A pilot study of the safety and effects of the matrix metalloproteinase inhibitor marimastat in gastric cancer.

The aim of this study was to evaluate the safety and tolerability of 4 weeks administration of marimastat, and to seek evidence of biological activity as observed by changes in the endoscopic appearance of the gastric tumours. 35 patients with advanced, inoperable gastric or gastro-oesophageal tumours were recruited. The dose of marimastat was reduced from the starting dose of 50 mg twice daily (6 patients) to 25 mg once daily (29 patients). 31 completed the 28 day study period. Marimastat was generally well tolerated, with the principal treatment-related toxicity being pain and stiffness of the musculoskeletal system. These symptoms occurred more frequently at the higher-dose, and increased to involve a total of 13 patients (37%) with longer-term treatment. The events were usually rapidly reversible on drug discontinuation. 3 patients receiving prolonged treatment experienced more severe symptoms, with the development of skin thickening and contractures in the hands. At endoscopy, 10 patients showed an increased fibrotic cover of the tumour, 8 had decreased haemorrhagic appearance, and in at least 2 cases where comparative tumour histology was assessable, there was evidence of increased stromal fibrotic tissue.

Gelatinase (MMP-2 and -9) expression in gastrointestinal malignancy.

The aim of the study was to investigate expression of the active and inactive gelatinases (MMP-2 and -9) in colorectal neoplasia and gastric cancer compared with normal mucosa. A total of 53 colorectal cancers and corresponding normal mucosa were studied using gelatin zymography as well as 15 colorectal adenomas and 13 gastric cancers with corresponding normal mucosa. Overexpression of all the gelatinases occurs in both colorectal and gastric cancer, with activation of MMP-2 appearing to be a feature of the malignant phenotype. Overexpression of MMP-9 occurs in colorectal adenomas. The gelatinases are overexpressed in gastrointestinal neoplasia, suggesting that these enzymes may have an important role in tumour invasion and metastasis.

Langerhans cells in normal and pathological vocal cord mucosa.

The Langerhans cells in samples of histologically normal and pathological vocal cord mucosa were counted after identification using S-100 polyclonal antibody. Langerhans cells were commonly seen in vocal cord polyp epithelium but were infrequent in normal cord mucosa. They were also identified in samples of squamous carcinoma, severe dysplasia and chronic inflammation.

Tumor-associated tissue eosinophilia and long-term prognosis for carcinoma of the larynx.

Tumor-associated tissue eosinophilia (TATE) has been previously reported to be associated with a favorable prognosis for a variety of carcinomas, including head and neck cancer. We have examined this relationship in a series of 104 patients with laryngeal cancer who were observed for 5 years or more. Patients were followed up for a minimum of 5 years or until death. The original laryngeal biopsy histologic slides were examined and graded for TATE. Tumor-associated tissue eosinophilia was absent in 73 of the 104 biopsy specimens, and 25 of these 73 patients died directly of laryngeal carcinoma within 5 years. In all, 31 patients had TATE and only 3 subsequently died of laryngeal carcinoma within 5 years (P = 0.009, chi-square test). It appears, therefore, that TATE is associated with a good long-term prognosis for laryngeal carcinoma.

Characterisation of complex carbohydrates in cervical glandular intraepithelial neoplasia and invasive adenocarcinoma.

Using a panel of 11 biotinylated lectins and an avidin-peroxidase detection system on paraffin sections of formalin-fixed material, the range of expression of N-linked glycoprotein backbone structures and glycoprotein terminal structures was established in normal cervical glands. A characteristic pattern of luminal lectin reactivity was noted in areas of glandular tubal metaplasia. A consistent change in the topographic distribution of carbohydrate structures in cervical glandular neoplasia was found. In contrast to cytoplasmic expression in normal glands, 90% of adenocarcinomas, 92% of cases of high-grade cervical glandular intraepithelial neoplasia, and 63% of cases of low-grade cervical glandular intraepithelial neoplasia showed novel linear expression along the lumina of glands. The results indicate molecular as well as morphological similarities between invasive adenocarcinomas and putative preinvasive glandular lesions.

Semiquantitative immunohistochemical studies of blood group antigen A, B, H, Le(a), Le(b) structures and Ii backbone chains in the normal human cervix and in cervical adenocarcinoma.

Epithelia frequently express blood group antigens and these are often perturbed in neoplasia. This study has characterized the range of expression of ABH and Lewis terminal structures and the Ii backbone chains in the normal human cervix by semiquantitative immunohistochemistry. Effects of the secretor gene were defined by determination of salivary secretor status. Modifications of blood group antigen expression in cervical adenocarcinoma were also addressed. Normal cervical squamous and glandular epithelia showed a range of expression of the antigens studied. Lewis-gene-negative cases showed no expression of Lewis antigens. Secretor status had no effect on ABH expression in squamous epithelium, but it did have a marked effect on ABH expression in glands and on Le(b) expression in both squamous and glandular epithelia. Patterns of expression of i chains in squamous epithelium suggest that these may be the carriers of ABH and Lewis antigens in a proportion of cases. Distinct patterns of expression were seen in glandular tubal metaplasia and in endothelium. Adenocarcinomas showed topographical rather than quantitative changes in blood group antigen expression with more extensive luminal expression of ABH, Lewis and Ii structures than that seen in normal glands. This change is distinct from those usually associated with malignancy.

Relationships between epithelial basement membrane staining patterns in primary colorectal carcinomas and the extent of tumour spread.

In colorectal cancer an association has been found between lack of epithelial basement membrane (EMB) immunostaining in the tumour centre and more extensive malignant spread. Interestingly, ultrastructural investigations suggest that EBM loss at the tumour periphery may be part of an invasive mechanism. To further assess the significance of EBM deficiencies in different tumour areas, we carried out a detailed study of the basement membrane laminin immunostaining patterns in 130 cases of colorectal carcinoma. We find that discontinuous EBM staining in the tumour centre is associated with poor tumour differentiation (p less than 0.005), presence of lymph-node metastases (p less than 0.02), and more advanced Dukes stage (p less than 0.02). The latter association is strengthened by excluding cases in which numerous polymorphonuclear leukocytes (PMNs) are present adjacent to EBM breaks, suggesting that these inflammatory cells are a confounding factor. Discontinuous EBM staining is more frequently observed in tumour deep to muscularis propria than in submucosal tumour (p less than 0.02), indicating intra-tumoral variation. At the tumour periphery, extensive EBM discontinuity shows no association with lymph-node involvement, but is linked with deeper local invasion (p less than 0.05). While EBM staining patterns around central and peripheral tumour glands are related (p less than 0.001), staining around peripheral glands is almost invariably more discontinuous. However, EBM lack at the tumour periphery is not as absolute as previously suggested, since in 18% of tumours fewer than 25% of peripheral tumour glands show EBM breaks. This appears consistent with the hypothesis that invasive changes at the tumour periphery are temporary and reversible.

Further studies of lectin binding by villous and extravillous trophoblast in normal and pathological pregnancy.

Examination of human pregnancy tissues with a panel of lectins provides the opportunity to probe different aspects of carbohydrate structure. Nine biotinylated lectins [concanavalin A (con A), wheat germ agglutin (WGA), Lens culinaris A (LCH-A), Pisum sativum (PSA), Phaseolus vulgaris (PHA-E and PHA-L), Ulex europaeus 1 (UEA1), Griffonia simplicifolia (GSI and GSII)] were used to investigate the lectin binding of human trophoblast in normal, tubal, and molar pregnancy. All lectins except UEA1 bound to normal villous syncytiotrophoblast. Binding of lectins to extravillous trophoblast was more restricted than to villous trophoblast, occurring predominantly with con A, PHA-E, PHA-L, WGA, GSI, and GSII. LCH-A reacted with cyto-trophoblastic columns but not with interstitial or endovascular trophoblast. Con A and GSII were the only lectins that bound to trophoblastic giant cells. GSI and GSII bound preferentially to extravillous trophoblast, showing only focal reactivity with villous trophoblast. Lectin binding in ectopic pregnancy was similar to that in normal first-trimester intrauterine pregnancy. Reactivity in molar pregnancy also generally mirrored that observed in normal pregnancy; however, reactivity of GSII with villous trophoblast was more consistent than that observed in normal pregnancy, and GSI showed uniform binding to proliferating syncytial areas. Thus, lectin binding studies allow definition of surface carbohydrates, which may play a role in the controlled trophoblast proliferation and invasion that occurs in normal pregnancy.

Demonstration of low frequency of human papillomavirus DNA in cervical adenocarcinoma and adenocarcinoma in situ by the polymerase chain reaction and in situ hybridization.

There is evidence that cervical adenocarcinoma is increasing in incidence, particularly in young women. In order to assess the possible role of human papillomaviruses in cervical glandular oncogenesis, 16 cases of invasive adenocarcinoma and eight cases of adenocarcinoma in situ have been examined by in situ DNA hybridization using biotinylated probes to human papillomavirus types 6, 11, 16, 18, and 31, and assessed by polymerase chain reaction analysis for human papillomavirus types 11, 16, and 18 sequences. Of the invasive adenocarcinomas, four of 16 contained human papillomavirus type 16 sequences and one of 16 contained type 18 sequences as assessed by polymerase chain reaction analysis. Five of eight cases of adenocarcinoma in situ contained human papillomavirus type 16 sequences by polymerase chain reaction analysis. Only one invasive adenocarcinoma and one case of adenocarcinoma in situ showed a positive in situ hybridization signal. The low rate of carriage of the human papillomavirus sequences examined suggests that these viral types may not play a major role in cervical glandular neoplasia.

Immunolocalization of alpha-amylase in ovarian mucinous tumours.

The distribution of alpha-amylase was studied immunohistochemically in 42 cases of ovarian mucinous tumour. Intense immunoreactivity for amylase was found in 6 of 8 cases of mucinous cystadenocarcinoma. In contrast, only 6 of 20 benign mucinous cystadenomas showed immunoreactive amylase, which was weak and patchy. Mucinous cystadenomas of borderline malignancy showed an intermediate degree of amylase immunoreactivity. The patterns seen are very similar to those reported in normal endocervix, cervical glandular atypia, and invasive adenocarcinoma and suggest molecular as well as morphologic similarities in neoplasia at these sites.

Demonstration of multiple HPV types in normal cervix and in cervical squamous cell carcinoma using the polymerase chain reaction on paraffin wax embedded material.

The prevalence of human papilloma virus (HPV) types 6, 11, 16 and 18 was investigated using the polymerase chain reaction on formalin fixed, paraffin wax embedded material in 19 cases of cervical squamous cell carcinoma and in 10 normal cervices. HPV DNA was detected in 16 of 19 carcinomas, with multiple types present in 11 of these. HPV 16 or 18, or both, were present in all cases in which HPV was shown. Six of 10 cases of normal cervix contained HPV; five of these contained two or more HPV types, including HPV 16 or 18, or both. This study shows the feasibility of using the PCR on paraffin wax embedded material and indicates a high rate of carriage of multiple HPV types in both normal and neoplastic cervix.

Minimal deviation adenocarcinoma (adenoma malignum) of the endocervix: a histochemical and immunohistochemical study of two cases.

The histopathological diagnosis of minimal deviation adenocarcinoma (adenoma malignum) of the endocervix may be difficult. Two cases of minimal deviation adenocarcinoma (MDA) were examined using mucin histochemistry and immunocytochemistry with antibodies to epithelial membrane antigens (HMFG1, Ep1), low-molecular-weight cytokeratins (CAM 5.2), carcinoembryonic antigen (CEA), and alpha-amylase. The results were compared with those for normal endocervical glands. Reactivity for CEA in MDA was focal and would be unreliable for biopsy diagnosis. Both cases of MDA contained abundant neutral mucins and sialomucins, whereas sulfomucins were rarely detected; this pattern contrasted with that of normal endocervix. Neoplastic glandular epithelial cells in MDA consistently showed both luminal and cytoplasmic reactivity with Ep1 and HMFG1, whereas normal cervix showed luminal labeling only. Thus, mucin histochemistry and immunohistochemical detection of epithelial membrane antigens may distinguish between extremely well differentiated neoplastic glands in MDA and normal endocervical glands, and hence may aid diagnosis in biopsy specimens.

Modulation of the antigenicity of amylase in cervical glandular atypia, adenocarcinoma in situ and invasive adenocarcinoma.

Using a polyclonal anti-salivary alpha-amylase antibody and a biotin-avidin-peroxidase detection system immunoreactive amylase was detected in 39/40 cases of invasive cervical adenocarcinoma, 6/7 cases of adenocarcinoma in situ, 6/6 cases of high-grade cervical glandular atypia and 12/14 cases of low-grade cervical glandular atypia. Expression was seen in only 2/18 normal endocervices and this was weak and patchy. In contrast, strong amylase activity was demonstrated in 18/18 normal endocervices using a starch film assay on frozen sections. It is proposed that antigenic modulation of amylase occurs during endocervical neoplastic transformation.

Patterns of reactivity with the monoclonal antibodies HMFG1 and HMFG2 in normal endometrium, endometrial hyperplasia and adenocarcinoma.

Using an indirect immunoperoxidase technique the expression of the epitopes in human milk fat globule (HMFG) membranes detected by the monoclonal antibodies HMFG1 and HMFG2 was studied in the normal endometrium and in cases of cystic glandular hyperplasia, glandular hyperplasia with architectural atypia (complex hyperplasia), glandular hyperplasia with cytological atypia (atypical hyperplasia) and invasive adenocarcinoma. Luminal reactivity with HMFG1 was seen in cases of normal endometrium, cystic glandular hyperplasia and glandular hyperplasia with architectural atypia. In contrast most cases of glandular hyperplasia with cytological atypia and invasive adenocarcinoma also showed areas of cytoplasmic reactivity. Reactivity with HMFG2 was scanty.

The distribution of beta-adrenoceptors in human cervix.

The distribution of beta-adrenoceptors in sections of human cervix taken at the proliferative phase of the menstrual cycle was studied using light-microscopic autoradiography. The radioligand [125I]iodocyanopindolol (125ICYP) was used to identify specific binding sites. Moderate density of labelling by 125ICYP was seen over smooth muscle and blood vessels. The most intense labelling, however, was seen over glands and surface columnar epithelium. The association of beta-adrenoceptors with glands and surface columnar epithelium suggests a possible adrenergic regulation of secretory function in the cervix.

A study of nucleolar organizer regions in cervical intraepithelial neoplasia and human papillomavirus infection.

A silver colloid technique for nucleolar organizer region-associated proteins (AgNORs) was applied to sections of cervix that comprised cervical intraepithelial neoplasia (CIN) grades I to III, koilocytosis indicative of human papillomavirus infection, squamous metaplasia, and basal cell hyperplasia. The AgNOR count was compared both within and between specimens. Proliferative activity was investigated by flow cytometric analysis. Significant differences in AgNOR numbers were found between normal internal controls and the lesions investigated. A correlation was found between the proliferative index (%S + G2) determined by flow cytometry and the number of AgNORs in cases of CIN.

Prognostic indicators in centroblastic-centrocytic lymphoma.

The prognostic importance of ploidy and proliferative index (%S + G2) assessed by flow cytometry, mitotic and centroblast counts, and histological growth pattern were evaluated in biopsy specimens taken before treatment from 60 cases of centroblastic-centrocytic non-Hodgkin's lymphoma. Cases with a high proliferative index (greater than or equal to 18%) or DNA aneuploidy showed significantly poorer survival than those with a low proliferative index (less than 18%). A high mitotic count was also associated with a poor prognosis. On multiple regression analysis the flow cytometric assessments and mitotic counts were significant predictors of survival. Assessments of proliferative activity clearly have prognostic potential in centroblastic-centrocytic lymphoma and may permit more accurate characterisation of individual tumours.

Cytoplasmic reactivity with the monoclonal antibody HMFG1 as a marker of cervical glandular atypia.

Cervical adenocarcinomas show cytoplasmic reactivity with the monoclonal antibody HMFG1 in contrast to the luminal border (cuticular) staining seen in normal endocervical glands and microglandular hyperplasia. HMFG1 was applied to 15 cases of cervical glandular atypia: thirteen showed cytoplasmic reactivity similar to that seen in overt endocervical neoplasia. This pattern of reactivity seems to be a useful marker of cervical glandular atypia and further supports its suggested place in the development of adenocarcinoma.