Jens Høstmark

Immunohistochemical markers in urinary bladder carcinomas from paraffin‐embedded archival tissue after storage for 5–70 years

To evaluate archival tissue specimens from bladder tumours and seek molecular changes in samples collected over seven decades previously, as although the frequencies of some cancer types have remained stable during the last 50 years, the incidence of others, including bladder tumours, has increased significantly, and molecular analyses of bladder cancer over periods with an increasing incidence are of interest as the findings might reflect varying external influences.

Ploidy Disturbance of Normal-Appearing Bladder Mucosa in Patients with Urothelial Cancer: Relationship to Morphology

A total of 4 patients with transitional cell carcinoma of the bladder underwent complete mapping of the mucosa and tumors with combined cytologic, histologic and flow cytometric evaluation of the extent of involvement of the neoplastic process. Flow cytometric measurement of the cellular deoxyribonucleic acid content in multiple cell samples taken at cystoscopy showed similar changes in the normal mucosa as in the tumors. These changes consisted of an increased fraction of cells with S-phase deoxyribonucleic acid content in 2 patients with grades 1 and 2 tumors, and the presence of extensive aneuploidy in 2 patients with World Health Organization grades 2 and 3 tumors. While grade 1 and some grade 2 tumors (World Health Organization) are composed only of diploid cells, some of the grade 2 and all grade 3 tumors consist of a mixture of diploid and aneuploid populations. Such aneuploid clones could be identified in normal-appearing mucosa and, thus, be a source of new occurrences. The impression of heterogeneity in histograms from different tumors within the same bladder is assumed to be caused by a variation in the ratio between aneuploid and diploid populations (high ratio in tumor and low in normal-appearing mucosa). This phenomenon may be the reason for variation in grading based on histological studies.

Is preoperative serum prostate-specific antigen level significantly related to clinical recurrence after radical retropubic prostatectomy for localized prostate cancer?

To evaluate the influence of preoperative serum prostate‐specific antigen (PSA) level and other clinicopathological variables on the probability of biochemical failure and clinical recurrence after radical prostatectomy (RP) for localized prostate cancer.

Nonadhesive Organ Culture of Human Exocrine Pancreatic Cells with Their Stroma

Introduction Studies using explanted tissue have shown that it is possible to keep adult human cells in organ culture with a preserved morphology for up to 1 month as spheres in a nonadhesive organ culture. Aims The current study was to determine whether human exocrine pancreatic cells also can be grown in this manner. Methodology Small tissue samples from organ donors and tumor-free resection rim from patients with pancreatic carcinoma were obtained (n = 16 adults). From each patient, fragments of approximately 300 &mgr;m in diameter were cultured and investigated with light microscopy and scanning and transmission electron microscopy at the time of explantation and after 5, 10, 20, 30, and 40 days of culture. Results Incubation of cultured fragments with vital dyes revealed a viable epithelium. At the time of explantation all the tissue fragments had a rough appearance with an uneven, torn periphery. During the first week of culture the fragments became rounder, with a smooth surface covering the whole circumference. This spheroid morphology persisted for the rest of the 6-week culture period. The fragments were within 1 week covered by a highly differentiated, polarized epithelium with secretory apparatus, apical secretion granules, and microvilli, as well as specialized cell junctions, with the same appearance as acinoductal pancreatic cells of the original tissue. The core of the fragments consisted of connective tissue with vascular elements, fibroblasts, leukocytes, and a few ductal and acinar elements. Transmission electron microscopy of the spheroids revealed a continuous basal lamina underneath the epithelium. Immunostaining for cytokeratin 5, 6, 7, 8, 17, and 18 was strongly positive in the epithelium. Conclusion These results show that normal exocrine pancreatic cells can be grown in vitro in a nonadhesive organ culture with their stroma.

Urokinase-type plasminogen activator receptor (uPAR) expression is associated with T-stage and survival in urothelial carcinoma of the bladder.

OBJECTIVES To evaluate the expression-and localization pattern of the urokinase-type plasminogen activator receptor (uPAR), focusing on its clinical implications in patients with urothelial neoplasia of the bladder treated with radical cystectomy. uPAR is a central molecule in tissue remodeling during cancer invasion and metastasis and is an established prognostic marker in cancer. The expression and localization of uPAR and its prognostic significance is only limitedly investigated in urothelial bladder neoplasia. MATERIALS AND METHODS The expression-and localization pattern of uPAR was investigated in formalin-fixed paraffin-embedded tumor tissue from 149 patients treated with radical cystectomy between 1988 and 2005. uPAR expression was determined by immunohistochemistry and scored as either negative or positive. Separate values were obtained for cancer cells, macrophages, and myofibroblasts at the invasive front and tumor core, respectively. Statistical analyses were performed to evaluate the association of uPAR localization and score with clinicopathologic covariates and survival. RESULTS uPAR positivity was seen in 122/137 (89%) and 118/149 (74%) of the neoplasias at the invasive front and tumor core, respectively. uPAR was primarily expressed by myofibroblasts and macrophages in the surrounding stroma as well as some cancer cells. A significant association between uPAR positivity and T-stage as well as grade was found for all 3 cell types in tumor core (P ≤ 0.04 for all comparisons). In univariate analysis, the uPAR positive group had a shorter survival than the uPAR negative group (hazard ratio = 2.39; 95% CI: 1.15-5.01; P = 0.020). CONCLUSIONS The expression of uPAR is a possible prognostic marker that could be useful in identification of patients with aggressive, highly invasive tumors that could benefit from additional chemotherapy or more intensive follow-up after cystectomy.

Local Chemotherapeutic Effects in Bladder Cancer Demonstrated by Selective Sampling and Flow Cytometry

A method is described by which the effect of intravesical chemotherapy can be monitored. Cytological samples obtained selectively during treatment were used for morphological and flow cytometric studies, and isoantigen (A, B and H) assessment in 2 patients with urothelial cancer. With flow cytometry even small aneuploid populations in the urothelium could be identified. From the histograms the urothelium was seen to contain 2 different cell populations: 1) diploid and 2) aneuploid. The ratio between aneuploid and diploid cells decreased significantly during treatment. Treatment was continued until no evidence of aneuploid cells could be identified in the histograms. Thus, it is demonstrated that intravesical chemotherapy for certain types of bladder cancer can eradicate the aneuploid cell population. A good correlation was found between cytological studies and flow cytometric measurements. Isoantigen assessment was done in the cell suspension used for morphological and flow cytometric studies. Isoantigen assessment also showed loss of antigens after completion of treatment, indicating that the diploid population was not normal biologically. Thus, 3 parameters can be correlated and related also to topography.

Seasonal variations of symptoms and occurrence of human bladder carcinomas.

A longitudinal study of transitional cell carcinomas of the urinary bladder has been performed in a region of Western Norway. Hematuria and infection were the first symptoms of neoplasia of the urinary bladder, showing marked and reproducible seasonal variations. Most of the tumours showed the first symptom at the end of the year. They also had a higher histological grade (WHO) than those occurring at other times of the year. This indicates that environmental factors dependent on the season may be of importance for the manifestation of malignant growth of the urinary bladder.

Mapping of Cell Cycle Distribution in Normal Human Urinary Bladder Epithelium

A method is described by which urothelial cells from well defined areas in the urinary bladder mucosa can be obtained. At cystoscopy epithelial cells are aspirated by means of a ureteral catheter. The cells obtained by this selective sampling technique are well suited for morphology and for quantitative single cell measurements by flow cytometry (FCM). In the present study the cell cycle distribution has been measured by FCM. Twenty-five patients (13 males and 12 females) had urothelial cells collected from well defined areas in the mucosa and the DNA content measured from different sites separately. The distribution of bladder mucosa cells in the different parts of the cell cycle (G1, S-phase and G2 + M) is reported. No significant differences were found with regard to sex and age of the patients. Also, a low regional variation was found in the bladder mucosa indicating that the method should enable the discrimination of even small abnormal cell populations in tumour disease. No evidence of polyploidy in the human urothelium was found.

Methylthioadenosine phosphorylase in human breast cancer

Methylthioadenosine (MTA) phosphorylase activity was measured in 47 biopsies from primary breast cancers (n = 34) and metastatic tumors (n = 13). Most specimens were also evaluated by DNA flow cytometry and determination of estrogen and progesterone receptor contents. Median MTA phosphorylase activity was 317 pmol/mg protein/min (range 50-1312 pmol/mg protein/min), but great variations were observed. Samples from four individuals had very low MTA phosphorylase activity (≤70 pmol/mg protein/min). No correlation with aneuploidy, receptor status, or the presence of metastases in the lymph nodes could be demonstrated. However, MTA phosphorylase activity showed a significant (p = 0.009) negative correlation with the fraction of cells in the S-phase of the cell cycle.

Flow Cytometric DNA Studies in Human Gastric Cancer and Polyps

Flow cytometric DNA studies were performed on cell suspensions of biopsy specimens from gastric tumors and neutral gastric mucosa in 18 patients with gastric cancer and 9 patients with gastric polyps. In cancer, aneuploidy was found in two tumors in the antral and five in the body part of the stomach (39%). The mean DNA index for aneuploid cancers was 1.57. In patients with aneuploid carcinomas three biopsy specimens from uninvolved mucosa also showed aneuploidy. In diploid carcinomas in the antral part of the stomach, the proliferative index (PI) was increased when compared with that of antral mucosa in controls (p less than 0.05). Increased PI was found in uninvolved mucosa in aneuploid carcinomas of the body part of the stomach when compared with that in diploid carcinomas (p less than 0.001). In uninvolved body mucosa in aneuploid carcinomas of the body part significantly reduced levels of acid-beta-glucuronidase (p less than 0.0001) were found when compared with diploid carcinomas. No polyps showed aneuploidy, and the PI in biopsy specimens from patients with gastric polyps did not differ from that in controls.