Showing papers in "Journal of Andrology in 2009"

Fifty-two-week treatment with diet and exercise plus transdermal testosterone reverses the metabolic syndrome and improves glycemic control in men with newly diagnosed type 2 diabetes and subnormal plasma testosterone.

Abstract: Men with the metabolic syndrome (MetS) and type 2 diabetes (T2D) often have low testosterone levels. Elevating low testosterone levels may improve features of the MetS and glycemic control. In a single blind, 52-week randomized clinical trial, the effects of supervised diet and exercise (DE 87.5% of them reached an HbA1c of less than 6.5%. Based on Adult Treatment Panel III guidelines, 81.3% of the patients randomized to D&E plus testosterone no longer matched the criteria of the MetS, whereas 31.3% of the D&E alone participants did. Additionally, testosterone treatment improved insulin sensitivity, adiponectin, and high-sensitivity C-reactive pro- tein. Addition of testosterone to supervised D&E results in greater therapeutic improvements of glycemic control and reverses the MetS after 52 weeks of treatment in hypogonadal patients with the MetS and newly diagnosed T2D.

Are Tests of Sperm DNA Damage Clinically Useful? Pros and Cons

Abstract: The advent of assisted reproductive technologies, particularly intracytoplasmic sperm injection (ICSI), has revolutionized the treatment of male-factor infertility. However, there are many unanswered questions regarding the safety of these techniques. These safety concerns are relevant because 1) these technologies often bypass the barriers to natural selection; 2) infertile men, particularly those with severe male-factor infertility, possess substantially more sperm DNA damage than do fertile men; and 3) experimentally, sperm DNA damage has been shown to adversely affect the developing embryo. This review discusses the etiology of sperm DNA damage, describes the individual tests of sperm DNA damage, and explores the relationship between sperm DNA damage and pregnancy outcomes. Based on a systematic review of the literature, sperm DNA damage is associated with lower natural, intrauterine insemination (IUI), and in vitro fertilization (IVF) pregnancy rates, but not with ICSI pregnancy rates. The literature also suggests that that sperm DNA damage is associated with an increased risk of pregnancy loss in those couples undergoing IVF or ICSI. Nonetheless, the true clinical utility of sperm DNA damage tests remains to be established, because the available studies are small and few in number and the study characteristics are heterogeneous. Although current data suggest that impaired sperm DNA integrity may have the greatest effect on IUI pregnancy rates and pregnancy loss by IVF and ICSI, further prospective studies are needed before testing should become a routine part of patient management.

The Dark Side of Testosterone Deficiency: III. Cardiovascular Disease

Abstract: A considerable body of evidence exists suggesting that androgen deficiency contributes to the onset, progression, or both of cardiovascular disease (CVD) The aim of this review is to evaluate the relationships between testosterone (T) deficiency and risk factors of CVD and to discuss the implications of androgen deficiency in men with cardiovascular risk factors The relationship between androgen deficiency and endothelial function, lipid profiles, inflammatory responses, altered vascular smooth muscle reactivity, and hypertension are discussed with regard to CVD A comprehensive literature search was carried out with the use of Pub Med from 1980 through 2009, and relevant articles pertinent to androgen deficiency and vascular disease were evaluated and discussed Low T, whether attributed to hypogonadism or androgen deprivation therapy, in men with prostate carcinoma, produces adverse effects on cardiovascular health Androgen deficiency is associated with increased levels of total cholesterol, low-density lipoprotein, increased production of proinflammatory factors, and increased thickness of the arterial wall and contributes to endothelial dysfunction Testosterone supplementation restores arterial vasoreactivity; reduces proinflammatory cytokines, total cholesterol, and triglyceride levels; and improves endothelial function but also might reduce high-density lipoprotein levels Testosterone is an anabolic hormone with a wide range of beneficial effects on men's health The therapeutic role of T in men's health, however, remains a hotly debated issue for a number of reasons, including the purported risk of prostate cancer In view of the emerging evidence suggesting that androgen deficiency is a risk factor for CVD, androgen replacement therapy could potentially reduce CVD risk in hypogonadal men It should be emphasized, however, that androgen replacement therapy should be done with very thorough and careful monitoring for prostate diseases

Urinary Metabolites of Di(2‐ethylhexyl) Phthalate Are Associated With Decreased Steroid Hormone Levels in Adult Men

Abstract: Experimental animal studies have demonstrated that exposure to some phthalates may be associated with altered endocrine function and adverse effects on male reproductive development and function, but human studies are limited. In the present study, urine and serum samples were collected from 425 men recruited through a US infertility clinic. Urinary concentrations of mono(2-ethylhexyl) phthalate (MEHP), the hydrolytic metabolite of di(2-ethylhexyl) phthalate (DEHP), and other phthalate monoester metabolites were measured, along with serum levels of testosterone, estradiol, sex hormone-binding globulin (SHBG), follicle-stimulating hormone, luteinizing hormone, inhibin B, and prolactin. Two oxidized urinary metabolites of DEHP were also measured in urine from 221 of the men. In multiple regression models adjusted for potential confounders, MEHP was inversely associated with testosterone, estradiol, and free androgen index (FAI). An interquartile range increase in MEHP was associated with 3.7% (95% confidence interval (CI), 26.8% to 20.5%) and 6.8% (95% CI, 211.2% to 22.4%) declines in testosterone and estradiol, respectively, relative to the population median hormone levels. There was limited evidence for effect modification of the inverse association between MEHP and FAI by the proportion of DEHP metabolites in the urine measured as MEHP (MEHP%), which is considered a phenotypic marker of less efficient metabolism of DEHP to its oxidized metabolites. Finally, the ratio of testosterone to estradiol was positively associated with MEHP (P 5 .07) and MEHP% (P 5 .007), suggesting potential relationships with aromatase suppres- sion. In conclusion, these results suggest that urinary metabolites of DEHP are inversely associated with circulating steroid hormone levels in adult men. However, additional research is needed to confirm these findings.

Genome-wide study of single-nucleotide polymorphisms associated with azoospermia and severe oligozoospermia.

Abstract: Male infertility affects a large proportion of the population. In spite of the magnitude of the problem and the considerable research effort that has been made to understand its causes, a large proportion of male infertility cases remain idiopathic in nature. This pilot genome-wide association study employed genotyping microarray technology to interrogate over 370 000 single-nucleotide polymorphisms (SNPs) in men with azoospermia and severe oligozoospermia, along with normozoospermic controls, in an effort to discover novel genetic variants significantly associated with male infertility. We found 20 SNPs significantly associated with azoospermia or oligozoospermia (P value for association of <1 × 10−5). Using a gene-centric approach evaluating SNPs associated with genes of known fertility function, we found 1 additional SNP with P < 1 × 10−4. Although additional studies with larger numbers of samples will be required to validate these results and to identify causal variants, this study represents an important first step in applying genome-wide approaches to identify the genetic causes of male infertility.

Boar fertility and sperm chromatin structure status: a retrospective report.

Abstract: Little information exists about boar sperm chromatin quality and fertility within a commercial setting. The objective of this report is to provide information about boar sperm chromatin integrity and its relationship to fertility. The sperm chromatin structure assay (SCSA) was used retrospectively to characterize sperm from 18 sexually mature boars having fertility information. Boar fertility was defined by farrow rate (FR) and average total number of pigs born (ANB) per litter of gilts and sows mated to individual boars. Fertility data was compiled for 1867 matings across the 18 boars. The SCSA uses flow cytometry to evaluate the structural integrity of sperm nuclear DNA. The SCSA parameters measured in this retrospective analysis were the percentage DNA fragmentation index (%DFI) and standard deviation of the DNA fragmentation index (SD DFI). The %DFI and SD DFI showed the following significant negative correlations with FR and ANB; %DFI vs FR, r = -0.55, P < .01; SD DFI vs FR, r = -0.67, P < .002; %DFI vs ANB, r = -0.54, P < .01; and SD DFI vs ANB, r = -0.54, P < .02. Although more information is required to better understand the relationship between DFI and boar fertility, this report suggests that the SCSA assay may be an important assay for identification of boars having potential for lowered fertility.

Semen quality decline among men in infertile relationships: experience over 12 years in the South of Tunisia.

Abstract: Concerns about the worldwide decline in semen quality over the past 50 years are increasing. Western countries have shown a decline in semen quality. However, in non-Western countries studies are sparse. We investigated trends in semen parameters between 1996 and 2007 in the Sfax area of southern Tunisia in a sample of 2940 men in infertile relationships. Age at semen collection, duration of sexual abstinence, volume of seminal fluid, the sperm count, percentages of motile and morphologically normal spermatozoa, and semen leukocyte concentration were determined. Linear regression was used to examine trends over time in sperm count, sperm motility, normal morphology, and semen leukocyte concentration. Mean age and semen volume did not change between 1996 and 2007. Data adjusted for age and abstinence showed a decreasing trend in sperm count and percentage of normal morphology over the last 12 years (R2 = 0.71, P = .0004, and R2 = 0.87, P < .0001, respectively). There was no significant change in sperm motility. However, semen leukocyte concentration increased significantly over time (R2 = 0.38, P = .03). These results coincide with the high prevalence of genital infectious diseases in the Sfax area, suggesting that infection may be a potential contributing factor in semen quality decline.

Measurement of Testicular Volume in Smaller Testes: How Accurate Is the Conventional Orchidometer?

Abstract: The aim of this study was to evaluate the accuracy of different methods, including the Seager orchidometer (SO) and ultrasonography (US), for assessing testicular volume of smaller testes (testes volume less than 18 mL). Moreover, the equations used for the calculations--the Hansen formula (length [L] x width [W](2) x 0.52, equation A), the prolate ellipsoid formula (L x W x height [H] x 0.52, equation B), and the Lambert equation (L x W x H x 0.71, equation C)--were also examined and compared with the gold standard testicular volume obtained by water displacement (Archimedes principle). In this study, 30 testes from 15 men, mean age 75.3 (+/-8.3) years, were included. They all had advanced prostate cancer and were admitted for orchiectomy. Before the procedure, all the testes were assessed using SO and US. The dimensions were then input into each equation to obtain the volume estimates. The testicular volume by water displacement was 8.1 +/- 3.5 mL. Correlation coefficients (R(2)) of the 2 different methods (SO, US) to the gold standard were 0.70 and 0.85, respectively. The calculated testicular volumes were 9.2 +/- 3.9 mL (measured by SO, equation A), 11.9 +/- 5.2 mL (measured by SO, equation C), 7.3 +/- 4.2 mL (measured by US, equation A), 6.5 +/- 3.3 mL (measured by US, equation B) and 8.9 +/- 4.5 mL (measured by US, equation C). Only the mean size measured by US and volume calculated with the Hansen equation (equation A) and the mean size measured by US and volume calculated with the Lambert equation (equation C) showed no significant differences when compared with the volumes estimated by water displacement (mean difference 0.81 mL, P = .053, and 0.81 mL, P = .056, respectively). Based on our measurements, we categorized testicular volume by different cutoff values (7.0 mL, 7.5 mL, 8.0 mL, and 8.5 mL) to calculate a new constant for use in the Hansen equation. The new constant was 0.59. We then reexamined the equations using the new 0.59 constant, and found that the equation Volume (V) = L x W(2) x 0.59 was the best for describing testicular volume among our subjects (difference between the new equation and the gold standard of water displacement = 0.19 mL, P = .726). We also found that US was more precise in measuring testicular dimensions. We propose a new formula, V = L x W(2) x 0.59, to assess the volumes of smaller testes.

Androgens play a pivotal role in maintaining penile tissue architecture and erection: a review.

Abstract: Androgens are essential for development, growth, and maintenance of penile structure, and regulate erectile physiology by multiple mechanisms. Here we provide a concise overview of the basic research findings pertaining to androgen modulation of erectile tissue architecture and physiology. A significant body of evidence exists pointing to a critical role of androgens in erectile physiology. Studies in animal models have provided fundamental knowledge on the role of androgens in modulating tissue architecture and cellular, molecular, and physiological mechanisms. Based on data from our laboratory and those reported by others, we believe that androgens play a pivotal role in maintaining the structure and function of the peripheral penile nerve network, the structural integrity of the corpora cavernosa, the tunica albuginea, and the endothelium of the cavernous spaces. Further, androgens play an important role in regulating the differentiation of precursor cells into trabecular smooth muscle. In this review, we will focus our discussion on findings pertaining to the role of androgens in regulating penile tissue architectural elements in modulating penile function. This knowledge has a profound impact on the potential use of androgens in the clinical setting to treat patients with erectile dysfunction.

Selenium Supplementation Does Not Affect Testicular Selenium Status or Semen Quality in North American Men

Abstract: Selenium (Se) is essential for sperm function and male fertility, but high Se intake has been associated with impaired semen quality. We reported previously a decrease in sperm motility in men fed high-Se foods, but we could not rule out the influence of other environmental and dietary factors. We now report on a randomized, controlled study on the potential adverse effects of Se supplementation on semen quality in 42 free-living men administered Se (300 microg/d) as high-Se yeast for 48 weeks. Semen analysis was performed 4 times before treatment began, then twice each week during treatment at 6, 12, 24, 36, and 48 weeks, and then after treatment at 72 and 96 weeks. Blood samples were collected 3 times before treatment and at each subsequent visit. Se concentration increased 61% in blood plasma and 49% in seminal plasma. However, Se supplementation had no effect on sperm Se, serum androgen concentrations, or sperm count, motility, progressive velocity, or morphology. We observed progressive decreases in serum luteinizing hormone, semen volume, and sperm Se in both the high-Se and placebo groups. Moreover, sperm straight-line velocity and percent normal morphology increased in Se-treated and placebo-treated participants. The lack of an increase in sperm Se suggests that testicular Se stores were unaffected, even though the participants' dietary Se intake was tripled and their total body Se approximately doubled by supplementation. These results are consistent with animal studies showing the Se status of testes to be unresponsive to dietary Se intake.

High Cholesterol Content and Decreased Membrane Fluidity in Human Spermatozoa Are Associated With Protein Tyrosine Phosphorylation and Functional Deficiencies

Abstract: Poor-quality sperm show reduced capacity to undergo capacitation-induced protein tyrosine phosphorylation and hyperactivation. Given that these deficiencies can be overcome by membrane-permeant stimulators of the cAMP-dependent kinase system, we hypothesize that the main defect underlying these deficiencies resides on the sperm plasma membrane. Spermatozoa from semen samples obtained from 15 consenting healthy donors were separated in 2 subpopulations, L45 (first interface) and L90 (pellet), using a 45:65:90 ISolate gradient centrifugation method. These sperm fractions were studied before and after a 6-hour capacitating incubation for sperm motion parameters (computer-assisted analysis), including hyperactivation, protein tyrosine phosphorylation (immunofluorescence), membrane fluidity (Laurdan fluorescence), and sterol and phospholipid content (high-performance thin-layer chromatography). In summary, data indicate that L45 (poor-motility) spermatozoa present an excess of cholesterol and desmosterol, which impairs the normal increase in membrane fluidity during capacitation and its consequent activation of protein tyrosine phosphorylation and hypermotility. Therefore, a defect in membrane composition and dynamics is underlying human sperm biochemical and functional deficiencies related to inadequate capacitation.

Medical management of Peyronie's disease.

Abstract: Peyronie’s disease (PD) is a fibrotic disorder of the tunica albuginea of the penis. It affects between 3% and 7.1% of adult men worldwide, making it a significant male health concern. Manifestations of PD include penile pain, varying degrees of curvature, penile plaques, and erectile dysfunction. Comprehensive assessment by subjective and objective evaluation is essential for both diagnosis and appropriate treatment. There are a variety of medical treatment options offering benefits of disease stabilization, pain reduction, plaque modulation, curvature correction, and improved sexual function. These medical treatment options include oral agents, intralesional injection therapy, topical drugs with or without iontophoresis, and combination therapy. While a plethora of medical treatments have been tried, no single medical management or common guideline has been proven to be superior. Radiation has been used to reduce penile pain associated with PD, but is not recommended because of theoretical cellular damage to cavernosal smooth muscle cells leading to erectile dysfunction. Surgical correction still remains the gold standard. This overview presents current information about the medical treatment of PD.

Single-nucleotide polymorphisms of the PRDM9 (MEISETZ) gene in patients with nonobstructive azoospermia.

Abstract: To investigate the possible association between variations in the PRDM9 (MEISETZ) gene and impaired spermatogenesis in humans, we screened for mutations in the human PRDM9 gene using DNA from 217 sterile male patients and 162 provenfertile male volunteers. Two single-nucleotide polymorphisms (SNPs), 17353G>T (Gly433Val) and 18109C>G (Thr685Arg), were identified, as well as an intronic SNP, 15549G>T. These SNPs were identified in the heterozygous state in separate patients who demonstrated azoospermia. Neither variant was identified in fertile subjects. Our results suggest that mutations in PRDM9 may cause idiopathic infertility in human males.

Occupational Exposures Obtained by Questionnaire in Clinical Practice and Their Association With Semen Quality

Abstract: In industrial countries, evidence suggests that semen quality has been steadily decreasing over the past 5 decades. We employed a short questionnaire to examine the association between self-reported physical or chemical occupational exposures and semen quality. The study included 402 men consulting for couple infertility (314 with oligospermia, asthenospermia, or teratospermia and 88 with normal semen; World Health Organization criteria). Exposure effects on global sperm quality and total sperm count, sperm motility, and sperm morphology were investigated. We found significant associations between semen impairment and occupational risk factors such as exposure to heavy metals (adjusted odds ratio [OR] = 5.4; 95% confidence interval [CI], 1.6-18.1), solvents (OR = 2.5; 95% CI, 1.4-4.4), fumes (OR = 1.9; 95% CI, 1.1-3.4), and polycyclic aromatic hydrocarbons (OR = 1.9; 95% CI, 1.1-3.5). Exposure to pesticides or cement was nearly significant (OR = 3.6; 95% CI, 0.8-15.8, and OR = 2.5; 95% CI, 0.95-6.5, respectively). Physical risk factors were associated with some sperm anomalies, such as mechanical vibrations with oligospermia and teratospermia as well as excess heat and extended sitting periods with impaired motility. Exposure to ionizing radiation and electromagnetic fields was not associated with semen impairment; these results, however, may be skewed, because very few subjects reported such exposure. Despite the small dataset, self-reported exposures were correlated with semen impairment. This approach may be recommended in routine clinical practice to seek relationships between occupational exposures to reprotoxic agents and impaired semen parameters. This knowledge would allow preventive measures in the workplace to be established and could be complemented by the use of biomarkers to better characterize exposure to chemical substances and their spermiotoxic effects.

The emerging link between hypogonadism and metabolic syndrome.

Abstract: The metabolic syndrome (MS) is comprised of various medical conditions that confer increased risk of diabetes and cardiovascular disease. The pathophysiologic components of MS include glucose abnormality, obesity or increased waist circumference, increased blood pressure, and hyperlipidemia. There is an increased risk of hypogonadism in men with MS and its individual components, including insulin resistance, considered by some to be at the core of MS. Hypogonadism may even predict MS. These factors are interwoven and impact overall health, including sexual dysfunction. One interesting and important question is whether treating hypogonadism with testosterone replacement will ameliorate the pathological components of MS.

Considerations in Evaluating Human Spermatogenesis on the Basis of Total Sperm per Ejaculate

Abstract: Total number of sperm per ejaculate (TSperm) is an important measure for clinicians to provide advice to patient couples. However, TSperm per hour of abstinence (TSperm/h) is a better measure for epidemiologist-andrologist teams or clinicians to evaluate spermatogenesis because it is a rate function. This review looks at the interplay and impacts of rate of sperm accumulation in the excurrent duct system, abstinence interval, sexual arousal, and masturbation vs intercourse on observed TSperm. It also examines why and when TSperm/h might provide a meaningful quantitative evaluation of spermatogenesis (ie, rate of sperm production). There is no doubt that TSperm increases with longer abstinence, and in different men plateaus after 2-9 days. Clinicians wishing to maximize number of fully functional sperm available during intercourse, or for artificial insemination, might wish to recommend 6-7 days of abstinence. Diagnostically, the important feature is TSperm/h. After abstinence interval exceeds 64-72 hours, TSperm/h has started to decline in most nonoligozoospermic men as rate of sperm accumulation in the excurrent ducts approaches zero; apparently increasingly more sperm are voided in urine. Clinicians or epidemiologist-andrologist teams wishing to have optimal distinction among individuals with high, typical, or low sperm production (ie, normal or abnormal spermatogenesis) should accurately measure TSperm/h for samples provided after 42-54 hours' abstinence (never 64 hours). Longer abstinence intervals reward men with poor sperm production, because sperm accumulate in the excurrent ducts for 7 days or more of abstinence, and penalize men with good sperm production, because after 3 days or less of abstinence their excurrent ducts probably are full.

Posttranslational Modification of Constitutive Nitric Oxide Synthase in the Penis

Abstract: Erectile dysfunction (ED) is a common men's health problem characterized by the consistent inability to sustain an erection sufficient for sexual intercourse. Basic science research on erectile physiology has been devoted to investigating the pathogenesis of ED and has led to the conclusion that ED is predominately a disease of vascular origin, neurogenic dysfunction, or both. The constitutive forms of nitric oxide synthase (NOS, endothelial [eNOS] and neuronal [nNOS]) are important enzymes involved in the production of nitric oxide (NO) and thus regulate penile vascular homeostasis. Given the effect of endothelial- and neuronal-derived NO in penile vascular biology, a great deal of research over the past decade has focused on the role of NO synthesis from the endothelium and nitrergic nerve terminal in normal erectile physiology, as well as in disease states. Loss of the functional integrity of the endothelium and subsequent endothelial dysfunction plays an integral role in the occurrence of ED. Therefore, molecular mechanisms involved in dysregulation of these NOS isoforms in the development of ED are essential to discovering the pathogenesis of ED in various disease states. This communication reviews the role of eNOS and nNOS in erectile physiology and discusses the alterations in eNOS and nNOS via posttranslation modification in various vascular diseases of the penis.

Comparison of different methods for assessment of sperm concentration and membrane integrity with bull semen.

Abstract: Assessing semen quality is crucially important for the exploitation of genetically superior sires in an artificial insemination (AI) program. In this study, we compare modern and conventional techniques to estimate bovine sperm concentration and membrane integrity. First, the NucleoCounter SP-100 was validated for sperm concentration and provided statistically reliable and repeatable estimates among aliquots and replicates of 25 fresh ejaculates. Sperm concentrations in 78 ejaculates were then determined with hemacytometer, flow cytometer, and NucleoCounter SP-100 and were significantly correlated (P < .001), with regression coefficients among these 3 techniques close to 1 (P < .01). However, the sperm concentration determined by hemacytometer was lower (P < .01) than by flow cytometer and NucleoCounter SP-100. Forty frozen-thawed semen samples were then assessed for sperm concentration and membrane integrity with hemacytometer, flow cytometer and NucleoCounter SP-100. Significant relationships were found for sperm concentration determined by hemacytometer and NucleoCounter SP-100 and for sperm membrane integrity determined by flow cytometer and NucleoCounter SP-100 (P < .01). Finally, the standard curves of sperm concentrations in 6 spectrophotometers, comparing optical density against counts drawn by hemacytometer and NucleoCounter SP-100 (n = 94 fresh ejaculates) showed different (P < .01) intercepts and regression coefficients (linear, quadratic, cubic). It was calculated that a breeding station can improve its production potential by 13% with the use of NucleoCounter SP-100 instead of hemacytometer for calibration of spectrophotometers. Flow cytometer and NucleoCounter SP-100 can be used with equal confidence to estimate sperm concentration and membrane integrity in domestic animals and human semen.

Improved Quality of Cryopreserved Cheetah (Acinonyx jubatus) Spermatozoa After Centrifugation Through Accudenz

Abstract: Sperm cryopreservation, in combination with assist- ed reproductive techniques, is a valuable tool for the genetic management of endangered felids. However, the acrosome of the cheetah spermatozoon is especially sensitive to cryopreservation, with approximately 40% of spermatozoa experiencing acrosomal damage immediately after thawing and then another approximately 15% loss during the next 4 hours in vitro. Additionally, thawing causes a reduction in sperm motility by approximately 20% with another decrease of approximately 12% during subsequent incuba- tion in vitro. We hypothesized that slow removal of glycerol from cryopreserved cheetah spermatozoa using an Accudenz gradient would improve acrosomal integrity, sperm motility longevity, and structural morphology. Accudenz was compared with traditional cheetah sperm processing methods for glycerol removal that involves washing, multistep resuspension, and swim-up processing. Electroejaculates (n 5 21 total from 8 males) were washed in Ham F10 medium, and sperm pellets were resuspended in TEST-yolk buffer with 0% glycerol. Samples were cryopreserved in straws in 4% final glycerol, thawed, and assessed for percent intact acrosomes (% IA), percent motility (% M), and forward progressive status (FPS; scale, 0-5). Sperm motility index (SMI) was calculated as (% M + (FPS 6 20)) 4 2. In study 1, glycerol removal by centrifugation through an Accudenz gradient (4%, 10%) was compared with traditional sperm washing (control) and multistep resuspension protocols. At each time after centrifugation (hourly for 4 hours), % IA was improved (P , .05) for Accudenz (range, 36%-39%) compared with control (30%-33%) and multistep (29%-33%) treatments. In study 2, a modified Accudenz protocol was compared with traditional washing and was found to improve (P , .05) SMI (range, 52-64) compared with controls (range, 41-52) at each time postthaw after centrifugation. In study 3, swim-up processed sperm were compared with those treated by centrifugation through Accudenz and traditional sperm washing for improving sperm morphology. The percentage of structurally-normal sperm recovered postthawing increased (P , .05) for both the Accudenz (38%) and swim-up (33%) treatments compared with controls (21%). Percent IA and SMI also were improved (P , .05) for Accudenz (range, 39%- 47% and 46-59, respectively) compared with controls (range, 26%- 33% and 40-53, respectively). Results indicate that using Accudenz for glycerol removal from cryopreserved cheetah sperm mitigates the significant loss in sperm quality that occurs after freeze-thawing. This alleviation of cellular damage resulting from cryopreservation contributes to a more than 10% improvement in overall sperm motility and, more importantly, allows retention of 40% or more of sperm with intact acrosomes.

The efficacy of recombinant human follicle-stimulating hormone in the treatment of various types of male-factor infertility at a single university hospital.

Abstract: The aim of the study was to prospectively investigate the efficacy of recombinant human follicle-stimulating hormone (rhFSH) in the treatment of various types of male-factor infertility at a single university hospital. The study included 61 infertile men receiving rhFSH because of various type of male infertility. Treatment included 100-150 IU of rhFSH 2-3 times/wk. All men were divided into 4 groups: hypogonadotropic hypogonadism (n = 21), isolated follicle-stimulating hormone (FSH) deficiency (n = 13), idiopathic oligoasthenospermia (n = 16) and maturation arrest on testicular biopsy (n = 11). Total motile sperm count (TMSC), serum FSH level, and testicular volume were compared before and after treatment in all groups. In the hypogonadotropic hypogonadism group, spermatozoa appeared in the ejaculate, with a mean TMSC of 6.67 +/- 1.57 million, in 15 of 21 patients (71.4%) who were totally azoospermic before the treatment. In the isolated FSH deficiency group, TMSC significantly increased from 6.64 +/- 3.27 to 32.4 +/- 9.09 million after the treatment (P = .003). TMSC did not significantly increase in the idiopathic oligoasthenospermia group. Two of the men with maturation arrest (18.1%) had spermatozoa in the ejaculate after the treatment. rhFSH therapy may be effectively used to improve sperm parameters in infertile men with hypogonadotropic hypogonadism and isolated FSH deficiency. In addition, rhFSH may effect some improvement by either providing sperm in ejaculate or increasing intracytoplasmic sperm injection success in infertile men with maturation arrest.

Analysis of risk factors for organic erectile dysfunction in Egyptian patients under the age of 40 years.

Abstract: Many risk factors have been implicated in the pathogenesis of erectile dysfunction (ED), but it is not clearly evident which of these factors are more relevant among the young population. The aim of this project was to find the most significant risk factors for this disease in young patients. We included 434 patients with organic ED younger than 40 years and 272 age-matched controls. All participants had their complete history taken (including the International Index of Erectile Function-5 [IIEF-5]) and underwent physical examination and some laboratory investigations. Univariate analysis was then applied to study the significance of the following factors in the predisposition of ED: smoking, use of recreational drugs, obesity, dyslipidemia, diabetes mellitus, hypertension, coronary heart disease, and chronic pelvic pain syndrome. This analysis showed that smoking, use of recreational drugs, dyslipidemia, hypertension, and obesity were the significant factors (P < .05 for each factor). When these significant factors were studied in the multivariate model, the only factors that sustained the statistical significance were smoking (P < .05; odds ratio [OR], 1.78; 95% confidence interval [95% CI], 1.16–2.72) and use of recreational drugs (P < .05; OR, 3.18; 95% CI, 1.15–8.82). In addition, a negative correlation was detected between the smoking index of the impotent patients and their IIEF-5 score (r2 = 0.67; P < .05). In conclusion, smoking and the use of recreational drugs are the most significant risk factors for organic ED in patients younger than 40 years.

Effects of Alfuzosin and Tamsulosin on Sperm Parameters in Healthy Men: Results of a Short‐Term, Randomized, Double‐Blind, Placebo‐Controlled, Crossover Study

Abstract: Ejaculation disorders are associated with tamsulosin treatment for lower urinary tract symptoms suggestive of benign prostatic hyperplasia (BPH). To assess whether tamsulosin has any effect on semen, sperm parameters were evaluated in healthy men receiving tamsulosin, alfuzosin, and placebo. Forty-eight healthy men received 5 days of tamsulosin 0.8 mg once daily (QD), alfuzosin 10 mg QD, and placebo in a randomized, double-blind, 3-way crossover study with a 10-14-day washout period between treatments. The changes (x +/- SE) from baseline in semen sperm concentration, semen sperm count, semen viscosity, semen fructose, sperm motility, and sperm morphology on day 5 of treatment were assessed. The change from baseline in semen sperm concentration was 3.1 +/- 8.3 million/mL with tamsulosin, 15.0 +/- 6.5 million/mL with alfuzosin, and 24.4 +/- 6.5 million/mL with placebo. The total sperm count in semen decreased from baseline with tamsulosin (-54.6 +/- 24.0 million) but not with placebo (81.5 +/- 18.8 million) or alfuzosin (46.2 +/- 19.0 million). The percentage of men with normal semen viscosity was lower with tamsulosin (65%) than with placebo (98%) or alfuzosin (92%). The change from baseline in semen fructose was comparable for all treatments. The percentage of motile sperm decreased 13.8% from baseline to day 5 of treatment with tamsulosin compared with decreases of 2.3% with placebo and 0.4% with alfuzosin. The percentage of abnormal sperm increased marginally with tamsulosin (0.6%) but not with placebo (-2.8%) or alfuzosin (-3.9%). The most common adverse events were dizziness (alfuzosin 11%, tamsulosin 14%, placebo 0%) and orthostatic hypotension (alfuzosin 25%, tamsulosin 11%, placebo 5%). The results suggest that tamsulosin has a negative effect on sperm in healthy men. Studies on the effects of alpha(1)-adrenergic blockers on sperm in men with BPH are warranted.

Increased aggressiveness of human prostate PC-3 tumor cells expressing cell surface localized membrane type-1 matrix metalloproteinase (MT1-MMP).

Abstract: Membrane type-1 matrix metalloproteinase (MT1-MMP) is a multidomain transmembrane endopeptidase with a major role in physiological and pathological processes through proteolysis of extracellular matrix and other pericellular proteins. We examined cell surface function of MT1-MMP in PC-3 human prostate tumor cells selected for metastasis in nude mice (PC-3-LN4), or transfected with the full-length wild-type (WT) MT1-MMP or with the mutant form lacking the cytoplasmic tail (ΔC-MT1-MMP). Enhanced cell surface MT1-MMP was determined by fluorescence-activated cell sorting analysis and evidenced mechanistically by increased activation of proMMP-2 and invasion into type-I collagen gels. PC-3 cells overexpressing MT1-MMP grew faster than mock-transfected control cells subcutaneously in nude mice. MT1-MMP localized in caveolae, as judged by immunofluorescence microscopy and sucrose-gradient, detergent-resistant cell fractionation. ΔC-MT1-MMP was strongly associated with caveolae, whereas the WT form was present in both caveolae and noncaveolae fractions. The role of plasma membrane MT1-MMP was supported by localization of MT1-MMP by immunofluorescence microscopy at the cell surface of tumor cells in primary prostate cancers. Increased plasma membrane localization of MT1-MMP, either in caveolae or in other lipid raft structures, is a mechanism to localize this proteinase in contact with extracellular matrix and other pericellular proteins, the cleavage of which can facilitate prostate cancer cell invasion and metastasis.

Hormonal regulation of total antioxidant capacity in seminal plasma.

Abstract: Infertility is associated with oxidative stress, normally counterbalanced by different antioxidant systems. In order to explore the hormonal control of seminal plasma total antioxidant capacity (TAC) we evaluated TAC and hormone patterns in a group of unselected infertile patients and control subjects. One hundred and ten infertile patients (divided into 3 groups: inflammation, varicocele, and other etiologies) and 31 fertile men were examined, evaluating blood serum gonadotropins, testosterone, estradiol, free tri-iodothyronine, free tetraiodothyronine (FT4), thyrotropin, prolactin (PRL), seminal parameters, and TAC. TAC was measured using the H2O2-metmyoglobin system, which generates the spectroscopically detectable radical cation of the chromogenous compound 2,2I-azinobis (3-ethylbenzothiazoline-6-sulfonate). The “lag time” of its appearance is proportional to the antioxidant activity. Lag phase was significantly higher in varicocele vs controls, whereas it was lower in patients with inflammation vs varicocele or other kinds of infertility. The correlation analysis between hormones and seminal parameters showed an inverse correlation between PRL and sperm motility, and a direct correlation of TAC with PRL and FT4, but not with gonadotropins or gonadal steroids. Our data suggest that systemic hormones may play a role in regulating seminal antioxidant capacity. This is interesting also because some hormones, such as thyroid and pituitary hormones, are not usually tested in the first-level evaluation of male patients with fertility problems.

Estrogen enhances recovery from radiation-induced spermatogonial arrest in rat testes.

Abstract: Irradiation of LBNF(1) rat testes induces spermatogonial differentiation arrest, which can be reversed by gonadotropin-releasing hormone (GnRH) antagonist-induced suppression of intratesticular testosterone (ITT) and follicle-stimulating hormone (FSH). Although exogenous estrogen treatment also enhanced spermatogenic recovery, as measured by the tubule differentiation index (TDI), it was not clear whether estrogen stimulated spermatogonial differentiation only by further suppressing ITT or by an additional independent mechanism as well. To resolve this question, we performed the following experiments. At 15 weeks after irradiation, rats were treated with GnRH antagonist; some also received 17beta-estradiol (E2) and were killed 4 weeks later. GnRH antagonist treatment increased the TDI from 0% to 8%, and addition of E2 further increased the TDI to 39%. However, E2 addition further reduced ITT from 7 ng/g testis, observed with GnRH antagonist to 3 ng/g testis, so decreased ITT levels might have contributed to recovery. Next GnRH antagonist-treated rats were given exogenous testosterone and flutamide to stabilize ITT levels and block its action. This increased TDI slightly from 8% to 13%, but the further addition of E2 significantly raised the TDI to 27%, indicating it acted by a mechanism independent of ITT levels. Plots of TDI for all treatment groups compared with ITT, FSH, or a linear combination of ITT and FSH showed that treatments including E2 produced higher TDI values than did treatments without E2. These results indicate that there was an effect of E2 on spermatogonial differentiation because of an additional direct action on the testis that is unrelated to its suppression of testosterone or gonadotropins.

Effects of an environmentally relevant organochlorine mixture and a metabolized extract of this mixture on porcine sperm parameters in vitro.

Abstract: Organochlorine chemicals are present in the envi-ronment worldwide; however, populations living in the Far North areparticularly at risk because their traditional diets are mainlycomposed of contaminated animals (fish, seals, whales, and polarbears). It has been suggested that male fertility is globally declining,possibly because of chronic, low-level exposure to environmentalcontaminants. This study was designed to assess the effects onfresh sperm fertility parameters using the porcine model of 1) anenvironmentally relevant mixture of 15 organochlorines and 2) themetabolized extract of this mixture. In the first experiment, theorganochlorine mixture (at relative concentrations of 10.5, 14.7, and21 mg/mL polychlorinated biphenyls [PCBs]) reduced sperm totalmotility, progressive motility, and viability, and increased capacita-tion, spontaneous acrosome reaction rates, and cytosolic calciumlevels, suggesting that the mixture alters the sperm membrane and isdetrimental to sperm function. In the second experiment, themetabolized extract of this organochlorine mixture (at relativeconcentrations of 0.9, 1.8, 2.7, 3.6, and 4.5 mg/L OH-PCBs) tendedto decrease only sperm total motility. In an in vitro porcine model, themixture of organochlorines, as found in the Arctic food chain, wasrapidly detrimental to sperm function at concentrations aboveenvironmental levels. In contrast, short and physiologically relevantexposure to the metabolized extract of this mixture induced onlylimited adverse effects on sperm motility.Key words: Metabolites, PCB, spermatozoa, DDE.J Androl 2009;30:317–324

Chemoprotective Effect of a Nuclear Factor-κB Inhibitor, Pyrrolidine Dithiocarbamate, Against Cisplatin-Induced Testicular Damage in Rats

Abstract: The objective of this study was to evaluate inducible nitric oxide synthase (iNOS) and nuclear factor-kappaB inhibitor (NF-kappaB) expression and the potential chemoprotective effects of an NF-kappaB inhibitor, pyrrolidine dithiocarbamate (PDTC), against cisplatin-induced testicular damage in rats. Rats were divided into 4 equal groups: group 1, control; group 2, injected with cisplatin (CIS) for 5 days (7 mg/kg/day intraperitoneally [IP]); group 3, injected with PDTC alone; group 4, injected with CIS plus PDTC (100 mg/kg IP). Body and testicular weights, plasma testosterone levels, and histopathologic structure of the testicular tissue were determined. The iNOS and NF-kappaB activity were evaluated immunohistochemically by staining p65 to define NF-kappaB activity. Malondialdehyde (MDA), reduced glutathione (GSH), and nitric oxide (NO) levels and glutathione peroxidase (GSH-Px) activity were assessed in testicular tissue. Body and testicular weights, plasma testosterone levels, activity of GSH-Px, and GSH levels were all significantly decreased, whereas the levels of MDA and NO were significantly increased in rats of the CIS group. PDTC treatment increased plasma testosterone levels. A significant increase in GSH levels and GSH-Px activity and a decrease in MDA and NO levels in testicular tissue were observed in the CIS + PDTC group. Immunohistochemically, there was a marked staining for iNOS and NF-kappaB/p65 expression in rats injected with CIS compared with the control (P < .001). CIS caused irregular seminiferous tubules, reduction of seminiferous epithelial layers, significant arrest of maturation, and perivascular fibrosis. Moreover, PDTC administration to CIS-treated rats significantly prevented these histopathologic chances, as well. CIS induces iNOS expression through activation of NF-kappaB/p65, and CIS-induced testicular toxicity may be prevented by PDTC, which is a selective NF-kappaB inhibitor.

Testicular function in poor-risk nonseminomatous germ cell tumors treated with methotrexate, paclitaxel, ifosfamide, and cisplatin combination chemotherapy.

Abstract: Our objective was to investigate the impact ofmethotrexate, paclitaxel, ifosfamide, and cisplatin (M-TIP) on long-term fertility in poor-risk nonseminomatous germ cell tumors(NSGCT). Thirty patients with poor-risk NSGCT (median age,29 years; range, 17–62 years) were treated with methotrexate250 mg/m 2 with folinic acid rescue (day 1) and paclitaxel 175 mg/m 2 (day 1), followed by ifosfamide 1.2 g/m 2 and cisplatin 20 mg/m 2 (days 2–6). Treatment consisted of 4 cycles of M-TIP administeredevery 3 weeks. Twenty-one patients were continuously disease-freeat a median follow-up of 5.3 years (range, 0.9–8.4 years). Spermcount and hormonal analyses were examined prechemotherapy (30patients) and postchemotherapy (21 patients). Counts were classi-fied as follows: lower than 1 6 10 6 /mL, azoospermia; 1–20 6 10 6 /mL, oligospermia (OS); higher than 20 6 10 6 /mL, normospermia(NS). Patients were followed for a median of 2.3 years (range, 0.9–3.8 years) postchemotherapy. The prechemotherapy median lutein-izing hormone (LH) serum levels were slightly above the uppernormal limit, whereas the serum levels of follicle-stimulating hormone(FSH) and testosterone (T) were within the reference interval. Eleven(52.3%) patients had NS prechemotherapy. Among the patients withNS, 72.7% still had NS following chemotherapy. Overall, 17 of 21(80.9%; 33.3% OS and 47.6% NS) patients had recovery ofspermatogenesis after treatment. The median FSH serum levelswere significantly elevated at least 1 year postchemotherapy whencompared with the pretreatment levels. Eighteen months after thecompletion of chemotherapy the median FSH levels had returned tothe reference limits. Serum LH and T levels were unaffected bychemotherapy. Prior to chemotherapy 4 of 30 patients had fathered 5children. Since completion of chemotherapy, 5 patients havefathered 5 children. The majority of men with poor-risk germ celltumors who were treated with the M-TIP regimen demonstratedrecovery spermatogenesis after treatment, and Leydig cell functionwas unaffected.Key words: Fertility, gonadal toxicity.JAndrol2009;30:280–286

Length of androgen receptor-CAG repeats in fertile and infertile Egyptian men.

Abstract: Androgens play key roles in spermatogenesis, and they exert their effect via the androgen receptor (AR). The AR gene has a repetitive DNA sequence in exon 1 that encodes a polyglutamine tract. Instability in the glutamine (CAG) repeat unit length is polymorphic across ethnic groups. Previous studies of the relationship between the repeat unit length and male infertility have been contradictory. To establish the range of wild-type alleles in Egyptian men, we determined the range of repeat lengths in a population of normally fertile, ethnically selected Egyptian men. We also investigated the association between trinucleotide repeat length within the AR gene and male factor infertility in a population of ethnically selected Egyptian infertile men, who were compared with fertile, ethnic group-matched and age-matched controls. The study included 129 clinically selected infertile Egyptian men who were scheduled for intracytoplasmic sperm injection and 52 ethnically matched fertile controls. The experimental population was grouped according to sperm counts ranging from nonobstructive azoospermia to normozoospermia. The CAG repeat N-terminal domain region of the AR gene was amplified in peripheral blood DNA, and allele size was determined by fragment analysis. Allele size and single-nucleotide polymorphism and mutation rates were determined by sequencing individual amplified alleles. The mean CAG repeat length in the azoospermia group was 18.55 +/- 2.0; in the severe oligozoospermia group it was 18.21 +/- 3.42; in the oligozoospermia group it was 18.27 +/- 2.93; and in the infertile with normal sperm count group it was 17.72 +/- 2.0. In the control group, the mean CAG repeat length was 18.18 +/- 3.63. No significant correlation was found between CAG repeat length and the risk of male factor infertility in an ethnically defined population of Egyptian men. However, a significant and positive correlation between CAG repeat length and serum testosterone concentration was demonstrated. This suggests the involvement of epigenetic regulation linked to this region.

Differential Gene Expression in the Testes of Different Murine Strains Under Normal and Hyperthermic Conditions

Abstract: Cryptorchidism and scrotal heating result in abnormal spermatogenesis, but the mechanism(s) prescribing this temperature sensitivity are unknown It was previously reported that the AKR/N or MRL/MpJ-+/+ mouse testis is more heat-resistant than the testis from the C57BL/6 strain We have attempted to probe into the mechanism(s) involved in heat sensitivity by examining global gene expression profiles of normal and heat-treated testes from C57BL/6, AKR/N, and MRL/MpJ-+/+ mice by microarray analysis In the normal C57BL/6 testis, 415 and 416 transcripts were differentially expressed (at least 2-fold higher or lower) when compared with the normal AKR/N and MRL/MpJ-+/+ testis, respectively The AKR/N and MRL/MpJ-+/+ strains revealed 268 differentially expressed transcripts between them There were 231 transcripts differentially expressed between C57BL/6 and 2 purported heat-resistant strains, AKR/N and MRL/MpJ-+/+ Next, the testes of C57BL/6 and AKR/N mice were exposed to 43 degrees C for 15 minutes and harvested at different time points for terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) studies and microarrays An increase of TUNEL-positive germ cell numbers was significant 8 hours after heat exposure in the C57BL/6 mouse However, this increase was not observed in the AKR/N mouse until 10 hours after heat exposure All tubules showed germ cell loss and disruption in C57BL/6 testis 24 hours after heat shock In contrast, although a number of seminiferous tubules showed an abnormal morphology 24 hours post-heat shock in the AKR/N mouse, many tubules still retained a normal structure Numerous transcripts exhibited differential regulation between the 2 strains within 24 hours after heat exposure The differentially expressed transcripts in the testes 8 hours after heat exposure were targeted to identify the genes involved in the initial response rather than those attributable to germ cell loss Twenty transcripts were significantly down-regulated and 19 genes were up-regulated by hyperthermia in C57BL/6 and did not show a parallel change in the AKR/N testis Conversely, heat shock resulted in 30 up-regulated transcripts and 31 down-regulated transcripts in AKR/N that were not similarly regulated in C57BL/6 A number of genes shared similar differential expression patterns and differential regulation by hyperthermia in both strains of mice Taken together, the results of the present study indicate that the diverse genetic backgrounds in the 3 strains lead to major differences in normal testis gene expression profiles, whereas the differences in heat shock responses involve a significantly smaller number of genes The data generated may provide insights regarding gene networks and pathways involved in heat stress and their relationship to spermatogenesis