Ovotesticular disorder of sexual development due to 47,XYY/46,XY/45,X mixed gonadal dysgenesis in a phenotypic male presenting as cyclical haematuria: clinical presentation and assessment of long‐term outcomes

Ovotesticular disorder of sexual differentiation (OTDSD) is a rare cause of disorder of sexual differentiation predominantly having 46,XX karyotype, female phenotype and ambiguous genitalia. We report a 15‐year‐old having male body habitus, axillary and pubic hair, well‐developed penis and right‐descended testis with history of penoscrotal hypospadias correction, presenting with three episodes of cyclical haematuria, who biochemically had normal serum testosterone (338 ng dl^?1) which increased following hCG stimulation (614 ng dl^?1), elevated estradiol (17.35 pg ml^?1) along with elevated luteinising hormone (11.3 mIU l^?1) and follicle‐stimulating hormone (31 mIU l^?1). Ultrasonography followed by micturating cystourethrogram and cystoscopy confirmed the presence of prostate, uterus, cervix and vagina draining into the urogenital sinus continuing till the penile urethra and left intra‐abdominal gonad. Patient underwent hysterectomy and left gonadectomy. Histopathologic study of resected gonad confirmed presence of ovotestis. Low estradiol (1.2 pg ml^?1) following gonadectomy confirmed the ovotestis origin of estradiol. Chromosomal analysis revealed complex karyotype predominant being 47,XYY (50%) followed by 46,XY (26%) and 45,X (24%). This is perhaps the first report of 47,XYY/46,XY/45,X causing OTDSD in a phenotypic male.     

Presence of a low capacity androgen receptor in the gubernaculum of the pig fetus

The aim of our study was to investigate the theory that the gubernaculum is an androgen-responsive structure responsible for fetal testicular descent. Using 3H-methyltrienolone (3H-R1881) as ligand, we performed a seven-point Scatchard analysis of androgen receptor binding in the gubernaculum, prostate, urethra and striated muscle tissue obtained from pig fetuses between 60 and 109 days of gestation. The gubernaculum demonstrated androgen receptor binding with an affinity and capacity significantly lower than that of fetal pig prostate and urethra, but not significantly different from that of striated muscle taken from both male and female fetuses. A marked increase in the size and mass of the gubernaculum of male (but not female) fetuses is known to accompany testicular descent. If the dramatic increase in the mass of the gubernaculum observed in male (but not female) fetuses were the result of androgen stimulation, it may be expected that the striated muscle mass (and therefore total body mass) of male fetuses should be greater than that of female fetuses, since the androgen receptor affinity and capacity of the gubernaculum is similar to that of striated muscle. However, we found no significant difference in total body mass between male and female pig fetuses at the same period of gestation. These findings raise doubt concerning the theory that growth of the gubernaculum during descent is the result of androgen stimulation.     

The hypophyseal-testicular axis and sex accessory glands following chemical sympathectomy with guanethidine of pre-pubertal to mature rats

Summary.  Selective chemical sympathectomy of the internal sex organs of prepubertal to mature male Wistar rats was performed by chronic treatment with low doses of guanethidine. Plasma testosterone and luteinizing hormone and the intratesticular level of testosterone were determined. The weight and fructose content of seminal vesicle and ventral prostate were also investigated. The results showed that sympathetic innervation is related to the control of the hypophyseal-testicular axis as well as to the growth and potential secretory activity of the male sex accessory glands.     

Prostate gland growth during development is stimulated in both male and female rat fetuses by intrauterine proximity to female fetuses

In rodents, steroid hormones are transported between adjacent fetuses, and male or female fetuses that develop in utero between female fetuses (2F males or 2F females) have higher serum levels of estradiol and lower serum levels of testosterone relative to siblings of the same sex that develop between two male fetuses (2M males or 2M females). The present study was prompted by the prior unexpected finding that as adults, 2F male mice have an enlarged prostate, and increased numbers of prostatic androgen receptors relative to 2M males. We examined prostate development in both male and female rat fetuses from different intrauterine positions using computer-assisted, 3-dimensional reconstruction of the urogenital complex. In males, this included the prostate, seminal vesicles and utricle (a remnant of the Müllerian ducts), while in females it included development of prostatic glandular buds. The mean cross-sectional area of developing prostatic epithelial buds, utricle and seminal vesicles was significantly increased in 2F male relative to 2M male fetuses. In female fetuses, prostatic bud development was significantly more likely to occur in 2F (67%) than in 2M (29%) animals. These findings suggest that the transport of a small supplement of estrogen from adjacent female fetuses enhances androgen-dependent accessory organ development. We also found that mRNAs encoding receptors for both estrogen and androgen were located in the mesenchyme of the developing male prostate. The localization of estrogen and androgen receptor mRNA in this region further suggests that the mesenchymal induction of prostatic epithelial growth involves both hormones. The cranial dorsolateral prostatic buds exhibited the greatest enlargement in 2F males. This region of the developing prostate in rats is comparable (that is the embryonic homologue) to the region exhibiting benign prostatic hyperplasia (BPH) during aging in men. We propose that the potential for pathological regrowth of the prostate during aging is imprinted by estradiol during fetal development.     

Sex differences in murine myocutaneous flap revascularization

Sex differences in susceptibility to ischemia/reperfusion injury have been documented in humans. Premenopausal women have a lower risk of ischemic heart disease than age‐matched men, whereas after menopause, the risk is similar or even higher in women. However, little is known about the effects of sex on myocutaneous ischemia/reperfusion. To explore sex differences in wound revascularization, we utilized a murine myocutaneous flap model of graded ischemia. A cranial‐based, peninsular‐shaped, myocutaneous flap was surgically created on the dorsum of male and female mice. Physiological, pathological, immunohistochemical, and molecular parameters were analyzed. Flaps created on female mice were re‐attached to the recipient site resulting in nearly complete viability at post‐operative day 10. In contrast, distal full‐thickness myocutaneous necrosis was evident at 10 days post‐surgery in male mice. Over the 10 day study interval, laser speckle imaging documented functional revascularization in all flap regions in female mice, but minimal distal flap reperfusion in male mice. Day 10 immunostained histologic sections confirmed significant increases in distal flap vessel count and vascular surface area in female compared to male mice. RT‐PCR demonstrated significant differences in growth factor and metabolic gene expression between female and male mice at day 10. In conclusion, in a graded‐ischemia wound healing model, flap revascularization was more effective in female mice. The recognition and identification of sex‐specific wound healing differences may lead to a better understanding of the underlying mechanisms of myocutaneous revascularization and drive novel discovery to improve soft tissue wound healing following tissue transfer for traumatic injury and cancer resection.     

Changes in the endocrine environment of the human prostate transition zone with aging: simultaneous quantitative analysis of prostatic sex steroids and comparison with human prostatic histological composition

BACKGROUND: It is well-known that the incidence of benign prostatic hyperplasia (BPH) increases with aging. The age-dependent changes in the ratio of serum sex steroid concentrations may play a role in BPH development. To clarify the relationship between the prostatic tissue concentrations of these steroids and age, we established a precise method of simultaneous quantitative analysis for prostatic sex steroids and used this method to investigate the tissue concentrations of three major sex steroids (testosterone, dihydrotestosterone, and estradiol) in the human prostate. METHODS: The methodology for the simultaneous quantitative analysis of prostatic sex steroids was established using castrated rat prostatic tissue, coupled with internal standards, for androgen-deprived medium, and the validation of the method was examined. Human prostatic tissues were collected during surgery and immediately frozen at -70 degrees C. Using our method, the steroidal fractions were extracted, purified, and quantified. The proportions of stroma, epithelium, and glandular lumen were measured on each histological specimen, using an image analyzer. RESULTS: The validation tests showed that our method of quantitative analysis was precise and sensitive enough for the quantification of testosterone, dihydrotestosterone, and estradiol in the prostate. In humans, the prostatic dihydrotestosterone concentration decreased with age, but the concentrations of testosterone and estradiol showed no relation with age. Therefore, the ratio of estradiol to dihydrotestosterone concentration (E2/DHT) in prostate increased with age. The E2/DHT ratio showed a significant positive correlation with the proportion of stroma. CONCLUSIONS: The age-dependent decrease in prostatic dihydrotestosterone and constant estradiol concentration lead to a relatively estrogen-dominant environment compared to that at younger ages. We assume that this relatively estrogen-dominant status induces stromal proliferation by some mechanism and leads to the development of BPH.     

Reduction in prostatic concentration of endogenous dihydrotestosterone in rats by hyperprolactinemia

Persistently high levels of circulating prolactin can be achieved in male rats when pituitaries from female donors are grafted under the renal capsule. Enhanced growth of the lateral prostate can be accomplished by an apparent synergistic effect of excess prolactin with subphysiologic doses of testosterone. To define this effect we measured the endogenous dihydrotestosterone (DHT) concentrations in rat prostate lobes. Twenty Sprague-Dawley male rats (250 g) were castrated and given subcutaneous implants of silastic tubing filled with 1 cm of crystalline testosterone. Ten of these rats simultaneously received two pituitary grafts under the renal capsule. The remaining ten rats received intrarenal skeletal muscle grafts as controls. Four weeks later, the weight (mg +/- SE) of the ventral and dorsal lobes were not significantly different between the two groups, while the lateral lobe was significantly (P less than 0.01) heavier in rats with pituitary grafts (121.4 +/- 7.5) than in controls (81.7 +/- 10.4). Endogenous DHT in the tissue extracts was determined by radioimmunoassay following KMnO4 treatment to eliminate testosterone. The concentration of DHT (ng/g tissue +/- SE) was significantly (P less than 0.01) lower in the lateral prostate of graft-bearing animals (4.40 +/- 0.29) than in the controls (5.98 +/- 0.34). These results indicate that, in the rat, hyperprolactinemia induced by pituitary grafts is associated with a heavier lateral prostate and a lower concentration of endogenous DHT in that tissue. These results also suggest that the action of prolactin in the rat prostate is not mediated through the action of androgen.     

Research progress on the relationship between sex hormone-binding globulin and male reproductive system diseases

Sex hormone-binding globulin, also known as testosterone–estradiol-binding globulin, is a multifunctional protein synthesised by hepatocytes. Sex hormone-binding globulin specifically binds and transports sex hormones to regulate plasma bioactive sex hormone levels and affects their bioavailability. As male sex hormone expression is dominated by testosterone, the binding of sex hormone-binding globulin with testosterone leads to the reduction in bioavailable testosterone, which cannot fulfil its physiological roles, thereby resulting in male infertility, erectile and gonadal dysfunction, prostate cancer and other male reproductive system diseases. Sex hormone-binding globulin may be involved in the pathogenesis of male reproductive system diseases, seriously affecting the quality of life of men. In this article, we review the association between sex hormone-binding globulin and male reproductive system diseases.     

Changes in testosterone and dihydrotestosterone levels in male rat accessory sex organs, serum, and seminal fluid after castration: establishment of a new highly sensitive simultaneous androgen measurement method

It is known that abnormal androgen dynamics in the tissues is a cause of androgen-dependent disorders. Investigation of tissue androgen levels could provide a clue to the elucidation of disorders. However, it is difficult to measure a trace amount of androgen in the tissues. We established a highly sensitive simultaneous quantification method of testosterone and dihydrotestosterone (DHT), which play the most important roles in the body among androgenic steroids in trace amounts, and investigated time course changes in testosterone and DHT levels in male accessory sex organs, serum, and seminal fluid after castration in rat models. In addition, changes in the testosterone/DHT ratio of male accessory sex organs and seminal fluid were observed. The simultaneous testosterone and DHT measurement method established by us was validated. Intra-assay variation and interassay precision and accuracy were all within +/-20%, and the quantification limits of testosterone and DHT were both 15.6 pg/g. With the use of this method, the testosterone and DHT levels in the prostate, seminal vesicles, and serum immediately after castration were similar to those previously reported. The testosterone and DHT levels were 350 pg/g and 605 pg/g, respectively; which showed dominance of DHT in seminal fluid, although it was not as marked as that in the male accessory sex organs. Androgens decreased with time after castration in the accessory sex organs, serum, and seminal fluid. In the prostate and seminal vesicles, testosterone and DHT decreased to about 50% and about 2% of the normal levels, respectively, 72 hours after castration. The serum levels were under the quantification limits 6 hours after castration and thereafter. In seminal fluid, the testosterone and DHT levels decreased to 49% and 35% of normal levels, respectively, 72 hours after castration. The testosterone/DHT ratio in the male accessory sex organs was lower in the prostate (0.06) than in the seminal vesicles (0.13) immediately after castration. In the seminal fluid, changes in the ratio were small compared with those in the accessory sex organs and serum. These results showed that our method was capable of measuring testosterone and DHT in very small amounts of samples such as prostate biopsy specimens, and it might provide a clue to the elucidation of the pathology of androgen-dependent disorders.     

Effect of age and sex on rat endocrine pancreas

Maximal glucose-stimulated insulin secretion was quantified in perfused pancreases of 11-wk-old and 12-mo-old female and male rats. In addition, measurements were made of body weight, total pancreatic weight, and percentage of the pancreas occupied by islet tissue. Body weight (mean +/- SE) of male rats was greater than that of female rats at both 11 wk (319 +/- 3 vs. 237 +/- 13 g) and 12 mo (684 +/- 17 vs. 376 +/- 13 g) of age. Pancreatic weight and percentage of the pancreas occupied by islet tissue were also greater in male rats and increased in approximate proportion to the gain in weight. The first phase and the second phase of maximal glucose-stimulated insulin secretion were both qualitatively and quantitatively similar in all four groups of rats. However, because islet cell mass increased with age, maximal glucose-stimulated insulin secretion declined with age in rats of both sexes when expressed per unit islet tissue. Although the fall in insulin secretion (per islet cell mass) with age was observed in perfused pancreases from both male and female rats, the pancreases of female rats contained relatively less islet tissue and secreted more insulin per unit islet cell mass than pancreases of male rats at either age. Thus, there are sex differences in both islet cell structure and function, but the effect of age on endocrine pancreatic function seems to be independent of sex.     

Benign periurethral mass lesions: the female prostate

Bladder outflow obstruction in the female is usually a functional complaint, but rarely may be of organic aetiology, due to peri-urethral mass lesions. These lesions are usually midline in location and present clinical and roentgenological features comparable with those of prostate in the male. Two such patients, one of pure fibroma and another of hyperplastic glandular tissue are presented and their management is discussed.     

Inhibition of vascular endothelial cell growth factor suppresses the in vivo growth of human prostate tumors

The LNCaP human prostate cancer cell line is androgenand stromal-dependent for in vivo growth. We co-inoculated LNCaP cells with human fetal fibroblasts, isolated from prostate, bone (male), and lung (male and female) derived from 18- to 22-week-old human fetal tissue, into non-castrate male nude mice. Co-inoculation of LNCaP with fetal prostatic fibroblasts resulted in high tumor take rates (27 of 30, or 90%) 6 to 8 weeks after subcutaneous co-inoculation. Serum prostate specific antigen (PSA) values correlated strongly with wet tumor weight (r=0.86). The fetal fibroblast enhancement of tumor take rates in vivo was neither gender- nor organ-specific. Fetal fibroblast-conditioned medium (CM) did not have a significant proliferative effect on LNCaP cell growth in vitro. Areas of angiogenesis were demonstrable in all tumors, with blood vessels arising at the interface between stromal and tumor cells. The fetal fibroblasts, but not the LNCaP cells, expressed significant amounts of the mRNA and protein for vascular endothelial cell growth factor (VEGF). Treatment of tumor-bearing animals with neutralizing antibodies to VEGF resulted in significant tumor growth suppression. These findings indicate that VEGF is an important mediator of stromal-induced enhancement of human prostate cancer cell growth in vivo.     

The role of a clinical score in the assessment of ambiguous genitalia

OBJECTIVE: To improve the initial assessment of ambiguous genitalia in infants. SUBJECTS AND METHODS: Using a specially devised scoring system, the external genitalia (external masculinization score, EMS, range 0-12) and internal reproductive structures (internal masculinization score, IMS, range 0-10) were assessed in 426 male newborns and 291 cases of ambiguous genitalia. RESULTS: In normal male newborns, the median (10th centile) EMS was 11 (10). In the affected infants, the sex of rearing was male in 202 and female in 89 cases, respectively. The median (10-90th centile) EMS in those cases reared male, at 3.5 (2-8), was significantly higher than in cases reared as females, at 2 (1-6) (P < 0.001). The median IMS in cases reared as males and females was the same, at 10, but the scatter of values was higher for males (10-90th centile, 4-10) than for females (0-10) (P = 0.01). Infants reared as females were more likely to have a micropenis, a uterus and/or a urogenital sinus, but there were 12 cases where the sex of rearing was male despite the presence of a uterus; five infants without micropenis were reared as female and 23 with a urogenital sinus were reared as male. CONCLUSION: The masculinization score provides a standardized format to summarize clinical features in newborn infants with ambiguous genitalia. Gender assignment does not solely depend on the appearance of the external genitalia and the nature of internal sexual organs.     

Endometriosis in the male

An 83-year-old man with an endometrioma of the lower abdominal wall has been reported. This occurred following the administration of 25 mg of TACE for a period of about 10 years for what was thought to be carcinoma of the prostate. A second transurethral resection done by Dr. R. C. Thompson proved to be adenocarcinoma. Subsequent to this he was continued on TACE. A review of the more commonly accepted theories of the development of endometriosis in the female has been presented. It is pointed out that the separation between the male and female urogenital systems occurs in the embryo between the eighth week and the fourth month. There is always a possibility for remnants of the opposite sex to remain in individuals. No such was seen in the case which is herein reported. Normal phenotype male was demonstrated in the chromosomal evaluation. A review of the literature on endometriosis in the male reveals several cases which have occurred; the origin of which is though to be from the prostatic utricle which is a remnant of the uterus existing in the male. After a prolonged course the patient reported was followed until he died in 1979. There was no recurrence of the abdominal wall mass but persistent low grade carcinoma of the prostate remained. The terminal process was related to cardiovascular disease and not carcinoma of the prostate. There was delay in publication of this unusual case. The original plan was to await final confirmation of the exact pathologic nature of this condition; unfortunately this was never done since a postmortem examination was not performed.(ABSTRACT TRUNCATED AT 250 WORDS)     

Characterization of the chondrogenic and osteogenic potential of male and female human muscle‐derived stem cells: Implication for stem cell therapy

People of all backgrounds are susceptible to bone and cartilage damage, and these injuries can be debilitating. Current treatments for bone and cartilage injuries are less than optimal, and we are interested in developing new approaches to treat these diseases, specifically using human muscle‐derived stem cells (hMDSCs). Our lab previously demonstrated that sex differences exist between male and female murine MDSCs; thus, this paper sought to investigate whether sex differences also exist in hMDSCs. In the present study, we characterized the chondrogenic and osteogenic sex differences of hMDSCs in vitro and in vivo. We performed in vitro osteogenic and chondrogenic differentiation using hMDSC pellet cultures. As demonstrated by microCT, histology, and immunohistochemistry, male hMDSCs were more chondrogenic and osteogenic than their female counterparts in vitro. No differences were observed based on the sex of hMDSCs in osteogenic and chondrogenic gene expression and cell surface markers. For our in vivo study, we transduced hMDSCs with lenti‐BMP2/GFP and transplanted these cells into critical‐sized calvarial defects in mice. MicroCT results revealed that male hMDSCs regenerated more bone at 2 weeks and demonstrated higher bone density at 4 and 6 weeks than female hMDSCs. Histology demonstrated that both male and female hMDSCs regenerated functional bone. Clinical relevance: These studies reinforce that stem cells isolated from male and female patients differ in function, and we should disclose the sex of cells used in future studies. Considering sex differences of hMDSCs may help to improve cell‐based therapies for autologous cell treatment of bone and cartilage damage. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 37:1339–1349, 2019.     

Sex determination in intersexuality in children

The authors performed the treatment of 124 patients with various patterns of androgyny: 51 with false female and 64 with false male hermaphroditism, and 9 patients with true hermaphroditism. Karyotyping of 31 studied patients revealed karyotype 46XY in 13 persons, mosaic type 45X0/46XY in 2 persons with false male hermaphroditism. In patients with false female hermaphroditism karyotype 46XX was identified in 12 persons. In patients with true hermaphroditism karyotype 46XX was identified in 2 subjects and 47XYY in 1 examinee. The authors supposed the term "gonadal" sex to be a determinant for the identification of the sex status in persons with various patterns of androgyny as it could define a "gamete" and "hormonal" sex. The key problem of androgyny is the detection of true causes of various sexual abnormalities and the technique of their prevention. A true "gonadal" sex should be identified during the first month of a child's life. It would permit one to start timely cortisone treatment for adrenopathic false female hermaphroditism an adequate registration of a child: proper name (documented sex), timely examination of its internal genitalia (removal of those sexual organs that contradict to the sex chosen) and planning the further scope and terms of necessary plastic surgery as well as alleviating severe emotional and psychic traumas of a child's parents.     

Sexing Bones: Improving Transparency of Sex Reporting to Address Bias Within Preclinical Studies

Despite knowledge that sexually dimorphic mechanisms regulate bone homeostasis, sex often remains unreported and unconsidered in preclinical experimental design. Failure to report sex could lead to inappropriate generalizations of research findings and less effective translation into clinical practice. Preclinical sex bias (preferential selection of one sex) is present across other fields, including neuroscience and immunology, but remains uninvestigated in skeletal research. For context, we first summarized key literature describing sexually dimorphic bone phenotypes in mice. We then investigated sex reporting practices in skeletal research, specifically how customary it is for murine sex to be included in journal article titles or abstracts and then determined whether any bias in sex reporting exists. Because sex hormones are important regulators of bone health (gonadectomy procedures, ie, ovariectomy [OVX] and orchidectomy [ORX], are common yet typically not reported with sex), we incorporated reporting of OVX and ORX terms, representing female and male mice, respectively, into our investigations around sex bias. Between 1999 and 2020, inclusion of sex in titles or abstracts was low in murine skeletal studies (2.6%–4.06%). Reporting of OVX and ORX terms was low (1.44%–2.64%) and reporting of OVX and ORX with sex uncommon (0.4%–0.3%). When studies were combined to include both sexes and OVX (representing female) and ORX terms (representing male), a bias toward reporting of female mice was evident. However, when the terms OVX and ORX were removed, a bias toward the use of male mice was identified. Thus, studies focusing on sex hormones are biased toward female reporting with all other studies biased in reporting of male mice. We now call upon journal editors to introduce consistent guidance for transparent and accessible reporting of murine sex in skeletal research to better monitor preclinical sex bias, to diversify development of treatments for bone health, and to enable global skeletal health equity. © 2022 The Authors. Journal of Bone and Mineral Research published by Wiley Periodicals LLC on behalf of American Society for Bone and Mineral Research (ASBMR).