Production of protein-associated DNA breaks by 10-[diethylaminopropylamino]-6-methyl-5H-pyrido[3',4':4,5]pyrrolo [2,3-g]isoquinoline in cultured L1210 cells and in isolated nuclei: comparison with other topoisomerase II inhibitors.

10-[3-Diethylaminopropylamino]-6-methyl-5H-pyrido[3',4':4,5] pyrrolo[2,3-g]isoquinoline (PZE) is an ellipticine derivative currently in clinical trials. PZE has been postulated to produce cellular DNA lesions by an uncommon mechanism. PZE-induced DNA damage was further investigated in L1210 cells in culture. PZE was highly cytotoxic for these cells (90% inhibitory concentration = 3.1 microM). The effects of PZE on cellular DNA were studied first by alkaline sucrose sedimentation, in comparison with those of 4'-(9-acridinylamino)methanesulfon-m-anisidide (m-AMSA). Like m-AMSA, PZE induced DNA strand breaks which were detected without a proteolytic treatment of the cell lysate. This result rules out the existence of covalent protein bridges sealing DNA termini at the break sites. PZE was less active than m-AMSA. DNA fragmentation was maximum at 5 microM and was lower at higher concentrations. The DNA effects of PZE were also studied by alkaline elution, and compared with those of Adriamycin and m-AMSA. Like Adriamycin, PZE induced single-strand breaks (SSBs) in a bell-shaped manner with respect to drug concentration. The maximum SSB frequency [1784 +/- 370 (SEM) rad equivalents)] was obtained at 16 microM. The kinetics of SSB reversion after drug removal was slower than in the case of m-AMSA. Similar bell-shaped curves were obtained for PZE-induced double-strand breaks and DNA-protein cross-links. PZE induced more double-strand breaks per SSB than did m-AMSA. However, as in the case of m-AMSA, PZE induced equal SSB and DNA-protein cross-link frequencies. These results suggest that PZE induces DNA breaks by inhibiting topoisomerase II as do other antitumor intercalators.

Interrelationship between affinity for DNA, cytotoxicity and induction of DNA-breaks in cultured L1210 cells for two series of tricyclic intercalators. Simplified analogues of ellipticine derivatives.

The interrelationship between affinity for DNA, cytotoxicity and induction of single-strand DNA breaks in cultured L1210 cells was studied for 21 compounds belonging to two series of tricyclic intercalators: 1-amino-substituted 4-methyl-5H-pyrido[4,3-b]indoles (gamma CARB) and 1-amino-substituted 4-methyl-5H-pyrido[3',4':4,5]pyrrolo[2,3-c]pyridines (PPP), which are simplified analogues of Ellipticine derivatives obtained by deletion of one cycle. Adriamycin, m-AMSA (4'-(9-acridinylamino) methanesulfon-m-anisidide), PZE (10-[diethylaminopropyl amino]-6-methyl-5H-pyrido[3',4':4,5]-pyrrolo[2,3-g] isoquinoline and RTE [( 1-(3-diethylaminopropylamino)-9-methoxy ellipticine, bimaleate) are used as reference compounds. The intercalation of these compounds into DNA was strongly suggested by three experimental observations: (i) the competitive inhibition of ethidium bromide intercalation, (ii) bathochromic and hypochromic effects on absorption spectra induced by DNA, and (iii) drug-induced increase of the DNA length, measured by viscosimetry. PPP derivatives are generally less cytotoxic and induce DNA breaks less efficiently than the gamma CARB ones, both in terms of maximum breakage frequencies and required drug concentrations. The most active compounds induced SSB in the DNA of L1210 cells, in a bell-shaped manner: the SSB frequency increased, rose to a maximum and then decreased as the drug concentrations increased. The maximum SSB frequencies induced by the most active compounds are of the same order as those of reference compounds Adriamycin and PZE. The structurally important requirements are essentially the same for both DNA breakage activity and cytotoxicity: (i) a N-CH3 in the 5-position, (ii) a CH3 in the 4-position, (iii) a hydroxy in the 8-position and (iv) the presence of an (aminoalkyl)amino side chain with preferentially a 3 carbon unit. There is no direct relationship between DNA affinity in vitro and induction of DNA breaks in cells, although a relatively high affinity seemed to be a necessary condition, since the most active compounds have the highest affinities and compounds having a very low affinity are totally inactive. The close correlation between cytotoxicity and extent of induction of DNA breaks suggests that these breaks may be in fact the lethal lesions responsible for cell death and thereby for the antitumor properties of these tricyclic intercalators.

Prenatal care access and pregnancy outcomes in Missouri.

Women's access to prenatal care and perception of barriers to obtaining care were acquired from data derived from the 1990 NICHD/Missouri Maternal and Infant Health Survey. Maternal surveys were available for 479 fetal deaths, 902 VLBW, 802 MLBW, and 919 normal birth weight infants. Very low birth weight mothers were less likely to receive prenatal care, perceived more barriers to obtaining prenatal care, and were more unhappy about their pregnancy than were the mothers of normal weight infants.

Missouri's parental consent law and teen pregnancy outcomes.

The Supreme Court decision of July 1989 upholding state regulation of abortion has led to numerous attempts to impose parental consent and/or parental notification legislation for females under the age of 18 seeking abortions. The effect of such legislation on teen pregnancy outcomes is hotly debated. Missouri vital statistics data from 1980 through 1992 are examined for the effect of such a law on pregnancy resolution choices among teens. The Missouri data suggest that since the enforcement of the parental consent statute in 1985 there has been a decrease in the selection of abortion as a pregnancy outcome, particularly among white teens. In addition there has been an increase in the percent of abortions among teens taking place in other states and an irregular but steady trend toward later abortions. The increasing number of births to unmarried mothers under the age of 18 suggest the need for specific services to help these young mothers cope.

PIP: Data on live births, fetal deaths, and induced abortions during 1980-92 were analyzed for Missouri residents to determine the potential effect on pregnancy outcomes of the 1985 enforcement of abortion parental consent and/or notification requirements for minors under 18. A comparison was made between data for females under 18 and those who were 18-19 when the pregnancy ended. It was found that the number of pregnancies in adolescents under age 18 declined from 8093 in 1980 to 5610 in 1992, while the number of live births increased from 2972 to 3508 (with a trend against marriage) and the number of abortions decreased about 33% (a 27% decrease was seen for the 18-19 year olds). No indication was found of an increase in the number of infants released for adoption. Since 1985, there has been a steady increase in the number of Missouri teens obtaining abortions in other states. An increase has also occurred in the percent of second semester abortions performed in both age groups both out-of-state and in Missouri. When analyzed by race, the data shows that the abortion rate among Whites under 18 decreased 46% since 1984 and that for Blacks decreased by 19%. Among the older teens, the abortion rate among Whites decreased 33% and that among Blacks increased. These data suggest that parental consent laws may affect the options for pregnant teenagers, especially for Whites. The laws may also delay the seeking of abortions. State, such as Missouri, that restrict access to abortions for minors should be prepared to address the needs of the increasing numbers of single mothers under the age of 18.

Sensitivity of fresh acute myeloid leukemia cells to etoposide: relationship with cell growth characteristics and DNA single-strand breaks.

In this study, we evaluated the individual in vitro sensitivity of fresh acute myeloid leukemia (AML) cells to VP-16, and attempted to correlate VP-16 cytotoxicity with AML cell growth characteristics and drug-induced DNA single-strand breaks (SSB). Primary (PE1) colony inhibition assays allowed us to characterize two distinct groups of AML: group I (patients 1 through 6), which displayed sensitivity to VP-16 similar to that of normal CFU-GM (IC90 of 20.52 +/- 2.44 micrograms/mL v 20.48 +/- 2.23 micrograms/mL after 1 hour drug exposure, respectively); and group II (patients 7 through 11), which was more sensitive to VP-16 (IC90 of 7.26 +/- 2.93 micrograms/mL, P = .004). Subsequently, groups I and II were termed normosensitive and hypersensitive, respectively. This objective VP-16 sensitivity classification, as determined by PE1, remained unaltered when assessed by secondary (PE2) colony inhibition assay (evaluating the self-renewal fraction of AML progenitors), or by cytofluorometric viability assay (evaluating the ultimately differentiated blast cell population). These findings would suggest that individual sensitivity to VP-16 of a particular cell population is maintained throughout CFU-AML differentiation. Finally, we report that sensitivity of AML cells to VP-16 did not correlate either with cell growth characteristics or with SSB generation. Indeed, AML cell sensitivity to VP-16 appeared more closely related to DNA repair kinetics after drug removal, ie, hypersensitivity being essentially characterized by a prolonged retention of SSB during the posttreatment period. Interestingly, the established HL-60 cell line, which presented greater sensitivity to VP-16 cytotoxicity than KG1, HEL, and K562, was also found to exhibit delayed DNA SSB repair kinetics, as compared with the other AML cell lines. These results suggest that hypersensitivity to VP-16 of some AML cells may be related to a deficient DNA-repair mechanism.