Alternative splicing determines the function of CYP4F3 by switching substrate specificity.

Diversity of cytochrome P450 function is determined by the expression of multiple genes, many of which have a high degree of identity. We report that the use of alternate exons, each coding for 48 amino acids, generates isoforms of human CYP4F3 that differ in substrate specificity, tissue distribution, and biological function. Both isoforms contain a total of 520 amino acids. CYP4F3A, which incorporates exon 4, inactivates LTB4 by omega-hydroxylation (Km = 0.68 microm) but has low activity for arachidonic acid (Km = 185 microm); it is the only CYP4F isoform expressed in myeloid cells in peripheral blood and bone marrow. CYP4F3B incorporates exon 3 and is selectively expressed in liver and kidney; it is also the predominant CYP4F isoform in trachea and tissues of the gastrointestinal tract. CYP4F3B has a 30-fold higher Km for LTB4 compared with CYP4F3A, but it utilizes arachidonic acid as a substrate for omega-hydroxylation (Km = 22 microm) and generates 20-HETE, an activator of protein kinase C and Ca2+/calmodulin-dependent kinase II. Homology modeling demonstrates that the alternative exon has a position in the molecule which could enable it to contribute to substrate interactions. The results establish that tissue-specific alternative splicing of pre-mRNA can be used as a mechanism for changing substrate specificity and increasing the functional diversity of cytochrome P450 genes.

Premature rupture of membranes at 34 to 37 weeks' gestation: aggressive versus conservative management.

OBJECTIVE: Our purpose was to compare induction of labor with preterm rupture of membranes between 34 and 37 weeks' gestation with expectant management. STUDY DESIGN: In this prospective investigation 120 gravid women at > or = 34 weeks 0 days and < 36 weeks 6 days of gestation were randomized to receive oxytocin induction (n = 57) or observation (n = 63). RESULTS: Estimated gestational age at rupture of membranes (34.3 +/- 1.4 weeks vs 34.5 +/- 1.4 weeks) and ultrasonographically estimated fetal weight (2230 +/- 321 gm vs 2297 +/- 365 gm) were equivalent between groups (not significant). Chorioamnionitis occurred more often (16% vs 2%, p = 0.007), and maternal hospital stay (5.2 +/- 6.8 days vs 2.6 +/- 1.6 days, p = 0.006) was significantly longer in the control group. Neonatal sepsis was also more common in the observation group (n = 3) than among induction patients (n = 0), but the difference was not statistically significant. CONCLUSION: Aggressive management of preterm premature rupture of the membranes at > or = 34 weeks 0 days of gestation by induction of labor is safe for the infant in our population and avoids maternal-neonatal infectious complications.

Laparoscopically directed ovarian cystectomy in premenopausal women. Impact of surgical experience on surgical time.

The purpose of this study was to correlate surgical experience with operating efficiency. A retrospective review of 303 operative laparoscopic procedures was performed during a 48-month interval and during the acquisition of surgical skills by one faculty member. Population demographics and surgery time were evaluated in 41 cases (13.5%) of ovarian cystectomy. Patients were categorized into fertility-related or gynecologic indications for surgery. Surgical time for successful laparoscopically directed resection of benign ovarian cysts was significantly reduced over the study interval (P = .008). Endometrioma was the most common pathologic finding in women with impaired fertility, and benign epithelial tumors were the lesion encountered most commonly in gynecologic patients. Benign teratoma occurred infrequently in both groups. No malignancy was observed in this group of premenopausal women, who had unilocular cysts less than 8 cm in diameter. Reduction in surgical time for laparoscopically directed ovarian cystectomy occurs after experience is gained by repeated application of the technique.