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.      

Early postnatal expression of L1 by retinal fibers in the optic tract and synaptic targets of the Syrian hamster

Previous immunohistochemical studies in mouse, rat, and chick have reported that the expression of the glycoprotein and cell adhesion molecule L1, a member of the immunoglobulin superfamily, shows regulation during development of retina and optic nerve. To extend our understanding of the role of L1 in developing neural circuitry, we have examined L1 expression in the optic tract and thalamic and midbrain synaptic targets of retinal fibers in the early postnatal Syrian hamster, a well-characterized developmental model of the primary visual projection. Metabolic labeling studies reveal that a synaptically targeted, sulfated, and glycosylated form of L1 undergoes rapid axonal transport from the retina. Retinofugal transport of L1 decreases commensurate with the decline in immunoreactivity of retinal fibers in the visual pathway. Retinal ganglion cell axons show intense L1 immunoreactivity as they navigate in highly fasciculated bundles in the optic tract overlying the lateral geniculate body and in the superior colliculus. We found no evidence of L1 immunoreactivity on retinal axon collaterals as they defasciculate from the optic tract and branch into target neuropils. L1 immunoreactivity wanes in optic tract as axon terminal arbors are elaborated in the lateral geniculate body and superior colliculus and as myelination in the visual pathway commences. This pattern of L1 expression suggests that, in the early postnatal period, L1 may support fasciculation of retinal fibers, maintaining them within the optic tract, and that subsequent down-regulation of L1 may facilitate their terminal arborization and myelination.     

Clinical Impact of 18F-FDG-PET in the Suspicion of Recurrent Ovarian Carcinoma Based on Elevated Tumor Marker Serum Levels

Purpose: To retrospectively evaluate the contribution of 18F-fluorodeoxy-glucose-positron emission tomography (FDG-PET) to the diagnosis and clinical management of patients who were suspected of recurrent ovarian carcinoma, based on elevated tumor markers levels with normal or equivocal computed tomography (CT) or nuclear magnetic resonance (NMR).Procedures: 20 patients with these characteristics underwent FDG-PET. PET findings were confirmed in 14, in 7 by surgery, and in the other 7 by clinical course.Results: Recurrence was confirmed in 12 patients, all with FDG-PET positive. In other 2, recurrence was rule out and in 1, FDG-PET was negative. FDG-PET accuracy was 93% with 4 surgeries avoided and guided other 6.Conclusions: FDG-PET is an useful technique for detecting recurrent ovarian carcinoma suspected by elevated tumor markers levels and normal or equivocal results in the morphologic imaging techniques and has an important clinical impact on the management of these patients.     

Exploring views about mindfulness groups for voice-hearing from the perspective of service users and staff: A Q-methodology study

Objectives: Despite prevailing beliefs about the potential benefits and harmfulness of mindfulness for people who hear voices, there is a paucity of research into staff and service user views. Q-methodology was used to explore views about mindfulness groups for voice-hearers. Method: Opportunistic sampling of mental healthcare staff (N?=?14) and service users with psychosis (N?=?17). Both samples were analysed using principle components factor analysis to identify the range of attitudes held by staff and service users. Results: Staff participants were particularly knowledgeable and interested in mindfulness. A single staff consensus factor was found suggesting mindfulness is helpful, and not harmful for mental health, but uncertainty surrounded its usefulness for voice-hearers. Service users held four distinct attitudes: (i) mindfulness helps to calm a racing mind; (ii) mindfulness helps to manage stress; (iii) mindfulness improves well-being, and does not alter the brain, reality beliefs, or cause madness; and (iv) mindfulness helps with managing thoughts, fostering acceptance, and is acceptable when delivered in a group format. Conclusions: Staff viewed mindfulness groups for psychosis as helpful, not harmful, but were uncertain about their utility. Consistent with previous research, service users viewed mindfulness groups as useful to promote well-being and reduce distress for individuals experiencing psychosis.