Showing papers in "International Journal of Andrology in 2008"
TL;DR: The acute toxicity of the polyfluorinated substances is moderate but some substances can induce peroxisome proliferation in rat livers and may change the fluidity of cell membranes, and hundreds of related chemicals, which potentially may degrade to perfluoroalkanoic (carboxylic) acids, are not regulated.
Abstract: In recent years, polyfluorinated chemicals (PFCs) have increasingly been used as surfactants in various industry- and consumer products, because of their unique properties as repellents of dirt, water and oils. The most well-known PFCs are perfluorooctane sulfonic acid (PFOS), perfluorooctanoic acid (PFOA) and their derivatives belonging to the group of perfluoroalkylated substances. The PFCs are very persistent in the environment, and some of them have been discovered as global pollutants of air, water, soil and wildlife and even found in remote polar areas. Bioaccumulation occurs also in humans, and everybody in our society has traces of these PFCs in their blood and internal organs such as the liver, kidneys, spleen, gall bladder and testes. In the blood, PFOS and PFOA are bound to serum proteins. The acute toxicity of the polyfluorinated substances is moderate but some substances can induce peroxisome proliferation in rat livers and may change the fluidity of cell membranes. Some of these PFCs, such as PFOS and PFOA, are potential developmental toxicants and are suspected endocrine disruptors with effects on sex hormone levels resulting in lower testosterone levels and higher oestradiol level. Other PFCs have oestrogenic effects in cell cultures. The industrial production of PFOS and its derivatives stopped in 2000, and the European Union has banned most uses from the summer of 2008. However, hundreds of related chemicals: homologues with shorter or longer alkyl chain, PFOA and telomers, which potentially may degrade to perfluoroalkanoic (carboxylic) acids, are not regulated.
366 citations
TL;DR: An animal model of exposure to diethylstilbestrol that has proven useful in studying mechanisms involved in abnormal programming of various oestrogen target tissues during differentiation is described, suggesting new targets of abnormal programming by endocrine disrupting chemicals.
Abstract: Environmental chemicals with hormone-like activity can disrupt the programming of endocrine signalling pathways that are established during perinatal life and result in adverse consequences that may not be apparent until much later in life. Increasing evidence implicates developmental exposure to environmental hormone mimics with a growing list of adverse health consequences in both males and females. Most recently, obesity has been proposed to be yet another adverse health effect of exposure to endocrine disrupting chemicals (EDCs) during critical stages of development. Obesity is quickly becoming a significant human health crisis because it is reaching epidemic proportions worldwide, and is associated with chronic illnesses such as diabetes and cardiovascular disease. In this review, we summarize the literature reporting an association of EDCs and the development of obesity, and further describe an animal model of exposure to diethylstilbestrol that has proven useful in studying mechanisms involved in abnormal programming of various oestrogen target tissues during differentiation. Together, these data suggest new targets (i.e. adipocyte differentiation and mechanisms involved in weight homeostasis) of abnormal programming by EDCs, and provide evidence that support the scientific term 'the developmental origins of adult disease'. The emerging idea of an association of EDCs and obesity expands the focus on obesity from intervention and treatment to include prevention and avoidance of these chemical modifiers.
307 citations
TL;DR: The concept of a cumulative TDI value may be more appropriate for the consideration of the overall exposure and the potential human health risks resulting from everyday and simultaneous exposure to several phthalates.
Abstract: Summary In human metabolism studies we found that after oral application of di(2-ethylhexyl) phthalate (DEHP), diisononyl phthalate (DiNP) and di(2-propylheptyl) phthalate (DPHP), at least 74, 44 and 34%, respectively, are excreted via urine. In contrast to the short chain phthalates, their oxidized products, not the simple monoesters, were found to be the main metabolites. Based on urinary phthalate metabolite concentrations we estimated in 102 German subjects between 6 and 80 years of age median daily intakes (μg/kg/day) of 2.7 for DEHP, 2.1 for di-n-butyl phthalate, 1.5 for diisobutyl phthalate, 0.6 for DiNP, and 0.3 for butylbenzyl phthalate. In general, children have higher exposures compared to adults and seem to have a more effective oxidative metabolism of phthalates. For individual phthalates tolerable daily intake (TDI) values have been deduced. However, in rats some phthalates have been shown to act as endocrine disrupters via a common mechanism of action in a dose-additive manner. Therefore, the concept of a cumulative TDI value may be more appropriate for the consideration of the overall exposure and the potential human health risks resulting from everyday and simultaneous exposure to several phthalates.
299 citations
TL;DR: Hormonal effects are of importance in risk assessment, and in some cases where effects are seen at low levels safety margins may be insufficient, and ways to reduce levels in major exposure routes considered.
Abstract: Brominated flame retardants (BFR) are endocrine disrupters in experimental systems, both in vitro and in vivo. Although BFR effects on thyroid hormones are well confirmed, studies of effects on oestrogen/androgen systems are fewer but today growing in numbers. The effects of BFR on other hormone systems are still unknown. Hormonal effect levels in animals start from ca 1 mg/kg b.w., but there are exceptions: effects on spermatogenesis, suggesting hormonal causes, have been observed at a low dose (60 μg/kg b.w.) of a polybrominated diphenyl ether (PBDE) congener, BDE-99. It could be concluded that hormonal effects are of importance in risk assessment, and in some cases where effects are seen at low levels safety margins may be insufficient. One additional uncertainty is the lack of reliable human data that could be used to support animal BFR observations. In spite of the recent regulation of PBDE production, levels of both PBDE and of other BFR groups are still present in environmental samples. Thus, we have to deal with the possible effects of human BFR exposure for times to come. In order to reduce BFR exposure, the routes of exposure should be carefully examined and ways to reduce levels in major exposure routes considered.
288 citations
TL;DR: Urinary concentrations of BP and metabolites of DEP and DBP after topical application are investigated and all 26 subjects showed increased excretion of MEP, MBP and BP following topical application.
Abstract: Diethyl phthalate (DEP), dibutyl phthalate (DBP) and butyl paraben (BP) are man-made chemicals used in personal care products, such as lotions and creams. Exposure to these chemicals causes a variety of adverse reproductive outcomes in animal studies. Humans can be exposed to these chemicals through dermal absorption, but there are no published data on absorption, metabolism, and excretion after dermal application. This study investigates urinary concentrations of BP and metabolites of DEP and DBP after topical application. In a 2-week single-blinded study, 26 healthy Caucasian male subjects were given a whole body topical application of basic cream 2 mg/cm(2) (control week) and then a cream containing 2% (w/w) of DEP, DBP and BP each (treatment week) daily. Twenty-four-hour urine samples were collected. Urinary total, and unconjugated BP, monoethyl phthalate (MEP) and monobutyl phthalate (MBP) metabolites were analysed by Liquid Chromatography-Tandem Mass Spectroscopy (LC-MS/MS). All 26 subjects showed increased excretion of MEP, MBP and BP following topical application. Total MEP, MBP and BP (mean +/- SEM) excreted in urine in the treatment week were, respectively, 41 +/- 1.9, 11.8 +/- 0.6 and 2.6 +/- 0.1 mg/24 h. On average 5.79, 1.82 and 0.32%, respectively, of the applied DEP, DBP and BP could be recovered in urine as MEP, MBP and BP. The concentration of the compounds peaked in urine 8-12 h after application. The fractions of unconjugated MEP, MBP, and BP were 78, 8.0 and 2.1%, respectively. Absorption of DEP, DBP and BP through skin could potentially contribute to adverse health effects. The three chemicals are systemically absorbed, metabolized and excreted in urine following application on the skin in a cream preparation. More DEP than DBP was absorbed, presumably because of a faster absorption rate for DEP.
251 citations
TL;DR: Assessing the health risks of thyroid disruption by environmental xenobiotics will need to include an improved understanding of how divergent mechanisms alter THs and consequent adverse impacts on nervous system development.
Abstract: Environmental contaminants are known to act as thyroid disrupting chemicals (TDCs). Broadly defined, TDCs are xenobiotics that alter the structure or function of the thyroid gland, alter regulatory enzymes associated with thyroid hormone (TH) homeostasis or change circulating or tissue concentrations of THs. For THs, homeostasis is defined as the normal range of THs and TSH in circulation and tissues. TDCs include a wide range chemical structures that act through a variety of mechanisms. Concern about TDCs has increased because of the critical role that thyroid hormones play in brain development. A major uncertainty regarding the endocrine disrupting potential of environmental xenobiotics is the potential for additive, antagonistic or synergistic effects following exposure to mixtures. In addition, there are a number of uncertainties in both interpretation and extrapolation of results from studies of TDC mixtures. Extrapolation of data from laboratory animals to humans is tempered by uncertainty in how the mechanism(s)-of-action of the TDCs may differ between species. The variety of mechanisms by which TDCs alter thyroid homeostasis also yields a difficulty in determining at what level of biological organization to cumulate effects. Should it be at the molecular level, which could be chemical class specific or at the level of a downstream consequence (e.g. circulating hormone levels, brain biochemistry and behaviour) which would be mechanism-independent? To date, the limited data from TDC mixture studies suggest that dose addition is reasonably accurate in predicting the effects on serum T4 concentrations. Assessing the health risks of thyroid disruption by environmental xenobiotics will need to include an improved understanding of how divergent mechanisms alter THs and consequent adverse impacts on nervous system development.
208 citations
TL;DR: It is concluded that a lack of knowledge about relevant exposure scenarios presents serious obstacles for better human risk assessment and epidemiology needs to abandon its focus on single endocrine disrupters and has to embrace the reality of endocrine Disrupter mixture effects by developing biomarkers that capture cumulative exposure to endocrinedisrupters.
Abstract: During the last years, a series of studies on combinations of oestrogenic, thyroid-disrupting and anti-androgenic chemicals at low doses have been published. The available experimental evidence shows that combination effects may result from endocrine disrupters that each produces very small effects, if they are present in sufficiently large numbers. This review examines the implications of these findings for chemicals risk assessment and epidemiology. It is concluded that a lack of knowledge about relevant exposure scenarios presents serious obstacles for better human risk assessment. Epidemiology needs to abandon its focus on single endocrine disrupters and has to embrace the reality of endocrine disrupter mixture effects by developing biomarkers that capture cumulative exposure to endocrine disrupters.
196 citations
TL;DR: It is demonstrated that doses well below the current lowest observed adverse effect level considered by the US-EPA, disrupt pancreatic beta-cell function producing insulin resistance in male mice, and altered blood glucose homeostasis by BPA exposure may enhance the risk of developing type II diabetes.
Abstract: The link between endocrine disruptors and altered blood glucose homeostasis has been recently suggested. Epidemiological studies have correlated levels of phthalates, dioxins and persistent organic pollutants with alterations of blood glucose homeostasis in humans. Environmentally relevant doses of the ubiquitous endocrine disruptor bisphenol-A (BPA) have profound effects on mice endocrine pancreas--an essential tissue involved in glucose metabolism. BPA exerts rapid non-genomic effects on insulin releasing beta-cells and glucagon releasing alpha-cells within freshly isolated islets of Langerhans. In vivo, a single BPA injection of 10 microg/kg rapidly increases plasma insulin and concomitantly decreases glycaemia. When mice were treated with BPA 100 microg/kg/day for 4 days, the environmental oestrogen produced an increase in beta-cell insulin content along with a post-prandial hyperinsulinaemia and insulin resistance. The results reviewed here demonstrate that doses well below the current lowest observed adverse effect level considered by the US-EPA, disrupt pancreatic beta-cell function producing insulin resistance in male mice. Therefore, this altered blood glucose homeostasis by BPA exposure may enhance the risk of developing type II diabetes.
191 citations
TL;DR: Results indicate that chemicals that disrupt foetal tissues during sexual differentiation act in a cumulative, dose-additive manner irrespective of the specific cellular mechanism of toxicity.
Abstract: To date, regulatory agencies have not considered conducting cumulative risk assessments for mixtures of chemicals with diverse mechanisms of toxicity because it is assumed that the chemicals will act independently and the individual chemical doses are not additive. However, this assumption is not supported by new research addressing the joint effects of chemicals that disrupt reproductive tract development in the male rat by disrupting the androgen signalling pathway via diverse mechanisms of toxicity [i.e. androgen receptor (AR) antagonism in the reproductive tract vs. inhibition of androgen synthesis in the foetal testis]. In this study, pregnant rats were exposed to four dilutions of a mixture containing vinclozolin, procymidone, linuron, prochloraz, benzyl butyl phthalate, dibutyl phthalate and diethylhexyl phthalate during the period of sexual differentiation and male offspring were assessed for effects on hormone sensitive endpoints including: anogenital distance, infant areolae retention and reproductive tract tissue weights and malformations. The ratio of the chemicals in the mixture was based upon each chemical's ED(50) for inducing reproductive tract malformations (hypospadias or epididymal agenesis). The observed responses from the mixture were compared with predicted responses generated with a toxic equivalency approach and models of dose addition, response addition or integrated addition. As hypothesized, we found that the mixture of chemicals that alter the androgen signalling pathway via diverse mechanisms disrupted male rat reproductive tract differentiation and induced malformations in a cumulative, dose-additive manner. The toxic equivalency and dose addition models provided the best fit to observed responses even though the chemicals do not act via a common cellular mechanism of action. The current regulatory framework for conducting cumulative risk assessments needs to consider the results, including those presented herein, which indicate that chemicals that disrupt foetal tissues during sexual differentiation act in a cumulative, dose-additive manner irrespective of the specific cellular mechanism of toxicity.
172 citations
TL;DR: Results indicate that doses of anti-androgens, which appear to induce no hypospadias when judged on their own, may induce a very high frequency of hypos padias when they interact in concert with other anti- androgenic chemicals.
Abstract: The incidence of hypospadias is increasing in young boys, but it remains unclear whether human exposure to endocrine disrupting chemicals plays a role. Risk assessment is based on estimation of no-observed-adverse-effect levels for single compounds, although humans are exposed to combinations of several anti-androgenic chemicals. In a mixture (MIX) study with three androgen receptor antagonists, vinclozolin, flutamide and procymidone, rats were gavaged during gestation and lactation with several doses of a MIX of the three chemicals or the chemicals alone. External malformations of the male reproductive organs were assessed on PND 47 using a score from 0 to 3 (normal to marked) for hypospadias. Markedly increased frequencies were observed after exposure to a MIX of the three chemicals compared to administration of the three chemicals alone. Anogenital distance at PND 1, nipple retention at PND 13, and dysgenesis score at PND 16 were highly correlated with the occurrence of hypospadias, and MIX effects were seen at doses where each of the individual chemicals caused no observable effects. Therefore, the results indicate that doses of anti-androgens, which appear to induce no hypospadias when judged on their own, may induce a very high frequency of hypospadias when they interact in concert with other anti-androgens.
151 citations
TL;DR: The sperm counts of the majority of 20 year old European men are now so low that the authors may be close to the crucial tipping point of 40 mill/mL spermatozoa, which would mean the possibility of more infertile couples and lower fertility rates in the future.
Abstract: Healthy men produce an enormous number of sperms, far more than necessary for conception. However, several studies suggest that semen samples where the concentration of sperms is below 40 mill/mL may be associated with longer time to pregnancy or even subfertility, and specimens where the concentration of sperms is below 15 mill/mL may carry a high risk of infertility. Historic data from the 1940s show that the bulk of young men at that time had sperm counts far above 40 mill/mL with averages higher than 100 mill/mL. However, recent surveillance studies of young men from the general populations of young men in Northern Europe show that semen quality is much poorer. In Denmark approximately 40 percent of the men have now sperm counts below 40 mill/mL. A simulation assuming that average sperm count had declined from 100 mill/mL in 'old times' to a current level close to 40 mill/mL indicated that the first decline in average sperm number of 20-40 mill/mL might not have had much effect on pregnancy rates, as the majority of men would still have had counts far above the threshold value. However, due to the assumed decline in semen quality, the sperm counts of the majority of 20 year old European men are now so low that we may be close to the crucial tipping point of 40 mill/mL spermatozoa. Consequently, we must face the possibility of more infertile couples and lower fertility rates in the future.
TL;DR: This study provides a quantitative estimate of the prevalence and main risk factors for infertility in Iranian couples and indicates that the prevalence of infertility falls within a relatively wide range being high in the Southern counties, and low in the Northern counties.
Abstract: To explore the prevalence and risk factors of infertility in Iran, a total of 12 285 ever-married women aged 15-50 years old and their husbands (if available) were interviewed by 82 female general practitioners and answered a self-administered questionnaire on several aspects of infertility. They were identified from the national population in 30 counties, and invited to a confidential interview. Data were obtained about their age, education, marital status, toxic habits, medical history, disabilities and illnesses, help-seeking, economy, ethnicity, geographic location, contraceptive use and age at which they had first intercourse. This study used the definition of childlessness proposed by World Health Organization: 'the woman has never conceived despite cohabitation and exposure to pregnancy for a period of 2 years'. The overall prevalence of infertility was 8% (95% CI: 3.2-15.0). The weighted national estimate of primary infertility was 4.6% (95% CI: 3.6-5.2). There was a pronounced regional pattern in the levels of primary infertility. The primary infertility increased significantly from 2.6 to 4.3 to 5.5% for the 1985-1989, 1990-1994 and 1995-2000 marriage cohorts. The prevalence of secondary infertility was 3.4% (95% CI: 2.4-5.1). Overall the prevalence of infertility falls within a relatively wide range being high in the Southern counties, and low in the Northern counties. The probability of first pregnancy at the end of 2 years of marriage was 0.78 for all ever-married women. The prevalence of infertility increased with age (linear chi-square 198.012, 1 d.f., p = 0.01). The age pattern of infertility also varies quite markedly across the counties analysed. No effect of race was detected; neither the intercept (analysis of covariance p = 0.36) nor the slope of the age relationship was influenced by race (analysis of covariance p = 0.41). Infertility were observed as significantly higher in the presence of history of tubo-ovarian surgery [odds ratio (OR): 1.43; 95% CI: 1.28-2.23; p = 0.01], salpingitis (OR: 2.34; 95% CI: 1.31-4.3; p = 0.016), ectopic pregnancy (OR: 2.45; 95% CI: 1.90-3.44; p = 0.04), varicocele (OR: 2.85; 95% CI: 1.61-5.20; p = 0.01) and cryptorchidism (OR: 3.81; 95% CI: 2.51-4.28; p = 0.031). This study provides a quantitative estimate of the prevalence and main risk factors for infertility in Iranian couples. Yet, further studies on the cause of primary and secondary infertility and geographical variations in the incidence and prevalence of infertility in Iran are needed.
TL;DR: Possible depression in males is related to decreased sperm concentration, and poor coping with stress is associated with increased occurrence of early miscarriage.
Abstract: In this study we sought to evaluate whether psychological factors in males affect semen quality and pregnancy. In 1076 men of infertile couples, psychological factors, i.e. exposure to acute stress, coping with stress, the WHO (five) Well-Being Index and the Zung's Anxiety Scale Inventory scores were assessed by a questionnaire at the time of semen analysis. Relationships between psychological factors and semen quality (sperm concentration, rapid and progressive motility and normal morphology) were assessed. In 353 men with infertility duration of or =5 x 10(6) sperm/mL and a female partner with a laparoscopically confirmed tubal patency, we looked prospectively for relations between psychological factors and the occurrence of a natural pregnancy at a 6-month follow-up (n = 124), and first-trimester loss (n = 18). Anxiety trait, found in 19% of men, was related to previous in vitro fertilization/intracytoplasmic sperm injection attempts (p = 0.014), cigarette intake (p = 0.006), alcohol intake (p = 0.026) and sexual difficulties (p < 0.001). Regression analyses indicated a significant positive relationship between the level of sperm concentration and the WHO (five) Well-Being Index score, each successive score number accounting for a 7.3% increase in sperm concentration (p = 0.039), whereas no correlation was found between psychological factors and sperm rapid progressive motility and normal morphology. Poorer coping with stress was related to the occurrence of a first-trimester miscarriage (p = 0.016) in the female partner. Possible depression in males is related to decreased sperm concentration, and poor coping with stress is associated with increased occurrence of early miscarriage.
TL;DR: Improved methods for the pre-fractionation of proteins, improved high speed, high resolution, high sensitivity mass spectrometry hardware and superior informatics/bioformatics software is heralding a new era in the authors' capacity to analyse the proteomic composition of human spermatozoa.
Abstract: The advent of enhanced methods for the pre-fractionation of proteins, improved high speed, high resolution, high sensitivity mass spectrometry hardware and superior informatics/bioformatics software is heralding a new era in our capacity to analyse the proteomic composition of human spermatozoa. Parallel improvements in our capacity to compare proteomic profiles from spermatozoa in different functional states (immature vs. mature, uncapacitated vs. capacitated, normal vs. defective) is also helping to define which specific elements of the proteome are of functional significance. The ultimate aim of such studies will be to integrate the proteome with the sperm metabolome so that we shall not only understand the cascade of post-translational modifications (e.g., phosphorylation, glycosylation, proteolytic cleavage) involved in generating a functional spermatozoon but also determine how these physiological changes are brought about. This fundamental information will then create a basis for identifying key points in the post-testicular maturation of spermatozoa that might be targeted for contraceptive purposes or implicated in the defective sperm function observed in a significant proportion of infertile males.
TL;DR: Overall the results show that many of the commonly used azole fungicides act as endocrine disruptors in vivo, although the profile of action in vivo varies.
Abstract: The endocrine-disrupting potential of four commonly used azole fungicides, propiconazole, tebuconazole, epoxiconazole and ketoconazole, were tested in two short-term in vivo studies. Initially, the antiandrogenic effects of propiconazole and tebuconazole (50, 100 and 150 mg/kg body weight/day each) were examined in the Hershberger assay. In the second study, pregnant Wistar rats were dosed with propiconazole, tebuconazole, epoxiconazole or ketoconazole (50 mg/kg/day each) from gestational day (GD) 7 to GD 21. Caesarian sections were performed on dams at GD 21. Tebuconazole and propiconazole demonstrated no antiandrogenic effects at doses between 50 and 150 mg/kg body weight/day in the Hershberger assay. In the in utero exposure toxicity study, ketoconazole, a pharmaceutical to treat human fungal infections, decreased anogenital distance and reduced testicular testosterone levels, demonstrating a demasculinizing effect on male fetuses. Tebuconazole, epoxiconazole and ketoconazole induced a high-frequency of post-implantation loss, and both ketoconazole and epoxiconazole caused a marked increase in late and very late resorptions. Overall the results show that many of the commonly used azole fungicides act as endocrine disruptors in vivo, although the profile of action in vivo varies. As ketoconazole is known to implicate numerous endocrine-disrupting effects in humans, the concern for the effects of the other tested azole fungicides in humans is growing.
TL;DR: The data indicate that the potential risk posed by endocrine active UV filters warrants further investigations, and expression and oestrogen sensitivity of oestrogens-regulated genes and nuclear receptor coregulator levels were altered at mRNA and protein levels in adult uterus, prostate and brain regions involved in gonadal control and sexual behaviour.
Abstract: Several ultraviolet (UV) filters exhibit estrogenic, some also anti-androgenic activity. They are present in waste water treatment plants, surface waters and biosphere including human milk, suggesting potential exposure during development. Developmental toxicity was studied in rats for the UV filters 4-methylbenzylidene camphor (4-MBC, 0.7, 7, 24, 47 mg/kg/day) and 3-benzylidene camphor (3-BC, 0.07, 0.24, 0.7, 2.4, 7 mg/kg/day) administered in chow to the parent generation before mating, during pregnancy and lactation, and to the offspring until adulthood. Neonates exhibited enhanced prostate growth after 4-MBC and altered uterine gene expression after both chemicals. 4-MBC and 3-BC delayed male puberty and affected reproductive organ weights of adult offspring. Effects on the thyroid axis were also noted. Expression and oestrogen sensitivity of oestrogen-regulated genes and nuclear receptor coregulator levels were altered at mRNA and protein levels in adult uterus, prostate and brain regions involved in gonadal control and sexual behaviour. Female sexual behaviour was impaired by both filters; 3-benzylidene camphor caused irregular cycles. Classical endpoints exhibited lowest observed adverse effect levels (LOAELs) and no observed adverse effect levels (NOAELs) of 7/0.7 mg/kg for 4-MBC and 0.24/0.07 mg/kg for 3-BC. Molecular endpoints were affected by the lowest doses studied. Our data indicate that the potential risk posed by endocrine active UV filters warrants further investigations.
TL;DR: It is suggested that sperm DNA damage as measured by the TUNEL assay may provide an indicator for patients with poor fertilization rates and/or those unable to achieve pregnancy following ART treatment.
Abstract: The literature suggests an association between sperm DNA damage and assisted reproductive technology (ART) outcomes. However, previous studies involved the transfer of multiple embryos, which has complicated the interpretation of the results. The aim of this study was to determine the relationship between the levels of sperm DNA damage and fertilization rate, embryo development as well as pregnancy outcome, following single embryo transfer. Patients (n = 113) undergoing in vitro fertilization (IVF) (n = 45) and intra-cytoplasmic sperm injection (ICSI) (n = 68) were assessed for their levels of sperm DNA damage in the sample used for insemination. DNA damage was determined using terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-nick end labelling (TUNEL). The relationship between DNA damage and outcomes were assessed using regression analysis. Overall data showed no association between sperm DNA damage and fertilization rate, or embryo development in vitro. However, when IVF was the insemination method, there was a significant negative correlation between fertilization rates and sperm DNA damage (p < 0.05). When ICSI was the insemination technique, low sperm DNA damage was associated with successful pregnancy (37.8 +/- 5.7% DNA damaged sperm) compared with failed implantation (52.9 +/- 3.9% DNA damaged sperm, p < 0.05). Our results suggest that sperm DNA damage as measured by the TUNEL assay may provide an indicator for patients with poor fertilization rates and/or those unable to achieve pregnancy following ART treatment.
TL;DR: As more and more molecular studies with anti-androgenic compounds are conducted, the number of mechanisms by which compounds can affect the androgen signalling pathway is likely to increase.
Abstract: Scientists have identified environmental chemicals that display anti-androgenic activity via multiple mechanisms of action. Early studies focused on pesticides acting as androgen receptor (AR) antagonists but it soon became apparent that was not the only endocrine mode by which compounds affected the androgen signalling pathway. Classes of chemicals currently known to interfere with the androgen signalling pathway include dicarboximide fungicides (e.g. vinclozolin), organochlorine-based insecticides (e.g. p,p'-DDT and -DDE), conazole fungicides (e.g. prochloraz), plasticizers (phthalates) and urea-based herbicides (linuron). Phthalate esters (PEs) and vinclozolin appear to act primarily via a single mechanism of action, while others such as linuron and prochloraz, appear to display dual mechanisms of action. Exposure to PEs decreases mRNA expression of key steroidogenic enzymes and also the peptide hormone insulin-like peptide 3 (insl3) from the foetal Leydig cells. Hence, both androgen- and inls3-dependent tissues are affected. Vinclozolin and procymidone act solely through binding to the AR as antagonists thus blocking the action of androgen at the cellular level but do not affect foetal testosterone synthesis or insl3 gene expression. The compounds linuron and prochloraz are AR antagonists but also inhibit foetal testosterone synthesis, although unlike the PEs, mRNA expression of steroidogenic enzymes and insl3 are not affected. All the above chemicals disrupt androgen signalling in the foetal male rat and produce some malformations in common, but the precise profiles of effects in the offspring are pathognomonic for each mode of action. For example, the 'phthalate syndrome' vs. the 'vinclozolin syndrome' each displays a profile of effects which is clearly different. In summary, as more and more molecular studies with anti-androgenic compounds are conducted, the number of mechanisms by which compounds can affect the androgen signalling pathway is likely to increase. Furthermore, the effects of mixtures of these compounds are just beginning to be explored.
TL;DR: It is hypothesized that MEHP is the bioactive toxicant and further metabolism to MEHHP/MEOHP may lower internal burden of MEHP and thus be protective from sperm DNA damage, and requires further exploration to confirm.
Abstract: Phthalates are a class of chemicals with widespread general population exposure. Some phthalates are reproductive and developmental toxicants in laboratory animals. Advances in the field of phthalate research in humans are dependent on the development and implementation of biomarkers to assess exposure and outcome, as well as potential markers that may be indicative of increased susceptibility. Recently, we incorporated a novel biomarker of potential 'susceptibility' into our study on the relationship of phthalates with semen quality and sperm DNA damage among men recruited from an infertility clinic. We measured urinary concentrations of three di(2-ethylhexyl) phthalate (DEHP) metabolites, mono(2-ethylhexyl) phthalate (MEHP) and two oxidative metabolites, mono-(2-ethyl-5-hydroxylhexyl) phthalate (MEHHP) and mono-(2-ethyl-5-oxohexyl) phthalate (MEOHP). We calculated the percent of DEHP excreted as the hydrolytic monoester (i.e., MEHP). We referred to this as %MEHP and considered it a phenotypic marker of the proportion of DEHP excreted in the urine as MEHP. In our sperm DNA study, we found novel results for the DEHP metabolites. Although MEHP was positively correlated with the oxidative metabolites, the association of sperm DNA damage with MEHP, as compared to MEHHP and MEOHP, were in opposite directions. We hypothesized that MEHP is the bioactive toxicant and further metabolism to MEHHP/MEOHP may lower internal burden of MEHP and thus be protective from sperm DNA damage. An alternative explanation may include that the relative percentage of DEHP excreted as MEHP was a surrogate for the function of phase I enzymes. Men with high %MEHP may have higher levels of sperm DNA damage because of poor metabolism (detoxification) of other genotoxic chemicals. Our hypothesis that %MEHP may represent a phenotypic marker of metabolism is novel but requires further exploration to confirm.
TL;DR: A large fraction of young German men raised in the former East Germany and West Germany seem to have impaired semen quality that may reduce their natural fertility, and to what extent poor semen quality contributes to the low German fertility rates is investigated.
Abstract: Population studies have shown that a high proportion of Nordic men may have so poor semen quality that they can be classified as sub-fertile according to international standards. A question is whether the Nordic data are specific for the Nordic countries or they should be seen as an expression of a general trend in Europe. We therefore carried out a prospective study of semen quality of young men raised in the former East Germany (Leipzig) and West Germany (Hamburg). To enable inter-regional comparisons, we utilized a common European research protocol previously used in studies in the Nordic-Baltic region. Three hundred and thirty-four young men representative of the general population from Hamburg, and 457 from Leipzig delivered semen samples, underwent physical examinations and provided information on life-style and reproductive health parameters. The study period in Hamburg was February 2003--July 2004, and in Leipzig July 2003--April 2005. No significant differences were observed in sperm concentration (median 46, 42, and 44 million/mL for men from Hamburg, Leipzig and the combined Hamburg-Leipzig group respectively) or total sperm count (154,141 and 149 million), whereas the differences for morphologically normal spermatozoa (9.4 and 8.4%) and motile spermatozoa (67 and 81%) were significantly different. Previously published studies have shown reduced fertility with decreasing sperm concentrations below 40-55 millions/mL and normal sperm morphology below 9-19%. Thus, a large fraction of young German men seem to have impaired semen quality that may reduce their natural fertility. However, it remains to be investigated to what extent poor semen quality contributes to the low German fertility rates.
TL;DR: This review of studies evaluating potential causes of abnormal protamine expression highlights studies evaluating the role of retained histones and DNA methylation in sperm, which may be affected in sperm with aberrant protamine replacement.
Abstract: During the elongating spermatid stage of spermatogenesis, there is a step-wise replacement of nuclear histones with protamines 1 and 2. In fertile men, the ratio of protamine 1/protamine 2 (P1/P2) is within the narrow range of 0.8-1.2. Ratios above or below that range are associated with infertility, exhibiting a wide range of defects including decreased sperm counts, morphology, fertilization ability, and embryo implantation capacity. In this review, we highlight studies evaluating potential causes of abnormal protamine expression, including the sequencing of genes relevant to protamine expression in both affected patients and controls. While the variants of the protamine genes themselves do not appear to be responsible for most observed defects, variants of the Contrin gene, a transcription factor and translation repressor, appear to be contributory to some cases of abnormal expression. Additionally, we explore the potential effects of abnormal protamine replacement on the epigenome of human sperm. Ongoing studies are evaluating the role of retained histones and DNA methylation in sperm, which may be affected in sperm with aberrant protamine replacement. This important area of epigenetic research has profound clinical implications.
TL;DR: It was concluded that the acceleration of rat sperm transit time appeared to harm normal sperm maturation, thus decreasing sperm quality and fertility capacity, in an androgen-dependent way.
Abstract: Summary
The epididymal sperm transit time seems to have an important role in the process of sperm maturation, and it seems that alterations to the transit can harm the process. The aim of the present work was to evaluate the influence of altered sperm transit time through the epididymis on sperm parameters and fertility of rats, as well as the role of testosterone in the alterations. Sprague–Dawley adult male rats were randomly assigned to four different groups and were treated for 12 days: (i) 10 μg/rat/day DES, to accelerate the transit; (ii) 6.25 mg/kg/day guanethidine sulphate, to delay the transit; (iii) same treatment as group 1, plus androgen supplementation; (iv) control animals received the vehicles. Guanethidine treatment delayed the sperm transit time through the epididymal cauda, provoking increased sperm reserves in this region. Animals exposed to DES showed an acceleration of sperm transit time in the epididymis, and consequently decreased sperm density in both epididymal regions, the caput-corpus and cauda, and diminished sperm motility. In both cases sperm production was not altered. Testosterone supplementation was able to restore the transit time to values close to normality, as they were higher than in the control rats. The same occurred in relation to sperm motility. Rats exposed to DES presented lower fertility after in utero artificial insemination using sperm collected from the proximal cauda epididymis. Therefore, it was concluded that the acceleration of rat sperm transit time appeared to harm normal sperm maturation, thus decreasing sperm quality and fertility capacity, in an androgen-dependent way.
TL;DR: Basic semen analysis and selected functional tests of sperm provide important information regarding male fertility status and a multiple regression model predicting fertility using selected parameters of semen analysis is developed.
Abstract: Accurate prognosis of male fertility based on semen measurements is still not straightforward. This study was designed to identify the best predictors of fertility and to develop a multiple regression model predicting fertility using selected parameters of semen analysis. The predictive value of standard semen parameters and selected functional tests were studied in 113 fertile men and in 109 subfertile men whose spouses had a normal infertility workup. Individual semen parameters were evaluated using the receiver operating characteristic curve. Logistic regression based on linear functions of analysed sperm parameters was used to predict the chance of spontaneous conception. Logistic regression modelling revealed that the best prediction of spontaneous conception was obtained using 12 semen parameters: sperm concentration, total progressive motility (A + B), motility grade C or D, normal sperm morphology, defects of: head, acrosome, midpiece and tail, spontaneous acrosome reaction, hypo-osmotic swelling (HOS) test and acid aniline blue test. This mathematical model reached 90.3% accuracy in predicting in vivo conception and 90.8% for its lack. A satisfactory prediction of male fertility was also obtained using only four semen measurements: sperm concentration, total progressive motility (grade A + B), normal morphology, and HOS test; this model correctly identified as fertile 84.1% of those who conceived and identified as subfertile 88.1% of those who did not achieve pregnancy. In conclusion, basic semen analysis and selected functional tests of sperm provide important information regarding male fertility status.
TL;DR: It is concluded that, although of interest clinically, the Percoll filtration technique in its present form is probably of limited value for cell biological and/or ultrastructural studies.
Abstract: Centrifugation of human spermatozoa on BWW- or HEPES-Krebs-Ringer-buffered media containing Percoll as a method to prepare pure gamete populations or subpopulations has been studied in detail. Application of this technique (Gorus & Pipeleers 1981) allowed us to obtain uncontaminated, motile spermatozoa and, after a subsequent Percoll gradient step, resulted in the enrichment of motile cells able to readily penetrate in vitro into AB-serum filled capillaries. The gradient sedimentation patterns of 25 different sperm samples with normal seminal parameters were found to be strongly influenced by the characteristics of the sample itself and also by the buffer used for filtration. However, the present study also revealed that ultrastructural changes and the release of intraspermatozoal enzymes were induced by the procedure. It is concluded that, although of interest clinically, the Percoll filtration technique in its present form is probably of limited value for cell biological and/or ultrastructural studies.
TL;DR: The exclusion of sperm presenting with those apoptosis-related features during assisted reproduction may improve success rates of procedures such as intrauterine insemination and in vitro fertilization.
Abstract: Human sperm have been documented to display apoptosis-like features such as externalization of phosphatidylserine (EPS), disruption of the transmembrane mitochondrial potential (MMP) and activation of caspases. Our aim was to evaluate possible association between activation of the apoptosis cascade in human sperm and its oocyte penetration capacity using the zona free hamster oocyte penetration assay (SPA). Semen specimens from 76 unselected donors were subjected to double density gradient centrifugation followed by incubation under capacitating conditions for 3 h and SPA. Apoptosis signalling was monitored by assessment of EPS, disruption of MMP and activation of caspase-3 by flow cytometry. Semen samples with subnormal SPA values ( 20% penetrated oocytes, p < 0.01). All three apoptosis markers showed a significantly negative correlation with the percentage of penetrated oocytes (p < 0.01). Apoptosis-related signalling appears to have a negative association with sperm-oocyte penetration. The exclusion of sperm presenting with those apoptosis-related features during assisted reproduction may improve success rates of procedures such as intrauterine insemination and in vitro fertilization.
TL;DR: It is found that younger cohorts had progressively lower TNCR and that in terms of their total fertility rate, the declining T NCR is compensated by an increasing use of ART, which may imply increasing reproductive health problems and lower fertility in the future.
Abstract: Recent findings of poor semen quality among at least 20% of normal young men in Denmark prompted us to use unique Danish registers on births and induced abortions to evaluate a possible effect of the poor male fecundity on pregnancy rates among their presumed partners--the younger cohorts of women. We have analysed data from the Danish birth and abortion registries as well as the Danish registry for assisted reproduction (ART) and defined a total natural conception rate (TNCR), which is equal to fertility rate plus induced abortion rate minus ART conception rate. A unique personal identification number allowed the linkage of these databases. Our database included 706,270 native Danish women born between 1960 and 1980. We used projections to estimate the fertility of the later cohorts of women who had not yet finished their reproduction. We found that younger cohorts had progressively lower TNCR and that in terms of their total fertility rate, the declining TNCR is compensated by an increasing use of ART. Our hypothesis of an ongoing birth cohort-related decline in fecundity was also supported by our finding of increasing and substantial use of ART in the management of infertility of relatively young couples in the later cohorts. Furthermore, the lower rates of induced abortion among the younger birth cohorts, often viewed as a success of health education programs, may not be fully explained by improved use of contraception. It seems more likely that decreased fecundity because of widespread poor semen quality among younger cohorts of otherwise normal men may explain some of the observed decline in conception rates. This may imply increasing reproductive health problems and lower fertility in the future, which is difficult to reverse in the short term. The current and projected widespread use of ART in Denmark may be a sign of such an emerging public health problem.
[...]
TL;DR: The azoospermia factor (AZF) locus in Yq11 is now functionally subdivided in three distinct spermatogenesis loci: AZFa, AZFb and AZFc, and a specific chromatin folding code in YQ11 is proposed, which controls the condensation cycle of the Y chromosome in the male germ line.
Abstract: The azoospermia factor (AZF) locus in Yq11 is now functionally subdivided in three distinct spermatogenesis loci: AZFa, AZFb and AZFc. After knowledge of the complete genomic Y sequence in Yq11, 14 Y genes encoding putatively functional proteins and expressed in human testis are found to be located in one of the three AZF intervals. Therefore, a major question for each infertility clinic performing molecular screening for AZF deletions has now raised concerning the functional contribution of the encoded AZF proteins to human spermatogenesis. Additionally, it has been shown that distinct chromatin regions in Yq11 overlapping with the genomic AZFb and AZFc intervals are probably involved in the pre-meiotic X and Y chromosome pairing process. An old hypothesis on the germ line function of AZF becomes therefore revitalized. It proposed a specific chromatin folding code in Yq11, which controls the condensation cycle of the Y chromosome in the male germ line. Thus, with the exception of AZF proteins functionally expressed during the pre-meiotic differentiation and proliferation of spermatogonia, the need for AZF proteins functionally expressed at meiosis or during the post-meiotic spermatid maturation process is difficult to assess before the identification of specific mutations in the corresponding AZF gene causing male infertility.
TL;DR: It is plausible that the dysgenesis, and hence the development of CIS cells, is a result of disturbed signalling between nurse cells and germ cells that allow embryonic germ cells to survive in the pre-pubertal and adult testis, which eventually results in theDevelopment of invasive testicular cancer in the adult.
Abstract: Recent increases in male reproductive disorders have been linked to exposure to environmental factors leading to the testicular dysgenesis syndrome (TDS). Testicular cancer is the most severe condition in TDS and studies have shown a clear correlation between risk of testicular cancer and other components of TDS and that the geographical location of the mother during pregnancy can be a risk factor. This suggests that the dysgenesis has its origin in utero and that TDS is initiated by environmental factors, including possibly hormone-disrupting compounds that act on the mother and the developing foetus, but the genetic background may also play a role. The morphological similarity of carcinoma in situ (CIS) cells (the precursor of the majority of invasive testicular cancers) with primordial germ cells and gonocytes, and overlap in expression of protein markers suggests an origin of CIS from primordial germ cells or gonocytes. CIS cells and germ cell-derived cancers of the human type have so far not been described in any animal model of TDS, which could be caused by species differences in the development of the male gonad. Regardless of this, it is plausible that the dysgenesis, and hence the development of CIS cells, is a result of disturbed signalling between nurse cells and germ cells that allow embryonic germ cells to survive in the pre-pubertal and adult testis. The post-pubertal proliferation of CIS cells combined with aberrant signalling then leads to an accumulation of genetic changes in the CIS cells, which eventually results in the development of invasive testicular cancer in the adult.
TL;DR: Foetal BPA exposure was sufficient to induce the development of oestrogen-sensitive pre-neoplastic and neoplastic lesions in the mammary gland in the absence of any additional treatment aimed at increasing tumour incidence.
Abstract: The hypothesis that prenatal exposure to endocrine disruptors might cause cancer arose from challenging two well-accepted notions: (i) mammalian development is merely the unfolding of a genetic programme and (ii) only mutagenic agents can cause cancer. This hypothesis required challenging genetic determinism. The ecological developmental biology (eco-devo) movement revitalized the concept of developmental plasticity through the occurrence of polyphenisms (a single genotype produces diverse phenotypes which are determined by environmental cues). Based on the principles of eco-devo and the tissue organization field theory of carcinogenesis and neoplasia, we tested the hypothesis that exposure to xenoestrogens during foetal development in rats increased the propensity to develop mammary cancer during adulthood. We chose exposure to bisphenol A (BPA) as a model for environmental oestrogen exposure. This endocrine disruptor induced the development of ductal hyperplasias and carcinoma in situ. These highly proliferative lesions contained an increased number of oestrogen receptor α-positive cells. Thus, foetal BPA exposure was sufficient to induce the development of oestrogen-sensitive pre-neo-plastic and neoplastic lesions in the mammary gland in the absence of any additional treatment aimed at increasing tumour incidence.
TL;DR: The results have shown that oxygen uptake by mitochondria of demembranated sperm cells from normozoospermic subjects is significantly stimulated by a large set of respiratory substrates and ADP, and a significant decrease in the sperm respiratory efficiency was found.
Abstract: In this work we report a relatively simple and fast method for analysing oxygen consumption and therefore mitochondrial functionality, in individual human ejaculates. This oxygraphic method requires a low number of cells, is highly reproducible and linearly correlates with sperm concentration. Our results have shown that oxygen uptake by mitochondria of demembranated sperm cells from normozoospermic subjects is significantly stimulated by a large set of respiratory substrates and ADP. The respiratory control ratio (RCR) values indicate a good coupling between respiration and phosphorylation by sperm mitochondria and thus a well preserved integrity of the mitochondria themselves. Interestingly, whereas the rates of oxygen uptake, as expected, changed with different sperm concentrations, the RCR values remained constant, thus demonstrating a linear response of the assay. In asthenozoospermic subjects, however, a significant decrease in the sperm respiratory efficiency was found. The results obtained suggest that this method, besides its potential clinical application, could be useful for a deeper understanding of the biochemical properties of sperm mitochondria and their role in ATP production in human spermatozoa.