You have accessJournal of UrologyInfections/Inflammation/Cystic Disease of the Genitourinary Tract: Interstitial Cystitis (PD01)1 Sep 2021PD01-12 PELVIC FLOOR MYALGIA AND TEMPOROMANDIBULAR JOINT DYSFUNCTION: A COMMON AND CLINICALLY IMPORTANT RELATIONSHIP8.6.1 Luke* Griffiths, Samir Derisavifard, Daniel Alaiev, Michael Funaro, Micah Levy, Jessica Kreshover, Robert Moldwin, and Sonia Bahani Luke* GriffithsLuke* Griffiths , Samir DerisavifardSamir Derisavifard , Daniel AlaievDaniel Alaiev , Michael FunaroMichael Funaro , Micah LevyMicah Levy , Jessica KreshoverJessica Kreshover , Robert MoldwinRobert Moldwin , and Sonia BahaniSonia Bahani View All Author Informationhttps://doi.org/10.1097/JU.0000000000001965.12AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Pelvic floor muscle tenderness/dysfunction (PFMD) is a cause of pelvic pain commonly occurring with other chronic pain syndromes. Temporomandibular Joint dysfunction (TMJD) is a similar disorder often associated with pain. The current study wished to determine the association between these two conditions. METHODS: Patients were prospectively enrolled in 2 study groups: those with and without exam-proven myofascial pelvic pain. Inclusion into the pain-free group was predicated on the absence of a urinary infection, >18-years-old, PFMD, and the presence of microscopic hematuria without associated pathology. Patients were surveyed using the TMJD screening questionnaire, the International Prostate Symptom Score (IPSS), and the Genitourinary Pain Index (GUPI). Univariate and Bayesian regression models were generated to predict the probability of a TMJD diagnosis. RESULTS: One-hundred patients were enrolled with 50 in each arm. Patients in the pain group were significantly more likely to have a history of TMJD than those without pain (38% vs. 14%, p=0.01). The pain group has a 94.6% posterior probability for higher chance of TMJD than the pain-free group after adjusting for age. Additionally, Patients in the pain group expressed higher rates of lower urinary tract symptoms (IPSS: 14.6 vs. 5.5, p<0.01) and poorer quality of life (GUPI: 26.7 vs. 5.2, p<0.01). CONCLUSIONS: Patients with PFMD associated pain are 94.6% more likely to suffer from TMJD than pain-free patients without PFMD. These data suggest TMJD and PFMD may be functionally related syndromes. Source of Funding: None. © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e35-e36 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Luke* Griffiths More articles by this author Samir Derisavifard More articles by this author Daniel Alaiev More articles by this author Michael Funaro More articles by this author Micah Levy More articles by this author Jessica Kreshover More articles by this author Robert Moldwin More articles by this author Sonia Bahani More articles by this author Expand All Advertisement Loading ...

You have accessJournal of UrologySexual Function/Dysfunction/Andrology: Medical and Non-surgical1 Apr 2014MP48-04 PREDICTORS OF POOR RESPONSE TO TRANSDERMAL TESTOSTERONE THERAPY IN MEN WITH METABOLIC SYNDROME Ashley Winter, Anthony Marte, Morgan Kelly, Michael Funaro, Peter Schlegel, and Darius Paduch Ashley WinterAshley Winter More articles by this author , Anthony MarteAnthony Marte More articles by this author , Morgan KellyMorgan Kelly More articles by this author , Michael FunaroMichael Funaro More articles by this author , Peter SchlegelPeter Schlegel More articles by this author , and Darius PaduchDarius Paduch More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2014.02.1478AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail Introduction and Objectives Testosterone replacement therapy (TRT) is approved for treatment of male hypogonadism, but TRT may be beneficial in hypogonadal men with metabolic syndrome. Anecdotal observations indicate that adiposity is associated with a reduced frequency of men reaching a eugonadal state. We have collected metabolic data on a cohort of men receiving transdermal TRT (T-TRT) to identify predictors of response to T-TRT. Methods We reviewed 58 consecutive patients with metabolic data and hypogonadism presenting to a male reproductive medicine practice. Patients received a physical examination and a full hormonal evaluation including total testosterone (TT) measured using liquid chromatography–mass spectrometry (LC/MS). All men initiated 50mg daily transdermal testosterone. Multivariate logistic regression assessed predictors of failure to reach eugonadal (TT=350ng/dL or free testosterone (FT) =65 pg/mL) status at most recent follow-up (median 21.2 months). Non-obese patients had BMI<30 (Group 1, n=32). Obesity was defined as BMI>30 (Group 2, n=26). Results Baseline age, primary diagnosis, and blood pressure were similar in each group (P>0.05). Initial FT, TT, follicle-stimulating hormone (FSH), cholesterol, and hemoglobin A1c (HbA1c) were also similar between groups (P>0.05). At the end of the study period, more non-obese men attained eugonadal testosterone levels than obese men (FT-81% vs 52% (P=0.03) and TT-81% vs 54% (P=0.018)). On multivariate logistic regression, age and obese BMI were predictive of final FT <65pg/mL (P=0.03 and 0.043). HbA1c >6.4 (diagnostic criteria for diabetes mellitus) predicted TT<350ng/dL (P=0.042). Response to therapy (final TT) was independent of baseline FT and TT in both univariate and multivariate models. Conclusions Hypogonadal men with diabetes and obesity are less likely to achieve normal free and total testosterone levels on topical testosterone therapy relative to non-obese, non-diabetic men. This relationship occurred independent of baseline hormone profile. The underlying mechanisms and optimal management of failure to respond to T-TRT needs to be determined. Health care practitioners and patients should aware of predictors of response. © 2014FiguresReferencesRelatedDetails Volume 191Issue 4SApril 2014Page: e528 Advertisement Copyright & Permissions© 2014Metrics Author Information Ashley Winter More articles by this author Anthony Marte More articles by this author Morgan Kelly More articles by this author Michael Funaro More articles by this author Peter Schlegel More articles by this author Darius Paduch More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Purpose 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. Materials and Methods 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. Results 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. Conclusions 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.

Diagnostic tests should detect disease, have prognostic value, and aid in clinical decision making. Nowhere else in laboratory medicine does one have to interpret a subject's results within the dynamic of a couple as in reproductive medicine. Abnormal markers of male reproduction do not necessarily mean sterility, but instead indicate problems with spermatogenesis, sperm maturation, transport through epididymis and ejaculatory duct, or abnormal ejaculatory function. Decades of research suggest that one test will never fit all scenarios and a battery of assays evaluating different aspects of male reproduction will likely have the best prognostic value. There is a strong need for standardization and harmonization of evolving assays to establish their clinical relevance. Next-generation genome sequencing and the discovery of small noncoding RNAs in sperm already are changing the field and permit further insight into the biology of male reproduction as well as offer new diagnostic tests.

Men with KS (47 XXY), frequently demonstrate resistance to exogenous testosterone. Posttranslational splicing of Filamin A (FlnA) into smaller fragments is a critical step for normal trafficking of activated AR and for activation of androgen response elements on DNA (1). The aim of this study was to show that abnormal expression and splicing of FlnA is present in KS and may explain retention of AR within cytoplasm and androgen resistance in KS. Tissue repository analysis study. 10 testicular biopsy samples were obtained from normal male patients (control); 7 testicular biopsy samples were obtained from men with KS. Tissue was prepared with Qiagen AllPrep DNA/RNA/Protein Mini kit. To quantify AR, nuclear and cytoplasmic protein fractions were extracted from snap-frozen testicular tissues. Amounts of AR and HSP-90 proteins were measured with ImageJ and after OD normalization expressed as a ratio of nuclear to cytoplasmic (N/C) AR. Quantitative real-time PCR (qRT-PCR) was performed using Roche Lightcycler 480. Gene expression was normalized to housekeeping gene HPRT. Western blots were run using novex 3-8% tris-acetate gels, Santa Cruz N20 AR, Millipore MAB1680 FlnA, and Sigma A1978 B-Actin antibodies. Expression was quantified using Li-Cor Odyssey. In patients with KS (N/C=2.1 ± 1.1) there was 3.5 times less AR in nucleus than in cytoplasm as compared to controls (N/C=7.1 ± 1.3) p=0.03, despite normal testosterone levels indicating retention of AR in cytoplasm in KS. qRT-PCR showed 3.4x higher expression of FlnA and elevated AR mRNAs expression in KS vs. normal men. Men with KS had an increase in whole FlnA protein with an 80% decrease in the expression of FlnA fragments (required to direct AR toward nucleus) as compared to controls.Table 1Expression of FlnA and AR mRNA and proteins.qRT-PCR Normalized ExpressionNormalKSAR (Target/Ref)16.5330.77FlnA (Target/Ref)11.8440.33Western Blot - Normalized Expression ratiosKS/NormalAR0.88FlnA (280)1.10FlnA (110)0.26FlnA (90)0.16 Open table in a new tab Men with KS have 3x higher retention of activated AR in cytoplasm, indicating a defect in trafficking of AR toward the nucleus. The 82% decrease in levels of FlnA 90 and 74% reduction in FlnA 110 observed in KS clearly support our hypothesis that abnormal trafficking of AR due to abnormal cleavage of FlnA is responsible for androgen resistance in KS. This study may give rise to new therapeutic avenues to treat men with hypogonadism not responding to treatment.

You have accessJournal of UrologyInfertility: Basic Research, Physiology, Pathophysiology1 Apr 20132037 AGE DEPENDENT DEGENERATION OF SERTOLI AND LEYDIG CELLS IN ADOLESCENTS AND MEN WITH KLINEFELTER SYNDROME: A HISTOLOGICAL ANALYSIS Michael Funaro, Akanksha Mehta, Russell Simons, Alexander Bolyakov, Anna Mielnik, Maria Shevchuk, Peter Schlegel, and Darius Paduch Michael FunaroMichael Funaro New York, NY More articles by this author , Akanksha MehtaAkanksha Mehta New York, NY More articles by this author , Russell SimonsRussell Simons New York, NY More articles by this author , Alexander BolyakovAlexander Bolyakov New York, NY More articles by this author , Anna MielnikAnna Mielnik New York, NY More articles by this author , Maria ShevchukMaria Shevchuk New York, NY More articles by this author , Peter SchlegelPeter Schlegel New York, NY More articles by this author , and Darius PaduchDarius Paduch New York, NY More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.2456AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Klinefelter syndrome (KS) is the most common chromosomal disorder leading to male infertility and hypogonadism. Limited clinical observations suggest that meiotic divisions during puberty lead to progressive loss of germ cells and testicular atrophy in KS. Sertoli (SC) and Leydig (LC) cells divide mitotically during puberty and are believed to be non-dividing cells in adulthood. Little is known about the effects of age on Sertoli and Leydig cell in KS patients after adolescence. This study aimed to correlate the relationship between patient age and testicular morphology on testis biopsy in 47,XXY men. METHODS Open testicular biopsies from 17 KS patients were preserved in Bouin's solution, embedded in paraffin, sectioned (0.5 um), and fixed on slides stained with H&E. The diameter of round seminiferous tubules (SFTs), LC internuclear distance, and number of SC per SFT were measured. At least 10 cross-sections of SFTs and 100 LCs were measured for each slide using light microscopy and morphometry software (Nikon NIS Elements®). RESULTS Testicular biopsies from six adolescents (mean age 15.8, 95%CI: 14.9-16.6), and 11 adults (mean age 34.7, 95%CI: 33.3-36) were available for analysis. There was an age-associated decrease in SFT diameter (R2=0.35, p=0.009), LC size (R2=0.28, p=0.03), and number of SCs per SFT (R2=0.53, p=0.0009) (Fig. 1). Mean number of SCs per SFT was lower in adults compared to adolescents: (8.5 vs. 35.6, p=0.0006). In men older than 20, decreases in number of SCs per SFT correlated with decrease in SFT diameter (R2=0.82, p<0.0001). LCs in adolescents were larger than in adults (19 um vs. 15.6 um, p<0.04). CONCLUSIONS Progressive testicular atrophy appears to correlate with loss in number of SCs in adult men with KS. Our findings strongly suggest that Sertoli cells undergo further morphological deterioration after puberty as opposed to Leydig cells, which, although smaller in adults, seem to be less affected by age. Age-related changes in testicular morphology may guide interventions for maximizing fertility potential in KS patients. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e836-e837 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Michael Funaro New York, NY More articles by this author Akanksha Mehta New York, NY More articles by this author Russell Simons New York, NY More articles by this author Alexander Bolyakov New York, NY More articles by this author Anna Mielnik New York, NY More articles by this author Maria Shevchuk New York, NY More articles by this author Peter Schlegel New York, NY More articles by this author Darius Paduch New York, NY More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Sperm chromatin damage has been associated with male infertility, increased risk for spontaneous abortion, and poor embryo development. Available methods for detecting chromatin damage render the sperm no longer suitable for clinical use. Early apoptotic events resulting in chromatin damage are associated with increased permeability of the cell membrane to large ions. We propose the use of a large fluorescent organic cation, proprietary fluorochrome (PF-1), for fluorescence-activated cell sorting (FACS) for negative selection of sperm without chromatin damage. Sperm with chromatin damage are PF-1 positive. Performance of cell sorting by PF-1 was verified with terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) after FACS on PF-1(+) and PF-1(-) subpopulations. Whereas 19.5% of PF-1 positive sperm were TUNEL positive only 1.5% sperm in the PF-1(-) fraction were TUNEL positive (p < 0.00001). TUNEL values below 1.9% were considered background fluorescence. Post-sorting motility and vitality were 49.4% (SD: 12.5) and 65.0% (SD: 14.99), respectively. Proprietary fluorochrome activated sperm sorting may decrease or most likely eliminate all of TUNEL positive sperm without adverse effects on viability, providing a new therapeutic avenue for men with a high percentage of TUNEL positive sperm. Further research is needed to determine if the reduction in TUNEL positive sperm using PF-1 will improve in vitro fertilization (IVF) outcomes.

Apoptosis has been implicated in sperm chromatin damage; it is unclear whether apoptosis occurs through cytoplasmic or mitochondrial pathways. Sperm has minimal volume of cytoplasm but prominent mitochondria. Propidium iodide (PI), annexin V (AV), DiIC1(5) and proprietary fluorochrome (PF-1) were used to investigate apoptosis activation in human sperm using multichannel flow cytometry. There was a time-dependent increase in staining of spermatozoa with both AV and PF-1 and decrease in mitochondrial staining with DiIC1(5). These results strongly suggest that the drop in mitochondrial potential precedes changes in membrane phospholipids, and thus suggest apoptotic activation through mitochondrial pathway in human spermatozoa.