Irma C. Oskam

Organochlorines Affect the Major Androgenic Hormone, Testosterone, in Male Polar Bears (Ursus Maritimus) at Svalbard

Normal sexual development and subsequent reproductive function are dependent on appropriate testosterone production and action. The regulation of steroid hormones, including androgens, can be influenced by both biological and environmental factors, including environmental chemicals. Concentrations of organochlorines are considerably greater in Svalbard polar bears than in polar bears from other regions. Between 1995 and 1998, samples were collected from 121 male polar bears (Ursus maritimus) from the Svalbard area. In this study, testosterone concentration variations were described for male polar bears during different seasons and for all age groups. To study possible relationships between plasma testosterone concentrations and biological factors, such as age, axial girth, and extractable plasma fat, and organochlorine contaminants including hexachlorocyclohexanes, hexachlorobenzene, chlordanes, p,p′–DDE, and 16 individual polychlorinated biphenyl (PCB) congeners, identical statistical analyses were performed on the total population and a subsample of reproductively active adults. Of the biological factors, axial girth showed a significant positive relationship and percentage extractable fat and a significant negative relationship with the testosterone concentrations. Both the Σpesticides and ΣPCBs made significant negative contributions to the variation of the plasma testosterone concentration. The continuous presence of high concentrations of organochlorines in male polar bears throughout their life could possibly aggravate any reproductive toxicity that may have occurred during fetal and early postnatal development.

Organochlorines Affect the Steroid Hormone Cortisol in Free-Ranging Polar Bears (Ursus maritimus) at Svalbard, Norway

Since the polar bear (Ursus maritimus) is among the most highly organochlorine-contaminated species of the Arctic mammals, there is growing concern that in addition to the natural stressors in the polar bears environment, several organochlorines (OCs) may be able to change basic endocrine pathways. Alterations in the hypothalamic–pituitary–adrenal (HPA) axis may affect plasma cortisol concentrations and inhibit physiological processes involved in the maintenance of homeostasis in a way that may endanger the animals health. Between 1995 and 1998, samples were collected from 121 male and 130 female free-ranging polar bears from the Svalbard area. The aim of the study was to investigate relationships between plasma cortisol concentrations, biological factors, and OCs. The variation in plasma cortisol concentrations was determined for the total sample. Axillary girth and body mass together with their interactions explained more than 50% of the variation in the plasma cortisol concentration. The sum of pesticides (Σpesticides) combined with the sum of polychlorinated biphenyls (ΣPCBs) and their interactions explained over 25% of the variation in the cortisol concentration. Although Σpesticides contributed negatively and ΣPCBs contributed positively to the variation in the plasma cortisol, the over-all contribution of the OCs to the plasma cortisol variation was negative. Despite the complexity on stress responses and the interactions with environmental factors, this study demonstrated that high concentrations of OCs in polar bears might alter plasma cortisol concentrations.

Endocrine Disruption Induced by Organochlorines (OCs): Field Studies And Experimental Models

Long-range transport of persistent organic compounds by air and ocean currents from industrialized areas resulted in high levels of these pollutants in food webs in the Svalbard area. With the aim to test if organochlorine (OC) exposure in free-living polar bears from Svalbard affected their plasma steroid hormone concentrations, it was found that polychlorinated biphenyls (PCBs) were associated with increased progesterone levels in females. The sum of pesticides (∑pesticides) and ∑PCBs contributed significantly negative to the variation of the plasma testosterone in males, and the overall contribution of the OCs to the plasma cortisol variation was negative. A second objective was to study the effects of selected OCs (i.e., PCB 153 and PCB 126) on animal health as a consequence of effects on endocrine-regulated functions such as reproduction and immunity in a goat model focusing on long-term and low-level exposure during the periods of fetal development and in the neonatal period. Additionally, acute exposure was studied in adult mice. The results indicated that exposure to low doses of PCB 153 in utero and in the suckling period influenced reproductive functions and both PCB 153 and PCB 126 exerted immunomodulatory effects on the offspring, whereas acute exposure of adult mice had minor effects on male reproductive function.

Altered Stress-Induced Cortisol Levels in Goats Exposed to Polychlorinated Biphenyls (PCB 126 and PCB 153) During Fetal and Postnatal Development

Short-term stress exposure is associated with activation of the hypothalamic–pituitary–adrenal (HPA) axis and a consequent rise in blood glucocorticoids and catecholamines, from the adrenal cortex and medulla, respectively. The HPA axis is a potential target for some persistent organic pollutants, among which polychlorinated biphenyls (PCB) were found to be modulators of the mammalian endocrine system. PCB are distributed globally in the environment, in food chains, and are transferred to the fetuses of pregnant animals and via mothers milk to suckling offspring. In the present study it was postulated that intrauterine and lactational exposure to either of two single congeners of PCB (PCB 153 and PCB 126, respectively) might affect basal cortisol concentrations, and also the cortisol response to short-term stress in adulthood. Thus, pregnant goats were orally exposed to one of these PCB congeners from d 60 of gestation until delivery, and their offspring studied. Low-dose exposure to PCB 153 and PCB 126 resulted in significantly lower mean basal cortisol concentrations in goat offspring during certain periods of pubertal development and their first breeding season. Male goat kids exposed to either PCB congener showed a greater and more prolonged rise in plasma cortisol levels than controls when animals were subjected to mild stress at 9 mo of age using frequent blood sampling. Neither the basal maternal cortisol plasma level nor goat kid adrenal masses were affected by PCB exposure.

Effects of hCG Stimulation on Hepatic Activities of Cytochromes P4502E1 and P4502A in Pubertal Male Pigs

The effects of human chorionic gonadotropin (hCG) stimulation on fat skatole concentrations and hepatic activities of cytochromes P4502E1 (CYP2E1) and P4502A (CYP2A) were studied in Landrace and Duroc breeds of entire male pigs. Pigs were divided into four groups: two control groups of each breed, without hCG stimulation (n = 20 for each breed), and two experimental groups of each breed, with hCG stimulation (n = 18 for each breed). Pigs were slaughtered 3 days after hCG stimulation and activities of CYP2E1 and CYP2A were measured in liver homogenate. Activities of both CYP2E1 and CYP2A were lower in hCG-stimulated pigs than control pigs for both Landrace (p = 0.005 for CYP2E1, p = 0.016 for CYP2A) and Duroc breeds (p = 0.003 for CYP2E1, p = 0.001 for CYP2A), and skatole concentrations in fat were higher in the hCG-stimulated pigs of both breeds (p < 0.01). For both control and hCG-stimulated groups, Duroc pigs had lower skatole concentrations than Landrace pigs (p = 0.001 for both groups). The activity of CYP2E1 did not differ significantly between breeds in either the control group or the experimental group (p = 0.233 for control pigs and p = 0.210 for experimental pigs). However, whereas CYP2A activity did not differ significantly between breeds in the control groups (p = 0.181 for CYP2A), in the hCG-stimulated groups, CYP2A activity was lower in Duroc pigs than in Landrace (p = 0.011). Based on these findings, we conclude that hCG stimulation can suppress hepatic CYP2E1 and CYP2A activities, probably through an increase in the levels of testicular steroids. Between-breed variations in skatole levels in fat were not related to the activities of CYP2E1 and CYP2A.

Bone marrow remission status predicts leukemia contamination in ovarian biopsies collected for fertility preservation.

Bone marrow remission status predicts leukemia contamination in ovarian biopsies collected for fertility preservation

Differences in testosterone, androstenone, and skatole levels in plasma and fat between pubertal purebred Duroc and Landrace boars in response to human chorionic gonadotrophin stimulation.

The concentrations of the boar taint compounds androstenone and skatole in plasma and fat, together with those of testosterone in plasma, were investigated in pubertal purebred Duroc and Landrace boars following stimulation with human chorionic gonadotrophin (hCG). Higher initial levels of androstenone and testosterone were found in Duroc than Landrace boars. Duroc boars, which were approximately ten days older than the Landrace boars, also showed a more advanced stage of spermatogenesis than Landrace boars. While Landrace boars had the highest skatole levels. Following stimulation with hCG the relative increases in testosterone, androstenone, and skatole concentrations were highest in Landrace boars. The level of androstenone in fat three days after hCG stimulation exceeded 1 microg/g fat in all stimulated boars. The decreases in plasma levels of androstenone and testosterone on Days 2 and 3 after hCG stimulation were more pronounced in Landrace than Duroc boars. However, unlike the plasma androstenone and testosterone levels, the plasma concentrations of skatole did not decrease on Days 2 and 3 following stimulation, but remained elevated on Day 3. These results indicate that the lower levels of testicular steroids in Landrace boars compared with Duroc boars was not due to a lower production capacity, but more likely to a faster disappearance of steroids in Landrace boars. In the present study, age, live weight, and testicular development did not significantly contribute to the variation in fat androstenone. The present data and previous reports on candidate genes related to androstenone biosynthesis and metabolism suggests that future selection against factors associated with boar taint remains a possible solution for the problem of boar taint in the swine industry.

Testicular germ cell development in relation to 5α-Androstenone levels in pubertal entire male pigs

Androstenone is a 16-androstene steroid pheromone produced in the Leydig cells in the testis, and considered to be one of the major compounds responsible for boar taint. In entire male pigs, progress of sexual maturation has been related to an increase in androstenone levels in fat. Onset of puberty and subsequent reproductive function involves genetic factors affected by the internal and external environment. In this study entire male cross-bred pigs were housed under two different light regimens in order to manipulate the onset of puberty. DNA flow cytometry (FCM) was used to study spermatogenesis and monitor the proportions of haploid (1n), diploid (2n), and tetraploid (4n) testicular cells, with conventional histological evaluation used as the reference technique. Agreement between these two methods was found to be good. The best fit model explained 34% of the variation in the androstenone concentrations. Sexual maturation in boars of 125-146 days of age, as assessed by DNA FCM, was not significantly associated with the variation in androstenone concentrations in adipose tissue when various independent variables (breed, age, light strategy, skatole concentrations in fat, and length of the bulbourethralis glands) were included in this model. These findings support the suggestion that selection against androstenone may be an option in the breeding of entire male pigs.

Effect of Previous Chemotherapy on the Quality of Cryopreserved Human Ovarian Tissue In Vitro

Background Cryopreservation of ovarian tissue has been widely accepted as an option for fertility preservation among cancer patients. Some patients are exposed to chemotherapy prior to ovarian tissue cryopreservation. Consequently, assessment of the developmental capacity of human ovarian tissue after chemotherapy is of primary importance. Materials In order to study the impact of previous chemotherapy on in vitro development and viability of ovarian follicles, quality control samples from 34 female cancer patients at median age of 15 years (range 1‒35), cryopreserved for fertility preservation before (n = 14) or after (n = 20) initiation of chemotherapy, were thawed and cultured for 7 days. The morphology and developmental stages of ovarian follicles were studied by light microscopy before and after culture. Possible associations between follicular densities, age and exposure to alkylating agents, expressed as cyclophosphamide equivalent dose (CED) were tested. Results Exposure to chemotherapy significantly impaired the survival and development of ovarian follicles in culture. After seven days, significantly higher densities of intermediary, primary and secondary follicles and lower densities of atretic follicles was detected in the samples collected before chemotherapy. Increasing dose of alkylating agents was identified by multivariate linear regression analysis as an independent predictor of a higher density of atretic follicles, whereas increasing age of the patient predicted a better outcome with less follicle atresia and a higher density of maturing follicles. Conclusion This study provides quantitative in vitro evidence of the impact of chemotherapy on developmental capacity of cryopreserved human ovarian tissue. The results indicate that fertility preservation should be carried out, if possible, before initiation of alkylating agents in order to guarantee better in vitro survival of ovarian follicles. In addition, ovarian samples from younger girls show lower viability and fewer developing follicles in culture.

Gene expression during testis development in Duroc boars.

Androstenone is a steroid pheromone occurring in the pubertal Leydig cells. Breeding against androstenone can decrease pheromone odour in swine meat but appears to cause unwanted side effects such as delayed onset of puberty. To study causality, global gene expression in developing boar testes at 12, 16, 20 and 27 weeks was investigated using a porcine cDNA microarray. The morphological status and androgenic levels of the same individuals have been described in a previous publication. In the present paper, expression of genes and pathways has been analysed with reference to these findings. Nine clusters of genes with significant differential expression over time and 49 functional charts were found in the analysed testis samples. Prominent pathways in the prepubertal testis were associated with tissue renewal, cell respiration and increased endocytocis. E-cadherines may be associated with the onset of pubertal development. With elevated steroidogenesis (weeks 16 to 27), there was an increase in the expression of genes in the MAPK pathway, STAR and its analogue STARD6. A pubertal shift in genes coding for cellular cholesterol transport was observed. Increased expression of meiotic pathways coincided with the morphological onset of puberty. Puberty-related change in Ca(2+) pathway transcripts, neurosteroids, neuronal changes and signalling in redox pathways suggested a developmental-specific period of neuromorphogenesis. Several growth factors were found to increase differentially over time as the testis matured. There may be interactions between MAPK, STAR and growth factors during specific periods. In conclusion, pathways for neurogenesis, morphological pathways and several transcripts for growth factors, which have known modulating effects on steroidogenesis and gonadotropins in humans and rodents, act at specific ages and developmental stages in the boar testis. The age dependency and complexity shown for development-specific testis transcripts must be considered when selecting phenotypic parameters for genetic selection for low androstenone. The results of selection based on measurement of phenotypic maturation and androstenone (or other steroid) levels at one specific age may differ depending on the age used. More research is necessary to find the optimal phenotype to use in order to reduce the unwanted side effects.