2040 A SYSTEMATIC REVIEW AND META-ANALYSIS OF ANEJACULATION IN SPINAL CORD INJURED PATIENTS IN FAVOUR OF A SPINAL GENERATOR OF EJACULATION

Vol. 189, No. 4S, Supplement, Wednesday, May 8, 2013

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.

THE JOURNAL OF UROLOGY姞

e837

lyzes. In the same way, epididymal, retroperitoneal and subcutaneous fat deposits were removed. Data were analyzed by one-way ANOVA and Bonferroni post-test, considering p ⬍ 0.05. RESULTS: Different high-fat diets promoted an increase in body mass when compared to SC group (p ⬍ 0.0001). Accordingly, epididymal, retroperitoneal and subcutaneous fat deposits were higher in high-fat groups (HF-S, HF-P, HF-SP) than SC group (p ⫽ 0.0004; p ⫽ 0.0001; p ⬍ 0.0001, respectively). As for serum parameters, the groups that received different high fat diets showed hyperglycemia, hyperinsulinemia, and hypercholesterolemia at the end of the experiment (p ⫽ 0.0055, p ⬍0.05, p ⫽ 0.0021, respectively). The levels of serum triglycerides did not differ among the groups. Regarding the testis, seminiferous epithelium height was less in HF-S group than SC group. However, HF-P group and HF-SP group presented an increase in this parameter in comparison to HF-S group resembling SC group (p⫽0.0003). Already cells proliferation was lower in HF-S group than SC group. However, only the HF-P group presented an increase in this parameter in comparison to HF-S group resembling SC group (p⫽0.0043). There was a decrease in seminiferous tubule diameter in the HF-SP group when compared SC group (p⫽0.0022). Among the groups, no difference was observed in the testicular volume. CONCLUSIONS: The high-fat diet administration, independent of the lipid quality, promoted overweight in adult rats. Nevertheless, diet rich in saturated fatty acids (lard) promoted an adverse remodeling upon the testicular morphology that was not observed in the group that received a diet rich in polyunsaturated fatty acids (canola oil). Source of Funding: FAPERJ, CAPES and CNPq.

2040 A SYSTEMATIC REVIEW AND META-ANALYSIS OF ANEJACULATION IN SPINAL CORD INJURED PATIENTS IN FAVOUR OF A SPINAL GENERATOR OF EJACULATION Source of Funding: 1. The Robert Dow Foundation 2. Mr. and Mrs. Howard and Irena Laks.

2038 DIFFERENTIAL DIAGNOSIS OF AZOOSPERMIA WITH PROTEOMIC BIOMARKERS QUANTIFIED IN SEMINAL PLASMA Andrei Drabovich*, Brendan Mullen, Eleftherios Diamandis, Keith Jarvi, Toronto, Canada Withdrawn

2039 DIET RICH IN POLYUNSATURATED FATTY ACID INDUCES OVERWEIGHT BUT IMPROVES THE TESTICULAR MORPHOLOGY IN ADULT RATS Pamella Silva, Ange´lica Furriel, Natha´lia Machado, Diogo Souza, Carla Gallo, Bianca Gregório*, Francisco Sampaio, Waldemar Costa, Rio de Janeiro, Brazil INTRODUCTION AND OBJECTIVES: The obesity and its metabolic complications affecting the endocrine system and multiple organs such as testis. Therefore, the objective of this study was to evaluate the effects of different high-fat diets on body weight and testicular morphology in rats at 7 months-old. METHODS: 39 male Wistar rats (3 months-old) were divided into 4 groups: SC (standard chow), HF-S (diet rich in saturated fatty acids), HF-P (diet rich in polyunsaturated fatty acids), HF-SP (diet rich in saturated and polyunsaturated fatty acids). During the experiment, food intake and body weight were measured daily and weekly, respectively. At 7 months-old, the animals were sacrificed. Next, blood samples were collected by cardiac puncture for analysis of serum glucose, insulin, total cholesterol and triglycerides. Testis were collected and processed for histomorphometrical and immunohistochemical ana-

Cle´ment Che´hensse, Montigny-le-Bretonneux, France; Ste´phane Bahrami, Pierre Denys, Garches, France; Pierre Cle´ment, Montigny le Bretonneux, France; Franc¸ois Giuliano*, Garches, France INTRODUCTION AND OBJECTIVES: After spinal cord injury (SCI) most men cannot ejaculate, although ejaculation can be obtained with medical assistance. There are gaps in the understanding of the neurophysiology of ejaculation and the pathophysiology of anejaculation in SCI males. Notably, the existence of a spinal generator of ejaculation, which has been identified in the rat, is not substantiated in the human. Studies reporting ejaculation after SCI have been reviewed in the light of recent advances on the physiology of ejaculation, especially regarding the spinal neurophysiology. METHODS: Using 4 databases since 1955, 513 studies were identified and 45 fulfilled the following inclusion criteria 1) characterisation of the SCI (completeness and either upper limit or somatic reflex function below this upper limit that allowed to estimate lower limit of SCI) and 2) antegrade ejaculation rate. A total of 3795 patients were pooled together. Subgroups analyses were performed depending on i) the method used to elicit ejaculation: ecological stimulation i.e. masturbation or coïtus, penile vibratory stimulation (PVS) with or without oral midodrine before stimulation and masturbation following acetylcholine esterase (AchE) inhibitors it prostigmine or sc physostigmine ii) SCI completeness according to ASIA Impairment Scale and iii) SCI upper and lower limit. RESULTS: Ejaculation rate in patients with complete SCI was 11.8%, 45.7% and 54.7% respectively during masturbation or coïtus, in response to penile vibratory stimulation (PVS) and during masturbation after AchE inhibitors. Complete injury of the sympathetic centres (spinal segments T12 to L2) or of the parasympathetic and somatic centres (spinal segments S2 to S4) totally disrupted ejaculation in response to PVS. AchE inhibitors followed by masturbation elicited ejaculation in 5 and 31% of men with complete injury of the T12 to L2 and S2 to S4 segments respectively. Controlling for the number of T12 to L2 segments completely injured, ejaculation rate was significantly lower when

e838

THE JOURNAL OF UROLOGY姞

Vol. 189, No. 4S, Supplement, Wednesday, May 8, 2013

injury extended to spinal segment L3 and below. Complete injury of segments S2 to S4 precluded the occurrence of rhythmic forceful ejaculation. CONCLUSIONS: These data confirm and reinforce the crucial role of the thoracolumbar sympathetic and sacral parasympathetic centres in emission and sacral somatic centre in expulsion. In addition this review of the literature strongly suggests that the segments L3 to L5 harbour a spinal generator of ejaculation in human. Source of Funding: French Society of Physical and Rehabilitation Medicine (SOFMER). Versailles Saint-Quentinen-Yvelines University (UVSQ).

2041 TESTICULAR MORPHOLOGY AND SPERMATOZOID PARAMETERS IN HYPERTENSIVE RATS TREATED OR NOT WITH ENALAPRIL Gustavo Bechara, Michele Simões, Bruno Felix-Patricio, Jorge Medeiros-Jr, Bianca Gregório, Diogo de Souza*, Waldemar Costa, Francisco Sampaio, Rio de Janeiro, Brazil INTRODUCTION AND OBJECTIVES: The target organ damage is well documented in arterial hypertension, but testicular damage is not clearly defined. This study evaluates the testicular morphology and spermatozoid parameters of spontaneously hypertensive rats (SHR) treated or not with enalapril. METHODS: 28 adult SHR animals were assigned into two groups, a hypertensive group (H, n⫽13), and an enalapril treated group (HE, n⫽15). Wistar-Kyoto rats (WKY, n⫽8) were used as controls. Animals of group HE received 30mg/Kg of enalapril daily by gavage while H and WKY animals received tap water. Arterial systolic pressure was weekly measured by plethysmography. After five weeks of treatment, all animals were killed by anesthetic overdose. The right epididimal tail was removed and used for spermatozoid collection. Sperm concentration, motility and viability (hypo-osmotic test) were assessed. The right testicles were fixed and routinely processed for obtaining hematoxylin and eosin stained histological sections. In these sections, the seminiferous tubule diameter, epithelial height and relative densities of the testicular structures were assessed by stereological methods. All data was compared by ANOVA with Tukey’s post test, considering p⬍0.05 as significant. RESULTS: Blood pressure in the HE group were similar WKY, and lower than H animals. Spermatozoid concentration had a 38% reduction in hypertensive rats when compared to controls while enalapril treatment restored normal concentration. Concerning spermatozoid motility and viability, no difference was found among the groups. Testicular vascular volumetric density was also altered in hypertensive group while HE animals were similar to controls. The seminiferous epithelium of HE animals showed the highest volumetric density, indicating a positive effect of enalapril in spermatogenesis. The analysis of seminiferous tubule diameter and epithelium height showed no significant difference among the studied groups. Table 1 presents all numerical data. CONCLUSIONS: In the SHR model, hypertension induces testicular alterations both in seminiferous tubule and intratesticular vessels resulting in low spermatozoid concentration. Enalapril treatement partially protected the testicle from the hypertension-induced alterations, restoring normal spermatozoid production.

Source of Funding: FAPERJ, CNPq and CAPES.

2042 DELETION AND MUTATION ANALYSIS OF RECOMBINATION GENES IN MEN WITH MEIOTIC DEFECTS Matthew Wosnitzer*, Ali Dabaja, Anna Mielnik, Peter Schlegel, Darius Paduch, New York, NY INTRODUCTION AND OBJECTIVES: Meiotic recombination pathway abnormalities are associated with non-obstructive azoospermia (NOA), affecting genome integrity, chromosome pairing, and gametogenesis. NOA patients have significantly lower recombination frequency (measured by MLH1 foci per pachytene cell) and decreased fidelity of chromosomal synapsis (% cells with ⱕ1 bivalent without MLH1 foci) compared to men with obstructive azoospermia and normal spermatogenesis. We sought to evaluate whether deletion or mutation of critical recombination pathway genes may underlie decreased meiotic recombination in oligospermic, NOA, or Klinefelter syndrome (KS) patients. METHODS: Genomic DNA from all subjects was extracted from peripheral blood leukocytes. Six patients with defective recombination (Gonsalves et al. 2004), 120 patients with severe oligospermia or NOA, and 34 KS patients underwent somatic screening for deletions of mismatch repair (MMR) genes encoding critical recombination machinery protein: MLH1, MLH3, SPO11, SYN1, FAM9 and SYCP3. Polymerase chain reactions were performed for each gene and amplified products were assessed by gel electrophoresis. Mutation analyses for SCP3 and MLH1 were conducted first in 6 patients with recombination defects and 8 control men. Following preliminary screening, 177 additional patients were screened for MLH1 exon 20 and 145 patients for SYCP3 exon 8. Denaturing high-performance liquid chromatography (HPLC) assessed single nucleotide polymorphisms and detected changes were sequenced. RESULTS: No deletions of recombination genes were identified in any patient. SYCP3 mutation analysis detected no alterations in any patient. Three heterozygous missense mutations were identified in MLH1 exon 20 (C718T, gene position 2152, 718 aa, histidine to tyrosine) in 1 patient with recombination defects, and in 2 patients with oligospermia/NOA (V716M, position 2146, 716 aa, valine to methionine in oligospermic patient, and R725H, position 2174, 725 aa, arginine to histidine in NOA patient). One patient with recombination defects had homozygous changes in flanking intronic regions of exons 16 and 19, while no homozygous point mutations were identified in controls. CONCLUSIONS: Detection of missense mutations in the DNA mismatch repair (MMR) protein MLH1 in men with impaired spermatogenesis and recombination defects suggests a role of this gene in early meiotic arrest and as a putative genetic cause of NOA. Patients with recombination defects appear more likely to exhibit MLH1 mutational aberrations than deletions. Further study of MLH1 mutations is needed for validation. Source of Funding: None