Quantitative assessment of long-term changes in insulin secretion after canine duct-obliterated pancreas transplantation.

In this study, we have quantitatively assessed the changes in insulin-secreting capacity after duct-obliterated segmental pancreatic autotransplantation in dogs. To this end, we have used a technique for the sampling of the complete and undiluted pancreatic venous blood with simultaneous flow measurement, by which means the actual insulin-secreting capacity was determined in a direct fashion. Histologic changes were analyzed in addition in order to address the underlying mechanisms of the changes in insulin-secreting capacity.

Type IV collagen immunoreactivity in colorectal cancer. Prognostic value of basement membrane deposition.

Using antibodies to type IV collagen, basement membrane (BM) deposition at the tumor-stromal border was studied in 163 cases of colorectal carcinomas. Immunoreactivity was scored semiquantitatively as moderate/extensive versus limited BM deposition and correlated with Dukes' stages and survival data. Cases with limited BM deposition showed an overall significant shorter survival and were overrepresented in Dukes' Stages C/D.

Culturing and xenografting of primary colorectal carcinoma cells: comparison of in vitro, and in vivo model and primary tumor.

In a series of 61 primary colorectal carcinomas, we attempted to determine which primary tumor characteristics correlated with the possibility to continuously maintain tumor cells in vitro or in vivo, and to what extent the characteristics of a primary tumor were maintained in vitro or as xenograft. Four continuous cell lines and 10 serially transplantable tumors were obtained. Only one cell line could be maintained both in vitro and in vivo.

Quantification of oestrogen receptors in breast cancer: radiochemical assay on cytosols and cryostat sections compared with semiquantitative immunocytochemical analysis.

A radiochemical oestrogen receptor assay on cytosol was correlated with a radiochemical and an immunohistochemical oestrogen receptor assay using cryostat sections from 50 breast cancer specimens. Oestrogen receptors were reliably quantitated in 6 micron cryostat sections with Scatchard analysis using radiolabelled oestradiol, and a good quantitative and qualitative relation with cytosol oestrogen receptor assay was found. Parallel sections were used for routine histological tissue verification and for direct comparison with immunohistochemistry for oestrogen receptor.

The prognostic value of morphometrical features and cellular DNA content in cis-platin treated late ovarian cancer patients.

In 73 CAP-1 treated stage III and IV ovarian cancers, the prognostic significance of morphometric features and cellular DNA content has been evaluated in comparison with histologic type, grade of differentiation and a number of clinical characteristics. Borderline tumours were excluded from the study. Median follow-up was 44 months, median survival time 36 months.

Progesterone receptor quantification with radiolabeled promegestone (R 5020) in frozen sections of endometrium and breast cancer tissue.

A technique for the determination of the progesterone receptor content at sections was developed. Series of coverglass-mounted unfixed frozen sections were incubated with [3H]R5020 only, to determine total binding, or with excess unlabeled R5020, to determine non-specific binding. Ligand binding in the tissue sections was measured by liquid scintillation counting after repeated washing of the coverslips.

Xenografting of normal colonic mucosa in athymic mice.

Colonic mucosa was implanted subcutaneously into nude mice in order to investigate the potential use of xenografts as an in vivo model system for the study of colonic epithelial proliferation and differentiation. The xenografts were followed for 5 weeks. Proliferation was studied by bromodeoxyuridine (BrdU) incorporation followed by visualization of the DNA-synthesizing cells with an anti BrdU monoclonal antibody.

A multivariate analysis of pathologic prognostic indicators in large bowel cancer.

A multivariate analysis of the pathologic data of 350 patients with primary colorectal cancer was performed. In addition to conventional parameters such as shape and size of the primary tumor, central node involvement, angioinvasive growth, grade, and stage, new variables such as the immunoreactivity patterns of carcinoembryonic antigen (CEA), CA 19-9, mucin, serotonin, secretory component (SC), and the DNA index were tested for their potential prognostic value. Every variable except CA 19-9, serotonin, and DNA showed significant prognostic information in univariate analysis.

Influence of implantation site on growth, antigen expression and metastatic potential of human colonic cancer HT29 and 5583 xenografts in nude mice.

In order to study the interaction between tumors and host environmental factors, we xenografted cells from the human colonic carcinoma cell lines HT29 and 5583-S in the subcutis, cecum, spleen and liver of nude mice and compared growth characteristics, metastatic potential and some phenotypic features of the xenografts in these sites. No remarkable differences were observed between the tumors at different inoculation sites in regard of their expression of carcinoembryonic antigen and secretory component or type of mucin produced. Also the proportion of DNA synthesizing cells as determined by bromodeoxyuridine incorporation appeared to be comparable in the studied implantation sites.

Monoclonal antibodies in immunocytochemistry.

In this review the application of monoclonal antibodies for immunocytochemical purposes is discussed. An important basic principle is that from a methodological point of view that there are no essential differences between monoclonal and polyclonal antibody based immunocytochemistry. Specificity of the primary antibody is the most important prerequisite for reliable immunocytochemistry.

Effect of tissue fixation on anti-bromodeoxyuridine immunohistochemistry.

We investigated the influence of various fixatives on monoclonal anti-BrdUrd antibody binding of BrdUrd-substituted DNA in tissue sections of routinely processed mouse small intestine after in vivo administration of BrdUrd. For denaturing fixatives such as ethanol or Carnoy's fluid, a standard denaturation protocol showed specific crypt cell labeling. With cross-linking agents such as formalin and glutaraldehyde, a remarkable increase in staining intensity was obtained after tissue digestion with pepsin before acid denaturation.

Mucosal alterations in the reservoir of patients with Kock's continent ileostomy.

Mucosal alterations in the reservoir of 10 patients with Kock's continent ileostomy were investigated and compared with normal ileal mucosa. Morphometric measurements showed a decreased villus length and an increased crypt length as compared with the control group. When the villus to crypt ratio was related to the time elapsed after the continent ileostomy operation, a decrease could be found.

Retrospective analysis of the prognostic significance of DNA content and proliferative activity in large bowel carcinoma.

In the present study we have evaluated the prognostic significance of ploidy levels and proliferative activity in 279 cases of large bowel carcinomas which were included in a surgical prospective randomized trial. Ploidy levels and proliferative activity were determined on nuclei isolated from paraffin-embedded tissues of 279 colorectal carcinoma patients, with a mean follow-up of 51.9 months.

Comparison of phenotypic and genotypic features in human primary large bowel carcinomas and lymph node metastases.

In order to test the contention that metastasis is a selective process and that therefore metastases might show a more restricted pattern of phenotypic and genotypic characteristics than primary tumors, we compared the expression of carcinoembryonic antigen, Ca 19-9, secretory component, serotonin, and mucin production as well as flow cytometric data on DNA content and percentage of S-phase cells in 87 primary large bowel carcinomas and their lymph node metastases. In a majority of the cases primary tumors and their metastases were largely identical with regard to the examined phenotypic features. In discrepant cases, however, metastases did not invariably show a more restricted pattern than primary tumors, indicating high differentiational plasticity of primary and metastatic colorectal cancer cells.

An improved method for the immunocytochemical detection of bromodeoxyuridine labeled nuclei using flow cytometry.

A new staining protocol is described for the immunocytochemical detection of BrdUrd labeled nuclei. Pepsin treatment of ethanol fixed cells or tissue, followed by DNA denaturation at low pH, resulted in increased sensitivity of BrdUrd staining comparable to the thermal denaturation protocol, and decreased background binding. This technique is applicable to cell suspensions, including cultured cells and bone marrow cells.

Primary malignant fibrous histiocytoma of the liver--a case report with immunocytochemical observations.

A case of a primary sarcomatous tumour in the liver of an elderly female is reported. The tumour consisted of bundles of spindle cells focally in a storiform pattern, intermingled with bizarre giant cells. Immunocytochemically, carcinoembryonic antigen, alpha-fetoprotein, keratin, desmin and type IV collagen could not be demonstrated.

Studies with anti-bromodeoxyuridine antibodies: II. Simultaneous immunocytochemical detection of antigen expression and DNA synthesis by in vivo labeling of mouse intestinal mucosa.

We developed a rapid and convenient immunocytochemical method for simultaneous detection of antigen expression and S-phase cells by means of anti-bromodeoxyuridine (BrdUrd) antibodies. Immunocytochemical detection of BrdUrd after in vivo administration in mice was compared with autoradiography using [3H]-BrdUrd. Both the sensitivity and specificity of the technique were high.

Human specific anti-type IV collagen monoclonal antibodies, characterization and immunohistochemical application.

This paper describes two new monoclonal antibodies reactive with human specific type IV collagen epitopes in frozen as well as routinely fixed and processed tissue sections. The antibodies (1042 and 1043) were raised against human placental type IV collagen and were shown by immunoblotting and ELISA tests to react exclusively with type IV collagen determinants. Extensive immunohistochemical survey studies on panels of tissues from various species, using unfixed cryostat sections, demonstrated that antibody 1042 reacted only with human type IV collagen whereas antibody 1043 in addition reacted with rabbit type IV collagen.

Integrated oncology course for first-year medical students.

In this report an oncology course for first-year medical students is described. The course is characterized by a high level of integration of the involved basic and clinical sciences and is designed according to the principles of problem-based learning. A limited number of lectures provides a frame of reference.

The establishment and characterization of two new cell lines derived from a single human colonic adenocarcinoma.

Two cell lines with different in vitro growth characteristics were established from a single mucinous colonic adenocarcinoma. Epithelial cells of the line 5583-E demonstrated anchorage-dependent growth while those of line 5583-S were anchorage-independent and grew as multicellular floating spheroids. Both cell lines shared common characteristics with respect to the expression of differentiation markers (secretory component, carcinoembryonic antigen), mucins and karyotype (trisomy 12 and 14, marker chromosome) but also showed consistent differences.