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.      

Factors Associated with High Myopia After 7 Years of Follow-up in the Correction of Myopia Evaluation Trial (COMET) Cohort

Purpose: To evaluate factors associated with the development of high myopia (worse than ?6.00 D) over 7 years of follow-up in the COMET cohort. Methods: COMET enrolled 469 ethnically diverse children (6–11 years) with myopia between ?1.25 and ?4.50 D. They were randomized to either progressive addition lenses (PALs) or single vision lenses (SVLs), and followed for 5 years in their original lens assignment and 2 additional years wearing either spectacles (PALs or SVLs) or contact lenses. Refractive error was measured annually by cycloplegic autorefraction and axial length by A-Scan ultrasonography. Myopia for each child was defined as the mean spherical equivalent refractive error (SER) of the 2 eyes. Analyses were based on 7 years of follow-up. Time to high myopia was analyzed by Cox proportional hazard models and linear regression. Parental refraction data were available from 240 COMET subjects. Results: Younger (6–7 years) versus older (11 years) age at baseline was a significant risk factor (adjusted hazard ratio (HR) = 6.6, 95% CI = 3.4, 12.7) for having high myopia within 7 years. More (SER from ?2.26 to ?4.50 D) vs. less (SER from ?1.25 to ?2.25 D) baseline myopia was also a significant risk factor for high myopia at 7 years (adjusted HR = 7.4, 95% CI = 4.4, 12.4). Gender, ethnicity, and treatment assignment were not associated with the risk of high myopia within 7 years. Increased number of myopic parents was associated with a significant risk of high myopia in the children (p = 0.008). Conclusions: Children who developed high myopia during 7 years of follow-up were younger and had more myopia at baseline. They also were more likely to have two myopic parents. These children may be at greater risk for sight-threatening conditions later in life.     

Mitoxantrone in the treatment of advanced non-squamous carcinoma of the cervix (A phase II trial of the gynecologic oncology group)

Twenty-five evaluable patients with advanced non-squamous carcinoma of the uterine cervix were treated with mitoxantrone 12 mg/m² every three weeks. All patients had good performance status and measurable disease and only 11 had received prior chemotherapy. One complete and one partial response were noted among 15 patients with no prior chemotherapy while no responses were seen in 11 previously treated patients. The major toxicity was myelosuppression; other toxicity was mild. The median progression-free interval was 2.1 months and median survival 4.3 months. Mitoxantrone displays minimal activity in patients with advanced non-squamous carcinoma of the cervix.     

Combination high-dose etoposide and vincristine infusion

Following the identification of a synergistic antitumor effect in a murine model, the combination of etoposide and vincristine has been explored in the clinic. Etoposide was given at 4 dose levels (250, 500, 750 or 1,000 mg/m²) with each dose given in 3 equal fractions daily for 3 days. The dose of vincristine was fixed (two 0.75 mg infusions over 22 hours each between doses of etoposide). A total of 26 patients were entered into study and 7, 11, 10 and 5 patients were treated at the 250, 500, 750, and 1,000 mg/m² dose levels, respectively. Myelosuppressioh was the principle side effect and Grade 4 WBC toxicity (<>³) developed in 14%, 27%, 40% and 40%, respectively, of the patients treated at each of these respective dose levels. Life-threatening infections occurred in 0%, 9%, 30% and 60% of the patients at these levels, respectively. Reversal of marrow toxicity was rapid with repeat courses given at 3-week intervals. Non-hematologic toxicity, including neurotoxicity, nausea, vomiting, and mucositis was generally mild when present. Objective responses were observed in 1 patient each with refractory Hodgkin's disease and immunoblastic lymphoma. Prolonged periods of stable disease occurred in 2 patients with adenocarcinoma of the lung and one patient with Hodgkin's disease. The starting dose of etoposide recommended for further trials of this agent in combination with infusion of vincristine is 500 mg/m² given in fractionated doses; dose escalation should be possible in many patients.     

Treatment effectiveness of hypnosis and behaviour therapy in smoking cessation: A methodological refinement

Studies in smoking cessation have generally failed to adequately control for active treatment effects and have assumed that measures of smoking behaviour (i.e., estimated smoking rate, self-monitoring and chemical analysis) are equally reliable measures. Sixty smokers were randomly assigned to one of four different smoking cessation treatment groups: hypnosis, focussed smoking, attention placebo and a waiting list control. Subjects were asked to estimate and monitor their own smoking behaviour. Blood samples were also taken for thiocyanate analysis before treatment. Smoking rates were similarly measured directly, at 3 months and 6 months after treatment. The results indicate that the three measures of smoking behaviour were all highly correlated. No significant differences were found between treatments, directly after treatment or at the 3- and 6-month follow-ups. These results suggest that active treatment effects may not be responsible for behavioural change in a smoking cessation program. The implications of these findings are discussed.     

Temporary and Permanent Hearing Loss

Long-term exposure to the dust of fiber glass used for insulation causes no demonstrable gross or microscopic pulmonary damage. This conclusion is based on the results of a postmortem study of the lungs of 20 fiber glass workers, who had had exposures to the dust of fibrous glass which ranged from slight to severe and for periods which ranged from 16 to 32 years, and a comparison of this study’s results with those obtained in a similar study of the lungs of 26 urban dwellers of both sexes, who presumably had not been exposed occupationally to fibrous glass dust, which showed that the average total amount of dust, the average total number of fibers per gram of dry lung, and the average dimensions of the fibers found in the lungs were not significantly different.