OBJECTIVE: The purpose of this study was to evaluate whether genetic polymorphisms in selected cytochrome P450 enzymes (CYPc17 alpha, CYP1A1, and CYP1B1), estradiol (E2) levels, and estrone levels were associated with hot flushes.METHODS: Women with hot flushes were those aged 45-54 years who reported ever experiencing hot flushes (n = 354). Women without hot flushes were those aged 45-54 years who reported never experiencing hot flushes (n = 258). Each participant completed a questionnaire and provided a blood sample for determination of genotypes, E2 levels, and estrone levels.RESULTS: Carriers of the CYP1B1 (Val432Leu) polymorphism were more likely to report having any hot flushes (risk ratio [RR] 1.16, 95% confidence interval (Cl) 0.98-1.37) and at least weekly hot flushes (RR 1.29, 95% Cl 0.98-1.70) than women without the polymorphism, although these associations were of borderline statistical significance. In addition, carriers of the CYP1B1 polymorphism were likely to have a statistically significant 30% increased risk of reporting moderate to severe hot flushes (RR 1.30, 95% Cl 1.02-1.67) and a statistically significant 27% increased risk of reporting hot flushes lasting a year or more (RR 1.27, 95% Cl 1.00-1.61) compared with women without the polymorphism. There were no associations between CYP1A1 or CYPc17 alpha polymorphisms and hot flushes. Low E2 and estrone levels were associated with hot flushes, but they did not alter the association between the CYP1B1 polymorphism and hot flushes.CONCLUSION: These data suggest that a CYP1B1 polymorphism may be associated with severe and persistent hot flushes, independent of E2 and estrone levels.

Aims: To examine the relation between current alcohol use, estradiol, estrone, and testosterone levels, and hot flashes in midlife women using a case-control study design. Methods: Cases were midlife women (45-54 years) who reported ever experiencing hot flashes. Controls were midlife women (45-54 years) who reported never experiencing hot flashes. Each participant completed a questionnaire and provided a blood sample that was used to measure estradiol, estrone, and testosterone levels by enzyme-linked immunosorbent assay. Results: The results indicate that current alcohol use (at least one day per month) was significantly associated with a reduced risk of hot flashes compared to non-use of alcohol, independent of age and smoking habits. The hot flashes experienced by current alcohol users were less severe and less frequent than those experienced by non-users of alcohol. Further, current alcohol users had similar levels of estradiol, estrone, and testosterone compared to non-users of alcohol. Conclusions: These data suggest that current alcohol use is associated with a reduced risk of any, severe, and frequent hot flashes in midlife women by a mechanism that may not include changes in sex steroid hormone levels.

ObjectiveGenetic polymorphisms in some CYP450 enzymes have been associated with elevated estrogen levels in breast cancer patients, but their role in regulating estrogen levels in healthy women is unknown. The objective of this work was to test the hypothesis that genetic polymorphisms in selected CYP450 enzymes (CYPc17, which is involved in estrogen synthesis and CYP1A1 and CYP1B1, which are involved in estrogen metabolism) are associated with altered estrogen levels in healthy women.DesignCross-sectional study of perimenopausal women aged 45–54 years.Materials and methods435 age-eligible women living in Maryland were recruited by direct mailing. Subjects completed extensive questionnaires regarding demographics, reproductive history, lifestyle variables, and medical history and provided a blood sample for measurement of genetic polymorphisms in CYP1A1, CYP1B1, and CYPc17 as well as estrogen levels. Genetic polymorphisms were measured by polymerase chain reaction and enzyme digests. Levels of estradiol (E2) and estrone (E1) were analyzed using estradiol enzyme immunoassays. Using SPSS software, groups were compared with ANOVA, t-tests, and logistic regression models where appropriate. A p value of <0.05 was considered to be significant.ResultsIn unadjusted models, levels of E1 and E2 were similar between the women without a genetic polymorphism and women who were heterozygous or homozygous for polymorphisms in each of the enzymes (CYP1A1, CYP1B1, and CYPc17). After adjusting for age, race, body mass index, smoking history, menstrual cycle day, and parity, genetic polymorphisms in CYP1B1 were associated with significantly increased E2 levels (p < 0.012), but not E1 levels (p = 0.978). CYP1A1 and CYPc17 were not significant predictors of either E1 or E2 levels.Conclusion ObjectiveGenetic polymorphisms in some CYP450 enzymes have been associated with elevated estrogen levels in breast cancer patients, but their role in regulating estrogen levels in healthy women is unknown. The objective of this work was to test the hypothesis that genetic polymorphisms in selected CYP450 enzymes (CYPc17, which is involved in estrogen synthesis and CYP1A1 and CYP1B1, which are involved in estrogen metabolism) are associated with altered estrogen levels in healthy women. Genetic polymorphisms in some CYP450 enzymes have been associated with elevated estrogen levels in breast cancer patients, but their role in regulating estrogen levels in healthy women is unknown. The objective of this work was to test the hypothesis that genetic polymorphisms in selected CYP450 enzymes (CYPc17, which is involved in estrogen synthesis and CYP1A1 and CYP1B1, which are involved in estrogen metabolism) are associated with altered estrogen levels in healthy women. DesignCross-sectional study of perimenopausal women aged 45–54 years. Cross-sectional study of perimenopausal women aged 45–54 years. Materials and methods435 age-eligible women living in Maryland were recruited by direct mailing. Subjects completed extensive questionnaires regarding demographics, reproductive history, lifestyle variables, and medical history and provided a blood sample for measurement of genetic polymorphisms in CYP1A1, CYP1B1, and CYPc17 as well as estrogen levels. Genetic polymorphisms were measured by polymerase chain reaction and enzyme digests. Levels of estradiol (E2) and estrone (E1) were analyzed using estradiol enzyme immunoassays. Using SPSS software, groups were compared with ANOVA, t-tests, and logistic regression models where appropriate. A p value of <0.05 was considered to be significant. 435 age-eligible women living in Maryland were recruited by direct mailing. Subjects completed extensive questionnaires regarding demographics, reproductive history, lifestyle variables, and medical history and provided a blood sample for measurement of genetic polymorphisms in CYP1A1, CYP1B1, and CYPc17 as well as estrogen levels. Genetic polymorphisms were measured by polymerase chain reaction and enzyme digests. Levels of estradiol (E2) and estrone (E1) were analyzed using estradiol enzyme immunoassays. Using SPSS software, groups were compared with ANOVA, t-tests, and logistic regression models where appropriate. A p value of <0.05 was considered to be significant. ResultsIn unadjusted models, levels of E1 and E2 were similar between the women without a genetic polymorphism and women who were heterozygous or homozygous for polymorphisms in each of the enzymes (CYP1A1, CYP1B1, and CYPc17). After adjusting for age, race, body mass index, smoking history, menstrual cycle day, and parity, genetic polymorphisms in CYP1B1 were associated with significantly increased E2 levels (p < 0.012), but not E1 levels (p = 0.978). CYP1A1 and CYPc17 were not significant predictors of either E1 or E2 levels. In unadjusted models, levels of E1 and E2 were similar between the women without a genetic polymorphism and women who were heterozygous or homozygous for polymorphisms in each of the enzymes (CYP1A1, CYP1B1, and CYPc17). After adjusting for age, race, body mass index, smoking history, menstrual cycle day, and parity, genetic polymorphisms in CYP1B1 were associated with significantly increased E2 levels (p < 0.012), but not E1 levels (p = 0.978). CYP1A1 and CYPc17 were not significant predictors of either E1 or E2 levels. Conclusion

Several studies indicate that demographic and cultural factors may be associated with the symptoms women experience in the perimenopausal period. However, few studies to date have examined whether impaired reproductive function is associated with an increased risk of perimenopausal symptoms. Thus, the objective of this work was to test the hypothesis that a history of self-reported infertility is associated with an increased risk of perimenopausal symptoms. Case-control study of women aged 45–54 years. Cases were defined as women who reported a history of attempting to conceive without success for at least one year; controls were women who reported that they had been able to become pregnant within one year of unprotected intercourse. 435 age-eligible women living in Maryland were recruited by direct mailing. Subjects completed extensive questionnaires regarding demographics, reproductive history including any history of inability to conceive for 1 year or longer, diet/lifestyle, and multiple perimenopausal symptoms. Each study subject provided a blood sample, which was subjected to estradiol enzyme immunoassay for measurement of estradiol (E2) and estrone (E1) levels. Using SPSS software, cases and controls were compared with Chi-Square tests and evaluated by multiple logistic regression analysis. Group means were compared with ANOVA and T-tests where appropriate. A p value of < 0.05 was considered to be significant. There were 119 cases (27.4%) and 316 controls (72.6%). Between cases and controls, there were no significant differences in mean age at menarche, body mass index (BMI), number of pregnancies or live births, history of oral contraceptive or tobacco use, level of physical activity, personal use of vitamins, herbal use, supplemental calcium use, level of education or household income. Cases had a significantly increased number of night sweats compared to controls, even though E1 and E2 levels were similar after adjusting for menstrual cycle day (E1: 145.2 ± 9.6 pg/ml for cases vs. 137.2 ± 6.2 pg/ml for controls, E2: 123.5 ± 10.3 pg/ml for cases vs. 119.7 ± 6.3 pg/ml for controls). There were no significant differences in other perimenopausal symptoms between cases and controls. After controlling for multiple potential confounders including estrogen levels, a history of self-reported infertility is associated with an increased risk of night sweats in women age 45–54 years. Future research is necessary to further delineate the mechanisms relating fertility and hypo-estrogenic symptoms.

ObjectiveGenetic polymorphisms in the CYP450 system have been associated with altered hormone levels, changes in the timing of menarche, and an increased risk of breast cancer in women. In contrast, the relation of genetic polymorphisms in the CYP450 system to infertility is not known. Thus, the objective of this work was to test the hypothesis that genetic polymorphisms in selected CYP450 enzymes (CYPc17, which is involved in estrogen synthesis, and CYP1A1 and CYP1B1, which are involved in estrogen metabolism) are associated with an increased risk of infertility in women.DesignCase-control study of women aged 45–54 years. Cases were defined as women who reported a history of attempting to conceive without success for at least one year; controls were women who reported that they had been able to become pregnant within one year of unprotected intercourse.Materials and methods435 age-eligible women living in Maryland were recruited by direct mailing. Subjects completed questionnaires regarding demographics, reproductive history including any history of inability to conceive for 1 year or longer, diet/lifestyle, and smoking history. Participants also provided a blood sample that was subjected to polymerase chain reaction and enzyme digests for measurement of selected genetic polymorphisms in CYPc17, CYP1A1, and CYP1B1. Cases and controls were compared with Chi-Square tests and logistic regression models using SPSS software. A p value of < 0.05 was considered to be significant.Results119 of the 435 participants (27.4%) reported a history of inability to conceive for at least one year. Cases were more likely to have genetic polymorphisms in CYP1A1 than controls (4.7% of cases were homozygous for the polymorphism vs. 0.9% of controls; p < 0.014 after adjustment for potential confounders including race, marital status, education level, income, smoking history, oral contraceptive use, and body mass index). There were no significant differences in the prevalence of genetic polymorphisms in CYPc17 between cases and controls (69.9% of cases vs. 63.5% of controls; p = 0.455). Similarly, there were no significant differences in the prevalence of genetic polymorphisms in CYP1B1 between cases and controls (51.4% of cases vs. 55.0% of controls; p = 0.345).ConclusionGenetic polymorphisms in CYP1A1, but not CYP1B1 or CYPc17, are associated with an increased risk of self-reported infertility in women. ObjectiveGenetic polymorphisms in the CYP450 system have been associated with altered hormone levels, changes in the timing of menarche, and an increased risk of breast cancer in women. In contrast, the relation of genetic polymorphisms in the CYP450 system to infertility is not known. Thus, the objective of this work was to test the hypothesis that genetic polymorphisms in selected CYP450 enzymes (CYPc17, which is involved in estrogen synthesis, and CYP1A1 and CYP1B1, which are involved in estrogen metabolism) are associated with an increased risk of infertility in women. Genetic polymorphisms in the CYP450 system have been associated with altered hormone levels, changes in the timing of menarche, and an increased risk of breast cancer in women. In contrast, the relation of genetic polymorphisms in the CYP450 system to infertility is not known. Thus, the objective of this work was to test the hypothesis that genetic polymorphisms in selected CYP450 enzymes (CYPc17, which is involved in estrogen synthesis, and CYP1A1 and CYP1B1, which are involved in estrogen metabolism) are associated with an increased risk of infertility in women. DesignCase-control study of women aged 45–54 years. Cases were defined as women who reported a history of attempting to conceive without success for at least one year; controls were women who reported that they had been able to become pregnant within one year of unprotected intercourse. Case-control study of women aged 45–54 years. Cases were defined as women who reported a history of attempting to conceive without success for at least one year; controls were women who reported that they had been able to become pregnant within one year of unprotected intercourse. Materials and methods435 age-eligible women living in Maryland were recruited by direct mailing. Subjects completed questionnaires regarding demographics, reproductive history including any history of inability to conceive for 1 year or longer, diet/lifestyle, and smoking history. Participants also provided a blood sample that was subjected to polymerase chain reaction and enzyme digests for measurement of selected genetic polymorphisms in CYPc17, CYP1A1, and CYP1B1. Cases and controls were compared with Chi-Square tests and logistic regression models using SPSS software. A p value of < 0.05 was considered to be significant. 435 age-eligible women living in Maryland were recruited by direct mailing. Subjects completed questionnaires regarding demographics, reproductive history including any history of inability to conceive for 1 year or longer, diet/lifestyle, and smoking history. Participants also provided a blood sample that was subjected to polymerase chain reaction and enzyme digests for measurement of selected genetic polymorphisms in CYPc17, CYP1A1, and CYP1B1. Cases and controls were compared with Chi-Square tests and logistic regression models using SPSS software. A p value of < 0.05 was considered to be significant. Results119 of the 435 participants (27.4%) reported a history of inability to conceive for at least one year. Cases were more likely to have genetic polymorphisms in CYP1A1 than controls (4.7% of cases were homozygous for the polymorphism vs. 0.9% of controls; p < 0.014 after adjustment for potential confounders including race, marital status, education level, income, smoking history, oral contraceptive use, and body mass index). There were no significant differences in the prevalence of genetic polymorphisms in CYPc17 between cases and controls (69.9% of cases vs. 63.5% of controls; p = 0.455). Similarly, there were no significant differences in the prevalence of genetic polymorphisms in CYP1B1 between cases and controls (51.4% of cases vs. 55.0% of controls; p = 0.345). 119 of the 435 participants (27.4%) reported a history of inability to conceive for at least one year. Cases were more likely to have genetic polymorphisms in CYP1A1 than controls (4.7% of cases were homozygous for the polymorphism vs. 0.9% of controls; p < 0.014 after adjustment for potential confounders including race, marital status, education level, income, smoking history, oral contraceptive use, and body mass index). There were no significant differences in the prevalence of genetic polymorphisms in CYPc17 between cases and controls (69.9% of cases vs. 63.5% of controls; p = 0.455). Similarly, there were no significant differences in the prevalence of genetic polymorphisms in CYP1B1 between cases and controls (51.4% of cases vs. 55.0% of controls; p = 0.345). ConclusionGenetic polymorphisms in CYP1A1, but not CYP1B1 or CYPc17, are associated with an increased risk of self-reported infertility in women. Genetic polymorphisms in CYP1A1, but not CYP1B1 or CYPc17, are associated with an increased risk of self-reported infertility in women.

The aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that binds various environmental contaminants. Despite our knowledge regarding the role of the AhR in mediating toxicity, little is known about the physiological role of the AhR. Previous studies indicate that the AhR may regulate folliculogenesis, because AhR-deficient (AhRKO) mice have fewer preantral and antral follicles than wild-type (WT) mice during postnatal life. Thus, the first objective of the present study was to test the hypothesis that AhR deficiency reduces the numbers of preantral and antral follicles by slowing growth and/or increasing atresia of follicles. Because alterations in follicular growth or atresia can affect the ability to ovulate, the second objective was to test whether AhR deficiency reduces the number of ovulated eggs. To test these hypotheses, follicular growth was compared in WT and AhRKO ovaries using morphometric techniques and by measuring the ability of the ovary and follicles to grow in response to eCG. Atresia was compared in WT and AhRKO ovaries using morphometric techniques, TUNEL assays, and 3'-end labeling of fragmented DNA. Ovulation was compared in WT and AhRKO mice by assessing the number of corpora lutea per ovary. The results indicate that follicular growth and ovulation were reduced in AhRKO ovaries compared to WT ovaries. The WT ovaries had a 1.5-fold increase in the number of preantral and antral follicles between Postnatal Days 32 and 45, were more responsive to eCG, and contained more corpora lutea than AhRKO ovaries. In contrast, no significant difference was observed in the incidence of atresia in WT and AhRKO ovaries. Taken together, these results suggest that the AhR may regulate growth, but not atresia, of preantral and antral follicles in the mouse ovary.

Although Smad 3 is known to serve as a signaling intermediate for the transforming growth factor beta (TGFbeta) family in nonreproductive tissues, its role in the ovary is unknown. Thus, we used a recently generated Smad 3-deficient (Smad 3-/-) mouse model to test the hypothesis that Smad 3 alters female fertility and regulates the growth of ovarian follicles from the primordial stage to the antral stage. In addition, we tested whether Smad 3 affects the levels of proteins that control apoptosis, survival, and proliferation in the ovarian follicle. To test this hypothesis, breeding studies were conducted using Smad 3-/- and wild-type mice. In addition, ovaries were collected from Smad 3-/- and wild-type mice on Postnatal Days 2-90. One ovary from each animal was used to estimate the total number of primordial, primary, and antral follicles. The other ovary was used for immunohistochemical analysis of selected members of the B-cell lymphoma/leukemia-2 family of protooncogenes (Bax, Bcl-2, Bcl-x), proliferating cell nuclear antigen (PCNA), and cyclin-dependent kinase 2 (Cdk-2). The results indicate that Smad 3-/- mice have reduced fertility compared with wild type mice. The results also indicate that Smad 3 may not affect the size of the primordial follicle pool at birth, but it may regulate growth of primordial follicles to the antral stage. Further, the results indicate that Smad 3 may regulate the expression of Bax and Bcl-2, but not Bcl-x, Cdk-2, and PCNA. Collectively, these data suggest that Smad 3 may play an important role in the regulation of ovarian follicle growth and female fertility.

Objective: Recent studies suggest that ovarian volume and antral follicle numbers may be sensitive, specific, and early indicators of menopausal status. The accuracy of these markers, however, has not been. compared directly to more traditional markers [age and follicle-stimulating hormone (FSH) levels]. Thus, the purpose of this study was to test whether ovarian volume and antral follicle counts are more sensitive and specific markers of menopausal status than age or FSH levels.Design: Premenopausal (n = 34) and postmenopausal (n = 25) women between 40 and 54 years old received a transvaginal ultrasound for determination of ovarian volume and antral follicle numbers, provided blood for measurement of FSH levels, and completed a questionnaire. FSH levels, age, ovarian volume, and antral follicle numbers were compared using t tests. Receiver operating characteristic curves were generated to evaluate the sensitivity and specificity of each marker.Results: Postmenopausal women had significantly higher FSH levels (p less than or equal to 0.0001), smaller ovarian volumes (p less than or equal to 0.002), and fewer antral follicles (p less than or equal to 0.002) than premenopausal women. Ovarian volume: and antral follicle numbers had similar sensitivity (27.3-100%) and specificity (3.4-92.9%) in indicating postmenopausal status as FSH levels and age.Conclusion: These data suggest that ovarian volume and antral follicle numbers may be useful indicators of menopausal status.

Little is known about the embryonic factors that regulate the size of the primordial follicle endowment at birth. A few studies suggest that members of the B-cell lymphoma/leukemia-2 (bcl-2) family of protooncogenes may be important determinants. Thus, the purpose of this study was to test whether bcl-2 regulates the size of the primordial follicle pool at birth. To test this hypothesis, three lines of transgenic mice (c-kit/bcl-2 mice) were generated that overexpress human bcl-2 in an effort to reduce prenatal oocyte loss. The overexpression was targeted to the ovary and appropriate embryonic time period with the use of a 4.8-kilobase c-kit promoter. This promoter provided two to three times more expression of bcl-2 in the ovaries with minimal or no overexpression in most nongonadal tissues. On Postnatal Days 8-60, ovaries were collected from homozygous c-kit/bcl-2 and nontransgenic littermates (controls) and processed for histological evaluation of follicle numbers. All lines of c-kit/bcl-2 mice were born with significantly more primordial follicles than control mice (P < or = 0.05). By Postnatal Days 30-60, however, there were no significant differences in follicle numbers between c-kit/bcl-2 and control mice. These results indicate that bcl-2 overexpression increases the number of primordial follicles at birth, but that the surfeit of primordial follicles is not maintained in postnatal life. These data suggest that it is possible that the ovary may contain a census mechanism by which excess numbers of primordial follicles at birth are detected and removed from the ovary by adulthood.