Biomedical subjects

L E Lee

Publications and source records attributed to L E Lee.

18 recordsLinked to original sources

PubMed1995

Improving the regulation of carcinogens by expediting cancer potency estimation.

The statutory language of the Safe Drinking Water and Toxic Enforcement Act of 1986 (Proposition 65; California Health and Safety Code 25249.5 et seq.) encourages rapid adoption of "no significant risk levels" (NSRLs), intakes associated with estimated cancer risks of no more than 1 in 100,000. Derivation of an NSRL for a carcinogen listed under Proposition 65 requires the development of a cancer potency value. This paper discusses the methodology for the derivation of cancer potencies using an expedited procedure, and provides potency estimates for a number of agents listed as carcinogens under Proposition 65. To derive expedited potency values, default risk assessment methods are applied to data sets selected from an extensive tabulation of animal cancer bioassays according to criteria used by regulatory agencies. A subset of these expedited values is compared to values previously developed by regulatory agencies using conventional quantitative risk assessment and found to be in good agreement. Specific regulatory activities which could be facilitated by adopting similar expedited procedures are identified.

Animals↗

PubMed1995

Development of a rainbow trout pituitary cell line that expresses growth hormone, prolactin, and somatolactin.

A cell line, RTP-2, has been developed from a normal-appearing pituitary of an adult rainbow trout. The cells grow in L-15 basal medium, supplemented with 2.5% to 10% fetal bovine serum, and have been passaged approximately 50 times over a 2-year period. At low density the cells have a stellate shape, whereas at confluency islands of polygonal cells appear among a tangle of bipolar cells. At the ultrastructural level, most cells contain numerous lysosomes, autophagic vacuoles, and intermediate filaments, but no obvious secretory granules. Reverse transcription-polymerase chain reaction (RT-PCR) with oligonucleotides specific for growth hormone (GH), prolactin (PRL), and somatolactin (SL) as amplification primers and Southern blot hybridization of the PCR products with probes specific for GH, PRL, and SL demonstrate that this cell line expresses GH, PRL, and SL. Digestion of the GH product of RT-PCR with restriction endonuclease SalI or KpnI confirms that both rainbow trout growth hormone genes are expressed in this cell line.

Animals↗

PubMed1994

Growth of fish cell lines in glutamine-free media.

The glutamine requirement for the in vitro proliferation of fish cells was investigated with cell lines from four different species and three tissues: goldfish skin (GFSk-S1), Chinook salmon embryo (CHSE-214), and rainbow trout liver (RTL-W1) and spleen (RTSp-W1). With a supplement of fetal bovine serum, the basal medium, Leibovitz's L-15, without glutamine supported the proliferation of all four cell lines as well, or nearly as well, as L-15 with 2 mM glutamine. This was true over short term assays of two to four weeks and for continuous propagation. CHSE-214 also grew as well with or without 2 mM glutamine in Minimum Essential Medium with fetal bovine serum. However, when the supplement was dialyzed fetal bovine serum, CHSE-214 grew much better in L-15 without glutamine. Therefore, glutamine was not required for growth in L-15, and in fact, was inhibitory in the absence of the dialyzable fraction of serum. By contrast, glutamine appeared to be important for growth in Minimum Essential Medium. When the supplement was dialyzed fetal bovine serum, CHSE-214 grew much better in Minimum Essential Medium with 2 mM glutamine. These results suggest that the glutamine requirement for the in vitro proliferation of fish cells is conditional and depends on the basal medium and serum supplement.

Animals↗

PubMed1991

Waterborne outbreak of Norwalk virus gastroenteritis at a southwest US resort: role of geological formations in contamination of well water.

From April 17 to May 1, 1989, gastroenteritis developed in about 900 people during a visit to a new resort in Arizona, USA. Of 240 guests surveyed, 110 had a gastrointestinal illness that was significantly associated with the drinking of tap water from the resort's well (relative risk = 16.1, 95% confidence interval 14.5 to 17.8) and this risk increased significantly with the number of glasses of water consumed (p less than 0.005). Three of seven paired sera tested for antibodies to the Norwalk agent had a four-fold or greater rise in titre. Water contaminated with faecal coliforms was traced back to the deep water well, which remained contaminated even after prolonged pumping. Effluent from the resort's sewage treatment facility seeped through fractures in the subsurface rock (with little filtration) directly into the resort's deep well. Although the latest technology was used to design the resort's water and sewage treatment plants, the region's unique geological conditions posed unexpected problems that may trouble developers faced with similar subsurface geological formations and arid climatic conditions in many parts of the world. In these areas, novel solutions are needed to provide adequate facilities for the treatment of sewage and supply of pure drinking water.

Cohort Studies↗

PubMed1989

The corticosteroid receptor and the action of various steroids in rainbow trout fibroblasts.

Corticosteroid binding sites with the characteristics of steroid receptors were detected with the synthetic corticosteroid, [3H]triamcinolone acetonide (TA), in monolayers of the rainbow trout fibroblast cell line, RTG-2. The sites had low capacity as saturation was achieved at approximately 5 nM. Scatchard plots of the data suggested a single population of high-affinity binding sites. The number of receptors per cell was approximately 20,000; the dissociation constant, 1 nM. Changes in [3H]thymidine incorporation and cellular morphology were monitored as potential corticosteroid-sensitive metabolic responses. Only cortisol and 11-deoxycortisol among 14 naturally occurring steroids and TA, fluocinolone acetonide, dexamethasone, and prednisolone among 6 synthetic corticosteroids inhibited [3H]thymidine incorporation and altered the morphology in RTG-2 cells. Two observations suggested that the corticosteroid receptor mediated these responses. The synthetic steroid, RU 38486, which is an antiglucocorticoid in mammals, did not elicit these responses, had a high affinity for the receptor, and blocked the ability of cortisol and TA to change [3H]thymidine incorporation and cellular morphology. Second, the affinity of various natural steroids for the receptor correlated with their ability to elicit a cellular response. Cortisol, and to lesser extent 11-deoxycortisol, showed strong affinity for the receptor. Cortisone, aldosterone, and the sex steroids had no affinity and did not elicit cellular responses.

Animals↗

PubMed1989

Action of cortisol on the proliferation of rainbow trout fibroblasts.

The effect of cortisol on the proliferation of the rainbow trout fibroblast cell line, RTG-2, was examined in synchronous and asynchronous cultures. When the transition from G1 to S was synchronized by restoring serum to serum-deprived cultures, the addition of cortisol at the time of serum restoration delayed the entry of cells into S phase. However, if cortisol was added 24 h after serum restoration, at the G1/S transition point, the subsequent peak of DNA synthesis was unaffected. In asynchronous cultures cortisol inhibited [3H]-thymidine and [3H]-uridine but not [3H]-leucine incorporation into acid-insoluble material. If the exogenous nucleoside concentration was raised, [3H]-thymidine but not [3H]-uridine incorporation continued to be inhibited by cortisol. This suggested that cortisol's effect on [3H]-thymidine incorporation reflected a change in entry into S phase and not just on thymidine uptake and metabolism. Cortisol inhibited the proliferation of RTG-2 in asynchronous cultures. At 1000 ng/ml of cortisol a reduction in cell number became apparent before the RTG-2 cultures were confluent, whereas at 100 ng/ml the reduction only became evident in confluent cultures. The synthetic antiglucocorticoid, RU 486, which acts at the level of the corticosteroid receptor, blocked the growth inhibition by cortisol. These results suggest that cortisol regulates rainbow trout fibroblast proliferation via the corticosteroid receptor and that the G1/S transition is one point at which this regulation occurs.

Animals↗

PubMed1988

Culture conditions for arresting and stimulating the proliferation of a rainbow trout fibroblast cell line, RTG-2.

Conditions for arresting and stimulating the proliferation of the rainbow trout fibroblast cell line RTG-2 have been examined and the time course of events after stimulation determined. Quiescent populations were achieved in two ways. Cultures grown to confluency without a medium change for at least 7 d had fewer than 5% of the cells in S phase and few mitotic figures. Cultures deprived of serum, which could be done for up to 3 d without a loss in cell number, also achieved quiescence. After 3 d without serum, less than 1% of cells were in S phase and mitotic figures were infrequent. Addition to these cultures of fresh serum-containing medium brought about the synchronous entry of cells into S phase and mitosis. For cultures in which either the medium had been changed after 7 d without a change or serum-containing medium had been added after 3 d of serum deprivation, DNA synthesis increased after a lag period of 20 to 24 h, was pronounced between 30 and 45 h, and then declined. This was followed by a peak in the mitotic index. These protocols for arresting and subsequently stimulating RTG-2 proliferation should allow the G1-S transition to be studied in a representative of teleosts.

Animals↗

PubMed1986

Preoperative radiation and surgery for cancer of the rectum. Veterans Administration Surgical Oncology Group Trial II.

In a prospective randomized trial, 361 male patients with histologically proven adenocarcinoma of the rectum, judged preoperatively to require abdominoperineal resection (APR), were treated by surgery alone or were given 3,150 rads of preoperative radiotherapy. Surgical resection was done on 320 patients, 262 having "curative" APR. Only moderate symptoms from radiotherapy were noted and postoperative complications and 30-day mortality were similar in both groups. Five-year survival for curative APR was the same in both groups (50% for both treated and control patients). The incidence of positive lymph nodes in the resected specimens was 35% in treated and 41% in controls. In the first preoperative radiotherapy trial conducted by the group, 5-year survival in patients undergoing "curative" APR was 47% in treated versus 34% in control groups. Additionally, the difference in positive lymph nodes in the resected specimens was substantially greater in the first trial (26% in treated versus 44% in controls).

Adenocarcinoma↗

PubMed1986

Effect of corticosteroids on the morphology and proliferation of two salmonid cell lines.

In medium with cortisol at 5 ng/ml and greater, cells of the rainbow trout cell line, RTG-2, lost their fibroblast-like morphology and became flattened and epithelial-like. This also occurred with dexamethasone triamcinolone acetonide, and fluocinolone acetonide but not with cortisone, progesterone, estradiol, or testosterone. Maintenance in medium with cortisol but not with cortisone caused these cells to adhere more tightly to the growth surface. The addition of cortisol at 100 ng/ml to RTG-2 cultures reduced the saturation densities that were achieved after monolayers had formed but had no effect on the times to reach confluency. Cortisone had no influence on cell population growth. Neither cortisol nor cortisone altered the morphology, adhesion, or proliferation of the Chinook salmon embryo cells, CHSE-214.

Adrenal Cortex Hormones↗

PubMed1976