CYP1A1 3801T>C polymorphism implicated in altered xenobiotic metabolism is not associated with variations in sperm production and function as measured by total motile sperm and fertilization rates with intracytoplasmic sperm injection.

OBJECTIVE: To evaluate the cytochrome P450 3801T>C polymorphism's frequency in relation to semen production, as determined by semen analysis parameters, and sperm function, as determined by fertilization rates with intracytoplasmic sperm injection (ICSI). DESIGN: Case-control study. SETTING: Academic-affiliated private practice. PATIENT(S): This study included patients undergoing IVF from 2004 to 2014 grouped into categories based on semen analysis parameters performed at a single andrology laboratory. Cases were patients with total motile sperm (TMS) counts of ≤20 × 10(6). Frequency-matched controls were selected with TMS of >20 × 10(6). INTERVENTION(S): The 3801T>C polymorphism was identified using DNA from serum samples with real-time quantitative polymerase chain reaction. MAIN OUTCOME MEASURE(S): CYP1A1 3801T>C polymorphism frequency in TMS groups and distribution in fertilization rate outcomes with ICSI. RESULT(S): A total of 460 cases were identified with ≤20 × 10(6) TMS, and 489 age-matched controls with >20 × 10(6) TMS were selected across the study time frame. For those with <5 × 10(6) vs. >20 × 10(6) TMS there was no difference when comparing heterozygous (odds ratio [OR] 0.96; 95% confidence interval [CI] 0.66-1.40) or homozygous mutant (OR 1.33; 95% CI 0.52-3.20) with the wild-type patients. Additionally, no difference was seen when analyzing subgroups <5 × 10(6), 5-20 × 10(6), and >20 × 10(6) TMS in a similar fashion. Receiver operating characteristic (ROC) curve analysis did not find a significant TMS count based on presence of the polymorphism (area under the ROC curve = 0.51). There were 460 patients who underwent IVF/ICSI, and fertilization rates did not differ with presence of the polymorphism (area under the ROC curve = 0.50). CONCLUSION(S): Allele frequency of the 3801T>C polymorphism does not correlate with semen production as determined by TMS counts or sperm function as determined by fertilization rates with ICSI. The use of neither semen analysis parameters nor fertilization rates with ICSI helps identify CYP1A1 polymorphism carriers.

Use of necrotic markers in the Drosophila ovary.

Necrosis is a form of cell death characterized by cytoplasmic and organelle swelling, compromised -membrane integrity, intracellular acidification, and increased levels of reactive oxygen species (ROS) and cytosolic Ca(2+). In the Drosophila ovary, two distinct forms of cell death occur naturally. In response to starvation, caspase-dependent cell death occurs during mid-oogenesis. Additionally, the nurse cells, which support the developing oocyte, undergo developmental programmed cell death during late oogenesis after they dump their contents into the oocyte. Evidence suggests that necrosis may be playing an important role during developmental programmed cell death of the nurse cells during late oogenesis. Here, we describe several methods to detect events associated with necrosis in the Drosophila ovary. Propidium iodide is used to detect cells with compromised membrane integrity, and H2DCFDA is used as an indicator of ROS levels in a cell. In addition, LysoTracker detects intracellular acidification and X-rhod-1 detects cytosolic Ca(2+). We also describe transgenic methods to detect Ca(2+) levels and expression patterns. These methods performed in the Drosophila ovary, as well as other tissues, may lead to a further understanding of the mechanisms of necrosis as a form of programmed cell death.