for both time to relapse and time to death, using the log rank test.

Patients with tumours demonstrating intense membrane staining had a poor prognosis compared with the rest, with a steeply sloped survival curve over the first 4 years; the survival difference was still evident at 12 years follow‐up (P < 0·001). The survival advantage for c‐erbB‐2 negative patients was maintained in lymph node negative patients (P < 0·001). However, c‐erbB‐2 status did not influence survival in the node positive group, where all patients had a uniformly poor outlook. These results applied to both time to relapse and time to death. In conclusion, c‐erbB‐2 status, determined using NCL‐CBll, is a powerful prognostic indicator, defining in particular node negative patients with a particularly poor prognosis, and for whom alternative therapeutic strategies may be appropriate.
Transforming growth factor alpha and epidermal growth factor in human pancreatic cancer
Authors: Barton Claire M. , Hall Peter A. , Hughes Christine M. , Gullick William J. , Lemoine Nicholas R.
Published: June 2005

Overexpression of the epidermal growth factor receptor (EGFR) has been reported as an important molecular abnormality in human pancreatic cancer. There is in vitro evidence that simultaneous overproduction of one of its ligands, transforming growth factor alpha (TGF‐alpha), might result in an autocrine loop with an increased proliferation signal. We analysed by immunocytochemical staining a retrospective series of human pancreatic cancers, chronic pancreatitis, and normal fetal and adult pancreatic tissues for the presence of TGF‐alpha and epidermal growth factor (EGF). Ductal epithelial cells showed TGF‐alpha immunoreactivity in both normal tissue and chronic pancreatitis, and 95 per cent of tumours showed strong immunoreactivity. In contrast, EGF immunoreactivity was not found in normal pancreas, but was expressed in 12 per cent of pancreatic carcinomas. Well‐defined areas of EGF immunoreactivity in exocrine ducts showing reactive changes in pancreatitis might represent a benign response to tissue damage similar to that previously described in the gastric mucosa.
Observer variation in the measurement of Breslow depth and Clark's level in thin cutaneous malignant melanoma
Authors: Colloby Peter S. , West Kevin P. , Fletcher Alan
Published: June 2005

We have assessed the degree of observer variation of both Breslow depth and Clark's level in a series of 50 thin malignant melanomas. Our findings are similar to those of previous international studies in that Breslow depth is the more reproducible measure. Significant intra‐ and inter‐observer variation exists and in some cases it was up to ±0·86 mm. Even small differences will potentially affect patient management at our centre and this was analysed using kappa statistics. Good agreement was found between observers and this could be improved by comparing the mean of two or more measurements. This removes larger errors, but smaller observer errors and differences in subjective interpretation of the deepest malignant cell mean that agreement will never be more than 90 per cent. This is high compared with studies of observer variation in other pathological conditions, e.g., dysplasia of the cervix, but where surgical management is potentially disfiguring it is not high enough. We conclude that Breslow depth and Clark's level should not be the sole basis of wide excision protocols.
The differential diagnosis of epithelial‐type mesothelioma from adenocarcinoma and reactive mesothelial proliferation
Authors: Soosay Geraldine N. , Griffiths Meryl , Papadaki Lucienne , Happerfield Lisa , Borrow Lynda
Published: June 2005

Three monoclonal antibodies (mAbs) have been applied to 14 adenocarcinomas, eight mesotheliomas, and nine reactive mesothelial proliferations. One of these, AUA1, is a novel antibody in this context. It was found in 13/14 of the adenocarcinomas but not in benign or malignant mesothelial tissue, making it a potential substitute for carcino‐embyronic antigen (CEA) in the differential diagnosis of mesotheliomas from adenocarcinomas. In addition, the lesions were stained using an argyrophil technique to identify nucleolar organizer regions. The results of counting these showed a small but significant overlap in their numbers between malignant and benign mesothelial proliferations. Although there was a difference overall in the numbers of nucleolar organizer regions in adenocarcinomas and mesotheliomas, on the one hand, and benign hyperplasias, on the other, this overlap considerably reduces their value in assessing individual difficult cases.
The expression of c‐fos protein in human breast
Authors: Walker Rosemary A. , Cowl J.
Published: June 2005

The expression of the c‐fos protein has been examined in non‐malignant and malignant human breast using immunohistochemistry. Since in some cell types expression is associated with differentiation, while in other cells induction is related to growth, the extent of reactivity in carcinomas has been related to tumour grade, oestrogen receptor, epidermal growth factor receptor, and Ki‐67 staining.

There was a greater degree of staining in the carcinomas than in the normal and benign tissue. In the latter, there was no relationship with menopausal status or with extent of proliferative changes. In all instances, staining was nuclear. Poorly differentiated carcinomas had a greater degree of reactivity, but the converse did not apply to the well differentiated tumours. There was no relationship with oestrogen receptor, epidermal growth factor receptor, or Ki‐67 labelling.

The greater expression observed in carcinomas correlates with previously reported RNA studies. There is no clear relationship between c‐fos protein detection and proliferation or differentiation.
Reduced apoptotic cell death in follicular lymphoma
Authors: Hollowood Kevin , Macartney James C.
Published: June 2005

It has been hypothesized that inhibition of germinal centre cell apoptosis may underlie the development of follicle centre cell lymphomas. We have performed a comparative, quantitative study of apoptotic cell death in germinal centres and in the neoplastic follicles of centroblastic–centrocytic, follicular, non‐Hodgkin's lymphoma (Cb/Cc NHL). Ten cases each of reactive follicular hyperplasia and Cb/Cc NHL were analysed. One‐micrometre‐thick resinembedded sections were examined at × 1000 magnification. The total numbers of cells, nuclear containing apoptotic bodies, and mitoses were counted in each follicle and the apoptotic and mitotic indices were derived. Transmission electron microscopy was performed on selected cases to confirm the typical features of apoptosis. The mean apoptotic (4·9 per cent) and mitotic indices (0·9 per cent) for the germinal centres were significantly higher (P < < 0·001) than those for the neoplastic follicles (0·9 and 0·14 per cent). These results lend support to the recent proposal that reduced apoptotic cell death may be important in the pathogenesis of follicular lymphomas.
Chronic pyelonephritis: The significance of renal renin and the vascular changes in the human kidney
Authors: Peh Suat Cheng , Lindop George B. M.
Published: June 2005

Hypertension complicates chronic pyelonephritis. Since arterial narrowing is common in the damaged kidney, activation of the renin–angiotensin system due to renal ischaemia has been suggested as a pathogenetic mechanism. We used an antiserum to human renin and an immunoperoxidase technique to study the anatomy of renin‐containing cells (RCC) in 18 kidneys removed for pyeloneophritis. We independently assessed the degree of arterial narrowing and correlated these variables with the clinical findings. There was histological evidence of hyperplasia of RCC in 5 of the 6 hypertensive patients and in 7 of the 12 non‐hypertensive cases. There was no difference in the apparent number or distribution of RCC between the hypertensive and the non‐hypertensive cases. Also, the degree of arterial narrowing did not correlate with either the hyperplasia of RCC or the blood pressure of the patients. Our results do not support the hypothesis that narrowing of the intrarenal arteries is important in the pathogenesis of hypertension in pyelonephritis. In our cases, the renal veins were more severely damaged than the arteries and their lumina were often obliterated by organized thrombus. We suggest that such widespread obliteration of the renal venous tree could impair blood flow and contribute to the tissue damage in the pyelonephritic kidney.
Simultaneous detection of fluorescent in situ hybridization and in vivo incorporated brdu in a human bladder tumour
Authors: van Dekken Herman , Vish Edward W. Scher , Pizzolo John G. , Fair William R. , Melamed Myron R.
Published: June 2005

We have used fluorescent in situ hybridization and simultaneous in vivo bromodeoxyuridine labelling of a solid bladder cancer to examine tumour cell subsets for possible proliferative growth differences. In this dual‐labelled preparation, most tumour cell nuclei exhibited monosomy 9, consistent with reported karyotypes of bladder cancer.1 Incorporated bromodeoxyuridine was visualized with a fluoresceinated antibody in 5–6 per cent of the tumour cells, concordant with S‐phase estimates by cell cycle analysis of the flow cytometric DNA histogram. A majority of the bromodeoxyuridine‐positive cells also carried the monosomy 9 chromosome abnormality. This is the first report to demonstrate the feasibility of combined in situ hybridization and detection of bromodeoxyuridine incorporated in vivo in human tumour cells in order to provide information on the growth rate of specific subsets of tumour cells identified by chromosomal constitution.
Malignant peripheral nerve sheath tumour with annulate lamellae mimicking pleomorphic malignant fibrous histiocytoma
Authors: Goodlad J. R. , Fletcher C. D. M.
Published: June 2005

The nature and nosologic status of malignant fibrous histiocytoma (MFH), or at least its most common pleomorphic variant, are a matter of controversy and it is possible that this entity represents the shared morphologic phenotype of a range of other dedifferetiated neoplasms. An unusual case of malignant peripheral nerve sheath tumour which very closely mimicked pleomorphic MFH is presented. Its true nature was only disclosed by ultrastructural examination. A further interesting and previously unreported feature in tumours of this type was the presence of annulate lamellae in many of the tumour cells. This case adds weight to the assertion that the light microscopic features of pleomorphic MFH may not be specific.
Late BCNU lung: A light and ultrastructural study on the delayed effect of BCNU on the lung parenchyma
Authors: Hasleton P. S. , O'Driscoll B. R. , Lynch P. , Webster A. , Kalra S. J. , Gattamaneini H. R. , Woodcock A. A. , Poulter L. W.
Published: June 2005

We describe eight patients who developed interstitial pulmonary fibrosis following BCNU (carmustine) therapy for cerebral tumours. The fibrosis presented 12–17 (mean 14) years after exposure to the drug. A distinctive pattern of pulmonary fibrosis with involvement of the apices and subpleural areas was seen in one patient dying of the disease. Light microscopy showed interstitial elastosis and intra‐alveolar fibrosis which was often focal with an associated mild lymphoplasmacytic infiltrate, intra‐alveolar oedema, macrophages, and some neutrophils. Ultrastructural studies showed electron lucency of type I pneumocytes, with breaks in the cytoplasmic membranes leaving a bare basement membrane. Degenerative change was also seen in endothelial cell cytoplasm along with lipofuscin deposition.

While BCNU pulmonary fibrosis has been described up to 2 years after treatment, this complication so late after therapy, though rare, has important implications for the follow‐up of patients receiving this drug.
Localization of immunoglobulin light chain mRNA expression in Hodgkin's disease by in situ hybridization
Authors: Ruprai Anita K. , Pringle J. H. , Angel Carole A. , Kind C. N. , Lauder I.
Published: June 2005

In situ hybridization techniques were used to detect immunoglobulin light chain messenger RNA (mRNA) in 28 formalin‐fixed, paraffin‐embedded samples of Hodgkin's disease. Cocktails of biotinylated oligonucleotide probes specific for the constant regions of kappa and lambda light chain mRNA were used. None of the Reed‐Sternberg cells or their variants in any of the cases studied showed positive staining with either probe, in contrast to normal plasma cells which showed strong staining in the same sections. It was concluded, therefore, that the cytoplasmic immunoglobulin frequently detected within these cells by immunocytochemistry is present not as a result of synthesis, but as a result of some other mechanism.
Measurement of the tissue distribution of immunoperoxidase staining with polyclonal anti‐BCG serum in lung granulomata of mice infected with Mycobacterium tuberculosis
Authors: Orrell J. M. , Brett S. J. , Ivanyi J. , Coghill G. , Grant A. , Beck J. Swanson
Published: June 2005

Mice inoculated with Mycobacterium tuberculosis, strain H37Rv were used as a model of human tuberculosis. The microanatomical location of immunoperoxidase staining with a polyclonal anti‐BCG serum was within macrophages and appeared granular rather than delineating whole bacilli. Immunoperoxidase staining appears to demonstrate degraded mycobacterial antigens from disrupted organisms and so reflects prior turnover of bacilli. On Ziehl–Neelsen staining, intact or almost intact bacilli are seen and so the extent of this form of staining reflects the current bacillary load. Both methods have limited sensitivity, but with larger mycobacterial loads the area of immunoperoxidase stain measured on a semi‐automated image analyser correlated with the numbers of bacilli observed. The immunoperoxidase method will be useful in the evaluation of residual antigen in studying the pathogenesis of experimental murine tuberculosis. In human mycobacterial granulomata, this immunohistochemical technique should provide an alternative method of estimating the extent of bacillary load: this approach may also provide evidence of mycobacterial infection from residual antigen deposits in the tissue when whole bacilli have been successfully cleared.
An assessment of MAC 387 antibody as a diagnostically useful marker of L1 epithelial antigen expression by lung tumours
Authors: Mayall Frederick G. , Gibbs Allen R.
Published: June 2005

A recent report describing the distribution of L1 epithelial antigen in lung tumours in relation to the histological type claimed that this antigen was a highly reliable marker of squamous cell carcinoma. Our study was designed to test this claim and to examine the potential of this antigen in the typing of lung tumours in biopsy specimens. A total of 143 lung tumours were typed according to the WHO classification and examined immunhistochemically for L1 epithelial antigen experession using commercially available monoclonal mouse anti‐human myeloid/histiocyte antigen (MAC 387). Positivity was found in 46 of 55 squamous cell carcinomas (84 per cent), 12 of 27 adenocarcinomas (44 per cent), 15 of 16 adenosquamous carcinomas (93 per cent), 10 of 15 large cell carcinomas (67 per cent), none of 20 small cell carcinomas, and none of 10 carcinoid tumours. Of those tumours expressing L1 epithelial antigen, most showed a patchy pattern of positivity. From this study it is clear that detection of L1 epithelial antigen, most showed a patchy pattern of positivity. From this study it is clear that detection of L1 epithelial antigen by MAC 387 antibody is not specific for squamous cell carcinomas, but it may have a limited use in the diagnosis of small cell carcinomas and carcinoid tumours as these are consistently negative.
Investigation of a quantitative post‐hybridization signal amplification system for mRNA‐oligodeoxyribonucleotide in situ hybridization
Authors: Hoyland Judith A. , Freemont Anthony J.
Published: June 2005

This study was undertaken to assess a post‐hybridization amplification system for increasing the sensitivity of in‐situ hybridization with oligodeoxyribonucleotide (oligonucleotide) probes.

The technique employs a multilayer streptavidin–biotinylated link protein amplification technique, similar to that used in the ABV histochemical amplification system, to attach increasing numbers of radiolabelled streptavidin molecules to a biotinylated ologonucleotide probe.

In tissue sections the amplification technique increases the signal‐to‐noise ratio dramatically, increasing the sensitivity of in situ hybridization and reducing the autoradiographic exposure time. On an artificial medium the amplification technique has been shown to increase the hybridization signal 100‐fold when compared with a directly radiolabelled oligonucleotide probe. The technique could be used to compare relative amounts of target molecule, there being a linear relationship between the detectable signal and the log of the concentration of the target molecule.
A histochemical and immunocytochemical study of hepatic copper and metallothionein in the pre‐ and post‐natal rat
Authors: Elmes Margaret E. , Haywood Susan , Jasani Bharat
Published: June 2005

Histochemical and immunocytochemical methods to demonstrate copper, copper‐associated protein, lipfuscin, and metallothionein were applied to the liver of fetal and neonatal rats (mid‐term, pre‐term, and 1,3, and 7 days post‐natal). Copper was detected only at 1 and 7 days post‐natal. No copper‐associated protein was found in any liver examined. Lipofuscin granules were present around term in small numbers but were most abundant at 7 days post‐natal when the majority of them were also positive for copper. Metallothionein staining, both nuclear and cytoplasmic, was maximal at mid‐term and becames less intense with maturity. Although nuclear staining was always present, cytoplasmic, staining varied between different areas of the liver. No marked difference in staining intensity was found between 3 and 7 days post‐natal. There are marked differences in copper metabolism in the maturing liver between the rat and man.
In vitro modulation of cellular localization of milk fat globule membrane antigens in human breast carcinomas
Authors: Corcoran David , Jones J. Louise , Walker Rosemary A.
Published: June 2005

Alterations in the cellular localization of cell surface components such as the milk fat globule membrane are a common feature of breast carcinomas and relate to the differentiation of a tumour. This study has examined the potential modulation of such components.

A group of carcinomas were cultured with and without insulin and/or hydrocortisone and the site of staining for milk fat globule membrane, as detected by the antibodies HMFG 1, HMFG 2, and NCRC 11, was assessed using light microscopic and electron microscopic immunohistochemistry. Modulation of localization, with a shift from cytoplasmic vesicle labelling to submembraneous vesicles/cell surface labelling and intracytoplasmic luminal labelling, was observed in 5 of 14 moderately differentiated and 8 of 11 poorly differentiated carcinomas. Two well differentiated carcinomas continued to show peripheral labelling; three poorly differentiated carcinomas showed no change from cytoplasmic labelling only; and the other carcinomas exhibited heterogeneous localization, making any change difficult to assess. Insulin was required for any change to be observed and it is suggested that this has an effect on the mechanisms for intracellular transport of membrane and secretory proteins.
Adhesion of normal and neoplastic lymphoid cells to fibroblasts
Authors: Jack A. S. , Chapman K. J.
Published: June 2005

Adhesion between lymphocytes and other cells is critical to many processes in the normal immune system. Alteration in the expression of cell adhesion molecules may be important in determining the behaviour of malignant lymphomas. In this study, the adhesion of normal lymphocytes to fibroblasts is compared with the adhesion of the T‐cell lymphomas J6 and Hut78 ICRF. J6 was significantly more adherent to fibroblasts than either Hut 78 or PBL despite the fact that J6 expresses almost no LFA‐1. Anti‐LFA‐1 had little effect on the basal adhesion of Hut 78 ICRF or PBL. Addition of anti‐CD2 caused enhanced adhesion of J6 and PBL but not Hut 78ICRF, which expresses little of this molecule. This enhancement was abrogated by anti‐LFA‐1. Anti‐CD45 also caused enhanced adhesion of PBL. This was largely due to LFA‐1‐mediated homotypic cell adhesion. The tumour cell lines display no such homotypic adhesion and the small enhancement of fibroblast adhesion was much less affected by the anti‐LFA‐1 antibody. These results show the complex interactions which occur between adhesion molecules and that differences in patterns of expression between normal and neoplastic cells could be a major determinant of tumour cell behaviour.
Cellular sources of matrix proteins in experimentally induced cholestatic rat liver
Authors: Abdel‐Aziz Gamal , Rescan Pierre‐Yves , Clement Bruno , Lebeau Guy , Rissel Maryvonne , Grimaud Jean‐Alexis , Campion Jean‐Pierre , Guillouzo André
Published: June 2005

Collagens (I, III, and IV), fibronectin, and laminin were localized using the indirect immunoperoxidase technique 14 days after bile duct ligation, i. e., when extensive fibrosis and numerous neoformed bile ducts were observed. Extensive fibrous septa in enlarged portal spaces were stained for collagens I, III and IV, fibronectin, and laminin. Collagen IV and laminin were abundant around proliferative bile ducts. In addition, collagen IV was nearly continuous in the sinusoids. At the ultrastructural leve, antigens were localized in the endoplasmic reticulum of several liver cell types. In portal spaces, bile duct cells and cells that form the transitional canal of Hering were strongly labelled for basement membrane components, particulariy laminin, but not for collagens I and III and fibronectin, which were abundant in fibroblast‐like cells. Inside the lobule, only Ito cells and, to a lesser extent, endothelial cells contained collagens, fibronectin, and laminin. Ito cells were found to be heavily stained for collagens III and IV, and laminin. Except for fibronectin, which was always abundant, precursors of extracellular matrix proteins were only slightly detectable in the endoplasmic reticulum of some heptocytes, particularly those located close to altered areas. This study demonstrates that experimental extrahepatic cholestasis in the rat induces periportal fibrosis and continuous deposition of collagen IV in the sinusoids. Several cell types participate in the formation of extracellular matrix components, particularly bile duct cells and Ito cells, with a possible involvement of hepatocytes, thus suggesting that cholestasis provokes changes in the pattern of matrix protein production in liver cells.
The latent membrane protein‐1 in Epstein‐Barr virus‐transformed lymphoblastoid cells is found with ubiquitin‐protein conjugates and heat‐shock protein 70 in lysosomes oriented around the microtubule organizing centre
Authors: Lázló Lajos , Tuckwell Jayne , Self Tim , Lowe James , Landon Michael , Smith Sharon , Hawthorne John N. , Mayer R. John
Published: June 2005

Immunofluorescence studies on Epstein‐Barr virus (EBV)‐transformed lymphoblastoid cells have previously shown that the latent membrane transforming protein (LMP‐1) is found in patch‐like inclusions which also immunostain for vimentin. We now show that EBV transformation causes a major reorganization of intermediate filaments, microtubules, mitochondria, and lysosomal elements, which generally become oriented around the microtubute organizing centre. Immunogold electron microscopy shows that LMP‐1 is primarily concentrated in secondary lysosomes together with ubiquitin‐protein conjugates and heat‐shock protein 70. Intermediate filament inclusion formation with the above characteristics may be a general response triggered by other membrane glycoproteins; as seen, for example, in major human neurodegenerative diseases such as diffuse Lewy body disease.
A series of 14 new monoclonal antibodies to keratins: Characterization and value in diagnostic histopathology
Authors: Bártek Jiří , Vojtěšek Bořivoj , Stašková Zdenka , Bártková Jiřina , Kerekés Zoltán , Rejther Aleš , Kovařík Jan
Published: June 2005

A series of 14 new mouse monoclonal antibodies (MAbs) to keratins is described and the data suggesting their potential value in the differential diagnosis of human tumours are reported. The specificities of individual MAbs of the ‘C‐series’ presented here range from monospecificity for keratin No. 7 (MAbs C‐18, C‐35, C‐62, and C‐68), keratin No. 8 (MAbs C‐15, C‐43, and C‐51), and keratin No. 18 (MAbs C‐04 and C‐08) up to the broadly reacting ‘pan‐keratin’ MAb C‐11, with the target epitopes of the remaining four MAbs being shared by different pairs of keratin polypeptides. The results of the biochemical characterization of the MAbs, together with their immunohistochemical staining patterns on frozen as well as on paraffin sections of normal human tissues, suggest that they represent a significant contribution to the growing list of anti‐keratin MAbs applicable in both research and routine diagnostic pathology. The immunohistochemical examination of a wide range of human neoplasms with the new MAbs not only confirmed their value in making distinctions between carcinomas, on the one hand, and lymphomas, melanomas, and gliomas, on the other, but also verified the possibility of more subtle subdivisions within the group of adenocarcinomas and their metastases. Furthermore, the identification of small subsets of breast carcinomas with decreased levels or apparent loss of the keratin No. 7 polypeptide and some cases of stomach carcinoma with apparently induced expression of this keratin suggests that such ‘exceptions’ must be considered when using keratin spectra as one of the criteria in differential diagnosis.
An outbreak of aflatoxicosis and boric acid poisoning in Malaysia: A clinicopathological study
Authors: Chao Tzee‐Cheng , Maxwell Sheila M. , Wong Su‐Yong
Published: June 2005

An outbreak of food poisoning resulting in 13 deaths in children occurred in Malaysia during the Chinese Festival of the Nine‐Emperor Gods in 1988. The offending food was a Chinese noodle called ‘Loh See Fun’ (LSF). The source was traced to a factory where a banned food preservative was added to make the LSF. The food poisoning was attributable to aflatoxins and boric acid. The clinical features included vomiting, pyrexia, diarrhoea, abdominal pain, anorexia, giddiness, seizures, and eventual coma. Initially, many presented with a Reye‐Like syndrome. Eleven post‐mortem examinations were performed. The pathological findings included extensive coagulative necrosis of the liver with proliferative ‘ductal/ductular metaplasia of the hepatocytes’. Giant cell formation, central vein sclerosis, bile stasis, and steatosis were also noted. There was presence of acute tubular necrosis, superficial upper gastrointestinal erosions, and ensuing encephalopathy. The eventual cause of death is acute hepatic and renal failure.
Immunocytochemical observations on macrophage populations in normal fetal and adult human liver
Authors: Bardadin Krzysztof A. , Scheuer Peter J. , Peczek Alicja , Wejman Jaroslaw
Published: June 2005

Phenotypic expression of macrophages was studied immunocytochemically in 25 human fetal livers at various stages of development and in 20 normal human adult livers. A panel of commercially available polyclonal and monoclonal antibodies (KP1, Mac387, LN3, CR3/43, and antibodies against muramidase, alpha‐1‐antitrypsin, and factor XIIIa) was applied to paraffin sections. From the seventh week of gestation macrophages in the fetal liver showed differences in distribution with the various antibodies. Macrophages in adult liver similarly varied in morphology and phenotypic expression. In the light of these results, we conclude that the population of human liver macrophages is heterogeneous from an early stage of fetal development and that this heterogeneity extends into adult life.
Thyroglobulin in human thyroid follicles with acid mucin
Authors: Harach H. Ruben
Published: June 2005

The minority of human thyroid follicles that contain acid mucin are often grouped in the C‐cell area of the gland, and are lined by abundant C cells. They are thought to be derived from the ultimobranchial contribution to the thyroid gland. This study demonstrates the presence of thyroglobulin reactive follicular cells and colloid as well as calcitonin and CGRP in these ultimobranchial‐related thyroid follicles. These findings add further support to the belief that the ultimobranchial body is a source of at least some C cells and follicular cells, and support the view that thyroid mixed medullary follicular neoplasia may originate from an ultimobranchial stem cell line.
Immunohistochemical and quantitative study of interstitial and intratubular leydig cells in normal men, cryptorchidism, and Klinefelter's syndrome
Authors: Regadera J. , Codesal J. , Paniagua R. , Gonzalez‐Peramato P. , Nistal M.
Published: June 2005

Testicular specimens from normal men and men with cryptorchidism (CR) or Klinefelter's syndrome (KS) were taken, processed for light microscopy, and stained with the avidin–biotin peroxidase complex method for immunohistochemical detection of testosterone. The Leydig cells were classified by their morphology (normal, multivacuolated, and pleomorphic Leydig cells) and by their staining affinity for anti‐testosterone antibodies (T−, T+, and T++ cells), and the average numbers of each cell type for each group of testes were calculated. Normal testes showed morpholigically normal interstitial Leydig cells (96·0 ± 10 per cent) and multivacuolated Leydig cells (4·0 ± 1 per cent). Cryptorchid testes showed normal Leydig cells (85·8 ± 11 per cent) and multivacuolated Leydig cells (14·2 ± 2·3 per cent). Men with KS showed normal Leydig cells (78·9 ± 9·1 per cent), multivacuolated Leydig cells (9·2 ± 1·2 per cent), and pleomorphic Leydig cells (11·0 ± 1·8 per cent). The percentage of T++ cells was higher in normal testes (29·4 ± 2·1 per cent) than in CR (11·4 ± 2·2 per cent) and KS testes (6·3 ± 0·7 per cent). This suggests reduced functional Leydig cell activity in CR and KS. Multivacuolated Leydig cells showed weaker immunostaining than did normal Leyding cells in all the testicular groups. No immunostaining was shown by pleomorphic Leydig cells. Intratubular Leydig cells were only found in CR and KS. Immunostaining was weaker in intratubular Leydig cells than in interstitial Leydig cells. This suggests that intratubular location reduces functional activity of Leydig cells.
Phenotypic analysis of malignant lymphoma in simian immunodeficiency virus infection using anti‐human antibodies
Authors: Ramsay Alan D. , Giddings June , Baskerville Arthur , Cranage Martin P.
Published: June 2005

Primates infected with simian immunodeficiency virus (SIV) develop a condition similar to the human acquired immunodeficiency syndrome (AIDS). The close resemblance between the simian acquired immunodeficiency syndrome (SAIDS) and the human disease has led to the widespread use of SIV‐infected monkeys as an animal model in the study of acquired immunodeficiency. We have investigated the use of standard anti‐human antibodies for the immunohistochemical analysis of formalin‐fixed, paraffin‐embedded tissues from monkeys with SAIDS. With the exception of antibodies UCHL1 (CD45RO), MT1 (CD43), 4KB5 (CD45RA), and Ber H2 (CD30), our routine (human) lymphoma panel of markers worked successfully on the animal tissues. Using the anti‐human antibodies, we were able to analyse the phenotypes of two cases of malignant lymphoma arising in a study group of 26 SIV‐infected rhesus monkeys. Both of the cases stained with the antibodies WR16(CD45RA) and L26 (CD20), and the B‐cell lineage of the lymphomas was confirmed by the detection of IgAδ immunoglobulin expression in one case, and IgM heavy chain in the other. We therefore report the successful use of anti‐human antibodies in the immunohistochemical analysis of lymphomas arising in non‐human primates infected with SIV.
Epstein–Barr virus and carcinomas: Undifferentiated carcinomas but not squamous cell carcinomas of the nasopharynx are regularly associated with the virus
Authors: Niedobitek Gerald , Hansmann Martin L. , Herbst Hermann , Young Lawrence S. , Dienemann Dieter , Hartmann Carl A. , Finn Teresa , Pitteroff Sibylle , Welt Anja , Anagnostopoulos Ioannis , Friedrich Reinhard , Lobeck Hartmut , Sam Choon K. , Araujo Iguaracyra , Rickinson Alan B. , Stein Harald
Published: June 2005

The Epstein–Barr virus (EBV) is consistently associated with undifferentiated nasopharyngeal carcinoma (NPC). There is, however, conflicting evidence as to whether squamous cell NPCs are also EBV associated. Moreover, it has been proposed that other epithelial tumours, particularly thymomas and thymic carcinomas, should be included in the group of EBV‐associated neoplasias. However, since the viral DNA in these studies was demonstrated only in extracted DNA, the cellular origin of the viral DNA is uncertain. We have therefore investigated 152 epithelial tumours from various sites for the presnece of EBV‐DNA by in situ hybridization with 35S‐labelled probes. Sixty‐eight of 77 undifferentiated NPCs showed an EBV‐specific autoradiographic signal, thus confirming the strong association of this tumour type with EBV even in geographical areas where undifferentiated NPC is not endemic. None of eight squamous cell NPCs showed an EBV‐specific signal. All of 15 carcinomas with a similar morphology to undifferentiated NPC but from different anatomic sites (thymus, tonsil, breast) were EBV‐negative as were 9 thymomas, 26 squamous cell carcinomas of the palatine tonsil, and 14 cervical carcinomas. Our results therefore suggest a unique association of EBV with undifferentiated NPC and support concepts assigning different biological properties to undifferentiated NPC as compared with squamous cell NPC.