Graduation date: 1992

Objective: Although associations between testosterone and cardiovascular (CV) morbidity in women have been proposed, no large prospective study has evaluated potential associations between testosterone and mortality in women. The objective was to determine whether baseline testosterone levels in women are associated with future overall or CV morbidity and mortality. Design: Prospective cohort study with a 4.5-year follow-up period. Methods: From a representative sample of German primary care practices, 2914 female patients between 18 and 75 years were analyzed for the main outcome measures: CV risk factors, CV diseases, and all-cause mortality. Results: At baseline, the study population was aged 57.96±14.37 years with a mean body mass index of 26.71±5.17 kg/m2. No predictive value of total testosterone for incident CV risk factors or CV diseases was observed in logistic regressions. Patients with total testosterone levels in the lowest quintile Q1, however, had a higher risk to die of any cause or to develop a CV event within the follow-up period compared to patients in the collapsed quintiles Q2–Q5 in crude and adjusted Cox regression models (all-cause mortality: Q2–Q5 versus Q1: crude hazard ratios (HR) 0.49, 95% confidence interval (CI) 0.33–0.74; adjusted HR 0.62, 95% CI 0.42–0.939; CV events: Q2–Q5 versus Q1: crude HR 0.54, 95% CI 0.38–0.77; adjusted HR 0.68, 95% CI 0.48–0.97). Kaplan–Meier curves revealed similar data. Conclusions: Low baseline testosterone in women is associated with increased all-cause mortality and incident CV events independent of traditional risk factors.

In the present study our objectives were (1) to follow the temporal patterns of testicular LH and FSH receptor (LH-R and FSH-R) concentrations and affinity (Ka) during sexual maturation in bulls, to see if such patterns could explain the control of rapid testicular growth that occurs after 25 weeks of age, when serum gonadotropin concentrations are low; (2) to see if transformation growth factors (TGF- alpha and beta 1, 2 and 3) and interleukins (IL-1 and IL-6) are produced in the developing bovine testis and if their concentrations change during development; (3) to see if the onset of puberty could be hastened by treating bull calves subcutaneously (sc) with 3 mg of bLH (n=6) or 4 mg of bFSH (n=6) once every 2 days, from 4 to 8 weeks after birth. Mean LH-R concentrations decreased from 13 to 25 weeks of age and increased to 56 weeks of age (P<0.05). LH-RKa decreased from 9 to 17 weeks of age, increased to 29 weeks and declined to 33 weeks of age (P<0.05). FSH-R concentrations declined from 17 to 25 weeks of age then increased to 56 weeks of age (P<0.05). FSH-RKa increased from 17 to 25 weeks of age (P<0.05). Testicular TGF-alpha concentrations increased from 13 to 17 weeks of age, decreased to 21 weeks and from 33 to 56 weeks of age (P<0.05). Testicular TGF-beta 1 concentrations decreased from 17 to 21 weeks of age, increased to 25 weeks and decreased from 25 to 29 weeks of age (P<0.05). Testicular TGF-beta 2 concentrations increased from 5 to 17 weeks of age, decreased to 21 weeks, increased to 25 weeks and decreased at 29 weeks of age (P<0.05). Testicular TGF-beta 3 concentrations increased from 13 to 17 weeks of age, decreased to 21 weeks of age and from 25 to 29 weeks of age (P<0.05). Mean testicular IL-1 alpha concentrations decreased from 5 to 9 weeks of age and 13 to 21 weeks of age (P<0.01) while mean testicular IL-1 beta concentrations decreased from 13 to 17 weeks and 29 to 33 weeks of age (P<0.01). Mean IL-1 bioactivity increased from 13 to 17 weeks of age, decreased to 21 weeks, increased to 25 weeks, decreased to 29 weeks and decreased from 33 to 56 weeks of age (P<0.05). Mean testicular IL-6 concentrations decreased (P<0.05) from 9 to 13 weeks of age, increased (P<0.05) to 21 weeks, decreased (P<0.05) to 25 weeks of age, increased (P<0.05) to 29 weeks and decreased (P<0.01) to 56 weeks of age. <p>We concluded that high concentrations of gonadotropin receptors might be critical to initiate postnatal testis growth and support it after 25 weeks of age in the face of low serum gonadotropin concentrations. Testicular TGF-alpha concentrations were higher in calves than adults while concentrations of TGF-beta and IL-1 were higher in the early postnatal period than the peripubertal period. The changes in testicular concentrations of TGFs and ILs led us to suggest a possible local regulatory role in development. Testicular IL-6 concentrations were higher in prepubertal calves than adults. Treatment of bull calves with bFSH from 4 to 8 weeks of age increased testicular growth (SC), hastened the onset of puberty (SC ≥ 28 cm), and enhanced spermatogenesis.

Androgens (such as testosterone) have been shown to increase lean body mass and reduce fat body mass in men through activation of androgen receptors (AR). While this suggests a potential clinical use for androgens, attempts at utilization of this class of hormones as a therapeutic are limited by side effects due to indiscriminate AR activation in various tissues. Thus, a greater understanding of the tissues and cells involved in promoting these changes would be beneficial. Here we show that selective overexpression of AR in muscle cells of transgenic (HSA-AR) rodents both increases lean muscle mass and significantly reduces fat mass in males. Similar effects can be induced in HSA-AR females treated with testosterone. Metabolic analyses of HSA-AR males show that these animals demonstrate increased O2 consumption and hypermetabolism. Thus, targeted activation of AR in muscle regulates body composition and metabolism, suggesting a novel target for drug development.

Androgens (such as testosterone) have been shown to increase lean body mass and reduce fat body mass in men through activation of androgen receptors (AR). While this suggests a potential clinical use for androgens, attempts at utilization of this class of hormones as a therapeutic are limited by side effects due to indiscriminate AR activation in various tissues. Thus, a greater understanding of the tissues and cells involved in promoting these changes would be beneficial. Here we show that selective overexpression of AR in muscle cells of transgenic (HSA-AR) rodents both increases lean muscle mass and significantly reduces fat mass in males. Similar effects can be induced in HSA-AR females treated with testosterone. Metabolic analyses of HSA-AR males show that these animals demonstrate increased O2 consumption and hypermetabolism. Thus, targeted activation of AR in muscle regulates body composition and metabolism, suggesting a novel target for drug development.

Social interactions can have profound effects on the behavior, physiology, and overall fitness of an individual. An example of this in Lythrypnus dalli is the removal of a male from a social group resulting in a dominant female fish changing sex. The dominant female's transformation involves a suite of changes including brain, behavior, morphology, and physiology. Following the social trigger (male removal), sex-changing individuals' morphology, steroid levels, and changes in the behavior were quantified in the field and compared to results found previously in the laboratory. There were lower rates of aggressive and courtship behavior in the field, but the change in behavior over time had a similar pattern and there were parallels in morphology and steroid levels between lab and field sex changers. The brains of dominant females also responded to social change. Aromatase, an enzyme that converts testosterone into estrogen, and oxytocin, a neuropeptide found in mammals, have been associated with vertebrate social and reproductive behavior. The fish homologue of oxytocin, isotocin, and aromatase are both found in L. dalli. Upon removal of a male from the social group, L. dalli dominant females experienced a decrease in the number of preoptic area isotocin-immunoreactive cells over the course of sex change (7-10 days) and a decrease in brain aromatase activity (bAA) levels within hours, but not minutes, of male removal, while gonadal aromatase activity (gAA) decreased at a much slower time scale (beyond a week). Hours, but not minutes, after male removal, the sex-changing individual's bAA correlated with aggressive behavior increases and not the amount of time following male removal. Males that had just changed from female had different gonadal allocation and higher bAA levels than established males. Subordinate females had high gAA, but their bAA was between that of males and sex changers. In conclusion, dramatic changes in anatomy and neuroendocrine function can occur in response to social cues, individuals with similar reproductive behavior and external morphology can have large neuroendocrine and internal morphologic variation, and social interactions can affect steroid metabolism locally on a short time scale independent of gonadal modulation of steroids.

I conducted an integrative field study on male cottonmouths (Agkistrodon piscivorus), a common pitviper of the southeastern United States, to investigate the evolution of contrasting mating patterns in North American pitvipers (bimodal and unimodal annual patterns) and resolve conflicting information about the pattern exhibited by the cottonmouth. I determined a unimodal late summer peak of testosterone (T) and a muted unimodal seasonal cycle of the sexual segment of the kidney (a secondary sexual characteristic), both of which were correlated with the single peak of spermatogenesis in late summer. I also conducted a study to determine diel and seasonal variation of corticosterone (CORT), the effect of captive handling on CORT, and the relationship between CORT and T after captive handling, and detected a significant elevation of CORT and a significant decrease of T after capture in male cottonmouths, as well as a significant negative correlation between CORT and T.

Declines in some fish populations in the Upper Danube River, Germany, have been reported during the past decades despite extensive stocking efforts. Many theories exist for why such declines have occurred including habitat change, dams, invasive species, disease and pollution. One of the factors of concern in the Upper Danube River is pollution because a number of studies have shown that sediments collected from this area were acutely and/or chronically toxic to fish. Although it can be difficult to link bioassay results to direct effects on the population level, bioassays can give us insight into the potential of exposure of wildlife including fish to sediment. In combination with other researchers a large battery of sediment testing on the Upper Danube River is being performed. Testing includes sediment testing of estrogen receptor mediated processes, dioxin-like responses and genotoxic effects. In this study, four sediment extracts from the Upper Danube River in Germany were used with a novel fractionation technique to characterize the sediment extracts and fractions for their ability to disrupt steroidogenesis, for their mutagenic activities and their teratogenic effects. Fractionation of each of the four sediment samples was performed by separating compounds according to their polarity, planarity, and the size of the aromatic ring system in an on-line fractionation procedure on coupled high performance liquid chromatography columns.<p> Mutagenic activity was measured in the raw sediment extracts and all 18 fractions using the Ames fluctuation assay and the Danio rerio embryo assay was used to assess lethal endpoints. Furthermore, disruptions of steroidogenesis were assessed by first establishing methods and a proof of concept of the H295R assay by exposing H295R cells to 7 model chemicals and measuring changes from a control in estradiol, testosterone and aromatase activity. Once methods were established all sediments and their fractions were analyzed using the Assay.<p> Specifically, in the <i>Danio rerio</i> assay, two raw sediment extracts killed 100% of <i>Danio rerio</i> embryos at a concentration of 33.3 mg sediment equivalents (SEQ)/ml, but none of the 18 fractions of these samples produced any measured toxicity at a concentration of 100 mg SEQ/ml. In the Ames fluctuation assay, significant mutagenic activity was measured in raw sediment extracts and in the fractions. Fraction 10 produced a significant mutagenic response in all sediment samples measured only in S9 bio-activated samples. Furthermore, fraction 15 produced a significant mutagenic response in all sediment samples measured only in non bio-activated samples.<p> All raw extracts tested in the H295R assay caused an increase in estradiol production up to 4-fold from controls. Testosterone production increased slightly from controls in only two of the raw extract samples. Of the 18 fractions, fractions 7, 10 and 15 increased estradiol in at least three of the samples studied (Sigmaringen2006, Opfingen2006, Lauchert2006 and Lauchert2004). Furthermore, fraction 7 significantly decreased testosterone production compared to controls in three of the four sediment samples.<p> Taken as a whole, these results show the value of using multiple bioassays and fractionation to characterize sediments that covers a variety of different biological endpoints. This study also demonstrates the usefulness of the H295R assay when combined with a new fraction technique to assess endocrine disrupting chemicals in sediment samples.

In the present study our objectives were (1) to follow the temporal patterns of testicular LH and FSH receptor (LH-R and FSH-R) concentrations and affinity (Ka) during sexual maturation in bulls, to see if such patterns could explain the control of rapid testicular growth that occurs after 25 weeks of age, when serum gonadotropin concentrations are low; (2) to see if transformation growth factors (TGF- alpha and beta 1, 2 and 3) and interleukins (IL-1 and IL-6) are produced in the developing bovine testis and if their concentrations change during development; (3) to see if the onset of puberty could be hastened by treating bull calves subcutaneously (sc) with 3 mg of bLH (n=6) or 4 mg of bFSH (n=6) once every 2 days, from 4 to 8 weeks after birth. Mean LH-R concentrations decreased from 13 to 25 weeks of age and increased to 56 weeks of age (P<0.05). LH-RKa decreased from 9 to 17 weeks of age, increased to 29 weeks and declined to 33 weeks of age (P<0.05). FSH-R concentrations declined from 17 to 25 weeks of age then increased to 56 weeks of age (P<0.05). FSH-RKa increased from 17 to 25 weeks of age (P<0.05). Testicular TGF-alpha concentrations increased from 13 to 17 weeks of age, decreased to 21 weeks and from 33 to 56 weeks of age (P<0.05). Testicular TGF-beta 1 concentrations decreased from 17 to 21 weeks of age, increased to 25 weeks and decreased from 25 to 29 weeks of age (P<0.05). Testicular TGF-beta 2 concentrations increased from 5 to 17 weeks of age, decreased to 21 weeks, increased to 25 weeks and decreased at 29 weeks of age (P<0.05). Testicular TGF-beta 3 concentrations increased from 13 to 17 weeks of age, decreased to 21 weeks of age and from 25 to 29 weeks of age (P<0.05). Mean testicular IL-1 alpha concentrations decreased from 5 to 9 weeks of age and 13 to 21 weeks of age (P<0.01) while mean testicular IL-1 beta concentrations decreased from 13 to 17 weeks and 29 to 33 weeks of age (P<0.01). Mean IL-1 bioactivity increased from 13 to 17 weeks of age, decreased to 21 weeks, increased to 25 weeks, decreased to 29 weeks and decreased from 33 to 56 weeks of age (P<0.05). Mean testicular IL-6 concentrations decreased (P<0.05) from 9 to 13 weeks of age, increased (P<0.05) to 21 weeks, decreased (P<0.05) to 25 weeks of age, increased (P<0.05) to 29 weeks and decreased (P<0.01) to 56 weeks of age. <p>We concluded that high concentrations of gonadotropin receptors might be critical to initiate postnatal testis growth and support it after 25 weeks of age in the face of low serum gonadotropin concentrations. Testicular TGF-alpha concentrations were higher in calves than adults while concentrations of TGF-beta and IL-1 were higher in the early postnatal period than the peripubertal period. The changes in testicular concentrations of TGFs and ILs led us to suggest a possible local regulatory role in development. Testicular IL-6 concentrations were higher in prepubertal calves than adults. Treatment of bull calves with bFSH from 4 to 8 weeks of age increased testicular growth (SC), hastened the onset of puberty (SC ≥ 28 cm), and enhanced spermatogenesis.

Declines in some fish populations in the Upper Danube River, Germany, have been reported during the past decades despite extensive stocking efforts. Many theories exist for why such declines have occurred including habitat change, dams, invasive species, disease and pollution. One of the factors of concern in the Upper Danube River is pollution because a number of studies have shown that sediments collected from this area were acutely and/or chronically toxic to fish. Although it can be difficult to link bioassay results to direct effects on the population level, bioassays can give us insight into the potential of exposure of wildlife including fish to sediment. In combination with other researchers a large battery of sediment testing on the Upper Danube River is being performed. Testing includes sediment testing of estrogen receptor mediated processes, dioxin-like responses and genotoxic effects. In this study, four sediment extracts from the Upper Danube River in Germany were used with a novel fractionation technique to characterize the sediment extracts and fractions for their ability to disrupt steroidogenesis, for their mutagenic activities and their teratogenic effects. Fractionation of each of the four sediment samples was performed by separating compounds according to their polarity, planarity, and the size of the aromatic ring system in an on-line fractionation procedure on coupled high performance liquid chromatography columns.<p> Mutagenic activity was measured in the raw sediment extracts and all 18 fractions using the Ames fluctuation assay and the Danio rerio embryo assay was used to assess lethal endpoints. Furthermore, disruptions of steroidogenesis were assessed by first establishing methods and a proof of concept of the H295R assay by exposing H295R cells to 7 model chemicals and measuring changes from a control in estradiol, testosterone and aromatase activity. Once methods were established all sediments and their fractions were analyzed using the Assay.<p> Specifically, in the <i>Danio rerio</i> assay, two raw sediment extracts killed 100% of <i>Danio rerio</i> embryos at a concentration of 33.3 mg sediment equivalents (SEQ)/ml, but none of the 18 fractions of these samples produced any measured toxicity at a concentration of 100 mg SEQ/ml. In the Ames fluctuation assay, significant mutagenic activity was measured in raw sediment extracts and in the fractions. Fraction 10 produced a significant mutagenic response in all sediment samples measured only in S9 bio-activated samples. Furthermore, fraction 15 produced a significant mutagenic response in all sediment samples measured only in non bio-activated samples.<p> All raw extracts tested in the H295R assay caused an increase in estradiol production up to 4-fold from controls. Testosterone production increased slightly from controls in only two of the raw extract samples. Of the 18 fractions, fractions 7, 10 and 15 increased estradiol in at least three of the samples studied (Sigmaringen2006, Opfingen2006, Lauchert2006 and Lauchert2004). Furthermore, fraction 7 significantly decreased testosterone production compared to controls in three of the four sediment samples.<p> Taken as a whole, these results show the value of using multiple bioassays and fractionation to characterize sediments that covers a variety of different biological endpoints. This study also demonstrates the usefulness of the H295R assay when combined with a new fraction technique to assess endocrine disrupting chemicals in sediment samples.