Notes: Summary Aphidicolin, a reversible inhibitor of DNA polymerase α and δ, has recently been reported to reverse the resistance to cisplatin (DDP) of an ovarian cancer cell line. We investigated the pharmacokinetics of aphidicolin in mice and examined its activity either alone or in combination with DDP in the DDP-sensitive M5076 (M5) murine reticular cell sarcoma as well as in a DDP-resistant subline (M5/DDP). The drug was cleared from plasma very rapidly (clearance, 41.6 ml min−1 kg−1), showing a half-life of 15 min. Aphidicolin concentrations in the tumor were approximately 50% of those found in plasma at steady state. Using several dose schedules and continuous infusions we failed to detect significant antitumor activity for aphidicolin glycinate. Potentiation of the activity of DDP by aphidicolin glycinate was moderate in mice bearing M5 tumor as well as in those bearing M5/DDP tumor. These data do not support the possible clinical use of aphidicolin in combination with DDP. However, further studies should be carried out in different tumor models before this possibility is conclusively ruled out.

Notes: Abstract Background: Conflicting data have been reported about the associationbetween glutathione S-transferase (GST), a family of proteins implicated indetoxification of cytotoxic drugs in human ovarian in vitro models, andresponse to chemotherapy and prognosis in ovarian cancer patients. The aim ofthis study was to analyze the possible clinical role of GST activity in alarge series of primary ovarian cancer patients. Patients and methods: The study included a large series of primaryuntreated ovarian cancer patients who underwent cytoreductive surgery andchemotherapy and who were followed up in a single institution. GST activitylevels were assessed in tumor extracts by using a biochemical assay. A cut-offof 250 units of enzymatic activity was chosen according to the receiveroperating characteristics (ROC) curve. Results: GST activity levels were distributed in an asymmetrical manner(median: 266 units; range: 4–918 units) and did not seem to beassociated with stage, histopathological grading, ascites, or residual tumorafter surgery. Higher GST activity levels were found in patients who respondedto chemotherapy (median: 298 units, range: 50–691) than in those whoresponded only partially (median: 227 units, range: 19–747) or not atall to chemotherapy (median: 246 units, range: 4–811) (H = 7.02, P =0.029). Moreover, the percentage of cases with 〉250 units was significantlyhigher among complete responders (66%) than partial responders(37%) or non-responders (48%) (χ2 = 7.32;P = 0.025). When multivariate analysis, including clinico-pathologicalparameters and GST activity status as predictors of response to chemotherapy,was carried out, residual tumor, stage and GST status retained independentpredictive value. Patients with high GST activity had more favourableprognosis than those with low GST activity. The median PFS was 42 months forpatients with high GST activity compared to 17 months for those with low GSTactivity (P = 0.037). The median overall survival was 72 months forhigh-GST-activity and42 months for low-GST-activity patients (P = 0.043). Substantially similarresults were obtained in the subgroup of stage II–III–IV ovariancancer patients. Multivariate analysis including the clinico-pathologicalparameters and GST activity status was performed in stage III–IV ovariancancer patients: Stage IV disease, residual tumor 〉2 cm, the presence ofascites and low GST activity status retained independent negative prognosticroles. Conclusion: A direct association between high GST activity and a betterclinical outcome in terms of response to chemotherapy and survival has beenobserved in a large series of primary untreated ovarian cancer patients. Theseresults, which are contrary to the expectations raised by in vitro studies,emphasize the need for caution when translating in vitro-generated hypothesesto the clinical setting.

Notes: Abstract Conceptions involving aging gametes are of relevance to natural family planning (NFP) because women using NFP to avoid pregnancy abstain from intercourse during the fertile time of the cycle. To help verify the safety of pregnancies occurring among NFP practitioners, our group has, since 1986, conducted a large cohort study involving six experienced NFP centers. Timing of conception was determined from NFP charts, in which women recorded days on which intercourse occurred. The number of days from the most probable conception intercourse to probable day of ovulation was first determined, and used as an estimate of the time gametes remained in the genital tract before fertilization. Several studies have already been completed, cohort as well as case-control in nature. 1. Spontaneous abortions. Three hundred and sixty-one conceptions occurred during the optimal time (day -1 or 0 relative to ovulation), and of these 33 resulted in spontaneous abortion (9.1%). Five hundred and seven conceptions occurred at non-optimal times during the cycle, and of these 55 resulted in spontaneous abortions (10.9%). These differences were not statistically significant (relative risk 1.19, 95% CI, 0.79-1.80) [1]. 2. Anomalies. Among 780 singleton births in 868 cohort pregnancies, 24 infants had major anomalies (3.1%) as of the present analysis [2]. This frequency is comparable to the general population. To further assess anomalies we employed a case-control approach. All consecutive births (live and stillborn) weighing 500 g or more taking place in 18 participating South American hospitals were examined for minor and major congential anomalies. Mothers of malformed and control infants did not differ with respect to the reported frequency of NFP use, which overall was 6.3% of the 10 642 mothers interviewed (5277 having a malformed infant; 5371 controls). Of 262 discordant pairs, there were 28 or 10.69% mothers within the Down syndrome case group vs. 16/262 or 6.11% among matched controls [3,4]. The odds ratio was 1.84; 95% CI, 0.99-3.96; however, even this non-significant difference narrowed substantially when adjusted for maternal age (OR, 1.78; 95% CI, 0.84-3.75); parity (OR, 1.68; 95% CI, 0.87-3.24); maternal educational level (OR, 1.71; 95% CI, 0.86-3.44); or all three together (OR, 1.74; 95% CI, 0.83-3.64). Conclusions. Our findings should be reassuring to natural family planning users. The overall rate of spontaneous abortion was not increased in NFP users who became pregnant, nor to date was the rate of anomalies. Any contribution to Down syndrome or abortion due to aging gametes would have to be small.

Notes: Abstract Although natural family planning (NFP) is a form of contraception without ostensible maternal risks (other than pregnancy), potential fetal risks could exist if aging gametes are involved in inadvertent fertilization. In the following report, we first review animal studies firmly establishing that aging sperm and aging oocytes (delayed fertilization) cause chromosomal abnormalities in mammals and other species. We next review human studies associating decreased coital frequency with trisomy and studies of NFP populations that generally show no increased frequency of anomalous offspring or spontaneous abortions. Our rationale for initiating an international cohort study is presented, along with the experimental design selected. Preliminary findings indicate that the experimental design chosen will indeed provide information allowing NFP safety to be assessed definitively.

Notes: Abstract Objective: Various birth defects and untoward perinatal outcomes have been claimed to be associated with pregnancies conceived by gametes aged in vivo before fertilization. Thus, these outcomes were systematically assessed in pregnancies occurring in natural family planning (NFP) users. Our international multicenter cohort study of NFP pregnancies (n = 877) is by far the largest systematic study designed to assess pregnancy outcome and is of sufficient power to allow us to address the concern of low birth weight (〈 2500 g) and preterm delivery (〈 37 weeks gestation). Study design: In addition to gathering baseline medical data, evaluation was performed at 16 weeks, 32 weeks and at term. Data were collected in a systematic cohort fashion, verified by the five collaborating international recruiting centers, and analyzed by investigators in the US. Most recruiting center principal investigators are obstetrician-gynecologists and, if not, have integral relationships with such specialists. Standard criteria could thus be applied within and among centers. In our cohort, birth weight was recorded accurately at delivery. Almost all of the deliveries occurred in hospitals; thus, data should be quite reliable. Neonatal examination for anomalies was usually conducted immediately after delivery, when birth weight was recorded. Results: Analysis of risk factors for low birth weight and pretern delivery showed that this population had a low risk profile. Low birth weight infants (〈 2500 g) and preterm deliveries were increased among women with a history of either prior low birth weight or preeclampsia in the index pregnancy. However, mean birth weight was unaffected by the timing of conception vis à vis ovulation or pregnancy history. Mean birh weight for the 877 singleton NFP pregnancies was 3349.6 g. The risk of preterm delivery was increased among older women who drank alcohol, but there were no significant effects of timing of conception vis à vis ovulation on preterm delivery. Results held when analysis was stratified according to whether NFP was being used for contraception or to achieve pregnancy. Conclusions: Our data do not appear to show striking differences between 877 NFP pregnancies and the general obstetric population. The timing of conception vis à vis ovulation does not exert significant effects on the birth weight or preterm delivery of resulting pregnancies, a reassuring finding for NFP users.

Notes: Abstract A multicenter cohort study was designed to assess pregnancy outcome among natural family planning (NFP) users, and provide the opportunity to address complications in NFP users by planning status and by timing of conception with respect to day of ovulation. There were 877 singleton births in this sample. Complications evaluated were abnormal vaginal bleeding, urinary tract infection, vaginal infection, hypertension of pregnancy, proteinuria, glycosuria, and anemia. There was no significant difference in the mean age, number of prenatal visits or birth weight among optimally and non-optimally timed pregnancies or for planned and unplanned pregnancies. There were higher incidences of "parity 2 or more" and current smokers in the non-optimally timed pregnancies and lower incidences of prior pregnancy loss and "currently employed" in the non-optimally timed pregnancies. There was little difference in pregnancy complications with respect to pregnancy timing, with the exception of a significant increased risk of vaginal bleeding late in pregnancy among non-optimally timed conceptions (11.5%) compared to optimally timed pregnancies (5.2%, RR = 2.2, 95% CI 1.3-3.7). More differences were observed in pregnancy complication rates by planning status. Unplanned pregnancies were associated with significantly more late pregnancy bleeding, vaginal infections, proteinuria, glycosuria and medication use than planned pregnancies. Unplanned pregnancies had lower incidences of maternal anemia. Complications of pregnancy were low in this NFP population, irrespective of planned versus unplanned status. Women with planned pregnancies had even fewer complications during pregnancy than women with unplanned conceptions, suggesting that women using NFP to plan their reproduction may be at particularly low risk.

Notes: The proton spectra of the four stereoisomers prepared by photocycloaddition of 2-cyclopentenone to cis and trans dichloroethylene have been analysed using tickling and INDOR techniques in order to obtain the sign of the cross-ring four-bond coupling constants in the cyclobutane ring. These parameters are correlated with the relative orientation of the protons involved. The complete NMR analysis of the seven-spin system of the cycloadduct endo-7-phenoxybicyclo[3.2.0]hept-2-en-6-one has been carried out and all the four-bond coupling constants have been obtained and discussed. The sign of the cross-ring coupling in the cyclobutanone ring has been determined.