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In this case report, a paraneoplastic syndrome caused by testicular neoplasia in a ten-year-old cryptorchid dog is described. Feminization and pancytopenia were observed, resulting from the testicular neoplastic production of estrogens. A diagnosis of testicular tumor and associated bone marrow suppression was made by ultrasonography and blood examination, with estrogen blood levels being severely elevated. Urinalysis revealed a urinary tract infection. Castration was performed together with a blood transfusion, and antibiotic treatment was started. After an initial improvement, the dog died suddenly after approximately three weeks. In this report, the importance is highlighted of identifying clinical signs associated with feminization in intact male dogs at an early stage, to avoid severe, potentially irreversible, hematological consequences due to bone marrow suppression. Elective orchidectomy of both testes is highly recommended in cryptorchid dogs as neoplastic transformation of the undescended testis may occur, with potentially fatal outcome.

Crustaceans are a morphologically, physiologically, and ecologically highly diverse animal group and correspondingly diverse are their reproductive characteristics. They have evolved structural specialties with respect to penis construction, sperm form, sperm storage, fertilization, and brood care. Unique in the animal kingdom are safety lines that safeguard hatching and first molting. Further curiosities are dwarf males in parasitic and sessile crustaceans and bacteria-induced feminization and gigantism of crustacean hosts. Record-breaking features are relative penis length, sperm size, clutch size, chromosome number, viability of dormant eggs, and fossil ages of penis, sperm, and brooded embryos. These examples from a single invertebrate subphylum and a single life history aspect illustrate that morphological solutions to functional requirements can be as spectacular as behavioral adaptations. They may provide valuable sources for comparative morphologists, ecologists, evolutionary biologists, and applied biologists to advance topical issues such as sperm competition, posthumous paternity, evolution of brood care, adaptation to freshwater, infectious feminization, sustainable male-based fishery, maintenance of genetic diversity under conditions of limited mating opportunity, and long-term impact of pollution on genotype and phenotype. J. Morphol. 277:1399-1422, 2016. © 2016 Wiley Periodicals, Inc.

Wolbachia infections have been described in several Drosophila species, but relatively few have been assessed for phenotypic effects. Cytoplasmic incompatibility (CI) is the most common phenotypic effect that has been detected, while some infections cause male killing or feminization, and many Wolbachia infections have few host effects. Here, we describe two new infections in a recently described species, Drosophila pandora, one of which causes near-complete CI and near-perfect maternal transmission (the "CI" strain). The other infection is a male killer (the "MK" strain), which we confirm by observing reinitiation of male production following tetracycline treatment. No incompatibility was detected in crosses between CI strain males and MK strain females, and rare MK males do not cause CI. Molecular analyses indicate that the CI and MK infections are distantly related and the CI infection is closely related to the wRi infection of Drosophila simulans. Two population surveys indicate that all individuals are infected with Wolbachia, but the MK infection is uncommon. Given patterns of incompatibility among the strains, the infection dynamics is expected to be governed by the relative fitness of the females, suggesting that the CI infection should have a higher fitness. This was evidenced by changes in infection frequencies and sex ratios in population cages initiated at different starting frequencies of the infections.

In order to elucidate the role of Cyp19a in sex differentiation of Schizothorax kozlovi, the full length cDNA of Cyp19a was cloned from the mature ovary of S. kozlovi by using rapid amplification of cDNA ends method, and then its relative mRNA expression levels among tissues and temperature groups were determined by using quantitative real-time PCR. The complete Cyp19a cDNA of 1795 bp of S. kozlovi was obtained, which encoded 517 amino acids and belonged to gonadal aromatase. Its deduced amino acid sequence had the above 70 % identity compared with gonadal aromatase genes of teleost fishes, but only 62–67 % when compared with brain aromatase genes of fishes. It was expressed only in heart and gonad, but no expression in other tissues, presenting relatively high tissue specificity. It also exhibited sex-specific expression pattern in gonads, but no sex differences in heart. Comparing with the Cyp19a expression levels at 12 days post hatching (dph), significant temperature effects were revealed in low temperature group (10 °C) at 18 dph, and in high temperature group (26 °C) at 40 dph. It suggested that gonadal aromatase Cyp19a gene may play important roles on the feminization or masculinization of S. kozlovi affected by temperature during the early developmental stage.

The endo-symbiotic relationship between microorganism and eukaryotes are very common and has been broadly investigated from all insect species. The genus Wolbachia are obligatory intracellular bacteria that induce evolutionary alterations and have been frequently reported in egg cytoplasm’s of various invertebrates including insects, spiders, mites, scorpions, crustaceans and nematodes, which exert a profound impact on host biology and behavior through a number of phenotypic alternations. Due to its ubiquitous phenotypic behavior, Wolbachia becomes a novel and promising natural micro-biocontrol agent to induce cytoplasmic incompatibility, killing of male embryos, parthenogenesis, and feminization. Autonomous transgenic capability, survival on several host species and the ability to modify the host reproductive systems makes it the most prominent and focusing scientific genomic research from last few decades. The aim of this review is to summarize the significance and recent discoveries of Wolbachia related to various arthropods which lead positive directions to be exploited in future for integrated pest management (IPM) strategies. Further, this review also discussed the biology, phylogeny, distributions, classifications and types of Wolbachia infections on insect orders and coleopterans in particular.

Sexual dimorphism in mouse reproductive tissues is observable in adult, post-natal, and embryonic stages. The development of sexually dimorphic tissues starts with an ambisexual structure. It is followed by sex-specific organogenesis as guided by different signaling pathways that occur from late embryonic stages. The measurement of the anogenital distance (AGD), and the observation of the external genitalia are practical ways to distinguish male and female pups at birth and thereafter. Careful observation of the morphological or histological features and the molecular signatures of the external genitalia and perineum enable identification of sex or feminization/masculinization of embryos. Aberrations in hormone signaling via castration or treatment with hormones or hormone disruptors result in dysmorphogenesis of reproductive tissues. Several hormone disruptors have been used to modulate different aspects of hormone action through competitive inhibition and exogenous hormone treatment. Concomitantly, the vast advancement of conditional mutant mouse analysis leads to the frequent utilization of Cre recombination technology in the study of reproductive/urogenital tissue development. Mouse Cre-lines that are tissue-specific and cell-specific are also effective tools in identifying the molecular mechanisms during sexually dimorphic development. Cre-lines applicable to different cell populations in the prostate, seminal vesicles, testis and ovaries, and mammary glands are currently being utilized. In the external genitalia and perineum, Cre lines that examine the signaling pathways of cells of endodermal, ectodermal, and mesenchymal origin reveal the roles of these tissues in the development of the external genitalia. The interaction of hormones and growth factors can be examined further through a variety of techniques available for researchers. Such cumulative information about various technologies is summarized.

Testosterone and its metabolites masculinize the brain during a critical perinatal window, including the relative volume of sexually dimorphic brain areas such as the sexually dimorphic nucleus of the preoptic area (SDN), which is larger in males than females. Serotonin (5HT) may mediate this hormone action, since 5HT given during the second week of life decreases (i.e., feminizes) SDN volume in males and testosterone-treated females. Although previous work indicates that the 5HT2A/2C receptor is sufficient to induce feminization, it is unclear whether other serotonin receptors are required and which subpopulation(s) of SDN cells are specifically organized by 5HT. Therefore, we injected male and female Sprague-Dawley rat pups with saline, a nonselective 5HTR agonist, a 5HT2A/2C agonist, or a 5HT2A/2C antagonist over several timecourses in early life, and measured the Nissl-SDN as well as a calbindin+ subdivision of the SDN, the CALB-SDN. When examined on postnatal day 18 or early adulthood, the size of the Nissl-SDN was feminized in males treated with any of the serotonergic drugs, eliminating the typical sex difference. In contrast, the sex difference in CALB-SDN size was maintained regardless of serotoninergic drug treatment. This pattern suggests that although gonadal hormones shape the whole SDN, individual cellular phenotypes respond to different intermediary signals to become sexually dimorphic. Specifically, 5HT mediates sexual differentiation of non-calbindin population(s) within the SDN. The results also caution against using measurement of the CALB-SDN in isolation, as the absence of an effect on the CALB-SDN does not preclude an effect on the overall nucleus. © 2016 Wiley Periodicals, Inc. Develop Neurobiol 76: 1241-1253, 2016.

Over the past 35 years, developmental geneticists have made impressive progress toward an understanding of how genes specify morphology and function, particularly as they relate to the specification of each physical component of an organism. In the last 20 years, male courtship behavior in Drosophila melanogaster has emerged as a robust model system for the study of genetic specification of behavior. Courtship behavior is both complex and innate, and a single gene, fruitless (fru), is both necessary and sufficient for all aspects of the courtship ritual. Typically, loss of male-specific Fruitless protein function results in male flies that perform the courtship ritual incorrectly, slowly, or not at all. Here we describe a novel requirement for fru: we have identified a group of cells in which male Fru proteins are required to reduce the speed of courtship initiation. In addition, we have identified a gene, Trapped in endoderm 1 (Tre1), which is required in these cells for normal courtship and mating behavior. Tre1 encodes a G-protein-coupled receptor required for establishment of cell polarity and cell migration and has previously not been shown to be involved in courtship behavior. We describe the results of feminization of the Tre1-expressing neurons, as well as the effects on courtship behavior of mutation of Tre1. In addition, we show that Tre1 is expressed in a sexually dimorphic pattern in the central and peripheral nervous systems and investigate the role of the Tre1 cells in mate identification.

The effects of pharmacological waste on aquatic ecosystems are increasingly highlighted in ecotoxicology research. Many of these products are designed for human physiology but owing to the conservative nature of vertebrate evolution they also tend to have effects on aquatic organisms and fishes in particular when they find their way into aquatic systems via wastewater effluent. One area of research has focused on reproductive control and the associated hormone treatments. Many of these hormones affect the reproductive physiology of fishes and may cause feminization of male reproductive traits. Alternative medicines have also been widely used particularly in traditional cultures but few of these alternative treatments have been assessed with respect to their potential impact on aquatic ecosystems. Rue (Ruta graveolens) has been used as a male contraceptive in traditional medicines but its effects on fish behavior and reproductive anatomy have yet to be established. Here we show that treating Siamese fighting fish, Betta splendens, with extract of rue has a significant effect on key aggressive/reproductive behaviors and the propensity to explore novel objects (boldness). In all cases the respective behaviors were reduced relative to controls and sham injected fish. Histological analysis of the testes revealed that rue exposure reduced the number of spermatozoa but increased the number of spermatocytes relative to controls.

Aging is associated with visceral adiposity, metabolic disorders, and chronic low-grade inflammation. 17α-estradiol (17α-E2), a naturally occurring enantiomer of 17β-estradiol (17β-E2), extends life span in male mice through unresolved mechanisms. We tested whether 17α-E2 could alleviate age-related metabolic dysfunction and inflammation. 17α-E2 reduced body mass, visceral adiposity, and ectopic lipid deposition without decreasing lean mass. These declines were associated with reductions in energy intake due to the activation of hypothalamic anorexigenic pathways and direct effects of 17α-E2 on nutrient-sensing pathways in visceral adipose tissue. 17α-E2 did not alter energy expenditure or excretion. Fasting glucose, insulin, and glycosylated hemoglobin were also reduced by 17α-E2, and hyperinsulinemic-euglycemic clamps revealed improvements in peripheral glucose disposal and hepatic glucose production. Inflammatory mediators in visceral adipose tissue and the circulation were reduced by 17α-E2. 17α-E2 increased AMPKα and reduced mTOR complex 1 activity in visceral adipose tissue but not in liver or quadriceps muscle, which is in contrast to the generalized systemic effects of caloric restriction. These beneficial phenotypic changes occurred in the absence of feminization or cardiac dysfunction, two commonly observed deleterious effects of exogenous estrogen administration. Thus, 17α-E2 holds potential as a novel therapeutic for alleviating age-related metabolic dysfunction through tissue-specific effects.

Estrogen, which is synthesized earlier in females than androgen in males, is critical for sex determination in non-mammalian vertebrates. However, it remains unknown that what would happen to the gonadal phenotype if estrogen and androgen were administrated simultaneously. In this study, XY and XX tilapia fry were treated with the same dose of 17α-methyltestosterone (MT) and 17β-estradiol (E2) alone and in combination from 0 to 30 days after hatching. Treatment of XY fish with E2resulted in male to female sex reversal, while treatment of XX fish with MT resulted in female to male sex reversal. In contrast, simultaneous treatment of XX and XY fish with MT and E2resulted in female, but withcyp11b2andcyp19a1aco-expressed in the ovary. Serum 11-ketotestosteron level of the MT and E2simultaneously treated XX and XY female was similar to that of the XY control, while serum E2level of these two groups was similar to that of the XX control. Transcriptomic cluster analysis revealed that the MT and E2treated XX and XY gonads clustered into the same branch with the XX control. However a small fraction of genes, which showed disordered expression, may be associated with stress response. These results demonstrated that estrogen could maintain the female phenotype of XX fish and feminize XY fish even in the presence of androgen. Simultaneous treatment with estrogen and androgen up-regulated the endogenous estrogen and androgen synthesis, and resulted in disordered gene expression and endocrine disruption in tilapia.

Environmental estrogens and androgens can be present simultaneously in aquatic environments and thereby interact to disturb multiple physiological systems in organisms. Studies on interaction effects in fish of androgenic and estrogenic chemicals are limited. Therefore, the aim of the present study was to evaluate feminization and masculinization effects in zebrafish (Danio rerio) exposed to combinations of two synthetic steroid hormones detected in environmental waters: the androgen 17β-trenbolone (Tb) and the oestrogen 17α-ethinylestradiol (EE2). Juvenile zebrafish were exposed between days 20 and 60 post-hatch to different binary mixtures of Tb (1, 10, and 50 ng/L) and EE2 (2 and 5 ng/L). The endpoints studied were whole-body homogenate vitellogenin concentration at 40 days post-hatch, and sex ratio including gonad maturation at 60 days post-hatch. The feminizing potency of 5 ng/L of EE2, alone as well as in combination with Tb, was clear in the present study, with exposures resulting in almost all-female populations and females being sexually immature. Masculinization effects with male-biased sex ratios were observed when fish were exposed to 2 ng/L of EE2 in combination with Tb concentrations. Intersex fish were observed after exposure to mixtures of 2 ng/L EE2 with 50 ng/L Tb. Sexual maturity generally increased among males at increasing concentrations of Tb. The results of the present study show that exposure to environmentally relevant mixtures of an oestrogen and androgen affects the process of gonad differentiation in zebrafish and lead to sexual disruption.

Introduction: In the decision-making process, the development of effective strategies to minimize vulnerability of young people to the human immunodeficiency virus (HIV), and the understanding of background determinants in which they interact, are important auxiliary tools for HIV prevention. Objective: To outline the contributing factors for individual vulnerability of young people to HIV infection. Material and Methods: This was a population-based survey with 417 individuals from elementary schools in the city of Joao Pessoa, PB, Brazil. A questionnaire was used to collect the data, which were analyzed through statistical inference using the Chi-square (χ2) test. Results: The results demonstrated statistical evidence of association of the sociodemographic variables (p-value

Background and Objectives: 46 XY disorders of sexual development (DSD) cover a wide spectrum of phenotypes ranging from unambiguous female genitalia to ambiguous male genitalia with hypospadias or dysgenetic gonads. Management of these patients depends on the cause of DSD, degree of feminization, age at presentation, and gender orientation. The aim of this study was to evaluate the presentation and management of patients with 46XY DSD at our center. Patients and Methods: All new and old patients of 46XY DSD attending the endocrine OPD in a period of 16 months were included in this study. Clinical, cytogenetic, hormonal, and radiological evaluation were done to identify the cause of DSD. Results: Among 19 patients, eight were diagnosed with disorders of gonadal development (one with complete gonadal dysgenesis, four with partial gonadal dysgenesis, two with congenital bilateral anorchia, and one with ovotesticular DSD) and eight with disorders of androgen synthesis and action (one with complete androgen insensitivity syndrome [AIS], three with partial AIS and four with 5α reductase deficiency). In three patients, a definitive diagnosis could not be made. Conclusions: Management of patients with DSD depends on etiology, gender assignment, gender orientation, hormonal treatment, genital surgery, and consequent psychosocial implications. Due to the overlapping clinical and biochemical parameters in different subsets of DSD, only a preliminary etiological diagnosis can be made in some cases. Genetic studies with long-term follow-up are required for an accurate diagnosis.

Body feminization, as part of gender affirmation process of transgender women, decreases the volume of their cortical and subcortical brain structures. In this work, we implement a rat model of adult male feminization which reproduces the results in the human brain and allows for the longitudinal investigation of the underlying structural and metabolic determinants in the brain of adult male rats undergoing feminization treatments. Structural MRI and Diffusion Tensor Imaging (DTI) were used to non-invasively monitor in vivo cortical brain volume and white matter microstructure over 30 days in adult male rats receiving estradiol (E2), estradiol plus cyproterone acetate (CA), an androgen receptor blocker and antigonadotropic agent (E2 + CA), or vehicle (control). Ex vivo cerebral metabolic profiles were assessed by 1H High Resolution Magic Angle Spinning NMR (1H HRMAS) at the end of the treatments in samples from brain regions dissected after focused microwave fixation (5 kW). We found that; a) Groups receiving E2 and E2 + CA showed a generalized bilateral decrease in cortical volume; b) the E2 + CA and, to a lesser extent, the E2 groups maintained fractional anisotropy values over the experiment while these values decreased in the control group; c) E2 treatment produced increases in the relative concentration of brain metabolites, including glutamate and glutamine and d) the glutamine relative concentration and fractional anisotropy were negatively correlated with total cortical volume. These results reveal, for the first time to our knowledge, that the volumetric decreases observed in trans women under cross-sex hormone treatment can be reproduced in a rat model. Estrogens are more potent drivers of brain changes in male rats than anti-androgen treatment., This work was supported by Grants (PGC2018-094919-B-C1 to AG; PGC2018-094919-B-C2 to RF and SAF-2017-83043-R to SC) from Ministerio de Ciencia, Innovación y Universidades (Spain).

Normally we encounter a myriad of chemicals in our daily lives. Endocrine disrupting chemicals [EDCs] are ubiquitous in our environment and upon bodily entry many be stored in adipose tissue and, in pregnant woman, can reach the developing fetal brain, disrupting normal fetal brain development. EDC-induced aberrant levels of sex hormones can alter sexual dimorphism [i.e. degree of feminization or masculinization; sex differences in brain and behavior] and may contribute to the differential susceptibility of males and females to autism. Fetal development is guarded by the placenta which expresses high levels of aromatase, an enzyme that converts testosterone [T] to estrogen [E], and it appears that this process prevents the transfer of maternal T to the vulnerable developing fetus. Many investigators have shown a clear association between exposure to high levels of T during the prenatal and early postnatal periods, and development of autism spectrum disorder [ASD]. Retinoic acid-related orphan receptor-alpha [RORA] is a transcription factor that regulates aromatase. We analyzed the effects of six commonly encountered EDCs on human progenitor neurons from both genders and evaluated the differential expression of both aromatase and RORA by real-time PCR using an in vitro approach. We also evaluated the effects of the EDCs on neurite formation. Male and female human neuroblastoma cell lines were used to evaluate the effects of EDCs on neurite formation and the expression of RORA and CYP19A1. The morphologic analyses showed significant neuromodifications in neurite formation and a significant differential downregulation of RORA in male, but not in female neurons. Dysregulation in CYP19A1 was not gender-associated but was dependent on the dose of EDCs. Our findings confirm previous reports of differential downregulation of RORA in male neurons while expanding the gene-environment connection where estrogenic or androgenic EDCs can profoundly dysregulate RORA and aromatase gene expression, leading to a cascade of a highly integrated gene-network, associated with ASD.

Summary: Osseous work in facial feminization has been largely developed by comparing the qualitative and quantitative anthropometric differences between male and female cadaveric skulls. While virtual surgical planning has allowed for improved accuracy and ease in osteotomies and recontouring procedures in facial feminization, ultimately, a subjective decision-making process is still required. In this work, we describe a novel method of simplifying facial feminization by virtually modeling all osseous surgical maneuvers for facial feminization with a reference female skull.

2,4-Dichlorophenol (2,4-DCP) is a ubiquitous contaminant of aquatic environments with an estrogenic effect on fish. However, the molecular mechanism underlying this effect remains elusive. To this end, the present study aimed to explore the effect of 2,4-DCP on sex differentiation and its relevant mechanism in zebrafish (Danio rerio). The results showed that a female-biased sex ratio was induced after exposing larval zebrafish to 2,4-DCP (0–160 μg/L) from 20 to 50 days post fertilization (dpf). The feminization of zebrafish was accompanied by decreased expression of male-related genes (sox9a, amh and dmrt1) under 2,4-DCP from 20 to 50 dpf. However, the expression of female-related genes (cyp19a1a, foxl2 and esr1) was also suppressed. Nevertheless, it is noteworthy that the methylation level of sox9a promoter was significantly increased, which may result in the significantly decreased expression of sox9a and ultimately the feminization effect of 2,4-DCP on zebrafish. In addition, 5-aza-2′-deoxycytidine (5-AZA), a methyltransferase inhibitor, significantly reduced the methylation level, increased the expression of sox9a, and partly impaired the feminization effect caused by 2,4-DCP, which further confirmed the importance of DNA methylation of sox9a in 2,4-DCP-induced feminization. These findings provide novel insights into the epigenetic mechanisms of DCP-induced estrogenic effect in fish.

In plants, dioecy characterizes species carrying male and female flowers on separate plants and occurs in about 6% of angiosperms. To date, the molecular mechanism(s) underlying sexual dimorphism is essentially unknown. The ability of gender-reversal by hormone application suggests that epigenetics might play an important role in sexual dimorphism. Proteome analysis of nuclei derived from flower buds of females, males and feminized males of the dioecious plantMercurialis annuarevealed differentially expressed proteins related to nucleic acid binding proteins, hydrolases and transcription factors, including floral homeotic genes. Further analysis showed that class B genes are mainly expressed in male flowers, while class D genes, as well asSUPERMAN-like genes, were mainly expressed in female flowers. Cytokinin-induced feminization of male plants was associated with down-regulation of male-specific genes concomitantly with up-regulation of female-specific genes. No correlation could be found between the expression of class B and D genes and their DNA methylation or chromatin conformation. Thus, our results ruled out epigenetic control over floral identity genes as the major determinants regulating sexual dimorphisms. Rather, determination of sex inM. annuamight be controlled upstream of floral identity genes by a gender-specific factor that affects hormonal homeostasis.HighlightsSex determination inMercurialis annuais not related to epigenetics of floral homeotic genes but appears to be modulated by an unknown gender-specific regulator(s) that affects hormonal homeostasis.

BACKGROUND Transgender patients may seek nonsurgical methods for facial masculinization and feminization as an adjunct or alternative to undergoing surgical procedures. OBJECTIVES The authors reviewed the existing literature regarding this topic and provided an overview of nonsurgical techniques for facial masculinization and feminization. METHODS A comprehensive literature search of the PubMed and MedLine databases was conducted for studies published through December 2017 for techniques and outcomes of nonsurgical facial masculinization and feminization. Keywords were used in performing the search. Data on techniques, outcomes, complications, and patient satisfaction were collected. RESULTS Four articles fit our inclusion criteria. Given the lack of published literature describing facial injectables in transgender patients, data from the literature describing techniques in cisgender patients were utilized to supplement our review. CONCLUSIONS Facial feminization can be achieved through injectables such as neurotoxin and fillers for lateral brow elevation, lip augmentation, malar augmentation, and improvement of rhytids. Facial masculinization can be achieved with injectables used for genioplasty, jawline augmentation, and supraorbital ridge augmentation. One must develop best practices for these techniques in the transgender patient population and increase awareness regarding nonsurgical options. LEVEL OF EVIDENCE: 4

Background: Wolbachia is the most common endosymbiotic bacteria in insectborne parasites and it is the most common reproductive parasite in the world. Wolbachia has been found worldwide in numerous arthropod and parasite species, including insects, terrestrial isopods, spiders, mites and filarial nematodes. There is a complicated relationship between Wolbachia and its hosts and in some cases, they create a mutual relationship instead of a parasitic relationship. Some species are not able to reproduce in the absence of infection with Wolbachia. Thus, the use of existing strains of Wolbachia bacteria offers a potential strategy for the control of the population of mosquitoes and other pests and diseases. Methods: We searched ten databases and reviewed published papers regarding the role of Wolbachia as a promising drug target and emerging biological control agents of parasitic diseases between 1996 and 2017 (22 years) were considered eligible. Also, in the current study several patents (WO008652), (US7723062), and (US 0345249 A1) were reviewed. Results: Endosymbiotic Wolbachia bacteria, which are inherited from mothers, is transmitted to mosquitoes and interferes with pathogen transmission. They can change the reproduction of their host. Wolbachia is transmitted through the cytoplasm of eggs and have evolved different mechanisms for manipulating the reproduction of its hosts, including the induction of reproductive incompatibility, parthenogenesis, and feminization. The extensive effects of Wolbachia on reproduction and host fitness have made Wolbachia the issue of growing attention as a potential biocontrol agent. Conclusion: Wolbachia has opened a new window to design a costly, potent and ecofriendly drug target for effective treatment and elimination of vector-borne parasitic diseases.

The widespread use of the synthetic estrogen 17 α-ethinylestradiol (EE2) has resulted in elevated levels in aquatic environments, where it is known to act as an endocrine disrupting chemical affecting fish and other aquatic organisms. Examining changes in the structure of the fish’ gonads and liver has proven to be an effective approach for assessing these impacts. While changes have been reported for various fish species, it is not clear whether impacts are equally severe in live-bearing fishes. The present study looked at gonadal and liver development in EE2-exposed least killifish (Heterandria formosa), a live-bearing Poeciliid. Exposures to 0, 5, or 25 ng/L EE2 began within six days of birth and continued until fish became sexually mature 12–23 weeks later. Exposure to 5 ng/L EE2 resulted in severe intersex in fish with external male characteristics, a slowdown of spermatogenesis in these intersex fish and a slowdown of oogenesis in the female fish. Moreover, these fish had a variety of liver injuries. Fish exposed to 25 ng/L EE2 exhibited intersex but at a lower frequency than occurred at 5 ng/L. In contrast, liver damage and slowdown of both oogenesis and spermatogenesis exhibited the typical dose-dependence. These findings illustrate the importance of including histological analyses when assessing endocrine disruption in fish, demonstrate that the live-bearing mode of reproduction appears to provide limited protection from the effects of waterborne EE2, and provide further evidence that EE2 has multiple impacts on fish health and reproduction that are severe enough to potentially affect fish populations.

In animals, sexual differences between males and females are usually determined by sex chromosomes. Alternatively, sex may also be determined by vertically transmitted intracellular microbial endosymbionts. The best known cytoplasmic sex manipulative endosymbiont is Wolbachia which can, for instance, feminize genetic males into phenotypic females in the terrestrial isopod Armadillidium vulgare. However, the molecular genetic basis of cytoplasmic sex determination is unknown. To identify candidate genes of feminization induced by Wolbachia strain wVulC from A. vulgare, we sequenced the genome of Wolbachia strain wCon from Cylisticus convexus, the most closely related known Wolbachia strain to wVulC that does not induce feminization, and compared it to the wVulC genome. Then, we performed gene expression profiling of the 216 resulting wVulC candidate genes throughout host developmental stages in A. vulgare and the heterologous host C. convexus. We identified a set of 35 feminization candidate genes showing differential expression during host sexual development. Interestingly, 27 of the 35 genes are present in the f element, which is a piece of a feminizing Wolbachia genome horizontally transferred into the nuclear genome of A. vulgare and involved in female sex determination. Assuming that the molecular genetic basis of feminization by Wolbachia and the f element is the same, the 27 genes are candidates for acting as master sex determination genes in A. vulgare females carrying the f element.

Summary Introduction A central ethical dilemma in management of the patient with a disorder of sex development (DSD) is the potential conflict between respect for the fundamental right of the child for physical and emotional integrity and self-determination, and the right of parents to serve as surrogate decision-makers and act in their child's best interest. Methods Over the past 2 years we have encountered three complex DSD cases on the spectrum of mixed gonadal dysgenesis to ovotesticular DSD in which gender assignment and therefore optimal surgical management was uncertain. All patients had mosaic karyotypes with Y chromosome, dysgenetic ovary and dysgenetic testis, a urogenital sinus, and prominent phallus. In all three cases a team approach was taken to assess functional potential and risks along either gender pathway and to develop a spectrum of treatment options for parental consideration, including: 1. masculinization with removal of dysgenetic ovary; 2. initial vaginoplasty but with retention of the phallus (±bilateral gonadectomy); 3. initial vaginoplasty with “burial” of corporal bodies (Pippi Salle procedure) (±bilateral gonadectomy); 4. full feminization: vaginoplasty and clitoroplasty (with bilateral gonadectomy); 5. no surgical intervention. Results In all three cases, after consideration of risks and benefits of all options, parents selected option 2: gonadectomy to eliminate tumor risk and vaginoplasty, taking advantage of the child's young age to exteriorize urinary and reproductive tracts to avoid incontinence and infection and supporting parental bias toward female gender, but preservation of phallic structures to ensure a male option should the patient later declare a male gender identity. Parents of the three patients were contacted post-operatively (at 7, 17, and 22 months) for follow-up. All (3/3) regarded their child's development and wellbeing positively, and their own decisions regarding gender assignment and surgical plan favorably. All (3/3) regarded the team advisory process as balanced and supportive. Conclusion In the setting of exposure of the neonatal brain to testosterone, vaginoplasty and phallic preservation afforded a balance between parental preferences and preservation of anatomic options, allowing potential reconstruction of male or female phenotype as gender identity is ascertained thereby respecting both parent and patient rights. Parents valued a spectrum of options, transparency, and the team decision-making process. Table . Patient characteristics. A B C Age at referral 14 months 23 months 6 weeks Karyotype 45,X/46,XY/47,XYY 45,X/46,XY 46XX/47XXY Gonads L Dysgenetic testis R Dysgenetic ovary L Dysgenetic testis R Dysgenetic ovary L Dysgenetic ovary R Dysgenetic testis Ductal structures Uterus R Fallopian tube Uterus L Fallopian tube Uterus UG sinus Mid confluence Low confluence Low confluence Phallus 4 × 1.5 cm 3 × 1 cm 2.7 × 1.3 cm Gender assignment Female Female Neutral

Background This study examined the clinical effects of leuprolide acetate in sexual offenders with paraphilic disorders evaluated by means of objective psychiatric assessment. Methods The subjects of this study were seven sexual offenders who were being treated by means of an injection for sexual impulse control by a court order. They had been diagnosed with paraphilia by a psychiatrist based on the Diagnostic and Statistical Manual of Mental Disorders, 5th edition (DSM-5) and had been put on probation by the Ministry of Justice between January 2016 and December 2016. Results After twelve months, we observed significant improvement in symptoms, as decrease of abnormal sexual interest and activity, sexual fantasy, Clinical Global Impression-Severity (CGI-S), and Clinical Global Impression-Impulsivity (GCI-I). There were a mild feminization of the body shape, feelings of fatigue, and mild hot flushes. No other adverse effect was reported. Conclusion These results suggested that the clinical effects of leuprolide acetate in sexual offenders might be an effective treatment and safety strategy., Graphical Abstract

Pharmaceutical pollution is becoming an unavoidable environmental issue of emerging concern. As forecasted, the consumption of medicinal drugs and their use in veterinary practice is expected to systematically increase over coming years, resulting in their increased discharge. The most commonly used pharmaceuticals include non-steroidal anti-inflammatory drugs (e.g., diclofenac, naproxen, ibuprofen), cardiovascular drugs (e.g., beta-blockers, diuretics, calcium channel blockers, lipid-regulating agents), antibiotics, oral contraceptives, anti-depressants, immunosuppressive drugs and cytostatics. Active pharmaceutical ingredients (APIs) are known to partially survive the conventional process of wastewater treatment. In freshwaters, they may undergo photodegradation, biodegradation, sorption to sediments and uptake by organisms. The latter results in metabolism or bioaccumulation, and potential toxicological effects and physiological responses. The magnitude of effects is largely modulated by the concentration of APIs, time of exposure and some environmental factors such as light and nutrient availability. The response to APIs in closely taxonomically related species may be significantly different. The concomitant presence of different APIs usually evokes potentiation of adverse effects. The most serious effects of pharmaceutical pollution evidenced so far for freshwaters include increase in antibiotic-resistant microorganisms, feminization, behavioral changes, and immunosuppression in fish. Beyond any doubt, it is imperative to support systematic research on API detection methods, to monitor the great number of APIs in wastewater, surface and groundwater, and tap water, and to assess the ecological risks arising from their increased presence in the freshwater environment.

Disorders of sex development (DSD) are a heterogenous group of “congenital conditions in which development of chromosomal, gonadal, or anatomical sex is atypical ( Lee et al., 2006 ).” Once the diagnosis has been established and metabolic abnormalities regulated, consideration must be given to gender assignment and subsequent gender confirming surgeries. The timing of surgery remains controversial, particularly in feminization surgery, but there is general consensus as to the goals of surgery. The fertility and malignancy potential of the gonads must also be addressed. Multidisciplinary teams are integral to the evaluation and life-long treatment of patients with DSD.

Feminization of the frontonasal-orbital complex is a critical aspect in surgical management of gender dysphoria. Here, we describe our utilization of a prefabricated cutting guide to facilitate frontal sinus setback.

Bisphenol F (BPF) has become a predominant bisphenol contaminant in recent years. It has significant estrogenic properties in both in vivo and in vitro studies. We have previously studied the disrupting mechanisms of BPF on the hypothalamic–pituitary–gonadal axis of adult zebrafish. However, the effects of BPF exposure on development and sexual differentiation of zebrafish embryos/larvae remain unclear. To determine the effects of BPF on the critical stage of sex differentiation in zebrafish, zebrafish embryos/larvae were exposed to 1, 10, 100, and 1000 μg/L BPF from fertilization to 60 days post-fertilization (dpf). Developmental malformations were induced by exposure to BPF from 2 h post-fertilization (hpf), with a LC50 of 10,030 μg/L at 96 hpf and 9391 μg/L at 120 hpf. Long-term exposure during sex differentiation tended to result in a female sex ratio bias. Histological analyses at 60 dpf indicated that the development of ovo-testes and immature ovaries was induced by 100 and 1000 μg/L BPF. Homogenate testosterone levels decreased and 17β-estradiol levels increased in zebrafish in a concentration-dependent manner. BPF exposure suppressed gene expression of double sex, Mab3-related transcription factor 1(dmrt1), fushi tarazu factor 1d (ff1d), sry-box containing gene 9a (sox9a) and anti-Mullerian hormone (amh); induced expression of the forkhead box L2 transcription factor (foxl2), leading to increased expression of aromatase (cyp19a1a), which promoted production of estrogens, and further caused phenotypic feminization of zebrafish. These results suggest that developmental exposure to BPF has adverse effects on sexual differentiation, and the results were useful for a BPF risk assessment.

Sexual dimorphism (SD) has evolved in response to selection pressures that differ between sexes. Since such pressures change across an individual's life, SD may vary within age classes. Yet, little is known about how selection on early phenotypes may drive the final SD observed in adults. In many dimorphic species, juveniles resemble adult females rather than adult males, meaning that out of the selective pressures established by sexual selection feminized phenotypes may be adaptive. If true, fitness benefits of early female-like phenotypes may constrain the expression of male phenotypes in adulthood. Using the common kestrel Falco tinnunculus as a study model, we evaluated the fitness advantages of expressing more feminized phenotypes at youth. Although more similar to adult females than to adult males, common kestrel fledglings are still sexually dimorphic in size and coloration. Integrating morphological and chromatic variables, we analysed the phenotypic divergence between sexes as a measure of how much each individual looks like the sex to which it belongs (phenotypic sexual resemblance, PSR). We then tested the fitness benefits associated with PSR by means of the probability of recruitment in the population. We found a significant interaction between PSR and sex, showing that in both sexes more feminized phenotypes recruited more into the population than less feminized phenotypes. Moreover, males showed lower PSR than females and a higher proportion of incorrect sex classifications. These findings suggest that the mechanisms in males devoted to resembling female phenotypes in youth, due to a trend to increase fitness through more feminized phenotypes, may provide a mechanism to constrain the SD in adulthood., This study was financed by the Spanish Ministerio de Economíıa y Competitividad (Projects: CGL2004-04479/BOS, CGL2007-61395/BOS, CGL2010-15726/BOS).

© 2015 American Chemical Society. Exposure to environmental estrogens in wastewater treatment works (WwTW) effluents induces feminized responses in male fish, including the development of eggs in male testes. However, the impacts on the offspring of exposed fish are not well understood. In this study, we examined whether roach (Rutilus rutilus) from mothers that had been exposed to an undiluted WwTW effluent from early life to sexual maturity had altered susceptibility to gonadal feminization and an impaired capacity to reproduce. For males from both WwTW effluent exposed mothers and dilution water exposed mothers, effluent exposure for up to 3 years and 9 months induced feminized male gonads, although the intersex condition was relatively mild. There was no difference in the severity of gonadal feminization in roach derived from either WwTW effluent exposed or dilution water exposed mothers. Furthermore, a breeding study revealed that roach with effluent-exposed mothers reproduced with an equal success as roach with mothers exposed to clean water. Roach exposed to the effluent for 3 years in this study were able to reproduce successfully. Our findings provide no evidence for impacts of WwTW effluent exposure on reproduction or gonadal disruption in roach down the female germ line and add to existing evidence that male roach with a mild intersex condition are able to breed competitively. Natural Environmental Research Council (NERC) grant nos. NE/G019355/1, NE/K004263/1; Department for Environment, Food and Rural Affairs, U.K.

Research on endocrine disruption in frog populations, such as shifts in sex ratios and feminization of males, has predominantly focused on agricultural pesticides. Recent evidence suggests that suburban landscapes harbor amphibian populations exhibiting similar levels of endocrine disruption; however the endocrine disrupting chemical (EDC) sources are unknown. Here, we show that sex ratios of metamorphosing frogs become increasingly female-dominated along a suburbanization gradient. We further show that suburban ponds are frequently contaminated by the classical estrogen estrone and a variety of EDCs produced by plants (phytoestrogens), and that the diversity of organic EDCs is correlated with the extent of developed land use and cultivated lawn and gardens around a pond. Our work also raises the possibility that trace-element contamination associated with human land use around suburban ponds may be contributing to the estrogenic load within suburban freshwaters and constitutes another source of estrogenic exposure for wildlife. These data suggest novel, unexplored pathways of EDC contamination in human-altered environments. In particular, we propose that vegetation changes associated with suburban neighborhoods (e.g., from forests to lawns and ornamental plants) increase the distribution of phytoestrogens in surface waters. The result of frog sex ratios varying as a function of human land use implicates a role for environmental modulation of sexual differentiation in amphibians, which are assumed to only have genetic sex determination. Overall, we show that endocrine disruption is widespread in suburban frog populations and that the causes are likely diverse.

Environmental chemicals can disrupt endocrine signaling and adversely impact sexual differentiation in wildlife. Bisphenol A (BPA) is an estrogenic chemical commonly found in a variety of habitats. In this study, we used painted turtles (Chrysemys picta), which have temperature-dependent sex determination (TSD), as an animal model for ontogenetic endocrine disruption by BPA. We hypothesized that BPA would override TSD and disrupt sexual development. We incubated farm-raised turtle eggs at the male-producing temperature (26°C), randomly assigned individuals to treatment groups: control, vehicle control, 17β-estradiol (E2, 20ng/g-egg) or 0.01, 1.0, 100μgBPA/g-egg and harvested tissues at hatch. Typical female gonads were present in 89% of the E2-treated "males", but in none of the control males (n=35). Gonads of BPA-exposed turtles had varying amounts of ovarian-like cortical (OLC) tissue and disorganized testicular tubules in the medulla. Although the percentage of males with OLCs increased with BPA dose (BPA-low=30%, BPA-medium=33%, BPA-high=39%), this difference was not significant (p=0.85). In all three BPA treatments, SOX9 patterns revealed disorganized medullary testicular tubules and β-catenin expression in a thickened cortex. Liver vitellogenin, a female-specific liver protein commonly used as an exposure biomarker, was not induced by any of the treatments. Notably, these results suggest that developmental exposure to BPA disrupts sexual differentiation in painted turtles. Further examination is necessary to determine the underlying mechanisms of sex reversal in reptiles and how these translate to EDC exposure in wild populations.

Endocrine-mediated effects in fish populations have been widely documented. Most attention has been focused on feminization caused by estrogenic substances, but this paper reviews evidence for the effects of a group of fungicides and pharmaceuticals, the azoles, which have been reported to cause masculinization in fish. The paper considers information from laboratory studies on the effects of azole compounds on fish endocrinology, and on the potential existence of such effects in wild fish. The occurrence of some azoles in effluents and surface waters has also been briefly reviewed. Under laboratory conditions, many azoles are able to cause masculinization or defeminization in fish by inhibition of the P450 enzyme aromatase (CYP19). However, in no case where such effects have been observed in the field has a link been established with this group of substances. In most instances, other more convincing explanations have been proposed. Peak concentrations of some azoles in surface waters can approach those which, under continuous long-term exposure in the laboratory, might lead to some aromatase inhibition. However, available data on exposure and effects provide reassurance that the concentrations of azoles found in surface waters are too low to cause adverse effects in fish by interference with their endocrine system. Compared to the widespread observations of feminization and estrogenic effects in (male) fish, there are relatively few papers describing masculinization or defeminization in (female) wild fish populations, suggesting that this is quite a rare phenomenon. The significance of this result is emphasized by the fact that fish are among the best studied organisms in the environment.

SUMMARYParasitic castration is a strategy used by parasites to minimize damage to the host by consuming its reproductive system, which results in the morphological alteration of the host. We determined that the forewing shape and density of the antennal sensilla of field-collected adult male mantids (Hierodula formosana), infected by horsehair worms (Chordodes formosanus) was partially feminized (intersexuality), and both male and female mantids infected by horsehair worms exhibited allometric changes in their wings and walking legs. In addition, the testes of most infected male adults disappeared or reduced in size, whereas the number of ovarioles in infected female adults was unaffected. The infection mainly influenced the structures related to host reproduction and locomotion, suggesting unbalanced energy exploitation and the reduction of parasitic virulence. In addition, the intersexuality of infected male adults indicated that sexual differentiation in insects, which researchers have considered to be an autonomous process, was influenced by the infection. The similarity of the antennae of infected male adults with those of last-instar female nymphs suggested that parasitic juvenilization may cause such feminization, but the mechanism of parasitic influence on insect sex characteristics should be studied further.

The intracellular alpha-proteobacteria Wolbachia pipientis infects about 40% of arthropod species from all major taxonomic groups; it also infects nematodes. Such a wide range of host species is unusual for highly specialized parasites, which include Wolbachia; it may be associated with the adaptation to the presence of germplasm in cells that exhibit minimal evolutionary variability in animals. Having type-IV bacterial secretion apparatus, Wolbachia has the ability to export proteins and ribonucleoprotein complexes to the cytoplasm of germline cells of the host organism, thus forming the basis for intervention at the earliest stages of ontogenesis. Different strains of Wolbachia induce parthenogenesis, male-killing, feminization, cytoplasmic incompatibility, and increased resistance to viruses in the hosts. All of these effects increase the reproductive success of infected females and ensure the preservation of Wolbachia in the host populations. In recent years, results from the studyof the insect-Wolbachia symbiotic systems made it possible to start developing methods of insect control and new methods of combating human diseases transmitted by insect vectors.

Runoff from lands fertilized with animal manure from concentrated animal feeding operations (CAFOs) is a source of hormones to surface water. In this study we tested the hypothesis that larval fathead minnows exposed to sex steroids singly or in a “typical” CAFO mixture during sex differentiation would respond with changes in the expression of a set of target genes, leading to gonadal abnormalities later in life. In the first experiment, a static daily-renewal system was used to expose larvae during the period of 10–20 days post-hatch (dph) to either 5 ng/L 17β-trenbolone (17β-TRB) or 5 ng/L 17α-ethinylestradiol (EE 2 ). In a second experiment, fish were exposed from 0 to 45 dph in a flow-through system to a CAFO mixture composed of steroids and degradates (2–16 ng/L), atrazine and degradates (15–250 ng/L), and nitrate (3–11 mg/L). In the single hormone experiment, expression of genes involved in steroidogenesis ( cyp19a, cyp17 , and star ) was decreased in females. In contrast, no differences in gene expression were observed in fish exposed to the CAFO mixture. However, the majority (84%) of treated males had testes containing an ovarian cavity, indicative of feminization, compared to 0% in the control males. Overall, our results show that: (1) changes in gene expression after single hormone exposures are sex-specific, with females more responsive than males; and (2) phenotypic alterations in testicular development can be elicited by a simulated “CAFO” mixture when fathead minnows are exposed during the first 45 days of development. More research is needed to further discern the complex response of fish to steroid mixtures, especially those associated with runoff from land-applied CAFO waste.

Horses are characterized as having a greater rate of chromosomal abnormalities than other species, which are mainly related to the sex chromosome pair and produce a series of different anomalies known as disorders in sexual development (DSD). In the present study, three Pura Raza Española (PRE) and one Menorquín (MEN) horses were studied and an incompatibility in their genetic and phenotypic sex were detected. Animals were karyotyped by conventional and molecular cytogenetic analyses and characterized using genomic techniques. Although all individuals, were totally unrelated, these animals had the same abnormality (64,XY SRY negative DSD) despite having an anatomically normal external mare phenotype. Therefore, this syndrome could remain undiagnosed in a large percentage of cases because the physiological and morphological symptoms are rare. In the present study, a slight gonadal dysgenesis was observed only in older individuals. Interestingly this chromosomal abnormality has been previously reported less than twenty times, and never in the PRE or MEN horses. With the present research, it is demonstrated that the use of genetic and cytogenetic diagnostic tools in veterinary practice could be an important complementary test to determine the origin of unexplained reproductive failures among horses.

In the article based on information from the years 1991–2011 the following hypotheses were subjected to the verification: 1. with time, we observe spatial standardizing of coefficient rate of the feminization of the demographic old age; 2. the spatial image of the rate of the feminization of the demographic old age did not undergo the change over time; 3. with time, the increases in the rate of the feminization of the demographic old age are more distinct in these areas, where its value was relatively high at the beginning of the analyzed period; 4. Different measures describing the spatial feminization of the demographic old age are leading to similar conclusions in the evaluation of its spatial convergence. Not all of the put forward theses were confirmed. W artykule na podstawie informacji z lat 1991–2011 weryfikacji poddano następujące hipotezy: 1. z czasem odnotowujemy przestrzenne ujednolicanie się wartości współczynnika feminizacji starości demograficznej; 2. przestrzenny obraz współczynnika feminizacji starości demograficznej nie uległ zmianie w czasie; 3. z czasem przyrosty współczynnika feminizacji starości demograficznej są wyraźniejsze na tych obszarach, gdzie jego wartość była relatywnie wysoka na początku badanego okresu; 4. różne miary opisujące przestrzenną feminizację starości demograficznej prowadzą do podobnych wniosków w ocenie jej przestrzennej konwergencji. Nie wszystkie z postawionych tez zostały potwierdzone.

Engaging debates around the status of unpaid reproductive labor, this chapter investigates Bernadette Mayer’s multifarious project Memory, which is simultaneously a performance, a conceptual work, an installation, and an epic poem. In attempting to document, down to the smallest detail, every aspect of her life for thirty days—using photographs, audio recordings, and written notation—Mayer effectively demonstrates the subsumption of the entirety of life by the protocols and routines of work as well as the transformation of the relationship between unpaid reproductive work and feminized wage labor. Mayer’s “total” artwork, which merges different technologies into a single apparatus, prefigures the reorganization of office work around the personal computer, a technology that has probably done more than anything else to ensure that work and home life are unified by enabling white-collar workers to accomplish tasks from home and, in that sense, never leave work.

Testis-ova differentiation in sexually mature male medaka (Oryzias latipes) is easily induced by estrogenic chemicals, indicating that spermatogonia persist in sexual bipotentiality, even in mature testes in medaka. By contrast, the effects of estrogen on testicular somatic cells associated with testis-ova differentiation in medaka remain unclear. In this study, we focused on the dynamics of sex-related genes (Gsdf, Dmrt1, and Foxl2) expressed in Sertoli cells in the mature testes of adult medaka during estrogen-induced testis-ova differentiation. When mature male medaka were exposed to estradiol benzoate (EB; 800ng/L), testis-ova first appeared after EB treatment for 14days (observed as the first oocytes of the leptotene-zygotene stage). However, the testis remained structurally unchanged, even after EB treatment for 28days. Although Foxl2 is a female-specific sex gene, EB treatment for 7days induced Foxl2/FOXL2 expression in all Sertoli cell-enclosed spermatogonia before testis-ova first appeared; however, Foxl2 was not detected in somatic cells in control testes. Conversely, Sertoli-cell-specific Gsdf mRNA expression levels significantly decreased after EB treatment for 14days, and no changes were observed in DMRT1 localization following EB treatment, whereas Dmrt1 mRNA levels increased significantly. Furthermore, after EB exposure, FOXl2 and DMRT1 were co-localized in Sertoli cells during testis-ova differentiation, although FOXL2 localization was undetectable in Sertoli-cell-enclosed apoptotic testis-ova, whereas DMRT1 remained localized in Sertoli cells. These results indicated for the first time that based on the expression of female-specific sex genes, feminization of Sertoli cells precedes testis-ova differentiation induced by estrogen in mature testes in medaka; however, complete feminization of Sertoli cells was not induced in this study. Additionally, it is suggested strongly that Foxl2 and Gsdf expression constitute potential molecular markers for evaluating the effects of estrogenic chemicals on testicular somatic cells associated with estrogen-induced testis-ova differentiation in mature male medaka.