The excitation and emission properties of several psoralen derivatives are compared using conventional single-photon excitation and simultaneous two-photon excitation (TPE). Two-photon excitation is effected using the output of a mode-locked titanium: sapphire laser, the near infrared output of which is used to promote nonresonant TPE directly. Specifically, the excitation spectra and excited-state properties of 8-methoxypsoralen and 4'-aminomethyl-4,5,8-trimethylpsoralen are shown to be equivalent using both modes of excitation. Further, in vitro feasibility of two-photon photodynamic therapy (PDT) is demonstrated using Salmonella typhimurium. Two-photon excitation may be beneficial in the practice of PDT because it would allow replacement of visible or UV excitation light with highly penetrating, nondamaging near infrared light and could provide a means for improving localization of therapy. Comparison of possible laser excitation sources for PDT reveals the titanium: sapphire laser to be exceptionally well suited for nonlinear excitation of PDT agents in biological systems due to its extremely short pulse width and high repetition rate that together provide efficient PDT activation and greatly reduced potential for biological damage.
Oocytes obtained from superstimulated rhesus monkeys are needed for in vitro fertilization experiments. The animals become refractory to repeated ovarian stimulation by pregnant mare serum gonadotropin (PMSG). Production of antibodies against PMSG was suspected. Antibodies were not detected by gel-diffusion tests, but were found using ELISA procedures in sera from all monkeys that had been treated with PMSG. Sera from seven of nine treated monkeys had antibodies that cross-reacted with FSH-P. An ovarian stimulation test using golden hamsters showed that sera from PMSG-treated rhesus monkeys completely blocked superstimulation of follicle development and ovarian weight gain by PMSG. This study demonstrates that the refractoriness of rhesus monkeys to repeated ovarian stimulation by PMSG is due to the production of nonprecipitating antibodies to PMSG that effectively block the activity of PMSG on the ovary. Formation of these antibodies may preclude use of other gonadotropin preparations in refractory monkeys. Alternative ovarian stimulation procedures avoiding use of gonadotropin preparations need to be developed to permit repeated treatment of rhesus monkeys for multiple oocyte collection.
Exposure to pesticides, dyes, and pollutants that mimic the growth promoting effects of estrogen may cause breast cancer. The pesticide DDT and the food colorant Red No. 3 were found to increase the growth of HTB 133 but not estrogen receptor (ER) negative human breast cells (HTB 125) or rat liver epithelial cells (RLE). Red No. 3, beta-estradiol, and DDT increase ER site-specific DNA binding to the estrogen response element in HTB 133 cells and increase cyclin-dependent kinase 2 activity in MCF-7 breast cancer cells. Site-specific DNA binding by p53 in RLE, HTB 125, HTB 133, and MCF-7 cells was increased when they were treated with Red No. 3, which suggests that cellular DNA was damaged by this colorant. Red No. 3 increased binding of the ER from MCF-7 cells to the estrogen-responsive element. Consumption of Red No. 3, which has estrogenlike growth stimulatory properties and may be genotoxic, could be a significant risk factor in human breast carcinogenesis.ImagesFigure 4. AFigure 4. BFigure 5. AFigure 5. BFigure 6.Figure 7. AFigure 7. BFigure 7. C
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