Journal of andrology最新文献

ABSTRACT: As the proportion of aged males attempting to reproduce continues to rise, so does the concern regarding the quality of spermatozoa from aged men. An imbalance between the generation of reactive oxygen species (ROS) and cellular antioxidant defenses, as occurs in aging, ultimately leads to decreased protein, lipid, and DNA quality. Spermatozoa are highly susceptible to oxidative damage, and thus an age-related shift in redox status may have serious implications for fertility. Therefore, we examined the effect of age on antioxidant enzymatic activity, ROS production, and extent of lipid peroxidation in both caput and cauda epididymal spermatozoa from young (4-month-old) and old (21-month-old) Brown Norway rats. Glutathione peroxidase (Gpx1, Gpx4) and superoxide dismutase (SOD) enzymes had decreased activity in aging spermatozoa. Immunofluorescence studies indicated that Gpx4 expression was decreased in both the head and midpiece regions of spermatozoa in aged animals. The decrease in nuclear Gpx4 points to a novel potential mechanism that may explain the previously noted decreased levels of protamine disulfide bonds in aged sperm nuclei. Further, hydrogen peroxide (H₂O₂) and superoxide (O₂•–) production were increased significantly in aging spermatozoa. Finally, lipid peroxidation was found to be drastically increased in aged spermatozoa. Taken together, these results suggest a decreased capacity for aged spermatozoa to handle oxidative stress and provide a potential basis for understanding the underlying cause of decreased quality of spermatozoa during aging.

Note: Postings to Androlog have been lightly edited before publication.

Patients who present with unexplained urologic pain often pose a substantial challenge in regard to both evaluation and treatment. It is safe to say that all clinically active urologists are quite familiar with patients presenting with chronic testicular pain, nonbacterial prostatitis, chronic epidiymitis, and the like. In this edition of Androlog, Dr Jay Sandlow describes a patient who presents with a more unusual complaint: perineal pain that occurs exclusively following ejaculation. The members of Androlog offer a variety of suggestions regarding the management of this condition.

Dr Jay Sandlow describes his patient:

Dr Richard Berger offers advice based upon his own clinical experience with patients such as this:

Dr Woet Gianotten offers his own perspective as a psychotherapist in the management of patients with complaints such as this:

Dr Toeman Kadiogly suggests ejaculatory duct reflux as a possible etiology:

Dr Michael Perelman joins the discussion, suggesting a specific evaluative approach aimed at determining the presence of a muscular component to this patient's pain:

Finally Dr Jorge Hallak suggests that acupuncture might be considered in the management of this patient:

ABSTRACT: Two novel surgical procedures that combine an autologous tunica vaginalis pedicle graft (TVG) with a subcapsular orchiectomy (SCOT) were evaluated in asymptomatic patients with rising prostate-specific antigen (PSA) values following radiation therapy, a radical retropubic prostatectomy, or a newly diagnosed prostatic cancer with bony metastasis. In the SCOT I procedure, the TVG was secured to the inner wall of the tunica albuginea. In the SCOT II procedure, the TVG was folded and secured to the external wall of the tunica albuginea. Between December 1, 1999, and July 1, 2000, 26 patients were offered hormonal therapy. Twelve patients selected the SCOT I procedure, 12 selected a luteinizing hormone-releasing hormone (LHRH) agonist, and 2 selected a bilateral total orchiectomy (BTO). Because the cosmetic outcome of the SCOT I procedure was less than ideal, this procedure was modified in December 2001. Between December 1, 2001, and July 1, 2002, 28 hormonal candidates were evaluated. Twelve patients selected the SCOT II procedure, 11 selected an LHRH agonist, and 5 selected a BTO. Preoperative measurements of the testicular area and PSA were obtained. During postoperative visits, the total testosterone, PSA, and testicular area were determined, and the Fugl-Meyer questionnaire (FMQ) and SCOT-specific questionnaire (SSQ) were completed. Between March 1, 2000, and December 1, 2002, 10 patients underwent a BTO. This group was the control for the postoperative SCOT total testosterone values. Sixty-three percent of the mean preoperative testicular area was preserved in the SCOT II group vs 43% in the SCOT I group at the 9- to 12-month visit (P < .01). The mean postoperative total testosterone values for the SCOT I, SCOT II, and BTO groups were in the castrate range. No statistically significant difference was noted between the preoperative and postoperative FMQ scores among the SCOT I and SCOT II groups. Eighty-three percent of the SCOT II patients experienced no change in masculine identity, and 58% noted no change in testicular size. One hundred percent of the SCOT I patients experienced no change in masculine identity and noted no change in testicular size. The SCOT II procedure preserved a greater testicular area than the SCOT I. Both SCOT procedures achieved castrate levels of total testosterone and maintained masculine identity in 83%–100% of patients.

ABSTRACT: This study evaluated the protective effect of butylated hydroxytoluene (BHT), a lipid-soluble antioxidant, against cryopreservation injuries to boar spermatozoa. In experiment 1, the lowest BHT concentrations able to reduce lipid peroxidation in boar spermatozoa were determined. Nine BHT concentrations (ranging from 0.025 to 3.2 mM) were evaluated, and the lowest (P < .05) production of malondialdehyde (MDA), as an indicator of lipid peroxidation, was obtained when BHT ranged from 0.2 to 1.6 mM. In experiment 2, sperm survivability was evaluated when BHT was added to a postthaw freezing extender by measuring the degree of sperm lipid peroxidation (using MDA production) and by measuring parameter such as motility, plasma membrane and acrosome integrity, and cell apoptosis. The ability of thawed spermatozoa to fertilize in vitro—matured oocytes and of embryos to develop to the blastocyst stage in vitro was also assessed. Pooled sperm-rich fractions collected from 3 mature Pietrain boars were frozen in 0.5-mL straws after dilution with lactose-egg yolk-glycerol-Orvus ES Paste extender supplemented with 0, 0.2, 0.4, 0.8, and 1.6 mM BHT. Postthaw sperm survival, evaluated 30 and 150 minutes after thawing, was higher in BHT-treated spermatozoa, being significant (P < .05) when the freezing extender was supplemented with 0.2, 0.4, and 0.8 mM BHT. The addition of BHT to the freezing extender resulted in a significant (P < .05) decrease in the MDA concentration in thawed spermatozoa, irrespective of the level of BHT used. BHT had no effect on oocyte cleavage rates, but the development to blastocyst was improved for embryos derived from spermatozoa frozen in extender supplemented with 0.4 mM BHT (16% vs 29% of blastocysts per total oocytes; P < .05). In conclusion, under the conditions tested in the present study, the addition of BHT to the freezing extender improved the overall efficiency of thawed boar spermatozoa.

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

ABSTRACT: The goal of this study was to perform 5-α-reductase type 2 gene (SRD5A2) analysis in a male pseudohermaphrodite (MPH) patient with normal testosterone (T) production and normal androgen receptor (AR) gene coding sequences. A patient of Chinese origin with ambiguous genitalia at 14 months, a 46,XY karyotype, and normal T secretion under human chorionic gonadotropin (hCG) stimulation underwent a gonadectomy at 20 months. Exons 1–8 of the AR gene and exons 1–5 of the SRD5A2 gene were sequenced from peripheral blood DNA. AR gene coding sequences were normal. SRD5A2 gene analysis revealed 2 consecutive mutations in exon 4, each located in a different allele: 1) a T nucleotide deletion, which predicts a frameshift mutation from codon 219, and 2) a missense mutation at codon 227, where the substitution of guanine (CGA) by adenine (CAA) predicts a glutamine replacement of arginine (R227Q). Testes located in the inguinal canal showed a normal morphology for age. The patient was a compound heterozygote for SRD5A2 mutations, carrying 2 mutations in exon 4. The patient showed an R227Q mutation that has been described in an Asian population and MPH patients, along with a novel frameshift mutation, Tdel219. Testis morphology showed that, during early infancy, the 5-α-reductase enzyme deficiency may not have affected interstitial or tubular development.

ABSTRACT: Increased DNA fragmentation is found in sperm from infertile men. Varicocele is an important cause of male infertility, even though it is present in 15% of men who father children. Semen analysis does not always identify infertility in these patients. Sperm motility is strongly correlated with male fertility potential. The goal of this study was to determine the correlation between apoptosis and kinematics in the ejaculated spermatozoa of patients affected by varicocele. Fresh semen samples were obtained from 30 patients with varicocele and 15 fertile controls. These samples were compared using computer-assisted semen analysis and were assayed to determine the degree of sperm apoptosis. The apoptotic index (AI) was calculated by dividing the number of terminal deoxynucleotidyl transferase—mediated deoxyuridine-5′-triphosphate nick end labeling (TUNEL) stained spermatozoa by the total number of Hoechst 33258-stained sperm cells for 300 sperm. Five microscopic fields were analyzed to obtain 5 AIs for each individual. Results demonstrated no significant difference in semen quality and sperm motion characteristics; however, a significantly higher AI (23.05% ± 4.07%: mean difference ± SE, 95% CI, 15.06%–31.03%, P < .0001) was identified in the varicocele group than in the fertile controls. We concluded that sperm apoptosis does not seem to correlate with semen quality and sperm kinematics and that apoptosis is increased in ejaculated spermatozoa in patients with varicocele compared to normal fertile men.

ABSTRACT: The etiologies of erectile dysfunction (ED) after nerve-sparing radical prostatectomy have not been clearly elucidated. The aim of this study was to evaluate the effects of cavernous nerve injury on cavernous fibrosis, and to consider measures to prevent irreversible damage to the cavernous tissues. Twenty male Sprague-Dawley rats constituted the study population. The animals were divided into 2 groups; group 1 consisted of sham-operated rats (n = 10), and group 2 consisted of rats that underwent incision of both cavernous nerves (n = 10). Three months later, all rats underwent intracavernous papaverine injection (300 and 600 mg), and intracorporal pressures were recorded. Transforming growth factor-β₁ (TGF-β₁) messenger RNA (mRNA) expression from rat penile tissue was measured using reverse transcriptase-polymerase chain reaction. Hypoxia-inducible factor-1α (HIF-1α), TGF-β₁, and collagen I and III protein expressions were determined by Western blot analysis and immunohistochemical staining. Erectile function as studied with intracavernosal papaverine injection and histological analysis of penile cross-sections at 3 months was similar in both groups. TGF-β₁ mRNA expression, HIF-1α, TGF-β₁, and collagen I and III protein expressions were significantly greater in the neurotomy group. Immunohistochemical staining for TGF-β₁, HIF-1α, and collagen III were qualitatively more positive in the neurotomy group, whereas collagen I staining was similar. This study demonstrates an increase in TGF-β₁, HIF-1α, and collagen III synthesis in rat cavernosal smooth musculature after cavernous neurotomies. In theory, cavernous fibrosis may be reduced by employing various vasoactive agents or interventions that increase oxygenation to the corporal tissues during the postoperative period.

ABSTRACT: Spermatozoa with deteriorated plasma membranes can be separated by magnetic-activated cell sorting (MACS) after binding superparamagnetic annexin V-conjugated microbeads (ANMBs) to membrane phosphatidylserine (PS). Semen samples from 15 donors and 25 infertile patients were divided into 2 spermatozoal fractions by annexin V-MACS. Activated caspases (aCPs), which mediate degradations of cell quality, were determined by CaspaTag in the 2 subpopulations. Spermatozoa from donors showed lower levels of bound annexin V (3.6% ± 0.5% vs 11.9% ± 1.1%; P < .01) and aCPs (21.8% ± 2.6% vs 43.2% ± 2.1%; P < .01) than did spermatozoa from infertile patients. MACS resulted in a decrease of spermatozoa with aCPs from 21.8% ± 2.6% (before separation) to 9.2% ± 1.4% (in the ANMB-negative fraction) in donors and from 43.2% ± 2.1% to 18.8% ± 2.6% in infertile patients (mean ± SEM; P < .01). Separation effects of the MACS technique were confirmed with flow cytometry using anti-annexin V antibodies and with electron microscopy. ANMB-MACS removes spermatozoa with PS-bound annexin V and produces a higher quality spermatozoal fraction. Spermatozoa with a deteriorated membrane are characterized by an increase in aCPs. A higher percentage of spermatozoa with ANMBs bound to PS and with aCPs were found in infertile patients.

Therapeutic cloning, or more accurately, as suggested by Vogelstein et al (2002), nuclear transplantation, is intended to “treat a specific disease of tissue degeneration.” As its name implies, the purpose of reproductive cloning is to replicate or duplicate a human. For the purpose of this article, the terms therapeutic cloning and reproductive cloning will be used because they are readily recognized and it suits the central question of this article, which is: Can cloning, whether therapeutic or reproductive, be objectively evaluated for use? This is a difficult question to address, especially when the issue being studied has an inexorable relationship with those making the evaluation (eg, clerics, politicians, and the lay community).

Cloning, or more generally, manipulation of the human reproductive process, is mired in politics, religion, and hyperbole. One central conundrum of the ethics of cloning is the relationship between the abstraction of when human life begins; that is to say, when the human embryo becomes distinct and separate from all other life forms during embryogenesis, and thus, the moral status of the human embryo. For some religious groups, human life becomes distinct at conception, and as such, the embryo merits the same rights and respect as you or me. In the reproductive sciences and for some countries in which in vitro fertilization is government-regulated, human life becomes distinct from other mammalian life forms at Day 14, when neural tube development begins. Can a globally acceptable definition for when human life begins be identified? The answer, clearly, is no. But why not?

The ability to clone embryos has caused a dramatic increase in the scrutiny of areas in science and medicine that are already the object of criticism. To make matters exponentially worse are the proclamations by individuals who are bent on cloning for the purpose of reproducing humans. One scientist, Dr Brigitte Boisselier (see http:www.clonaid.com), is linked to a cult of people, the Raelians (http:www.rael.org), who identify with extraterres-trials. Reports state that the daughter of Dr Boisselier has volunteered to act as a gestational surrogate for such experiments. Two other scientists, Drs Severino Antinori and Panayiotis Zavos, are pursuing cloning for the purpose of human duplication under the pretext of remedying infertility. These few renegade scientists are in the minority relative to the vast majority of scientists on a global scale who oppose cloning for purposes of human propagation.

In the United States, reproductive and therapeutic cloning have become inexorably linked. This link has been reinforced through the media-covered side-circus provided by the Raelians, Drs Antinori and Zavos, and through publicized debate on the topics in the US Congress. Arguably, the inability to distinguish between reproductive and therapeutic cloning was exacerbated on Thursday, August 9, 2001, when President Bush th