J. O'd. McGee

Secretion of epidermal growth factor by macrophages associated with breast carcinoma

By means of a cytokine release assay we have shown that in cell populations derived from primary breast carcinoma, epidermal growth factor (EGF) is secreted by cells with the characteristic morphological and immunophenotypic profile of activated macrophages (positive for CD68, CD16, CD25). EGF secretion was observed in 11 (31%) of 35 primary tumours. Secretion of EGF by normal or malignant epithelial cells was not observed. We found no association between EGF secretion by the primary tumour and recognised clinical indices of prognosis.

A new marker for human cancer cells, 2 immunohistochemical detection of the Ca antigen in human tissues with the Ca1 antibody.

The Ca1 antibody has been used in an immunohistochemical procedure to detect the Ca antigen in sections of tissues routinely embedded in paraffin wax. A representative sample of benign and malignant tumours from all the systems of the human body has been examined. The majority of malignant tumors express the Ca antigen. The exceptions are: prostatic carcinomas, testicular teratocarcinomas and seminomas, some sarcomas, some lymphomas, malignant brain tumours, neuroblastomas, and melanomas. The antigen is least readily detected in epithelial malignancies of the alimentary system, particularly of the colon. The Ca1 antibody does not react with any benign tumour. The only normal tissues that react specificity with this antibody are the epithelium of the fallopian tube and the transitional epithelium of the urinary tract. The Ca1 antibody also readily distinguishes malignant cells in smears of malignant effusions. These findings indicate that the Ca1 antibody may be useful in the diagnosis of malignancy in routine clinical practice where the morphological interpretation of the biopsy or cytological smear is in doubt.

Diclofenac associated hepatitis

Diclofenac is a widely used non-steroidal anti-inflammatory drug, being the most commonly prescribed of its kind in the world. This paper describes five cases of hepatitis with clinical features indicating a direct link with diclofenac. All the patients presented with an acute hepatitis, three being jaundiced. They gave a history of taking diclofenac up to the time of presentation, four of the five having started the drug within the previous 3 months. There were no other features in the histories to suggest alternative causes for the liver dysfunction. Liver function tests were grossly abnormal in all cases, showing a hepatitic picture. A liver biopsy was performed in 4 cases, and showed features of an acute hepatitis with inflammation and hepatocyte damage dominating. The liver dysfunction returned to normal on drug withdrawal in four of the five cases, with full recovery by 3 months. One patient developed steroid-responsive chronic active hepatitis. Hepatotoxicity associated with diclofenac is documented, but previously only a few isolated cases have been described. The occurrence of five cases in one gastroenterology unit over a 12-month period suggests that hepatitis associated with diclofenac may be commoner than previously supposed.

New sites of cellular vitronectin receptor immunoreactivity detected with osteoclast-reacting monoclonal antibodies 13C2 and 23C6.

The immunohistochemical profile of osteoclast-reacting monoclonal antibodies, 13C2 and 23C6, known to detect the alpha-chain of the vitronectin receptor, is described. Both antibodies reacted with several cell types apart from osteoclasts including megakaryocytes, smooth muscle cells, endothelial cells, thyroid follicular epithelium, renal glomeruli and tubular epithelium, myoepithelial and epithelial cells in the breast and prostate, and both cytotrophoblast and syncytiotrophoblast. In addition, macrophage polykaryons, synovial lining cells, a small number of mononuclear cells in buffy coats, and a few macrophage-like cells in the stroma of various tissues were also stained. The epitopes recognized by these antibodies are thus not osteoclast-specific and are present on other cells of the mononuclear phagocyte system. The implications of these results for osteoclast ontogeny, the nature of the antigens described and the question of osteoclast-specific antibodies are discussed.

A NEW MARKER FOR HUMAN CANCER CELLS. 3. IMMUNOCYTOCHEMICAL DETECTION OF MALIGNANT CELLS IN SEROUS FLUIDS WITH THE Cal ANTIBODY

The Ca1 antibody has been used in an immunocytochemical procedure on smears of cells recovered from 50 effusions from the pericardial, peritoneal, and pleural cavities. In 25 samples containing malignant cells, as assessed by conventional morphological criteria, this antibody distinguished malignant from non-malignant cells in 21 samples. The exceptions were 2 lymphomas and 2 out of 6 carcinomas of the lung. The Ca1 antibody also reacted with malignant mesothelial cells from 2 mesotheliomas, but not with reactive mesothelial cells. This antibody thus provides a new method for the identification of malignant cells in pericardial, peritoneal, and pleural effusions.

Human tumour-associated macrophages differentiate into osteoclastic bone-resorbing cells

Macrophages are commonly found within osteolytic secondary carcinomas in bone, but the manner in which these cells contribute to malignant bone resorption is uncertain. Macrophages isolated from primary breast carcinomas were co‐cultured for up to 21 days with UMR 106 rat osteoblast‐like cells on bone slices and glass coverslips in the presence and absence of 1,25‐dihydroxyvitamin D3 [1,25(OH)2D3] and human macrophage colony stimulating factor (M‐CSF). Cell cultures were then assessed for the presence of phenotypic markers of macrophage and osteoclast differentiation. Isolated cells were negative for osteoclast markers including tartrate‐resistant acid phosphatase (TRAP), vitronectin receptor (VNR), and the ability to carry our lacunar bone resorption, but were positive for CD11b and CD14, macrophage markers which are not present on osteoclasts. In 21‐day co‐cultures of breast carcinoma tumour‐associated macrophages (TAMs) and UMR 106 cells, incubated in the presence of 1,25(OH)2D3 and M‐CSF, numerous TRAP‐ and VNR‐positive multinucleated cells capable of extensive lacunar resorption were formed. Contact with UMR 106 cells and the presence of 1,25(OH)2D3 and M‐CSF were absolute requirements for differentiation of human breast carcinoma TAMs into mature functional osteoclasts. TAM–osteoclast differentiation may represent an important cellular mechanism of osteolysis in metastatic skeletal carcinomas.

Robotic telepathology: efficacy and usability in pulmonary pathology

Robotic telepathology is well established in the USA as a method of case referral, but is less frequently used in the UK. Using cases covering a broad spectrum of pulmonary pathology, this study assessed its application in primary diagnosis and its functionality in terms of accuracy of diagnosis and time per case, for both small biopsies and open lung biopsies/resections. Forty cases (20 bronchoscopic and 20 surgical lung biopsy/resection specimens) were reviewed in blinded fashion by a single pathologist using robotic telepathology. Connection between the John Radcliffe and Royal Brompton Hospitals was via 10 Mb/s LAN to the Internet (supported by the Joint Academic Network). The cases were then randomized and reviewed a second time with conventional light microscopy. Diagnosis, initial time to reach diagnosis, and overall time per case were recorded. In two bronchoscopic biopsy cases, there were clinically significant differences between telepathology and conventional light microscopy, one probably attributable to user inexperience and the other to either speed of image capture or digital image quality. In the surgical lung biopsies and resections, there was one variation of opinion: with telepathology a case was considered to be probably mesothelioma, whereas this was thought less likely on light microscopy. In both instances, immunohistochemistry was requested prior to clinical management. Telepathology was 14 times slower than conventional light microscopy when examining bronchoscopic biopsies. The average time spent per slide was 7 min 21 s, compared with 32 s per slide with conventional light microscopy. When assessing open lung biopsies and resections, telepathology was five times slower, at 6 min 13 s compared with 1 min 10 s with conventional light microscopy. This study showed that robotic telepathology is accurate for primary diagnosis in pulmonary histopathology, but modifications in both laboratory protocols and telepathology hardware are needed to decrease the time difference between telepathology and conventional light microscopy, for telepathology to be usable within the framework of a busy referral practice. Copyright

SKIN PROLYL HYDROXYLASE IN PATIENTS WITH OBSTRUCTIVE JAUNDICE

Abstract Prolyl hydroxylase was estimated in the skin of eight jaundiced and nine control patients. The activity of this enzyme was considerably reduced (approximately 70%) in the skin of patients with obstructive jaundice as compared to control patients.

The discrimination of high-risk HPV types by in situ hybridization and the polymerase chain reaction

SummaryThe parameter Tmt has been defined by non-isotopic in situ hybridization and describes the tissue melting temperature (Tmt) of human papillomavirus (HPV) DNA sequences. In this study, multiple in situ hybridization signals for HPV types 16, 31 and 33 in individual archival biopsies hybridized with genomic probes are shown by polymerase chain reactions to be due to cross-hybridization of probe sequences to a single tissue target. Tmt is independent of viral type but depends on the homology between probe and target when using nick-translated whole genomic probes. The difference between Tm and Tmt is not due to the presence of viral capsid protein. Multiple HPV signals in archival material should not therefore be interpreted as indicative of multiple HPV infection unless adequate stringency conditions have been employed or they are present in morphologically distinct areas of the biopsy.Furthermore, extrapolation of calculated DNA homologies to non-isotopic in situ hybridization analysis may not be appropriate. A hybridization signal does not imply probe and target identity: this has implications for HPV typing in clinical material.

Epidermal growth factor, insulin-like growth factor-1 and basic fibroblast growth factor modulate the cytostatic effect of tumour necrosis factor-α on the breast cancer cell line, T47D

The effect of epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and insulin-like growth factor-1 (IGF-1) on the cytostatic activity of tumour necrosis factor-alpha (TNF-alpha) was studied on the breast cancer cell line, T47D. TNF-alpha completely blocked the potent growth-promoting activity of all three factors on T47D cells. EGF, bFGF and IGF-1 partially antagonized the inhibition of cell growth and thymidine incorporation induced by TNF-alpha. These data suggest that the growth of these breast cancer cells is regulated, in part, by the antagonistic interaction between the cytostatic effect of TNF-alpha and the growth-promoting activity of factors such as EGF, IGF-1 and bFGF.