bluuebunny/crossref_metadata_2025 huggingface dataset:
Vasopressin Receptors in Human Seminal Vesicles: Identification, Pharmacologic Characterization, and Comparison with the Vasopressin Receptors Present in the Human Kidney
Authors: MAGGI MARIO , BALDI ELISABETTA , GENAZZANI ALESSANDRO DAVID , GIANNINI STEFANO , NATALI ALESSANDRO , COSTANTINI ALFIERO , RODBARD DAVID , SERIO MARIO
Published: December 2015

Because of the presence of a high density of vasopressin receptors in the epithelial cells of porcine seminal vesicles similar to the V2 vasopressin receptors of renal tubules, human seminal vesicles and kidney were investigated using quantitative binding and adenylate cyclase studies. Tissues were obtained at surgery from 17 patients with urologic diseases. A homogeneous class of vasopressin binding sites have been found in both seminal vesicles and renal medulla. However, the vasopressin receptors present in these tissues are different in terms of ligand specificity and adenylate cyclase activation. In seminal vesicles, the V1 vasopressin antagonist d(CH2)5 TyrMeAVP is 36‐fold, more potent than the V2 agonist dVDAVP in displacing [3H]AVP binding, while in the medullopapillary portion of kidney dVDAVP is 24‐fold, more selective than d(CH2)5 TyrMeAVP for the arginine vasopressin binding site. Furthermore, arginine vasopressin induces a dose‐dependent increase in adenylate cyclase activity in renal membranes, while it was ineffective in seminal vesicle membranes. These results indicate that a very high affinity (0.2 nM), low capacity (14 fmoles/mg protein) class of vasopressin receptors is present in human seminal vesicles, having pharmacologic characteristics similar to the V1 subtype of vasopressin receptors. The presence of a high affinity (1.6 nM), high capacity (350 fmoles/mg protein) V2 subtype of vasopressin receptors in human renal membranes is also confirmed. The density of the vasopressin receptors present in human seminal vesicles is inversely correlated with patient age, consistent with a physiologic role for vasopressin in the regulation of accessory sex gland activity.
Testicular Dysfunction in the Adjuvant‐Induced Arthritic Rat
Authors: CLEMENS JEFFREY W. , BRUOT BRENT C.
Published: December 2015

Adjuvant‐induced arthritis, an autoimmune disease similar to rheumatoid arthritis, was used to investigate possible mechanisms of immune system modulation of the reproductive system. This laboratory previously reported that arthritic male rats have reduced serum testosterone and elevated serum LH concentrations. In the experiments described here, serum prolactin levels were not significantly different in arthritic animals compared with non‐injected control animals. Neither reduced food consumption of arthritic rats nor the injection vehicle appear to cause a reduction of serum testosterone. Serum corticosterone was significantly elevated in the arthritic group compared with both the non‐injected or the vehicle‐injected control animals. Testicular cells from arthritic animals secrete significantly less testosterone in vitro compared with cells from non‐injected control animals, both basally and in response to dbcAMP and hCG. In summary, the reduced serum testosterone of arthritic animals appears to be the result of a testicular dysfunction.
The Differential Effects of the Indazole‐Carboxylic Acid Derivative, Tolnidamine, on Sertoli Cell Protein Secretion
Authors: ROSSI VILMA , CHENG C. YAN , GUNSALUS GLEN L. , BARDIN C. WAYNE , SPITZ IRVING M.
Published: December 2015

The indazole‐carboxylic acid derivative tolnidamine (TOL) has marked antispermatogenic activity in rats. Previous morphological and biochemical studies indicate that Sertoli cells are one of the targets of this compound. The aim of this study was to assess the effect of TOL on the in vitro secretory functions of primary Sertoli cell‐enriched cultures prepared from rats of different ages by monitoring the changes of three known Sertoli cell proteins, androgen binding protein (rABP), transferrin (rTF), and testibumin (rTB). The addition of TOL at the beginning of the culture period reduced the plating efficiency of Sertoli cells; however, TOL did not induce a significant change in cell number if it was added 24 h after plating of the cells. Sertoli cell‐enriched cultures prepared from testes of 10‐day‐old rats were highly sensitive to TOL as evidenced by a marked inhibition in secretions of rABP, rTB, and rTF in all experiments. In cultures prepared from 15‐ and 20‐day‐old rats, TOL had no apparent effect on rABP secretion, but reduced rTF and increased rTB secretion. Thus, TOL has a differential effect on the secretion of individual proteins in Sertoli cells cultured from rats between 10 and 20 days of age. This phenomenon is presumably a consequence of the progressive maturation of Sertoli cells in the seminiferous epithelium.
Stimulation of the Proliferation and Differentiation of Leydig Cell Precursors after the Destruction of Existing Leydig Cells With Ethane Dimethyl Sulphonate (EDS) Can Take Place in the Absence of LH
Authors: TEERDS KATJA J. , de ROOIJ DIRK G. , ROMMERTS FOCKO F.G. , VAN DER HURK ROBERT , WENSING CEES J.G.
Published: December 2015

In hypophysectomized rats, 2 days after the administration of the cytotoxic drug ethane dimethyl sulphonate (EDS), the proliferative activity of Leydig cell precursors increased six‐fold. Thus, factors other than LH act locally to stimulate the proliferation of precursor cells after EDS. Twenty‐six days after EDS administration, neither cells with the morphological characteristics of Leydig cells nor histochemical enzyme activities, such as 3 beta‐HSD and alpha‐naphtyl esterase, could be detected in testis tissue. In hypophysectomized rats treated daily with hCG (100 iu) for 7 days, starting at 26 days after EDS, the number of Leydig cells was increased to 48 ± cells (per 1000 Sertoli cells), which is approximatley 4.5% of the intact control level. 3 beta‐HSD and alpha‐naphtyl esterase activity could be detected, and plasma testosterone levels had increased 15‐fold compared with the hypophysectomized controls. These results show that proliferation and some differentiation of precursor cells along the Leydig cell lineage can occur independent of LH, but the final stages of the differentiation process require hCG stimulation.
Prolonged Suppression of Rat Testis Function by a Depot Formulation of Zoladex,® a GnRH Agonist
Authors: WARD JERRY A. , FURR BARRINGTON J.A. , VALCACCIA BARBARA , CURRY BRENDA , BARDIN C. WAYNE , GUNSALUS GLEN L. , MORRIS IAN D.
Published: December 2015

A sustained‐release formulation of a potent gonado‐tropin‐releasing hormone (GnRH) agonist, Zoladex® (D‐Ser(But),6 Aza Gly10‐GnRH; ICI 118,630; goserelin), was administered subcutaneously (3.6 mg/depot) to male rats once every 28 days for 2–24 wk to determine the extent to which pituitary‐testis function could be suppressed and whether suppression was maintained throughout the period of treatment. Administration of Zoladex resulted in sustained decreases in weight of the testis, epididymis, seminal vesicles and prostate gland. The decreases were apparent within 2 wk of initiating treatment. Patchy degeneration of the seminiferous tubules and atrophy of the Leydig cells were observed, but did not progress beyond the degree observed after 1 month of treatment. Serum and testis testosterone were markedly depressed after 2 wk of treatment, as was testis [125I]hCG binding. Serum gonadotropins were also reduced by treatment. Serum androgen binding protein (ABP) was elevated, testis ABP content remained unchanged, and epididymal ABP content was reduced. The changes are consistent with the hypothesis that this compound affects both the anterior pituitary gland and the testis. These findings indicate that depot delivery systems are a convenient way to administer GnRH analogs for sustained treatment schedules.
Quantitative Assessment of Nocturnal Penile Tumescence and Rigidity In Normal Men Using a Home Monitor
Authors: BURRIS ALLEN S. , BANKS STEVEN M. , SHERINS RICHARD J.
Published: December 2015

Current methods now permit the measurement of nocturnal penile tumescence and rigidity (NPTR) in men with erectile dysfunction. But the relationship of rigidity to tumescence and the changes in rigidity with age have not been defined in nornal men. Accordingly, the authors assessed NPTR in 47 normal men using a portable, take‐home monitor (Rigiscan). Penile tumescence time was found to decrease with advancing age (p < 0.05), whereas the number of erectile episodes and penile rigidity did not significantly change with age for men in the third through sixth decades (p < 0.05). Using area‐under‐the‐curve as an integrated measure of amplitude and duration, significant correlations between tumescence and rigidity (p < 0.001), and between tip and base measurements (p < 0.001) were found. With these normative data, prospective studies should determine the sensitivity and specificity of various NPTR parameters in the diagnosis of erectile dysfunction.
Underestimation of Subfertility Among Relatives When Using a Family History: Taboo Bias
Authors: van, Der Avoort Irene A.M. , van Golde Ron J.T. , Tuerlings Joep H.A.M. , Kiemeney Lambertus A. , Meuleman Eric J.H. , Braat Didi D.M. , Kremer Jan A.M.
Published: December 2015

ABSTRACT: Family history is widely used in clinical practice and research in order to study genetic aspects of disorders in general, and is recommended as a tool in the assessment of male subfertility. Unfortunately, little is known about the validity of this tool. In this survey, we sent questionnaires to 474 randomly selected men aged 25–40 years in order to collect data on subfertility among them and their relatives. A nonresponder study was also conducted in order to evaluate selection bias. A personal interview was also performed with some respondents in order to gauge how well the data corresponded with questionnaires that were returned. Two hundred forty‐three men (51.3%) completed the questionnaire. The responders reported a significantly lower prevalence of subfertility among their relatives than among themselves. Among brothers, the reported prevalence was about 5 times lower (ie, 3.6%) than among responders (15.3%). The nonresponder study and personal interviews showed that these differences were not caused by a selective response to the survey or by the use of a questionnaire instead of a personal interview. We conclude that subfertility among relatives is severely underestimated through the use of family history, probably because of the taboo of discussing subfertility. Knowledge of subfertility may spread selectively within families, causing substantial misclassification. Therefore, researchers and clinicians should be aware that an inquiry of family history is likely to lead to underestimation of subfertility among relatives.
Rapid Disappearance of Spermatozoa After Vasal Occlusion in the Dog
Authors: Schiff Jonathan D. , Li Philip S. , Schlegel Peter N. , Goldstein Marc
Published: December 2015

ABSTRACT: The reproductive system of the male dog is unusual in that the seminal vesicles and bulbourethral glands are absent. Therefore, we chose the dog as a model to evaluate the effect of seminal vesicles on clearance of spermatozoa from the male reproductive tract after vasal occlusion. Thirty adult male beagle dogs with ejaculates containing at least 500 × 106 sperm with greater than 90% motility were used for this study. The dogs' vasa were occluded percutaneously using a Vasocclude clip‐applying device through a small scrotal puncture site. Dogs were ejaculated and semen analysis was performed before and after vas occlusion. The first 24 dogs were completely azoospermic 1 week following vas occlusion. In order to explain these unanticipated results, an additional 6 dogs were evaluated to determine the specific time course of sperm disappearance from the ejaculate at 1, 3, 5, and 7 days after vas occlusion. The results revealed that spermatozoa were almost completely absent within 1 day after vas occlusion (99.9% reduction, × = 1.0 + 1.1 × 106 sperm per milliliter at 1 day, P < .0005 vs prevas occlusion). The rapid elimination of spermatozoa after vas occlusion indicates that sperm transit rapidly through the vas deferens in dogs. Therefore, the delayed clearance of spermatozoa from the ejaculate in humans may be due to sperm storage in the seminal vesicles.
Effects of Medical or Surgical Castration on Erectile Function in an Animal Model
Authors: Traish Abdulmaged M. , Munarriz Ricardo , O'Connell Luke , Choi Seong , Kim Soo Woong , Kim Noel N. , Huang Yue‐Hua , Goldstein Irwin
Published: December 2015

ABSTRACT: The goal of this study was to investigate the effects of medical castration (luteinizing hormone‐receptor hormone [LH‐RH] agonist treatment) or surgical castration on erectile function in an animal model. New Zealand White male rabbits were either kept intact (control); surgically orchiectomized; or treated for 2, 4, or 8 weeks with the LH‐RH agonist leuprolide acetate (107 μg/kg/mo). At 2 weeks, plasma testosterone levels of orchiectomized and leuprolide acetate—treated animals were 12.8% and 57.4% of intact control animals, respectively. Erectile function was assessed by continuously recording systemic arterial pressure (SAP) and intracavernosal blood pressure (ICP) and determining the ICP:SAP ratios in response to electrical stimulation of the pelvic nerve at varying frequencies (2.5–32 Hz). Androgen deprivation by surgical (orchiectomy) or medical (leuprolide acetate) castration reduced ICP at all frequencies tested but did not alter SAP. Administration of the phosphodiesterase type 5 inhibitor vardenafil (10 μg/kg) did not enhance ICP in surgically orchiectomized or leuprolide acetate—treated animals. Nitric oxide synthase and arginase activities in the corpus cavernosum were not significantly altered by surgical or medical castration. Further, Masson trichrome staining of erectile tissue from androgen‐ablated animals showed a reduction in smooth muscle content. These data demonstrate that androgen deprivation achieved by surgical or medical castration adversely affects penile hemodynamics and erectile function without producing significant changes in the activities of nitric oxide synthase or arginase. We conclude that androgen deprivation produces structural alterations in the corpus cavernosum leading to corporal veno‐occlusive dysfunction.
Osmotic Tolerance Limits and Properties of Rhesus Monkey (Macaca mulatta) Spermatozoa
Authors: Rutllant Josep , Pommer Angela C. , Meyers Stuart A.
Published: December 2015

ABSTRACT:Fundamental cryobiological characteristics of rhesus spermatozoa must be determined for successful cryopreservation techniques to be established. The main objectives of the present study were to determine the osmotic behavior and osmotic tolerance limits of rhesus macaque spermatozoa. Cell volume changes over anisotonic conditions were assessed using an electronic particle counter and sperm motility was evaluated with a computer‐assisted sperm analysis system. Analysis of membrane integrity and mitochondrial membrane potential was performed using flow cytometry. Rhesus monkey spermatozoa behave as linear osmometers in the osmotic range tested (75–900 mOsmol kg−1), as shown by the Boyle van't Hoff plot (r2= .99). Rhesus spermatozoa have a mean cell volume of 36.8 ± 0.5 μm3at 22°C, with 77.2% of the intracellular volume being osmotically inactive. Results regarding sperm tolerance to osmotic stress showed that sperm motility was more sensitive than membrane integrity to deviations from isotonicity and, in addition, that rhesus sperm motility and membrane integrity were more sensitive to hypertonic than hypotonic conditions. Mitochondrial membrane potential did not explain the lack of sperm motility observed under anisosmolal conditions in our study. Although most spermatozoa were able to recover initial volume after osmotic stress, they were not able to recover initial motility.
Immunolocalization of the Yb1 Subunit of Glutathione S‐transferase in the Adult Rat Epididymis Following Orchidectomy and Efferent Duct Ligation
Authors: Andonian Sero , Hermo Louis
Published: December 2015

ABSTRACT: In addition to the maturation of sperm, the epididymis also serves to protect sperm from harmful reactive oxygen species. To this end, various antioxidant enzymes are produced by the epididymis, such as glutathione S‐transferases (GSTs), a family of dimeric proteins that catalyze the conjugation of glutathione to various electrophilic compounds, thus providing cellular detoxification. In the present study, the regulation of the Yb1 subunit of GST was examined in Bouin‐fixed epididymides of adult control, orchidectomized (O) rats with or without testosterone (T) supplementation and efferent duct—ligated (EDL) rats using light microscope immunocytochemistry with an anti‐Yb1—GST antibody. The intensely reactive ciliated cells of the efferent ducts and principal cells of the epididymis showing a checkerboard staining pattern were unaltered in their expression of Yb1‐GST after all experimental procedures, suggesting their regulation by factors other than of testicular origin. On the other hand, the intense reaction of narrow/apical cells and moderate reaction of basal cells of the proximal initial segment of control animals became negligible in O rats and was not restored with T supplementation. As staining was also absent after EDL, the data suggest that a luminal testicular factor(s), other than androgens, regulates expression of Yb1‐GST in narrow/apical and basal cells of the proximal initial segment. Although basal cells of the caput and cauda epididymidis were unreactive after all experimental protocols, as also noted in controls, the intensely reactive basal cells of the corpus epididymidis of control animals became unreactive in O animals. However, Yb1‐GST expression was restored to these cells with T supplementation, and as there was no effect on Yb1‐GST expression after EDL, the data suggest that circulating testosterone or one of its metabolites regulates expression of Yb1‐GST in basal cells of the corpus region. Taken together, these data indicate a differential regulation with respect to the expression of Yb1‐GST in the various cell types and regions of the epididymis.
A Castrated Mouse Model of Erectile Dysfunction
Authors: Palese Michael A. , Crone Julie K. , Burnett Arthur L.
Published: December 2015

ABSTRACT: To establish a mouse model for the study of venoocclusive erectile dysfunction, we investigated erectile function in wild‐type (WT), castrated (CAST), and castrated mice receiving immediate testosterone replacement (TEST). Adult C57BL6 mice (∼30 g) underwent electrical stimulation of the cavernous nerve in vivo (parameters: 16 Hz frequency, 5 ms duration, 4V stimulatory voltage) with intracavernosal pressure (ICP) monitoring. A total of 55 mice (5 WT, 25 CAST, and 25 TEST) were evaluated. CAST and TEST (5.0 mg/pellet, 60‐day release) mice were divided into groups of 5 and evaluated at 24 hours, 72 hours, 1 week, 2 weeks, and 4 weeks. Penile tissue was immunohistochemically stained for α‐actin (marker for smooth muscle cells) and CD‐31 (marker for endothelial cells). Stained slides were analyzed using Image Pro‐plus software. In secondary studies, a Doppler flow meter was employed to evaluate penile blood flow. ICP measurements (mm Hg) were significantly decreased in CAST mice at 24 hour‐, 72 hour‐, 1 week‐, 2 week‐, and 4‐week time points compared with WT mice (41.9 ± 14.9, 19.1 ± 4.2, 17.5 ± 8.2, 14.2 ± 4.4, and 10.0 ± 3.8, respectively, vs 50.2 ± 2.8), but TEST animals maintained or had an increase in ICP in comparison with WT mice (48.0 ± 1.4, 52.3 ± 1.3, 60.8 ± 7.6, 80.5 ± 2.1, and 81.5 ± 1.2, respectively). Mean systemic arterial pressure remained approximately 80 mm Hg irrespective of treatment. CAST mouse penis specimens revealed decreased α‐actin and CD‐31 immunoreactivity only at the 4‐week interval, compared with WT and TEST specimens. Doppler ultrasound flow rates (centimeter per second), taken before, during, and immediately after cavernous nerve stimulation, were WT 45.4 ± 7.3, 30.6 ± 5.2, 55.3 ± 8.2 vs CAST (2 weeks) 22.2 ± 2.5, 25.0 ± 1.5, 23.1 ± 2.0 vs TEST (2 weeks) 30.5 ± 6.5, 25.7 ± 2.0, 45.2 ± 4.5. This prominently showed that intrapenile flow was not reduced normally during erectile stimulation in CAST mice. This is the first described mouse model of castration‐induced veno‐occlusive erectile dysfunction. Erectile response abnormalities as measured by ICP and Doppler ultrasound studies in CAST mice may be attributed to hypogonadal effects on erectile tissue function. Morphologic changes in the cavernosal tissue of CAST mice coincide with these abnormalities to some extent. This study defines an androgen‐dependent mechanism of veno‐occlusive erectile function in the mouse. The castrated mouse model can be applied in future studies of veno‐occlusive erectile dysfunction.
Effect of Transient Embryonic In Vivo Exposure to the Endocrine Disruptor Methoxychlor on Embryonic and Postnatal Testis Development
Authors: Cupp Andrea S. , Uzumcu Mehmet , Suzuki Hiroetsu , Dirks Kristen , Phillips Brigette , Skinner Michael K.
Published: December 2015

ABSTRACT: The current study was designed to examine the effects of a transient embryonic exposure to the pesticide methoxychlor, an endocrine disruptor, on in vivo rat testis development and function. Gestating female rats were transiently administered methoxychlor (MXC) from embryonic day 7 (E7; EO = plug date) through E15. Embryonic testes were collected at E16 and postnatal (PO = day of birth) testes at P4, P10, P17–20, and P60. Seminiferous cords formed in testes from MXC exposed males. However, at E16, there was a decrease in the area of cords and an increase in interstitial area in MXC exposed testes when compared with controls. At all postnatal ages collected, there did not appear to be differences in seminiferous cord/tubule area, interstitial area, or number of seminiferous cords/tubules between untreated controls and males exposed to MXC. Exposure to the endocrine disruptor also had no effect on the postnatal organ weights of a variety of different organs, nor were testosterone levels altered. Interestingly, there were reductions in the number of germ cells in testes from MXC‐exposed males at P17‐P20 when compared with untreated controls. Furthermore, there was a twofold increase in apoptotic cells in tubules from pubertal P17‐P20‐MXC exposed males when compared with untreated controls. Testes were collected from adult P60 males to determine if early embryonic and postnatal alterations in germ cell numbers or testis cellular composition had compromised spermatogenesis. In adult P60 MXC exposed testes there were no gross morphological changes in testis structure or cellular composition over that of controls. However, there was an increase in apoptotic cell number in elongating spermatids in MXC exposed testes. Four P60 males that were exposed to MXC during gestation and 4 control males were bred with unexposed females to determine their ability to produce offspring. All MXC exposed males were capable of impregnating females and had normal litter size and pup weights. Combined observations demonstrated that exposure to MXC during gestation at a critical stage of testis development (ie, sex determination) affects embryonic testis cellular composition, germ cell numbers, and germ cell survival. While alterations in these parameters does not affect the ability of males to produce offspring, there appears to be a reduced spermatogenic capacity associated with MXC treatment. Therefore, transient embryonic exposure to an endocrine disruptor (methoxychlor) during gestation can influence the germline and fertility in adult males.
A Transgenic Analysis of Mouse Lactate Dehydrogenase c Promoter Activity in the Testis
Authors: Kroft Tim L. , Li Siming , Doglio Lynn , Goldberg Erwin
Published: December 2015

ABSTRACT: Transcription of the mouse testis‐specific lactate dehydrogenase c (mldhc) gene is limited to cells of the germinal epithelium. Cloning and analysis of the mldhc promoter revealed that a 100‐bp core promoter was able to regulate testis‐specific transcription in vitro and in transgenic mice. Surprisingly, expression of the reporter in transgenic testes was limited to pachytene spermatocytes, whereas native LDH‐C4 was detected in pachytene and all later germ cells. To locate additional regulatory sequence that could recapitulate the native LDH‐C4 distribution pattern, we investigated the contribution of 5′ and 3′ flanking sequences to the regulation of LDH‐C4 expression. We found that transcription factor YY1 binds to the mldhc promoter, that the mldhc 3′ untranslated sequence does not permit a postmeiotic expression of a β‐galactosidase reporter in transgenic mice, and that native mldhc mRNA is predominately meiotic, with only a low level of postmeiotic distribution. Our results suggest that the high level of LDH‐C4 in postmeiotic cells results from mRNA and protein stability.
The Ratio of Second to Fourth Digit Length in Azoospermic Males Undergoing Surgical Sperm Retrieval: Predictive Value for Sperm Retrieval and on Subsequent Fertilization and Pregnancy Rates in IVF/ICSI Cycles
Authors: Wood Simon , Vang Ellen , Manning John , Walton Julie , Troup Steven , Kingsland Charles , Lewis‐Jones Iwan David
Published: December 2015

ABSTRACT: The differentiation of the urogenital system and the appendicular skeleton in vertebrates is under the control of Homeobox (Hox) genes. It has been shown that this common control of digit and gonad differentiation has connected the pattern of digit formation to spermatogenesis and prenatal hormone concentrations in males. We wished to establish whether digit patterns, particularly the ratio between the lengths of the second and fourth digit in males (2D : 4D), was related to spermatogenesis and, more specifically, the presence of spermatozoa in testicular biopsies from azoospermic men undergoing surgical sperm retrieval. Forty‐four men were recruited, of whom 16 were diagnosed with nonobstructive azoospermia and 4 with congenital bilateral absence of the vas deferens, and 24 previously fertile men were azoospermic after previous vasectomy. Our results show that men with previous fertility or of an acquired form of azoospermia had significantly lower 2D : 4D ratios than men with nonobstructive azoospermia. In nonobstructive azoospermia, there was a significantly lower 2D : 4D ratio on the left side in men who had successful retrieval than those with unsuccessful retrieval. For these men who had a successful retrieval, none had a 2D : 4D ratio more than 1 on the left side, whereas 4 of 7 men in whom sperm was not found had a 2D : 4D ratio greater than 1. On successful sperm retrieval, subsequent fertilization and clinical pregnancy rates were unaffected by 2D : 4D ratios.
Chromosomal Abnormalities and Y Chromosome Microdeletions in Infertile Men With Varicocele and Idiopathic Infertility of South Indian Origin
Authors: Rao Lakshmi , Babu Arvind , Kanakavalli Murthy , Padmalatha Venkata , Singh Amarpal , Singh Prashant Kumar , Deenadayal Mamata , Singh Lalji
Published: December 2015

ABSTRACT: Various factors cause spermatogenesis arrest in men and, in a large number of cases, the underlying reason still remains unknown. Little attention is paid to determining the genetic defects of varicocele‐related infertility. The objective of our present study was to investigate the chromosomal abnormalities and Y chromosome microdeletions in infertile men of South Indian origin with varicocele and idiopathic infertility. Metaphase chromosomes of 251 infertile men with varicocele and unexplained infertility were analyzed using Giemsa‐Trypsin‐Giemsa (GTG) banding and fluorescence in situ hybridization (FISH). The microdeletions in 6 genes and 18 sequence‐tagged‐sites (STS) in the Yq region were screened using polymerase chain reaction (PCR) techniques. Out of 251 infertile men, 57 (22.7%) men were with varicocele, of which 8.77% were azoospermic, 26.31% were severely oligozoospermic, 21.05% were mildly oligozoospermic, and 43.85% were oligoasthenoteratozoospermic (OAT), and 194 (77.29%), with idiopathic infertility, of which 51% were azoospermic, 13.40% were severely oligozoospermic, 19.07% were mildly oligozoospermic, and 16.4% were with OAT. Genetic defects were observed in 38 (15.13%) infertile individuals, including 14 (24.56%) men with varicocele and 24 (12.37%) men with idiopathic infertility. The frequencies of chromosomal defects in varicocele and idiopathic infertility were 19.3% and 8.76%, respectively, whereas Y chromosome microdeletions were 5.26% and 3.60%, respectively. Overall rate of incidence of chromosomal anomalies and microdeletions in 251 infertile men were 11.5% and 3.98%, respectively, indicating a very significant higher association of genetic defects with varicocele than idiopathic male infertility. Our data also demonstrate that, among infertile men with varicocele, severely oligozoospermic and OAT men with varicocele have higher incidences of genetic defects than mildly oligozoospermic and azoospermic men.
Cryopreservation of Ram Semen Facilitates Sperm DNA Damage: Relationship Between Sperm Andrological Parameters and the Sperm Chromatin Structure Assay
Authors: Peris Soliman I. , Morrier Ariane , Dufour Maurice , Bailey Janice L.
Published: December 2015

ABSTRACT:We hypothesized that cryopreservation and incubation in conditions that mimic the female genital tract following insemination increases the susceptibility of ram sperm DNA to denaturation. Ram sperm samples (n = 12) underwent the sperm chromatin structure assay (SCSA) and semen quality tests, including motility parameters, viability, and chlortetracycline fluorescence (CTC) patterns. We also assessed correlations between SCSA variables and semen quality parameters. Analyses were performed for both fresh and cryopreserved samples at 0, 3, and 20 hours of incubation in synthetic oviductal fluid (SOF; 39°C, 5% CO2). The SCSA variables, mean alpha t (Xαt) and standard deviation of alpha t (SDαt), were higher because of cryopreservation (P < .05, P < .001, respectively) after 20 hours in SOF. For both fresh and frozen spermatozoa, SCSA values (Xαt, SDαt, and the percentage of cells outside the main population of αt[%COMPαt]) increased during incubation in SOF. Motility was negatively correlated with both SDαtand %COMPαt, ranging from −0.39 (P < .01) to −0.59 (P < .001) for both fresh and cryopreserved semen; viability also was negatively correlated with Xαt, SDαt, or %COMPαt(−0.36; P < .05, –.40 and ‐.46; P < .01, respectively) in fresh semen. The %COMPαtwas positively correlated to the percentage of CTC pattern AR (P < .001) and negatively correlated to the percentages of patterns F and B (−0.33 to −0.60, P < .05 to P < .001). Variation among ejaculates within ram was observed (P < .01). Cryopreservation clearly facilitates DNA damage in physiological conditions. The low to moderate correlations between SCSA variables and classical semen quality parameters indicate that the SCSA provides additional information to standard tests for evaluating ram sperm quality.
Role of Nitric Oxide Concentrations on Human Sperm Motility
Authors: Balercia Giancarlo , Moretti Simona , Vignini Arianna , Magagnini Mattia , Mantero Franco , Boscaro Marco , Ricciardo‐Lamonica Giuseppe , Mazzanti Laura
Published: December 2015

ABSTRACT: Nitric oxide (NO) is a free radical generated from the oxidation of l‐arginine to l‐citrulline by 3 isoforms of reduced nicotinamide adenine dinucleotide phosphate (NADPH)‐dependent NO synthases. Several data suggest a relevant role in sperm cell pathophysiology, but any conclusive data on its role in spermatozoa motility are still lacking. In the present study, we have correlated NO concentration in semen and kinetic features of sperm cells from normozoospermic fertile donors and infertile patients affected by idiopathic asthenozoospermia. Normozoospermic fertile men exhibited NO concentrations that were significantly lower than those of asthenozoospermic infertile men. A significant linear negative correlation was evident between NO concentration and percentage of total sperm motility. A further significant linear negative correlation was found between NO concentration and spermatozoa kinetic characteristics determined by a computerized analysis (curvilinear and straight progressive velocity). These data suggest that the overproduction of this free radical and the consequent excessive exposure to oxidative conditions have a potential pathogenetic implication in the reduction of sperm motility. The positive role played by NO in spermatozoa capacitation leads us to speculate that such paradoxical involvement in both pathologic and physiologic processes depends on the alternative redox state and relative level of NO.
Expression of a Novel RAD23B mRNA Splice Variant in the Human Testis
Authors: Huang Xiaoyan , Wang Hui , Xu Min , Lu Li , Xu Zhiyang , Li Jianmin , Zhou Zuomin , Sha Jiahao
Published: December 2015

ABSTRACT: A gene coding a novel human RAD23B protein named RAD23‐like protein B, presumably involved in spermatogenesis, was identified and characterized using a complementary DNA (cDNA) microarray. In the human testis, its expression was 2.33 times higher than it was in the embryo testis, with a particularly high expression in ejaculated human spermatozoa. The full length of this gene is 1548 bp, and the putative protein is 338 amino acids long. This protein is homologous to RAD23B, which is one of two human homologs of Saccharomyces cerevisiae RAD23, and is involved in both nucleotide excision repair (NER) and ubiquitin (Ub)‐dependent proteolysis. However, RAD23‐like protein B lacks the Ub‐like (UbL) domain that functions as a proteasome localization signal. Multiple‐tissue expression profile of the messenger RNA (mRNA) that encodes the RAD23‐like protein B also showed that it is highly expressed in the human testis and in ejaculated spermatozoa. Our present study indicates that this novel alternative splicing form of RAD23B is correlated with human spermatogenesis.
Effect of Chronic Ischemia on Constitutive and Inducible Nitric Oxide Synthase Expression in Erectile Tissue
Authors: Azadzoi Kazem M. , Master Tara A. , Siroky Mike B.
Published: December 2015

ABSTRACT: Arterial occlusive disease is one of the leading causes of organic erectile dysfunction (ED). Recent studies have shown that the incidence of cardiovascular disease closely correlates with the prevalence of ED. Also, ED is thought to be an early signal of impending cardiovascular problems. We previously found that the atherosclerosis of iliohypogastric arteries in the rabbit causes ED, down‐regulates cavernosal neuronal nitric oxide synthase (nNOS) gene expression, and impairs NO synthesis. The goal of this study was to determine the effect of atherosclerosis‐induced ischemia on cavernosal nNOS, endothelial NOS (eNOS), and inducible NOS (iNOS) expression and NO‐mediated smooth muscle relaxation in the rabbit. Our study showed that iliac artery blood flow, intracavernosal blood flow, and intracavernosal oxygen tension were unchanged 4 weeks after the induction of arterial atherosclerosis, whereas they were significantly diminished at weeks 8 and 16. Erectile responses to nerve stimulation and cavernosal smooth muscle relaxation were unchanged at week 4 and were significantly diminished at weeks 8 and 16 after the induction of atherosclerosis. Western blotting showed that cavernosal nNOS and eNOS protein levels were unaffected at week 4 but were significantly decreased at weeks 8 and 16 after the induction of atherosclerosis. iNOS protein, however, markedly increased during the course of the induced arterial disease. Immunohistochemical staining showed no change in cavernosal eNOS or nNOS expression at week 4. A dramatic decrease in both was evident at 8 and 16 weeks. iNOS expression progressively increased between 4 and 16 weeks of atherosclerosis. Down‐regulation of nNOS and eNOS, along with up‐regulation of iNOS, may explain ischemic cavernosal smooth muscle relaxation impairment in the rabbit. Ischemically altered NOS expression may be of great pathophysiologic importance in atherosclerosis‐induced ED. These data may provide further insight into the mechanism of arteriogenic ED.
Evaluation of ICSI‐Selected Epididymal Sperm Samples of Obstructive Azoospermic Males by the CKIA System
Authors: Ramos Liliana , de Boer Peter , Meuleman Eric J. H. , Braat Didi D. M. , Wetzels Alex M. M.
Published: December 2015

ABSTRACT: The objective of this study was to evaluate nuclear normality in intracytoplasmic sperm injection (ICSI)‐selected epididymal sperm from obstructive azoospermic (OA) patients. We evaluated whether the selection criteria used in routine ICSI (morphology and motility at a magnification of 400×) is adequate for selecting “normal” sperm from epididymal samples. Surgically retrieved spermatozoa from the caput epididymis of 15 OA patients and ejaculated sperm samples from 9 normospermic donors were evaluated with a DNA‐specific stain (Feulgen) and in combination with the computerized karyometric image analysis (CKIA) system. Original (unselected) samples and ICSI‐selected sperm were compared in donor and patient samples. In the original fraction, a larger variation in almost all measured parameters was found in epididymal sperm than in ejaculated sperm. After sperm selection, the morphometry was comparable between epididymal and ejaculated sperm. However, for those parameters related to the DNA stainability and chromatin texture (nuclear condensation), significant differences between patients and donors were observed. This result suggests that the size and form of the sperm do not necessarily hold similar internal structures. Thus, the frequency of “normal” sperm significantly increased after ICSI selection, but the improvement was more marked in donor than in OA patients' samples. In conclusion, at least a twofold increase in the number of normal spermatozoa was achieved after ICSI selection. The heterogeneity in the stainability and chromatin condensation of epididymal samples from OA patients indicates that some of the selected spermatozoa have a hypocondensed or hypercondensed chromatin. Even in the best of donor cases, no more than 55% of the selected sperm scored normal with CKIA, indicating that the present routine ICSI selection criteria are not sufficient for selecting normal condensed nuclei.
Calcification of the Epididymis and the Tunica Albuginea of the Corpora Cavernosa in Patients on Maintenance Hemodialysis
Authors: Guvel Sezgin , Pourbagher Mir Ali , Torun Dilek , Egilmez Tulga , Pourbagher Aysin , Ozkardes Hakan
Published: December 2015

ABSTRACT: The aims of this study were to determine the incidence rates of genital calcification in male hemodialysis patients based on ultrasonography findings and to identify risk factors for this condition. Twenty‐three male end‐stage renal disease (ESRD) patients (mean age, 51.4 ± 12.1 years) who were on maintenance hemodialysis underwent penile and scrotal ultrasonography. For each case, we recorded the underlying renal disease and measured serum levels of phosphorus, intact parathormone, and calcium × phosphorus product. Patients were also questioned about erectile dysfunction. The control group consisted of 22 consecutive patients (mean age, 51 years) with type 2 diabetes mellitus with normal renal function who underwent penile and scrotal ultrasonography for various reasons. In the ESRD group, ultrasound revealed calcification of the tunica albuginea of the corpora cavernosa in 15 patients (65%) and calcification of the epididymis in 16 patients (70%; 14 bilateral and 2 unilateral cases). Twenty patients (87%) showed calcification of the epididymis and/or the tunica, and 10 (43%) showed calcification of both these tissues. The rates of epididymal and penile calcification in the ESRD patients and the controls were significantly different (P < .001 for both). There were no significant differences between patients with and without penile and epididymal calcification with respect to age, hemodialysis duration, frequencies of elevated serum phosphorus, elevated serum intact parathormone, elevated calcium × phosphorus product, and frequency of erectile dysfunction (ED) (P > .05 for all). Ultrasonography revealed high rates of penile (tunica albuginea of the corpora cavernosa) and epididymal calcification (65% and 70%, respectively) in the ESRD patients studied, but no association was found between risk factors such as age, underlying renal disease, hemodialysis duration, frequencies of elevated serum phosphorus, elevated serum intact parathormone, and elevated calcium × phosphorus product.
Detection of a Short CCR5 Messenger RNA Isoform in Human Spermatozoa
Authors: Januchowski Radosław , Bręborowicz Andrzej K. , Ofori Harold , Jedrzejczak Piotr , Pawelczyk Leszek , Jagodzinski Paweł P.
Published: December 2015

ABSTRACT: It has recently been reported that the Regulated upon Activation of Normal T‐cells Expressed and Secreted (RANTES) chemokine may exhibit a chemotactic effect on sperm. The RANTES chemokine acts on target cells by binding to the CCR5 receptor, which is present on the surface of various cells. Spermatozoa contain a complex repertoire of messenger RNAs (mRNAs) that may provide an insight into past events of spermatogenesis. The type and amount of CCR5 chemokine receptor transcript were investigated in spermatozoa that were isolated by the swim‐up method from semen samples of men with normozoospermia. Using reverse transcription and real‐time quantitative polymerase chain reaction (RQ‐PCR) analysis, we found that the CCR5 mRNA isoform in human spermatozoa consists of exons 3 and 4, and is shorter than the transcript in leukocytes. This CCR5 transcript may represent a more stable mRNA isoform; one that is used to biosynthesize the CCR5 receptor in spermatogenesis or the early stages of embryonic development.
Cinnoxicam andl‐Carnitine/Acetyl‐l‐Carnitine Treatment for Idiopathic and Varicocele‐Associated Oligoasthenospermia
Authors: Cavallini Giorgio , Ferraretti Anna Pia , Gianaroli Luca , Biagiotti Giulio , Vitali Giovanni
Published: December 2015

ABSTRACT:The objective of this study was to detect a therapy for idiopathic and varicocele‐associated oligoasthenospermia (OAT). Idiopathic and varicocele OAT patients were randomized into 3 groups. Each group was composed of varying degrees of left varicoceles (graded into 5 grades with echo‐color Doppler) and of idiopathic OATs. Group 1 used a placebo, group 2 used orallcarnitine (2 g/d) + acetyl‐lcarnitine (1 g/d), group 3 usedlcarnitine/acetyl‐l‐carnitine + 1 × 30‐mg cinnoxicam suppository every 4 days. Drugs were administered for 6 months. The groups were composed as follows: group 1, 71 varicoceles and 47 idiopathic OATs; group 2, 62 varicoceles and 39 idiopathic OATs; group 3, 62 varicoceles and 44 idiopathic OATs. Sperm concentration, motility, and morphology before during and after treatments were assessed. Pregnancy rates and side effects were recorded. Group 1 did not have modified sperm patterns during treatment. Group 2 had significantly increased sperm patterns at 3 and 6 months into therapy in idiopathic patients and in patients with grades I, II, and III varicocele, but not in grades IV and V. Group 3 had significantly increased sperm parameters in all patients, with the exception of grade V varicocele. Group 3 sperm patterns proved significantly higher during therapy than group 2. All sperm patterns fell to baseline after therapy suspension. Minor side effects occurred. Pregnancy rates were 1.7% (group 1), 21.8% (group 2), and 38.0% (group 3) (P < .01).l‐Carnitine/acetyl‐l‐carnitine + cinnoxicam suppositories proved a reliable treatment for low‐grade varicoceles and idiopathic OATs.
Vasectomy‐Dependent Dysregulation of a Local Renin‐Angiotensin System in the Epididymis of the Cynomolgus Monkey (Macaca fascicularis)
Authors: Saez Fabrice , Légaré Christine , Laflamme Julie , Sullivan Robert
Published: December 2015

ABSTRACT: The mammalian epididymis is a fundamental organ for sperm cell maturation; it allows mammals to acquire their fertilizing ability. We have previously shown that during obstruction in cases of vasectomy, gene expression profiles were modified in human and cynomolgus monkey epididymides. Paracrine factors thus appear to be key elements in local gene expression along the epididymis. Local renin‐angiotensin systems (RAS) have been described in many other organs as paracrine regulators of gene expression. This work demonstrates the presence of a local RAS in the epididymis of the cynomolgus monkey and investigates the vasectomy‐dependent changes occurring in this system. After unilateral vasectomy in 4 monkeys (two for 3 days and two others for 7 days), the presence of two major components of the RAS (ie, angiotensinogen [ANG] and the type 1 receptor to angiotensin II [AT‐I]) was evaluated in the vasectomized and the normal controlateral epididymides of each monkey. We also show by in situ hybridization that the principal cells of the epididymis express ANG and AT‐I mRNAs and immunohistochemistry permitted to verify the co‐localization of the AT‐I protein and mRNA. Quantitative comparisons of individual variations in the mRNA and protein profiles for ANG and AT‐I revealed that vasectomy altered the RAS expression profiles in an individual manner, thus confirming its role as a local system. This study provides a good basis for further investigation of the possible implications of the RAS in the physiology of the epididymis. Furthermore, the individual dependent modifications are in accordance with the very fluctuating results obtained in the fertility status of human patients undergoing a vasectomy reversal. The variations observed in the RAS expression profiles may be a good model to study the causes of the overall epididymal gene expression dysregulation that follows vasectomy and potentially affects fertility.
In Vivo Effects of Histone‐Deacetylase Inhibitor Trichostatin‐A on Murine Spermatogenesis
Authors: Fenic Irina , Sonnack Violetta , Failing Klaus , Bergmann Martin , Steger Klaus
Published: December 2015

ABSTRACT: The acetylation state of core histones is controlled by two classes of enzymes, histone acetyl transferases (HATs) and histone deacetylases (HDACs). HDAC inhibitors, such as trichostatin‐A (TSA), are able to induce cell cycle arrest by stimulating transcription of genes that negatively regulate cell growth and survival. However, little is known about the effect of HDAC inhibitors on spermatogenesis. TSA treatment of cultured murine germ cells from whole testes resulted in an increase of histone H4 acetylation in round spermatids, suggesting that a hypoacetylated state of these cells is important for their normal differentiation. In the present study, the in vivo effects of TSA on murine spermatogenesis were investigated. Subcutaneously applied TSA resulted in a dose‐dependent decrease in relative testis weight due to impaired spermatogenesis. No obvious toxic effects of TSA treatment could be found. A second animal experiment confirmed that male mice receiving TSA under the same conditions as in the first experiment became infertile. This phenomenon was completely reversible. No evidence of histone H4 hyperacetylation in round spermatids could be found; however, the number of spermatids significantly decreased with increasing TSA concentrations. Additionally, a dramatic loss of pachytene‐diplotene spermatocytes due to increased apoptosis was observed. This suggests that TSA was mainly effective at the level of meiosis. The other male reproductive organs showed no morphological changes compared to controls, suggesting that TSA action on the testis was not mediated by sex hormones.