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ABSTRACT: Male infertility is a frequent medical condition, compromising approximately one in twenty men, with infections of the reproductive tract constituting a major etiological factor. Bacterial epididymo-orchitis results in acute inflammation most often caused by ascending canalicular infections from the urethra via the continuous male excurrent ductal system. Uropathogenic Escherichia coli (UPEC) represent a relevant pathogen in urogenital tract infections. To explore how bacteria can cause damage and cell loss and thus impair fertility, an in vivo epididymo-orchitis model was employed in rats by injecting UPEC strain CFT073 into the vas deference in close proximity to the epididymis. Seven days post infection bacteria were found predominantly in the testicular interstitial space. UPEC infection resulted in severe impairment of spermatogenesis by germ cell loss, damage of testicular somatic cells, a decrease in sperm numbers and a significant increase in TUNEL (+) cells. Activation of caspase-8 (extrinsic apoptotic pathway), caspase-3/-6 (intrinsic apoptotic pathway), caspase-1 (pyroptosis pathway) and the presence of 180 bp DNA fragments, all of which serve as indicators of the classical apoptotic pathway, were not observed in infected testis. Notably, electron microscopical examination revealed degenerative features of Sertoli cells (SC) in UPEC infected testis. Furthermore, the passive release of high mobility group protein B1 (HMGB1), as an indication of necrosis, was observed in vivo in infected testis. Thus, necrosis appears to be the dominant cell death pathway in UPEC infected testis. Substantial necrotic changes seen in Sertoli cells will contribute to impaired spermatogenesis by loss of function in supporting the dependent germ cells.
PLoS ONE 01/2013; 8(1):e52919. · 4.09 Impact Factor
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ABSTRACT: There is no consensus for the best testicular sperm extraction (TESE) technique in patients with "low-chance" nonobstructive azoospermia (NOA).
To determine sperm retrieval rates in an intraindividual comparison using three locations of the testicle with and without the assistance of a microscope (microsurgical TESE [M-TESE]).
A series of 65 patients with low-chance NOA presenting with low testicular volume (<8 ml) and high serum follicle-stimulating hormone (FSH) (>12.4 IU/l) underwent trifocal-TESE plus M-TESE bilaterally (four biopsies per testis).
Sperm retrieval was performed as trifocal-TESE (upper, middle, and lower testicular pole) with and without the assistance of a microscope in the middle incision. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: The number of evaluated tubules, the mean spermatogenetic scores, and the sperm retrieval rates were evaluated to determine retrieval locations and the use of the microscope. The Friedman and Cochrane Q tests were applied to determine statistical differences. Receiver operating characteristic curves were used for the analysis of serum FSH and testicular volume as preoperative prognostic factors.
The sperm retrieval success of 66.2% using the combined technique, meaning the percentage of patients with at least one tubule containing elongated spermatids, was the highest in the combination of trifocal- and M-TESE (p<0.01), indicating this technique as optimal for patients with low-chance NOA. M-TESE and trifocal-TESE alone were not significantly better. The mean spermatogenetic score giving the number of tubules with elongated spermatids in relation to all tubules was significantly higher in M-TESE versus conventional TESE (p<0.01), indicating the superior quality of the tissue harvested using the microscope. These results are limited by the definition of "success" using "one" spermatid/tubule. Preoperatively, high serum FSH and low testicular volumes did not exclude successful sperm retrieval.
The combination of trifocal- and M-TESE is the best technique to reach high sperm retrieval rates in patients with low-chance NOA.
European urology 03/2012; 62(4):713-9. · 7.67 Impact Factor
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ABSTRACT: The morphology and size of spermatozoa hinder the study of the functional properties of the spermatozoa plasma membrane. However, some studies have revealed the presence of a number of ion channels in this cell. We set out to measure the endogenous currents and to study the effect of the venom of the Chilean black widow spider (Latrodectus mactans). By patch-clamping bovine spermatozoa our results indicate the presence of an outwardly rectifying current, sensitive to changes in K(+) concentration (30-140 mM) and to tetraethylammonium (TEA, 10-100 mM). The application of the venom (7.5 microg/ml) blocks these K+ currents and then alters the passive properties of the plasma membrane. This leads to the entry of Ca(++), reflected by a change in basal fluorescent units (5+/-2 at 35+/-10 FAU). The Ca(++) influx follows a reduction in the membrane conductance (control 22+/-2; venom 10+/-1 pS), as calcium channels open in accord with voltage dependence.
Systems biology in reproductive medicine 02/2010; 56(1):37-43. · 0.80 Impact Factor
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European urology 07/2009; 56(1):215-6. · 7.67 Impact Factor
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ABSTRACT: It is debated whether chronic urogenital inflammations and infections may trigger the formation of antisperm antibodies (ASA) in semen.
To evaluate the formation of ASA in defined chronic inflammatory and infectious diseases of the male reproductive tract (MRT).
Three hundred sixty-five patients retrospectively enrolled in a single center were categorized as having National Institutes of Health (NIH) category II chronic prostatitis (n=38), NIH category IIIa chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) (n=59), NIH category IIIb CP/CPPS (n=213), chronic epididymitis (n=34), and chronic urethritis (n=21). Forty-five age-matched men served as controls.
All subjects underwent microbiologic and cytologic analysis for common bacteria, yeasts, and mycoplasma using the four-glass test. Urine samples, ejaculates, and urethral swabs were analyzed with polymerase chain reaction (PCR) for Chlamydia trachomatis and Neisseria gonorrhea. Semen analysis followed World Health Organization (WHO) standards. ASA in seminal plasma were analyzed using the mixed agglutination reaction (MAR) test.
The overall positive detection rate of clinically significant levels (> or = 50% of spermatozoa coated by ASA) of IgG and IgA antibodies was 1.8% and 0.8%, respectively, in the patient group. No clinically significant levels of ASA were detected in the control group, and no statistically significant difference was observed between controls and patients (IgG, p=1.0; IgA, p=1.0). No difference was found between the different inflammatory and infectious diseases and the control group in the detection rate of ASA, even when the cut-point value was lowered to > or = 1% (IgG, p=0.4; IgA, p=0.3). Moreover, in one selected subgroup of patients (n=26) with persistent increased inflammatory parameters (peroxidase-positive leukocytes [PPL] > or = 1 x 10(6)/ml and elastase > or = 230 ng/ml), no significant difference in the levels of ASA was observed compared with the controls (IgG, p=0.1; IgA, p=0.8).
There is no association between chronic inflammatory or infectious diseases of the MRT and the presence of ASA in semen.
European urology 09/2008; 56(4):708-15. · 7.67 Impact Factor
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ABSTRACT: o p e a n u r o l o g y s u p p l e m e n t s 7 (2 0 0 8) 7 0 3 – 7 1 4 a v a i l a b l e a t w w w . s c i e n c e d i r e c t . c o m j o u r n a l h o m e p a g e : w w w . e u r o p e a n u r o l o g y . c o m Article info Abstract Context: Sperm retrieval in combination with IVF/ICSI is the only medical procedure for an azoospermic man to father a child. Different techniques, especially testicular sperm extraction (TESE), have evolved over time and have dramatically improved the outlook for men with testicular azoospermia. However sperm retrieval rates are associated not only with the operation proposed but especially with a distinct pattern of prognostic factors that must be effectively managed for all these infertile patients for their best benefit. Objectives: To review the etiology, clinical work-up including operative techniques, and prognostic factors for testicular sperm retrieval in azoospermic men to maximin clinical benefit by these procedures. Evidence Acquisition: Data from basic and clinical studies with a defined, standar-dized approach pre-and postoperatively were analyzed. Evidence Synthesis: Different standardized surgical techniques can be offered to extract spermatozoa of azoospermic men from either the epididymis and/or the testis for ICSI. Sperm retrieval offers a treatment for both patients with testicular azoos-permia and men with obstructive azoospermia in cases where microsurgical referti-lization is not an option or has already failed. Among surgical techniques testicular sperm extraction (TESE) and microsurgical epididymial sperm aspiration (MESA) have become the most popular techniques. However, also percutaneous techniques are employed due their easy feasibility and low costs. By utilizing these techniques together with kryopreservation of extracted spermatozoa a single surgical interven-tion is able to provide spermatozoa for several ICSI attempts. Extensive surgical interventions in the testis of azoospermic patients have raised concerns about the potential influence on the endocrine compartment of the testis, particularly in patients with small testes and low levels of testosterone. Conclusions: Testicular sperm retrieval is a feasible and successful procedure. Testicular spermatozoa can be retrieved from the testis in up to 70% of patients, even in cases with testicular azoospermia and severe disorders of spermatogenesis. However, surgical damage of the testis might also compromise the interstitial compartment of the testis with testosterone deficiency as a consequence. Conclu-sively, endocrine follow-up can be considered mandatory.
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ABSTRACT: Ion channels have been assigned a pivotal importance in various sperm functions and are therefore promising targets for contraceptive development. The lack of data on channel functionality and pharmacology has hampered this goal. This is a consequence of technical problems of applying electrophysiological techniques to spermatozoa due to their small size and form. By using a laminin coating to increase adherence of spermatozoa and nystatin in the patch pipette for pore formation, we have adapted the whole-cell recording technique to study currents in mature uncapacitated bovine spermatozoa. Employing these conditions, in the head region, patched spermatozoa could be transferred into the whole-cell configuration. For the first time we document an outward rectifying current in mature bovine spermatozoa was blocked by tetraethyl ammonium (TEA) chloride. The observation of a shift in the reversal potential as a response to changes in the extracellular concentration of K(+) ions allowed us to identify this current as K(+) selective. This result shows that K(+) channels in the head region of mature uncapacitated bovine spermatozoa can be suitably investigated using the whole-cell recording patch-clamp technique.
Systems biology in reproductive medicine 54(6):231-9. · 0.80 Impact Factor
