[Show abstract][Hide abstract] ABSTRACT: The aim of this study was to design a molecular assay for the diagnosis of Klinefelter syndrome (KS), based on the detection of supernumerary X-chromosomes (X-chs). DNA was extracted from peripheral blood samples of twenty-six 47,XXY males; two 46,XY/47,XXY males; twenty-two 46,XY males; and 15 females; and deaminated. Methylation-specific quantitative polymerase chain reaction (MS-qPCR) was performed using primers for unmethylated and methylated copies of the X-ch inactive-specific transcript (XIST-U and XIST-M) gene. X-ch disomy was determined on the basis of XIST methylation status. Degree of mosaicism in the 46,XY/47,XXY males was compared with karyotype and fluorescent in situhybridization (FISH) results. Data analysis was performed using the Roche; LightCycler software V. 3.5.3., including determination of crossing points (CPs) by fit-point analysis and melting curve analysis. X-ch disomy was detected in all female controls and KS patients; male controls expressed XIST-M only. CPs ranged from 29.5 to 32.5 (standard deviation (s.d.) 0.8) for XIST-U and from 29 to 31 (s.d. 0.6) for XIST-M. Limit of detection of mosaicism was 1%. Based on XIST-U/XIST-M ratios for the two 47,XXY/46,XY patients, the calculated degree of mosaicism (1.8% and 17.8%) was comparable to FISH results (2.3% and 15%, respectively). Turnaround time from DNA deamination to final data analysis was under 9 h. We conclude that MS-qPCR is a sensitive, specific and rapid test for the detection of X-ch disomy, with applicability for the screening and diagnosis of KS, even in the setting of low grade 47,XXY/46,XY mosaicism.
Asian Journal of Andrology 05/2014; · 2.53 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Purpose
We investigated the safety and tolerability of testosterone replacement therapy in adolescents with Klinefelter syndrome.
Materials and Methods
We reviewed the medical records of all consecutive adolescents with Klinefelter syndrome evaluated between 2007 and 2012. Patients receiving testosterone replacement and aromatase inhibitor therapy were identified. Data on demographics, physical characteristics, medical history and serum hormone concentrations were collected for each patient. We evaluated longitudinal changes in serum testosterone, luteinizing hormone and follicle-stimulating hormone as well as changes in body mass index after the initiation of testosterone replacement therapy.
We identified 151 adolescents with Klinefelter syndrome. Mean age at presentation was 11.6 years. Testosterone replacement therapy and aromatase inhibitors were initiated in 110 and 75 patients, respectively, at an average age of 13 to 14 years. Topical testosterone replacement therapy was used in 95% of patients with good clinical efficacy and compliance based on serial serum testosterone values. After the initiation of testosterone replacement therapy average serum testosterone improved from 240 to 650 ng/ml. Serum luteinizing hormone and follicle-stimulating hormone increased with the progression of puberty from 2.6 to 16.6 and 7 to 42 mIU/ml, respectively. No adverse outcomes related to testosterone replacement therapy were reported.
Hormone supplementation with testosterone and aromatase inhibitors in adolescents with Klinefelter syndrome appears to be safe and effective for maintaining serum testosterone within the normal range. Compliance with topical formulations is high. Topical testosterone replacement therapy is not associated with the suppression of endogenous serum luteinizing hormone or follicle-stimulating hormone.
The Journal of urology 05/2014; · 3.75 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Estradiol (E2) modulates testicular functions including steroidogenesis, but the mechanisms of E2 signaling in human testis are poorly understood. GPER-1 (GPR30), a G protein-coupled membrane receptor, mediates rapid genomic and non-genomic response to estrogens. The aim of this study was to evaluate GPER-1 expression in the testis, and its role in estradiol dependent regulation of steroidogenesis in isolated rat Leydig cells and human testis.
Isolated Leydig cells (LC) from adult rats and human testicular tissue were used in this study. Expression and localization studies of GPER-1 were performed with qRT-PCR, immunofluorescence, immunohistochemistry and Western Blot. Luteinizing Hormone (LH) -stimulated, isolated LC were incubated with estradiol, G-1 (GPER-1-selective agonist), and estrogen receptor antagonist ICI 182,780. Testosterone production was measured with radioimmunoassay. LC viability after incubation with G-1 was measured using 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay.
GPER-1 mRNA is abundantly expressed in rat LC and human testis. Co-localization experiments showed high expression levels of GPER-1 protein in LC. E2-dependent activation of GPER-1 lowers testosterone production in isolated rats LCs and in human testis, with statistically and clinically significant drops in testosterone production by 20-30% as compared to estradiol-naïve LC. The exposure to G-1 does not affect viability of isolated LCs.
Our results indicate that activation of GPER-1 lowers testosterone levels in the rat and human testis. The expression of GPER-1 in human testis, which lack ERα, makes it an exciting target for developing new agents affecting testosterone production in men.
PLoS ONE 01/2014; 9(4):e92425. · 3.53 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: To evaluate surgical sperm retrieval rates in adolescents with Klinefelter syndrome and testosterone replacement therapy (TRT).
Academic medical center.
Ten patients with Klinefelter syndrome, aged 14-22 years, treated with testosterone replacement and aromatase inhibitor therapy for a period of 1-5 years before surgical sperm retrieval.
Microsurgical testis sperm extraction with cryopreservation of harvested tissue.
Presence of spermatozoa within testis tissue.
Successful sperm retrieval in 7/10 patients (70%).
Use of topical TRT did not appear to suppress spermatogenesis in adolescents with KS. It is uncertain whether sperm retrieval rates would be higher or lower without testosterone replacement in these young males. Sperm cryopreservation should be discussed in all KS adolescents who are either receiving or considering initiating TRT.
Fertility and sterility 07/2013; · 3.97 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: PURPOSE:: To evaluate the effect of daily peri-operative celecoxib on patient-reported pain control and opioid use following testicular surgery. MATERIALS AND METHODS:: Men scheduled to undergo elective, outpatient microsurgical testicular sperm extraction were prospectively randomized to receive 200 mg of celecoxib or placebo b.i.d., initiated the night prior to surgery, and continued for six days thereafter. Using an 11-point visual analog scale, participants self-reported their post-operative pain level, and use of acetaminophen/hydrocodone for supplemental pain control. Differences in pain scores and opioid use between the two groups of patients were compared using Student's t-test. A p-value <0.05 was considered significant. RESULTS:: At one-year interim analysis, 35/78 (45%) of eligible participants had returned study questionnaires; 34 were included in the final analysis. Patients given celecoxib (16/34) had a significantly lower post-operative opioid use compared to patients given placebo (6 vs. 16 pills, p=0.02). A statistically significant difference in POD #1 and #2 patient-reported pain scores (4 vs. 6, p<0.05 and 3 vs. 5, p=0.03) and opioid use (1 vs. 5 pills, p<0.01 and 2 vs. 4 pills, p=0.02), was seen between the celecoxib and placebo groups, respectively. No study complications were identified. The trial was terminated early based the results of the interim analysis. CONCLUSIONS:: Twice daily celecoxib use, started pre-operatively, significantly reduces patient-reported post-operative pain and opioid use, especially in the early post-operative period. A short course of celecoxib is well-tolerated, and may be effectively used as part of a multimodal post-operative analgesia in patients undergoing testicular surgery for sperm retrieval.
The Journal of urology 04/2013; · 3.75 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Men presenting with chronic pelvic pain syndrome (CPPS) frequently report concomitant erectile dysfunction (ED), but the underlying cause of ED in this patient population has not been previously studied. This study prospectively investigated the aetiology of ED in men with CPPS. The study population comprised 46 men with penile pain or dysorgasmia, and concomitant ED. All participants completed the NIH-CPSI and international index of erectile function- erectile function domain (IIEF-EFD) questionnaires, and underwent penile duplex Doppler ultrasonography (DUS), following intracavernosal trimix injection, to evaluate erectile hemodynamic parameters. Pearson's correlation between NIH-CPSI and IIEF-EFD scores, and between NIH-CPSI score and the erectile response to trimix injections was investigated. The prevalence of mild, moderate and severe CPPS symptoms was 26, 48 and 26% respectively. The severity of ED was mild, moderate or severe in 15, 61 and 24% of men respectively. NIH-CPSI and IIEF-EFD scores were negatively correlated (r = -0.32, p = 0.002). Peak systolic velocity (PSV) and end-diastolic velocity (EDV) were normal in 96 and 100% of men respectively. The majority of men (78%) required ≥2 trimix injections to attain an adequate erection for DUS. NIH-CPSI scores and the number of trimix injections needed were positively correlated (r = 0.22, p = 0.035). The aetiology of erectile dysfunction in men who present with CPPS and concomitant ED is almost always psychogenic. Penile DUS in this population of men is fraught with the potential for error, and frequently necessitates more than one dose of a vasoactive agent.
[Show abstract][Hide abstract] ABSTRACT: To investigate the impact of early hormone replacement therapy (HT) on sperm retrieval rates in patients with Klinefelter syndrome (KS).
A systematic review of the relevant literature using the PubMed NLM database.
There are no randomized controlled trials evaluating the impact of HT on sperm retrieval or reproductive outcomes in men with KS. On average, surgical sperm retrieval rates in men with KS are around 51%, with a range of 28%-69%. Young patient age is the most consistent positive predictor of sperm retrieval. Lower retrieval rates have been reported in a small subset of KS adults who previously received exogenous T, although the nature, duration, and reason for such therapy in these patient subsets are unknown.
Early HT is recommended in patients with KS, but its effect on fertility potential has not been definitively studied. Larger studies are needed to better answer this question. Cryopreservation of sperm-containing semen or testicular tissue from a significant proportion of affected adolescents is possible, even when containing very low numbers of spermatozoa, and should be considered to maximize future fertility potential.
Fertility and sterility 06/2012; 98(2):274-83. · 3.97 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Study Type - Therapy (case series) Level of Evidence 4 What's known on the subject? and What does the study add? Climacturia is present in ∼20-40% of men after radical prostatectomy, and adversely affects sexual satisfaction. Although several strategies have been proposed for the treatment of climacturia, none have been systematically studied to date. This observational study shows that use of a penile variable tension loop can significantly reduce the degree and frequency of orgasm-associated incontinence, and the associated distress experienced by patients and partners. Climacturia resolves completely in half the patients, and occurs occasionally or rarely in the remainder. OBJECTIVE: • To define the impact of the use of a penile variable tension loop on climacturia and on the distress level experienced by patients and their partners as a result of climacturia. PATIENTS AND METHODS: • All patients presenting for sexual function assessment after radical prostatectomy (RP) were questioned regarding climacturia. • The study population consisted of men who had undergone RP < 6 months before initial evaluation, had reported having climacturia on ≥25% attempts where orgasm was achieved, had agreed to use the variable tension loop consistently, and continued to have climacturia when not using the loop at follow-up. • Patients were interviewed regarding the frequency and degree of climacturia, and their own and their partner's distress levels secondary to climacturia, both with and without the use of the variable tension loop. RESULTS: • The study population comprised 124 men. • At baseline, the degree of climacturia was small, moderate, and large in 16%, 72%, 12% of patients, respectively, and 28%, 26% and 0%, respectively, at follow-up (all P < 0.01). • Climacturia occurred rarely, occasionally, most of the time, or always in 15%, 48%, 16% and 21% of cases, respectively, at baseline, and 48% of patients experienced no climacturia with use of the variable tension loop. • Distress was experienced by 14% and 61% of patients and partners at baseline, and 2% and 11% of patients and partners at follow up (P < 0.01). • Severity of distress was significantly lower at follow-up for both partners and patients (P < 0.01). CONCLUSIONS: • Climacturia is a common complication of radical prostatectomy. • Application of the variable tension loop can result in a significant decrease in the frequency and volume of climacturia. • Use of the variable tension loop is a simple and non-invasive strategy for relieving the distress associated with climacturia in patients who have undergone RP and their partners.
[Show abstract][Hide abstract] ABSTRACT: Klinefelter syndrome (KS) remains the most common, yet often undiagnosed, chromosomal aberration in men. Early diagnosis and treatment can improve the health of patients with KS. The aim of this study was to evaluate the inactivation pattern of supernumerary X chromosomes. The secondary aim was to design a reliable and cost-effective molecular test for detection of X chromosome disomy. Methylation-specific polymerase chain reaction (M-PCR), with primers for familial mental retardation (FMR1) and X chromosome inactive-specific transcript (XIST) genes, was used to detect the presence of X chromosome disomy in men. Seventeen fertile males, 12 females, and 35 males with KS (28 with 47,XXY karyotype, and 7 with 47,XXY/46,XY mosaics) were included in the study. Results of the karyotype were compared with the results of semiquantitative M-PCR. Inactivation of X chromosomes was measured by XIST/FMR-1 methylation ratio. Differences in the methylation patterns of FMR1 and XIST genes between 46,XY men and men with X chromosome disomy allowed for rapid detection of the presence of an additional X chromosome, achieving 100% sensitivity and specificity using M-PCR. The methylated:unmethylated FMR1 amplicon ratio allowed the detection of 1 additional X chromosome per 100 normal XY cells (1% of XX/XY mosaicism). In our series, 50% of 47,XXY men showed skewed inactivation of the X chromosome. Men with KS can have incomplete inactivation of supernumerary X chromosomes. M-PCR is a sensitive, specific, fast, and relatively inexpensive test for the diagnosis of X chromosome disomy.
Journal of Andrology 03/2012; 33(5):955-62. · 3.37 Impact Factor