Isolation and characterization of propagable cell lines (HUNC) from the androgen-sensitive Dunning R3327H rat prostatic adenocarcinoma

The Dunning H rat prostate tumor (R3327H) is a widely used experimental model of human prostatic adenocarcinoma (CaP). The Dunning H tumor has been characterized as androgen-sensitive, androgen-receptor (AR) positive, prostate-specific antigen and prostatic acid phosphatase (PAP) positive. To date, the tumor has been maintained by serial passage in vivo because of the lack of an in vitro cell line that retains the characteristics of the in vivo tumor. The objective of the present study was to establish a propagable cell line from R3327H adenocarcinoma that maintained androgen sensitivity and expression of AR, PSA and PAP. Tissue harvested from an in vivo R3327H tumor was dissociated with collagenase and placed into Richter's improved media (with supplements). A cytokeratin-positive epithelial cell line (HUNC- E) and a vimentin-positive stromal cell line (HUNC-S) were generated from the primary culture, subcultured continuously for >300 days, and passaged >50 times. Survival of the HUNC-E cell line in vitro depended on several media supplements, including nicotinamide, insulin, transferrin, selenium and epidermal growth factor (EGF). HUNC-E cells expressed AR and produced PSA and PAP throughout the culture period, as confirmed by immunocytochemistry and Western blot analyses. Addition of 14 nM testosterone (T) or dihydrotestosterone (DHT) to HUNC-E cells, stimulated DNA synthesis as well as anchorage-independent growth and PSA production, which demonstrated the androgen-sensitive nature of the cells in vitro. When HUNC-E and HUNC-S cells were combined in a 3:1 ratio and introduced subcutaneously into syngeneic male hosts, tumors formed in 2/3 animals with an average latency of 7 months. RT-PCR and immunocytochemical characterization of the HUNC cell lines revealed that the cells expressed several growth factors and their cognate receptors, including HGF, TGF-alpha and the TGF-betas, indicating the establishment of potential autocrine loops in the neoplastic cells. The HUNC-E and HUNC-S CaP cell lines, which retain the characteristics of the epithelial and stromal components of the in vivo R3327H tumor, will allow a more thorough and informative molecular and biological analysis of prostatic adenocarcinoma.      

Zinc replenishment reduces esophageal cell proliferation and N- nitrosomethylbenzylamine (NMBA)-induced esophageal tumor incidence in zinc-deficient rats

Previous work has shown that sustained increased and decreased cell proliferation, induced by dietary zinc deficiency and caloric restriction respectively, influence the course of N- nitrosomethylbenzylamine (NMBA)-induced esophageal carcinogenesis in rats. The present study considered whether the increased cell proliferation and esophageal tumor incidence induced by zinc deficiency are reversed upon zinc replenishment. Weanling rats were maintained initially on a deficient diet containing 4 p.p.m. zinc. After 5 weeks, carcinogen-treated animals were given six intragastric doses of NMBA (2 mg/kg twice weekly). Controls were untreated. After the second NMBA dose, the rats were divided into three dietary groups. One group was continued on the deficient diet, while the other two groups were switched to diets containing either 75 or 200 p.p.m. zinc, with half of the members in each group fed ad libitum and half pair-fed with deficient rats. NMBA-untreated controls were similarly replenished. At various time points, esophageal cell proliferation was assessed in five animals from each group by immunohistochemical detection of cells in S phase, with in vivo 5-bromo-2'deoxyuridine labeling. At 11 weeks after the first dose, esophageal tumor incidence was greatly reduced, from 100% in the deficient group to 26 and 14% respectively in the replenished groups fed ad libitum 75 and 200 p.p.m. zinc and to 14 and 11% respectively in the replenished groups pair-fed 75 and 200 p.p.m. zinc. In addition, the number of tumors per esophagus was reduced from 9.93 +/- 4.25 in deficient rats, to a range of 0.11 +/- 0.31-0.30 +/- 0.54 in replenished animals. Following zinc replenishment, esophageal cell proliferation, as measured by labeling index (LI), the number of labeled cells and the total number of cells, was markedly decreased in NMBA-untreated and -treated esophagi as compared with those in corresponding deficient esophagi. Thus, the esophageal cell proliferation induced by zinc deficiency is reversed by zinc replenishment and replenished animals have a markedly lower incidence of esophageal tumors.      

A model for faculty mentoring in academic radiology

RATIONALE AND OBJECTIVES: The purpose of this report is to describe the development and implementation of a faculty mentoring program in radiology designed to promote the career development of junior faculty and enhance communication in the department. MATERIALS AND METHODS: The mentoring program was implemented in five stages: organizational readiness, participant recruitment, mentor matching and orientation, implementation, and evaluation. Evaluations were based on Likert scale ratings and qualitative feedback. A retrospective analysis was also conducted of the annual performance reviews of junior faculty in the areas of research, teaching, patient care, and overall performance. RESULTS: An average of 83% (19 of 23) of the junior faculty participated in the pilot phase of the mentoring program. During five rounds of testing, the median rating (1 indicates not important; 10, extremely important) from responding junior faculty was 10 for overall value of individual mentoring meetings; the median rating for the mentors responding was 8.75. Research and academic development were identified as the areas of greatest importance to the faculty. Research and patient care were most improved as assessed by faculty peers during performance reviews. The schedule of semiannual formal mentoring meetings was reported to be optimal. CONCLUSION: The program was implemented to the satisfaction of junior faculty and mentors, and longitudinal performance suggests positive effects. Issues to be contended with include confidentiality and the time needed for mentoring beyond already saturated schedules. Overall, the authors propose that mentoring programs can be an asset to academic radiology departments and a key factor in maintaining their vitality.     

Integration of magnetic resonance imaging into radiation therapy treatment planning: I. Technical considerations

This paper presents the results of a feasibility study specifically addressing the technical and operational difficulties in making quantitative use of Magnetic Resonance Imaging (MRI) in radiation therapy treatment planning (RTTP). Selected radiotherapy patients have been studied with both CT and MRI as part of the treatment planning process. Both sets of images, along with mechanically-obtained external contour and simulator film data, are entered into the treatment planning system. All of the capabilities of the fully three dimensional planning system U-MPlan are available to both the CT and MRI images, in which any image can be used as the backdrop for interactive beam positioning, beam portal simulation, and dose distribution displays for external beam and brachytherapy applications in both 2- and 3-dimensionally-oriented displays. The study has shown that to use MRI data for RTTP, one must (a) use careful patient positioning and marking, (b) transfer information from CT to MRI and vice versa, (c) determine the geometrical consistency between the CT and MR data sets, (d) investigate the unwarping of distorted MR images, and (e) have the ability to use non-axial images for determination of beam treatment technique, dose calculations, and plan evaluation.