Effects of Ca2+ or Na+ on metallothionein expression in tilapia larvae (Oreochromis mossambicus) exposed to cadmium or copper.

The objectives of this study were to try to determine the reasons of the external Ca(2+) and Na(+) enhancement of Cd(2+) and Cu(2+) resistance in fish. Tilapia larvae at 3 days posthatch were exposed to (A) 0 (control), 40 microg/L Cd(2+), 40 microg/L Cd(2+) + 2 mM Ca(2+) (Cd/hyper-Ca), and 2 mM Ca(2+) or (B) 0 (control), 75 microg/L Cu(2+), 75 microg/L Cu(2+) + 0.52 mM Na(+) (Cu/hyper-Na), and 0.52 mM Na(+). After 48 hours, results indicated that (1) Cd/hyper-Ca and Cu/hyper-Na treatments showed decreased growth inhibition induced by the metals; (2) metal accumulation in Cd/hyper-Ca-treated larvae was lower compared with those exposed only to Cd; and (3) metallothionein (MT) expression was significantly higher in Cu/hyper-Na-treated larvae than in the group treated with Cu only. Taking all of this into account, either supplementary Ca(2+) or Na(+) in ambient water may help fish to maintain Ca(2+) or Na(+) homeostasis, which could decrease metal accumulation and its detrimental effects. Consequently, the fish increase MT expression and retard the growth inhibition caused by metals.

Synthesis, characterization and preclinical formulation of a dual-action phenyl phosphate derivative of bromo-methoxy zidovudine (compound WHI-07) with potent anti-HIV and spermicidal activities.

In a systematic effort to develop a microbicide contraceptive capable of preventing transmission of human immunodeficiency virus (HIV), as well as providing fertility control, we have previously identified novel phenyl phosphate derivatives of zidovudine (ZDV) with 5-halo 6-alkoxy substitutions in the thymine ring and halo substitution in the phenyl moiety respectively. Here, we describe the synthesis, characterization, and successful preclinical formulation of our lead compound, 5-bromo-6-methoxy-3'-azidothymidine-5'-(p-bromophenyl) methoxyalaninyl phosphate (WHI-07), which exhibits potent anti-HIV and sperm immobilizing activities. The anti-HIV activity of WHI-07 was tested by measuring viral p24 antigen production and reverse transcriptase activity as markers of viral replication in HIV-1 infected human peripheral blood mononuclear cells (PBMC). WHI-07 inhibited replication of HIV in a concentration-dependent fashion with nanomolar IC50 values. The effects of WHI-07 on human sperm motion kinematics were analysed by computer-assisted sperm analysis (CASA), and its effects on sperm membrane integrity were examined by confocal laser scanning microscopy (CLSM), and high-resolution low-voltage scanning electron microscopy (HR-LVSEM). WHI-07 caused cessation of sperm motility in a concentration- and time-dependent fashion. The in-vitro cytotoxicities of WHI-07 and nonoxynol-9 (N-9) were compared using normal human ectocervical and endocervical epithelial cells by the MTT cell viability assay. Unlike N-9, WHI-07 had no effect upon sperm plasma and acrosomal membrane integrity. N-9 was cytotoxic to normal human ectocervical and endocervical cells at spermicidal doses, whereas WHI-07 was selectively spermicidal. The in-vivo vaginal absorption and vaginal toxicity of 2% gel-microemulsion of WHI-07 was studied in the rabbit model. The sperm immobilizing activity of WHI-07 was 18-fold more potent than that of N-9. Over a 10 day period, there was no irritation or local toxicity to the vaginal epithelia or systemic absorption of WHI-07. Therefore, as a potent anti-HIV agent with spermicidal activity, and lack of mucosal toxicity, WHI-07 may have the clinical potential to become the active ingredient of a vaginal contraceptive for women who are at high risk for acquiring HIV by heterosexual vaginal transmission.