A differential cytokine expression profile before and after rFSH treatment in Sertoli cell cultures of men with nonobstructive azoospermia.

To evaluate the effects of follicle-stimulating hormone (FSH) treatment on cytokine gene expression in cultured Sertoli cells from men with nonobstructive azoospermia, a total of 15 azoospermic men diagnosed as obstructive azoospermia (OA) (n = 5) and nonobstructive azoospermia (NOA) (n = 10) were included in the study. NOA patients were split into two further subgroups: nFSH and hFSH serum FSH levels. Expression of cytokine gene panel (88 genes), FSHR and ABP was evaluated by real-time PCR array analysis. FSHR protein level was measured by the Western blot. In primary cultures of Sertoli cells, seven genes were found to be increased and 13 were decreased in NOA group, when compared to OA (p < .05). When rFSH was introduced into the culture media, expression of 12 genes in the NOA group restored a comparable level to those of the control OA group. Sertoli cells in all groups responded rFSH administration with increased expression of ABP. Our results suggest that FSH treatment may have positive effects on Sertoli cells of nonobstructive azoospermic patients via changing the expression levels of certain genes and restoring their levels in normal Sertoli cell population. Some cytokine levels can be considered as a potential candidate for detecting NOA patients. ABP is a good marker for cell viability and functionality in primary Sertoli cell culture.

Role of ADAMTS1 and ADAMTS5 in male infertility.

The aim of this study was to investigate the relationship of infertility with metalloproteinases ADAMTS1 and ADAMTS5, which are known to be responsible for the degradation of extracellular matrix (ECM) proteins associated with many diseases. ECM is the noncellular component that provides structural and biochemical support to the surrounding cells required for tissue morphogenesis, differentiation and homoeostasis. Sixty infertile individuals and 10 healthy semen donors were included in this study. The infertile individuals were classified as normozoospermia (NS; n = 20), oligozoospermia (OS; n = 20), azoospermia (AS; n = 20) groups. ADAMTS1 and ADAMTS5 protein levels in semen were analysed by Western blot. ADAMTS1 protein level was 3.0-, 3.3- and 1.6-fold lower in the OS, AS and NS groups, respectively, than in the control group (P < 0.001). ADAMTS5 protein level was 3.2-, 2.7- and 1.4-fold lower in the OS, AS and NS groups, respectively, than in the control group (P < 0.001). Sperm count and sperm motility showed a negative correlation with the levels of ADAMTS1 and ADAMTS5 protein expression: r = -0.477, r = -0.470; and r = -0.332, r = -0.275 respectively (P < 0.001). In conclusion, ADAMTS1 and ADAMTS5 protein expressions in semen are significantly related with sperm production. It is very important to understand molecular function and organisation of ADAMTSs which will be significant in enlightening the process of spermatogenesis in male infertility.

Association of MDR1 C3435T and C1236T single nucleotide polymorphisms with male factor infertility.

Infertility affects 1 in 6 couples and approximately 1 in 25 men. Male factor infertility is a major cause of spermatogenic anomalies, the causes of which are largely unknown. Impaired repro-ductive functions in men might result from physiological, genetic, and/or environmental factors such as xenobiotics. The multi-drug re-sistance1 (MDR1) gene encodes a P-glycoprotein which has a role in the active transport of various substrates providing protection of somatic cells from potentially toxic substances, including xenobi-otics. MDR1 is highly expressed at the luminal surface of capillary endothelial cells, and is expressed in Leydig cells, testicular mac-rophages, and Sertoli cells. We performed genotype and haplotype analyses of MDR1 in 192 infertile and 102 fertile Turkish men for the genetic markers C1236T and C3435T, using polymerase chain reaction-restriction fragment length polymorphism analysis. In the overall population, correlations were analyzed in all genotype mod-els. We found that the C3435T polymorphism TT vs CT genotypes showed statistically significant differences in their association with infertility (P = 0.045), and that the CT genotype was associated with high sperm DNA damage (P = 0.02), suggesting that the CT genotype might be a susceptibility factor for infertility. Additionally, the T-T haplotype was significantly more frequent in the control group (13.2 vs 6.5%; odds ratio = 0.459, 95%CI = 0.259-0.814, P = 0.006). This study showed that MDR1 might have a role in male infertility. Fur-ther research in large cohorts with different populations is required to clarify the role of MDR in male fertility.

Correlation of seminal parameters with serum lipid profile and sex hormones.

The phospholipids and fatty acid composition of sperm are altered under the influence of serum lipids on impaired seminal parameters in infertile men. We studied the correlation of the serum lipid profile and sex hormone results of a group of infertile men with sperm characteristics of 18 infertile men. Semen samples were collected and analyzed. Serum fasted and post-meal glucose levels, insulin, total cholesterol, HDL, LDL, VLDL, triglyceride, FSH, LH, and testosterone measurements were performed. The increased serum VLDL, total triglyceride, and testosterone values were significantly correlated with decreased sperm motility. Serum FSH values were also inversely correlated with normal sperm morphology. The increased VLDL impaired seminal parameters; additionally, increased triglycerides may have deleterious effects on spermatogenesis. Deterioration is related with increased serum VLDL and triglyceride levels.

Effect of sperm chromatin damage on fertilization ratio and embryo quality post-ICSI.

A total of 56 semen samples was collected from men in the ICSI program. The female partners of all men were under 35 years of age. Sperm chromatin integrity was evaluated by staining the spermatozoa with acridine orange. > 56% red fluorescence was taken as abnormal chromatin status. A significant negative association was found between the percentage of sperm with DNA fragmentation and fertilization rate. Increased red fluorescence indicated impaired fertilization outcome. Good quality embryo rates were significantly lower in abnormal AO stained group. Chromatin damage precedes the loss of fertilization potential and poor embryo quality.

The impact of the source of spermatozoa used for ICSI on pronuclear morphology.

BACKGROUND: The aim of this prospective study was to find out whether the source of spermatozoa used for intracytoplasmic sperm injection (ICSI) has an impact on the morphological features of pronucleate zygotes, which make up the basis of a pronuclear scoring system for the selection of the most viable embryos for transfer. METHODS AND RESULTS: The study group consisted of 194 two pronucleate (2PN) ICSI zygotes, of which 144 originated from ejaculated (ES) and 50 from testicular spermatozoa (TS). At 18 h postinjection, 2PN zygotes were assessed for pronuclear alignment, polarity in nucleoli and cytoplasmic appearance; all of which were found to exhibit similar patterns of distribution between the ES and TS groups (P = not significant). At 25 h, the presence of first cleavage was similar for both groups; 11% of zygotes in the ES and 10% of those in the TS group underwent early cleavage (P = not significant). At 48 h, a quality score was obtained for cleaving embryos by multiplying the number of blastomeres with the grade of the embryo. Pronuclear scoring in both groups of spermatozoa correlated with embryo quality score at 48 h postinjection. There was a trend for a higher incidence of early cleavage and a lower incidence of pronuclear arrest with better pronuclear scoring embryos for both types of spermatozoa. CONCLUSION: The morphological features of pronucleate zygotes at 18 h after microinjection with ES and TS are similar to each other.

Nicotine toxicity to the ultrastructure of the testis in rats.

OBJECTIVE: To investigate the influence of nicotine exposure on the ultrastructure of the rat testis. MATERIALS AND METHODS: Twenty rats were injected with nicotine at a dose of 0.4 mg/100 g body weight daily for 3 months; a group of 20 control rats matched for weight and age were injected with saline only for the same duration. The testes were then harvested and examined by transmission electron microscopy. RESULTS: Rats given nicotine showed: thickening of the tunica propria, caused by an increase in the collagen fibres under the irregular basal lamina; degeneration of junctional specializations between the Sertoli cells, with malformed nuclei showing condensed chromatin; Sertoli cells with numerous polymorphic mitochondria with irregular cristae and an electron-dense matrix. The germ cells were degenerated, spermatids retained excess cytoplasm and accumulated electron-dense lipid droplets in the cytoplasm. The acrosomes were irregular and abnormally configured. CONCLUSION: There were ultrastructural alterations in rats exposed to nicotine that could be attributed to the detrimental effects of nicotine on germ cells, peritubular structures and Sertoli cells.

Ultrastructural effects of cigarette smoke on rat testis.

The purpose of this study was to investigate the ultrastructural changes taking place in the testicular tissue of adult male Wistar rats following exposure to cigarette smoke. Twenty rats were exposed to smoke in a smoking machine for 2 h daily for 60 days; 10 control rats were placed in the machine for the same amount of time but were exposed to room air. After ether anesthesia and dissection of the animals, the testes were removed and fixed in 2.5% glutaraldehyde. The sections were examined under a transmission electron microscope. Evident degeneration and dissociation of the spermatogenetic cells were common findings. Basal lamina of the tubuli seminiferi contorti was thickened and significantly irregular. Cigarette smoke inhalation damaged the seminiferous epithelium. The primary spermatocytes as well as Sertoli cells appeared to be targets for the smoke. The results indicated that cigarette smoke inhalation could cause specific lesions in the development of spermatozoon, and it might be either directly or indirectly toxic to spermatogenesis.

Penile implants in erectile impotence. The importance of clinical experience on outcome.

PURPOSE: We assessed maximum patient and partner acceptance and satisfaction by informed preoperative prosthesis selection. MATERIALS AND METHODS: Six months to 5 years after penile prosthesis implantation, 71 men and partners were interviewed personally. Evaluation was focused on delineating not only the etiologic factors but also factors most likely to affect prosthesis recommendation and contribute to prognosis. RESULTS: Overall, 94% of the patients and 95% of the partners were satisfied, and 90% of these believed that they had equal erections to previously without the implant, and 81% reported an improvement in nonsexual relations with their partners after the device was implanted. 83 and 17% of the patients said they had increased or had no change in sexual desire, respectively. No one had a decrease in sexual desire. 86% of all the patients believed that their orgasm was improved. 89% of the patients were completely satisfied with the fit of their clothes after prosthesis implantation. 93% of the patients reported good to excellent emotional adjustment to the prosthesis and the prosthesis produced a positive change in self-esteem in 94% of all the patients. CONCLUSIONS: A detailed assessment of patient-partner expectations is essential in order to be able to predict more accurately which patients will be best suited to the implant and which may need either concurrent counseling or preimplant education. A team was formed in which urologists and psychologists worked together, provided comprehensive information on several personality variables of interest in treatment planning and postoperative satisfaction. Our results clearly demonstrate that not only patient preference but also clinical data are important factors in enhancing patient and partner satisfaction.

Comparative study of long-term stone recurrence after extracorporeal shock wave lithotripsy and open stone surgery for kidney stones.

BACKGROUND: Extracorporeal shock wave lithotripsy (ESWL) has become the treatment of choice for most calculi of upper urinary tract and the need for open stone surgery (OSS) have considerably reduced. However, stone recurrence is often encountered as a long-term problem requiring re-treatment. METHODS: In the present retrospective study, the recurrence rates of ESWL and OSS were compared in the treatment of kidney calculi. During the 1 year period of the present study, 43 patients were treated by OSS and 400 underwent ESWL (the Dornier MPL 9000 lithotriptor). The recurrence of stone was defined as reappearance of the stone on plain abdominal radiography during the follow-up period. RESULTS: Complete removal of all stone fragments was achieved in 51.2% of patients (n = 22) treated with OSS. In the ESWL group, the stone free rate was 56.7% (n = 237). The recurrence rate was 31.8% within a mean of 40 months (range 32-48 months) in the OSS group, whereas this figure was 13.9%, with a mean period of 46 months (range 42-48 months) in the ESWL group (P < 0.05). The mean periods of recurrence in ESWL and OSS groups were 20 (range 6-42 months) and 11 months (range 8-44 months), respectively (P < 0.05). However, the mean stone burdens of both groups were different (2.9 +/- 0.8 vs 1.4 +/- 1.1 cm). For comparable stone burdens, the recurrence rate was similar. In the ESWL group, stones in the lower calyx, multiple stones and larger stones showed a higher recurrence rate. The recurrence after OSS was also influenced by stone burden. CONCLUSIONS: The results of the present study demonstrate that stone burden may be the primary risk factor for stone recurrence after ESWL and OSS.

Clinical experience with Dynaflex penile prostheses in 120 patients.

OBJECTIVES: To present our experience with the Dynaflex (American Medical Systems) self-contained inflatable penile prosthesis and define specific complication and patient dissatisfaction rates. METHODS: From May 1990 through January 1998, 120 men underwent implantation of a Dynaflex prosthesis. The follow-up period ranged from 2 to 80 months (mean 42). RESULTS: Specific complication rates observed after penile prosthesis implantation were as follows: prosthetic infection, 4.16%; mechanical device failure, 7.5%; and patient dissatisfaction because of inability to work the pump, 16.66%. After intensive teaching, the patient dissatisfaction rate dropped to 0.83%. The overall complication rate was 14.15%. CONCLUSIONS: With several types of penile prostheses available, Dynaflex can be regarded as an appropriate alternative with a relatively low mechanical failure rate. However, careful preoperative assessment and patient education in the use of the pump mechanism are essential to obtain a successful result.

Calyceal stones: fate of shock wave therapy with respect to stone localization.

In a retrospective analysis the results of extracorporeal shock wave lithotripsy (ESWL) treatment were evaluated in patients with renal stones according to calyceal localization of treated stones. The 198 patients who underwent ESWL with the Dornier MPL 9000 were analyzed for success rate, complication rate, residual fragments, regrowth and recurrence rates. Totally 210 calyceal calculi located in different portions of the kidney have been comparatively evaluated. No major complications were noted during or after ESWL. Some minor complications such as flank pain, renal colic, haematuria were observed. Flank pain was observed during ESWL treatment especially in patients with upper calyceal stones. Although stone-free and residual fragment rates were similar in pelvic, upper and middle calyces, patients with lower calyceal and pelvicalyceal stones had high residual fragment rate and lower stone-free rate. Patients with stones in the lower calyces or pelvicalyces had high recurrence and regrowth rates (p < 0.05). ESWL has been considered as the optimal treatment modality for most upper urinary tract calculi. It is especially effective in patients with pelvic, upper and middle calyceal stones. Patients with lower calyceal stones often failed to eliminate the fragments, hence had high recurrence and regrowth rates.

Testicular effects of vasectomy in rats: an ultrastructural and immunohistochemical study.

OBJECTIVES: The correlation between infertility and morphofunctional alterations following vasal occlusion is not clearly understood. Although a correlation has been found between the expression of a high titer of antisperm antibodies and the status of infertility, the relationship between the immunoglobulin (Ig) depositions in the testis and ultrastructural alterations of the peritubular structures has not been clearly established. The objective of this study is to explain the etiopathogenesis of diffuse and focal testicular lesions caused by vasal obstruction. METHODS: Unilateral vasectomy was performed on adult male rats. Ultrastructural changes of testicular tissues and immunostaining affinity of peritubular structures with anti-actin, anti-vimentin, anti-laminin, and anti-immunoglobulin G (anti-IgG) antibodies were analyzed. RESULTS: In an ultrastructural study, vasectomized animals presented abundant collagen fibril accumulation in the peritubular area. In testis with intense IgG antibodies, staining revealed weak immunostaining of peritubular myoid cells with anti-actin antibodies, but intense immunostaining with anti-vimentin antibodies. The tubules of rats with no IgG deposition on peritubular structures revealed intense immunostaining with anti-actin antibodies but scant immunostaining with anti-vimentin antibodies. Anti-laminin deposits decreased severely in most of the tubules demonstrating intense IgG deposition. CONCLUSIONS: Our findings suggest that spermatogenesis deteriorates more severely in testes with dense IgG deposition. The sclerosis of the lamina propria in cases with vasal occlusion is secondary to alterations in the peritubular myoid cells. With the progressive changes that occur in the peritubular myoid cells or differentiation of the peritubular myoid cells that acquire active cell characteristics, collagen accumulation adjacent to these cells increases markedly. The alterations of the peritubular myoid cells reported here may be caused by alterations in basement membrane structures.

Analysis of the relationship between histologic alterations and the generation of reactive oxygen species in vasectomized rat testes.

OBJECTIVES: To evaluate the effects of vasal obstruction on testicular structure, to determine if tissue and/or cell damage can cause significant reactive oxygen species (ROS) generation, and to correlate the histologic alterations to the measured levels of ROS products. METHODS: To evaluate the effects of ROS generation in vasectomized testes, unilateral vasectomy was performed on 17 rats and tissue samples were examined by light microscopy. The histologic alterations were correlated to the measured tissue malondialdehyde levels by thiobarbituric acid analysis as an indicator of the ROS-induced tissue damage. RESULTS: Unilateral vasectomy significantly affected the weights of the ipsilateral testes, but not the contralateral testes. No significant changes were noted in plasma estradiol, follicle-stimulating hormone, luteinizing hormone, and testosterone levels. The mean values of malondialdehyde were found to be significantly higher in vasectomized rats than in control rats (0.025 +/- 0.008 versus 0.018 +/- 0.007 nmol/mg protein) (P < 0.01). The malondialdehyde level was a mean of 28% lower in the contralateral testes tissue as compared to that found in ipsilateral testes (P < 0.01). Malondialdehyde output from vasectomized rats had a parallel correlation to tissue destruction. CONCLUSIONS: Our results indicate that overproduction of ROS may be involved in vasectomy-induced testicular damage.

The role of cigarette smoking in prostatic enlargement.

OBJECTIVE: To investigate the association between cigarette smoking and sex hormone concentrations in men with benign prostatic hyperplasia (BPH) and thus provide some understanding of the underlying mechanism of the effects of cigarette smoking on prostatic enlargement. PATIENTS AND METHODS: The association between cigarette smoking and prostatic volume was investigated in 68 men with BPH (mean age 59 years, range 52-74) by assessing changes in serum levels of oestradiol, testosterone, dihydroepiandrosterone (DHEA) and dihydroepiandrosterone sulphate (DHEA-S). Possible enzymatic mechanisms in the prostate gland that might be affected by cigarette smoking were assessed. RESULTS: The mean prostate volume was greater in non-smokers than smokers. Current cigarette smokers had significantly higher mean serum oestradiol levels than did the non-smokers (33.8 and 26.7 pg/mL, respectively, P < 0.01). Cigarette smoking was inversely but not significantly related to testosterone level. These differences persisted after adjusting for body mass index. There was no significant difference in serum DHEA and DHEA-S levels between smokers and non-smokers. There was a weak correlation between the degree of prostatic enlargement, the presence of obstructive symptoms and urinary flow rates. CONCLUSION: These results indicate that cigarette smoking may affect the size of the enlarging prostate, but through indirect effects of smoking on factors provoking the development of BPH.

Is deep dorsal vein arterialization an alternative surgical approach to treat venogenic impotence?

OBJECTIVE: Several surgical techniques have been applied for the treatment of cavernosal venous leakage, sufficient enough to create erectile dysfunction. In light of the variable success rates, we report our experience with deep dorsal penile vein arterialization for the management of severe cavernosal venous leakage. PATIENTS AND METHODS: Twenty-four impotent patients with venous leakage were treated by the Virag-II type of end-to-end anastomosis of the inferior epigastric artery to the deep dorsal penile vein and ligation of the vein proximally. Diagnostic evaluation included color flow Doppler sonography before and after intracavernous 60 mg papaverine injection and dynamic cavernosometry/ cavernosography, which indicated veno-occlusive dysfunction in 28 patients. Revascularization was technically possible in 24 of the patients, although in 4 anastomosis could not be achieved due to the poor quality of the inferior epigastric artery. RESULTS: Five patients had early occlusion in the immediate postoperative period and 4 had late occlusion within 8 months. Potency was improved initially in 9 (38%) of the 24 patients in whom successful anastomosis had been achieved, with longer term improvement in 6 (25%) of 24 who realized restoration of erectile potency as defined by clinical investigations. The mean follow-up was 24 months (range 3-36 months). CONCLUSION: We believe that anastomosis of the inferior epigastric artery to the deep dorsal penile vein and ligation of the vein proximally in cases of venous leakage results in a low success rate due probably to a pancavernosal alteration in corporal tissue compliance.

Effect of obesity on prostatic hyperplasia: its relation to sex steroid levels.

In 68 men with benign prostatic hyperplasia, we evaluated the association between obesity and prostatic enlargement, as well as changes in serum levels of oestradiol, testosterone, dihydroepiandrosterone and dihydroepiandrosterone sulphate. Despite the larger adenomas, no increase in the symptom score for BPH was observed with increasing obesity. Average specimen weights increased with increasingly obesity and increasing host age from 46 to 80 g. We also found the serum oestradiol level significantly elevated in obese men who were 140% or over recommended weight compared to underweight men younger than 60 years (51.3 pg/ml versus 26.8 pg/ml, p < 0.01). This pattern was present in all age groups. These results indicate that obesity is a risk factor for prostatic enlargement but not for obstruction. Also the degree of obesity appears to have a direct effect on oestradiol levels through transformation of androgens in adipose tissue to oestrogens. In conclusion, further studies to evaluate the pathogenesis, pathophysiology, natural history and symptomatology of BPH would be of great interest and should help to define better the associations that we have recognized.

Evaluation and surgical treatment of patients with penile curvature.

In patients with adult penile curvatures the aim of treatment is to have a penis straight enough and firm enough to allow satisfactory sexual function. Several operative approaches have been used to correct this condition. Herein, we describe our experience with the evaluation and surgical treatment of 37 patients with penile curvature. When we have decided on surgery, we either preferred to elongate the ventral corpora cavernosa by interposition of a piece of dura mater (2 patients) or in most cases (the remaining 35 patients) only excised wedges of tunica albuginea at the arcs of the maximum curvature located with artificial erection (Nesbit's operation). In conclusion, our results showed that Nesbit's operation is a versatile procedure that proved to be useful for correction of penile curvatures in patients with Peyronie's disease. However, further studies and clinical trials are needed to determine if the dura mater is an alternative in the treatment of penile curvatures due to congenital aetiology.

Clinical experience with inflatable and malleable penile implants in 104 patients.

Penile prosthesis was implanted for erectile impotence of mainly organic origin in 104 patients. The AMS Dynaflex penile prosthesis was inserted in 39 cases, the AMS Malleable 600 prosthesis in 61 and the AMS Ultrex Plus prosthesis in 4. In 1 patient receiving a malleable prosthesis both rods had to be removed owing to erosion into the urethra and reimplantation was performed at the same operation. Only 1 patient who underwent implantation of the Dynaflex device had mechanical failure that necessitated surgical revision. After implantation of the new prosthesis both patients had satisfactory intercourse. One Dynaflex penile prosthesis, implanted into a juvenile-onset diabetic, became infected and required removal. Another complication was secondary to spontaneous erosion (noninfected) in 1 patient with malleable prosthesis who suffered loss of only one rod and who is still satisfied with the result. The overall complication rate in our series has been approximately 4%. The rest of the patients report satisfaction with the ability to move the penis voluntarily permitting normal sexual activities as well as normal appearance in the flaccid position. According to our experience, careful preoperative assessment from the views of both patient and device selection along with patient education, and strictly obeying the rules of sterility during implantation and applying systemic and local antimicrobial prophylaxis are essential in obtaining a successful result in prosthesis implantation for the individual patient.

Ultrastructual changes of corpora cavernosa in vascular erectile dysfunction.

Corpora cavernosa of 4 impotent patients with arterial pathology and 6 with venous insufficiency were studied by electron microscopy. The findings in all of the smooth muscle samples were the following: pronounced thickening of the basal lamina; marked reduction of contractile myofilaments and electron dense bodies; finger-like cytoplasmic projections; increase in mitochondria with swelling and aggregation; huge protrusions of nuclear membrane into the cytoplasm; increased proportion of interstitial matrix to smooth muscle cells. These were more marked in the dark than in the light cells, which was considered as the beginning of degeneration. Another finding was degenerative changes in the endothelial cell lining of the sinusoids, especially denudation, fragmentation and marked thickening of the basal lamina. Severity of the symptoms did not change in any of the patients with different pathologies. So, we can assume that the pathological alterations may be due to chronic penile vascular insufficiency or vice versa.