Edward F. Krieg

Methods of monitoring menstrual function in field studies : efficacy of methods

Efficacy of methods for monitoring female reproductive potential under field study conditions was evaluated. Women (n = 10) were recruited to participate for two menstrual cycles on the bases, in part, of not seeking fertility assistance, working full-time but not in the medical field, and having less than one year of college education. Luteinizing hormone (LH), estrone-3-glucuronide, and pregnanediol-3-glucuronide were measured in daily morning urine and normalized to creatinine concentrations. These urinary measures were parallel to serum LH, estradiol, and progesterone profiles. Based on these urinary measures, 6 of 19 cycles were judged to be atypical. Transvaginal ultrasonography provided insights into ovarian activity during the atypical cycles. Of 13 LH surges detected by radioimmunoassay, 7 were not detected by a semiquantitative dipstick (OvuSTICK), perhaps due to that methods sensitivity to loss of LH immunoactivity caused by sample freezing. While intervals from salivary and vaginal mucous electrical resistance signals to the LH surge during typical cycles were similar to those reported previously, they were not predictive of ovulatory status during atypical cycles. Fifty-three percent of the cycles were misclassified on the basis of the basal body temperature rise. Cervical mucous color, amount, and consistency were not predictive of ovulation under these study conditions. The results from these 19 menstrual cycles provide information about the efficacy of various methods for characterizing menstrual function under field study conditions. In this regard, urinary endocrine measures are the most informative or practical.

Stability of urinary female reproductive hormones stored under various conditions.

Urinary reproductive hormones afford specific and sensitive evaluation of female reproductive potential in epidemiologic and clinical settings. The goal of this study was to characterize the stability of urinary luteinizing hormone, follicle stimulating hormone, estrone 3-glucuronide, pregnanediol 3-glucuronide, and creatinine during storage as functions of time, temperature, and additives. After 2 weeks with no additives, activity of the four analytes, relative to initial concentrations, ranged from 91.9 to 102.8% at 4 degrees C, 35.1 to 89.6% at 25 degrees C, and 7.5 to 66.9% at 37 degrees C. Antimicrobial additives did not consistently improve stability. Analyte activity for samples stored with no additives for 24 weeks at -80 degrees C ranged from 69.0 to 101.2%. Glycerol and bovine serum albumin improved analyte stability; activity ranged from 91.1 to 106.3%. Other additives were ineffective. These results reveal conditions for storing reproductive hormone analytes in urine during epidemiologic field studies.

Validations of time-resolved fluoroimmunoassays for urinary estrone 3-glucuronide and pregnanediol 3-glucuronide

Competitive time-resolved fluoroimmunoassays (FIAs) were developed for measuring 1,3,5(10)-estratrien-3-ol-17-one glucosiduronate (estrone 3-glucuronide, E(1)3G) and 5 beta-Pregnane-3 alpha,20 alpha-diol 3-glucosiduronate (pregnanediol 3-glucuronide, Pd3G) in unextracted urine. The assays are specific, detect 0.98 ng E(1)3G/mL and 0.035 microgram Pd3G/mL, measure 102.8 +/- 2.0% of E(1)3G and 93.6 +/- 2.9% of Pd3G added, and exhibit between and within assay coefficients of variation, respectively, of 5.3% and 7.1% for E(1)3G and 6.8% and 7.8% for Pd3G. The urine matrix does not interfere with the assay. Urinary steroid glucuronide profiles measured by these FIAs conform to those of urinary steroid glucuronides and serum estradiol and progesterone measured by other established immunoassays. These FIAs afford the advantages of non-radioisotopic procedures and urine sample collection (convenience, non-invasiveness, integration of pulsatile secretion) to evaluate menstrual function in epidemiological, medical, and athletic populations.

Time-resolved immunofluorometric assays for urinary luteinizing hormone and follicle stimulating hormone

Abstract The goal of this effort was to develop and validate non-radioisotopic immunoassays for measuring luteinizing hormone and follicle stimulating hormone in unextracted urine. Towards this goal, commercial time-resolved immunofluorometric assays (IFMAs) were modified. Validation demonstrated that the resultant assays were specific, sensitive, accurate, and precise. Urine matrix was shown not to interfere with the assay. Gonadotropin profiles generated using these assays conformed to those measured in urine and serum by other established immunoassays. These IFMAs afford the collective advantages of non-radioisotopic procedures and urine sample collection (convenience, noninvasiveness, integration of pulsatile secretion), plus the superior sensitivity and specificity of IFMAs. Applications include epidemiology and medicine.

Detection of DNA damage in workers exposed to JP-8 jet fuel.

The genotoxicity of jet propulsion fuel 8 (JP-8) was assessed in the leukocytes of archived blood specimens from U.S. Air Force personnel using the comet assay. No differences in mean comet assay measurements were found between low, moderate, and high exposure groups before or after a 4h work shift. Before the work shift, mean tail DNA and mean tail (Olive) moment increased as the concentration of benzene measured in end-exhaled breath increased, indicating that prior environmental or work-related exposures to benzene produced DNA damage. The number of cells with highly damaged DNA decreased as the pre-shift benzene concentration in breath increased. It is not clear why the decrease is occurring. Mean tail DNA and mean tail (Olive) moment decreased as the concentrations of benzene and naphthalene measured in breath immediately after the work shift increased. These inverse relationships may reflect a slower rate of absorption or a faster rate of expiration of benzene in the lung. The number of cells with highly damaged DNA increased as the concentration of urinary (2-methoxyethoxy)acetic acid (MEAA) increased. This relationship was not seen in urinary MEAA adjusted for creatinine. MEAA is a metabolite of the deicing agent 2-(2-methoxyethoxy)ethanol contained in JP-8. MEAA or a component of JP-8 correlated with MEAA may have a toxic effect on DNA.

Methods of monitoring menstrual function in field studies: Attitudes of working women

This study was designed to determine the attitudes and compliance of working women toward methods being evaluated for use in the assessment of the effects of toxicants on reproductive potential. Women such as the highly motivated fertility patients and nurses, who are typically familiar with the methods and procedures of fertility assessment and the value of medical research, have been used to validate such methods in a clinical setting. However, the attitudes of a general working female population toward these methods are unknown. Nine participants were selected on the bases, in part, of not seeking fertility assistance, working full-time but not in the medical field, and having less than one year of college education. Attitudes were also evaluated for 193 non-participating women to whom the procedures had been verbally described. Participants measured basal body temperature and salivary and vaginal mucous electrical resistance, evaluated cervical mucus manually (CME), and collected the first morning urine for two menstrual cycles. Blood, saliva, and transvaginal ultrasonograms (US) were obtained at a fertility clinic 6 to 9 days per cycle. Participants brought urine to the laboratory every 3 days. All participants performed all methods. Participants were paid

The relationships between blood lead levels and serum follicle stimulating hormone and luteinizing hormone in the National Health and Nutrition Examination Survey 1999-2002.

400; nonparticipants were not compensated. Only 3% of the respondents objected to the proposed methods: principally to CME, US, and giving blood samples. No respondent perceived the study as unimportant.(ABSTRACT TRUNCATED AT 250 WORDS)

A mathematical model of performance on a simple reaction time test

The relationships between blood lead levels and serum follicle stimulating hormone and luteinizing hormone were assessed in a nationally representative sample of women, 35-60 years old, from the National Health and Nutrition Examination Survey 1999-2002. The blood lead levels of the women ranged from 0.2 to 17.0 μg/dL. The estimated geometric mean was 1.4 μg/dL, and the estimated arithmetic mean was 1.6 μg/dL. As the blood lead level increased, the concentration of serum follicle stimulating hormone increased in post-menopausal women, women who had both ovaries removed, and pre-menopausal women. The concentration of luteinizing hormone increased as blood lead level increased in post-menopausal women and women who had both ovaries removed. The lowest concentrations of blood lead at which a relationship was detected were 0.9 μg/dL for follicle stimulating hormone and 3.2 μg/dL for luteinizing hormone. Lead may act directly or indirectly at ovarian and non-ovarian sites to increase the concentrations of follicle stimulating hormone and luteinizing hormone.

Blood lead, serum homocysteine, and neurobehavioral test performance in the third National Health and Nutrition Examination Survey.

A nonlinear function with components for learning and fatigue was used to model individual performance on a simple reaction time test. The relationships between the parameters of the model and the mean and variance of the reaction times are discussed. The function is used to analyze data from a field study of agricultural workers exposed to organophosphate pesticides. Exposure had a significant effect on the relationships between education level and initial performance, age and fatigue rate, and age and performance variability. Parameter estimates from the model were able to distinguish between effects that the mean and standard deviation of the reaction times were unable to distinguish.