Azoospermic Semen Research Articles

Techniques – Tension-relieving microdot vasovasostomies and longitudinal intussuscepted vasoepididymostomy vasectomy reversals

Introduction: Tension and malalignment of vasectomy reversal (VR) anastomoses are hypothesized to contribute to failure. We report VR outcomes using a novel technique introducing a tension-reliving hitch in the multi-layer microdot vasovasostomy (VV) and longitudinal intussuscepted vasoepididymostomy (LIVE; VE). Methods: All vasectomy reversal patients between May 2019 and September 2023 from a single surgeon were reviewed. Patients were included if they underwent a VR, with at least one semen analysis within six months of surgery and a minimum of six months of followup after the surgery to deem a failure. The primary outcome was patency, which was defined as 1) any sperm in the ejaculate; and B) functionally as at least two million motile sperm. Late failure was defined as an azoospermic semen analysis result after previously documented presence of sperm. Results: A total of 159 patients were evaluated, of which 136 patients met the inclusion criteria. The patency rate among all VRs was 97.7 %, with an overall functional patency rate of 93.1%. One hundred and one patients underwent bilateral VVs, with a 99% patency rate and 95.5% functional patency rate. Twenty-three patients underwent a mixed VV/VE with a patency rate of 100% and a functional patency rate of 88.8%. Finally, 12 patients underwent bilateral VE, with a patency rate of 83.3% and a functional patency rate of 77.7%. Among these patients, four VV patients were identified to have a late failure. Conclusions: The combination of tension-relieving stitches for VVs and VEs, along with attention to symmetrical and precise stitch placement, results in high patency rates.

P-040 The characteristic of human seminal plasma extracellular vesicle (SPEVs) profiles with different sperm features in a large cohort of 309 Taiwanese men

Abstract Study question To investigate human seminal plasma extracellular vesicle profiles in different sperm characteristics and their potential clinical application. Summary answer Normozoospermic semen has significantly higher SPEV concentration than asthenozoospermic or oligozoospermic samples. Moreover, SPEV concentration was highly correlated with total motile sperm count. What is known already Seminal plasma extracellular vesicles (SPEVs) are cell-derived membrane structures that carry proteins, nucleic acid, and lipids, which can modify the constitution and biological function of sperm. However, the characteristics of SPEVs in different sperm features remained unclear. Study design, size, duration From January 2021 to September 2023, 309 men who visited the NCKUH andrology clinic agreed to collect semen samples for this study, which the Institutional Review Board has approved. Participants/materials, setting, methods Semen parameters were measured by the X1 CASA system (Bonraybio, Taiwan). SPEVs were isolated by size exclusion chromatography column and ExoQuick™. Next, the SPEVs’ concentration and size distribution were performed with nanoparticle tracking analysis. The Kruskal-Wallis test calculated comparison in multiple groups, and Dunn’s multiple comparison tests accessed group-group comparison. The Pearson correlation coefficient r of exosome concentration with individual sperm parameters was calculated. A p-value less than 0.05 was set as statistical significance. Main results and the role of chance Among them, 59 semen samples were presented with normozoospermia, 36 were oligozoospermia, 73 were asthenozoospermia, and 141 samples had no sperm in semen. Among these 141 azoospermic men, 89 men are characterized by non-obstructive azoospermia (NOA), while the other 52 men had a vasectomy. Our data showed that normozoospermic semen has significantly higher SPEV concentration than asthenozoospermic or oligozoospermic semen samples (26 x109 [IQR 17.25 x109-56.25 x109] versus 12 x109 [IQR 5.6 x109-19.5 x109] versus 15.5 x109 [IQR 8.575 x109-26.75 x109], p = 0.0001). Nevertheless, azoospermic semen has the lowest SPEV concentrations. The concentration of SPEVs from infertile men diagnosed with NOA is slightly lower than vasectomized men (3.9 x109 [IQR 1.875 x109-7.675 x109] versus 5.4 x109 [IQR 2.9 x109-8 x109], p = 0.0001). The Pearson r coefficient of SPEV concentration with sperm concentration, progressive motility, and total motile sperm count was 0.238, 0.303, and 0.451, respectively. (P < 0.0001) Limitations, reasons for caution Because our data were collected from Taiwanese men, it is not yet known whether SPEV profiles differ among races. Wider implications of the findings Human SPEV concentration and its bio-content may be a potential non-invasive biomarker to predict the male reproductive outcome. Trial registration number not applicable

MP30-06 A UNIQUE EPIGENETIC-BASED TEST TO ACCURATELY PREDICT SUCCESSFUL TESE AND mTESE OUTCOMES

You have accessJournal of UrologyCME1 Apr 2023MP30-06 A UNIQUE EPIGENETIC-BASED TEST TO ACCURATELY PREDICT SUCCESSFUL TESE AND mTESE OUTCOMES Tim Jenkins, Ryan Barney, Kelaney Stalker, Ryan Miller, Kristin Brogaard, Jordan Kassab, and Larry Lipshultz Tim JenkinsTim Jenkins More articles by this author , Ryan BarneyRyan Barney More articles by this author , Kelaney StalkerKelaney Stalker More articles by this author , Ryan MillerRyan Miller More articles by this author , Kristin BrogaardKristin Brogaard More articles by this author , Jordan KassabJordan Kassab More articles by this author , and Larry LipshultzLarry Lipshultz More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003258.06AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Affecting 1% of men, non-obstructive azoospermia (NOA) is the absence of sperm in the ejaculate, and is the most severe form of male infertility. Currently, men with NOA have decreased chances of conceiving a child of their own. Despite recent progress in reproductive medicine, men with NOA have essentially one invasive and expensive procedure to isolate sperm (microdissection testicular sperm extraction or mTESE). Unfortunately, mTESE has a ∼50% failure rate. Here we present the use of an innovative DNA methylation-based technology to non-invasively predict the presence of sperm in the testicle of men with NOA. METHODS: At a single academic center biopsies (N=18) and azoospermic semen samples (N=5) were procured from consented men who had undergone a testicular aspirate, biopsy, or mTESE procedure. Utilizing new epigenetic technology, the biopsies and semen samples were analyzed for sperm-specific DNA. Specifically, these samples were analyzed for a sequence of DNA that can distinguish sperm cells from all other somatic cells via a unique methylation pattern. The quantity of sperm-specific DNA can be used to determine the absence (men with <0.25 ratio of sperm to somatic DNA) or presence (>0.5 ratio of sperm to somatic DNA) of sperm in the testicles. These ratios were defined for each sample and compared to clinical outcomes of the TESA, TESE or mTESE procedures. RESULTS: Out of the 18 biopsies, 13 had >0.5 ratios of sperm DNA to somatic cell DNA in the testicular biopsies. All of these men were found to have sperm in their biopsies as defined from the clinical notes. 3 of the 18 men had <0.25 ratios of sperm DNA to somatic cell DNA. Again, consistent with previous analysis, none of these men had sperm in their biopsies. The remaining 2 men had ratios between 0.26 – 0.49, with one having sperm in his biopsy and one without sperm. Of the 5 azoospermic semen samples with associated TESA results, 3 has ratios of <0.25 and all 3 had no sperm in the corresponding TESA samples. 2 semen samples had ratios of >0.5 and were confirmed to have sperm in the TESA sample. CONCLUSIONS: In the analysis of testicular biopsies and semen samples, the sperm-specific DNA methylation ratios of <0.25% or >0.5% were 100% predictive of either the absence or presence of sperm in the testicle. With these data the next steps of our work will focus on continued analysis of sperm-specific cfDNA in seminal plasma to predict the presence or absence of sperm prior to a testicular extraction procedure. The ultimate goal will be to develop a non-invasive and cost-effective way to avoid unnecessary surgical procedures. Source of Funding: Venture Capital © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e393 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Tim Jenkins More articles by this author Ryan Barney More articles by this author Kelaney Stalker More articles by this author Ryan Miller More articles by this author Kristin Brogaard More articles by this author Jordan Kassab More articles by this author Larry Lipshultz More articles by this author Expand All Advertisement PDF downloadLoading ...

MicroRNAs expression in semen and testis of azoospermic men.

MicroRNAs are involved in the regulation of spermatogenesis, are detected in semen and may be useful as molecular markers for predicting residual complete spermatogenesis in azoospermic men. To study the biomarker potential of microRNAs that are detected in semen and testicular tissue. MicroRNA profiles were analyzed in semen fractions of normozoospermic (n=3) and azoospermic (n=6) men by small RNA deep sequencing. Specific microRNAs were further analyzed by reverse transcription and quantitative polymerase chain reaction in eight testicular samples and 46 semen supernatants. The semen supernatant samples included 18 normozoospermic and 28 azoospermic men with various pathologies. The sequenced microRNA profiles of semen supernatant fraction samples were distinct from the other fractions. Significant expression differences were observed between the semen supernatant of normozoospermic and azoospermic men. Further analysis by reverse transcription and quantitative polymerase chain reaction revealed that expression of miR-202-3p was considerably reduced (undetectable in most samples) in the azoospermic semen supernatants. The expression of miR-202-3p was significantly lower in the azoospermic specimens than in the normozoospermic specimens and a trend was observed for miR-629-5p (p=0.03 and 0.06, respectively). Differences in expression levels in the semen supernatant were observed among the various pathologies but not to a level of significance, possibly because of the small subgroups. miRNA-370-3p was significantly higher in semen supernatant samples from azoospermic men without sperm cells in testis (p=0.05). In testes, the three microRNAs were expressed at higher levels in the obstructive and spermatocyte maturation arrest pathologies than in mixed atrophy and Sertoli cell only. miR-202-3p was detected in all testicular samples. MicroRNA expression profiles in semen were distinguishable between azoospermic and normozoospermic men. The microRNA profile also diverged among azoospermic men subdivided according to their testicular pathologies. The levels of specific microRNAs in testis and in the semen supernatant were not directly correlated.

Comparison between MACSprep™ forensic sperm microbead kit and Erase Sperm Isolation kit for the enrichment of sperm fractions recovered from sexual assault samples

Sexual assault samples often contain mixtures of cells coming from at least two donors. Ideally, one would need to separate the cells into two cellular fractions: one consisting of the alleged aggressor’s spermatozoa (the sperm fraction) and the other containing the victim’s epithelial cells (the non-sperm fraction). This separation increases the probability of obtaining the alleged offender’s autosomal DNA profile. However, spermatozoa are often collected along with an excess of biological material originating from the victim, and with unfavorable male:female biological material ratios, the absence of separation could result in the PCR amplification of the victim’s DNA profile only. Several approaches are available to enrich/purify the spermatozoa present on sexual assault samples. In this paper, we compare a new method, the MACSprep™ Forensic Sperm MicroBead Kit (MACSprep, based on microbeads conjugated with antibodies bound to spermatozoa and their retention within a magnetic column) with the Erase Sperm Isolation Kit (Erase, a standard differential lysis separation procedure combined with a specific removal of free DNA) routinely used in our lab. The performance of both kits was tested using sets of vaginal and buccal swabs loaded with different dilutions of sperm, or azoospermic semen, representing a total of 120 independent samples. For the samples containing undiluted sperm, an average recovery of 58% was observed for the MACSprep’s sperm fractions and 43% for Erase’s. Significantly better recovery of azoospermic semen was observed in MACSprep’s non-sperm fractions (~ 85%) compared to Erase (~ 28%). Erase performed significantly better than MACSprep in terms of recovery for diluted sperm samples (1:10 to 1:800 sperm dilutions) in the presence of vaginal cells, while the purities of the achieved sperm fractions were in favor of MACSprep for the highest sperm dilutions tested. Similar trends were observed with buccal swabs loaded with 1:200 sperm dilutions. Increased sperm dilutions on vaginal swabs resulted in higher variability in the male material recovered, whatever the separation method used. Both methods were easy to perform and resulted in male DNA extracts ready to use in less than 2 h. Both kits showed their specificities in terms of recovery efficiency and purity of the sperm fractions. Ideally, additional experiments should be performed in different laboratories, using workflow and chemistries different than ours, to better define the peculiarities observed with MACSprep for high dilutions. Improving the recovery of MACSprep for diluted samples, in addition to its better purity observed in the experiments performed, could make it a method of choice for laboratory workflow, despite MACSprep’s current price per sample being about twice the price of Erase’s.

Proposed age-stratified reference intervals of FSH derived from normozoospermic men

Objective: To demonstrate that serum follicle-stimulating hormone (FSH) in men rises with age, and to explore FSH reference intervals of age-related partitioning. Methods: Men aged 20-50 years (n=1 190) underwent semen analysis according to World Health Organization (2010) methods. Serum was frozen prior to measurement of FSH by using the Siemens ADVIA Centaur® XP immunoassay system. FSH central 95% intervals after logarithmic transformation based on age were derived from 1 037 normozoospermic men. These were then applied to oligozoospermic and azoospermic men. Men producing azoospermic semen samples were further classified as having non-obstructive azoospermia by clinical diagnostic criteria, including genetic analysis and surgical exploration. Results: Serum FSH in normozoospermic men increased with age (P&lt;0.05), and reference intervals were determined with 10-year brackets: 21-30 years [(1.0-8.2) IU/L], 31-40 years [(1.4-9.5) IU/L], 41-50 years [(1.9-12.0) IU/L]. The proportion of oligozoospermic men with normal FSH concentrations was less than the normozoospermic men, which in turn was lower among azoospermic men (both P&lt;0.01). The azoospermic men were further broken down according to the nature of the azoospermia as either obstructive or non-obstructive azoospermia, and 86.4% (38/44) men with non-obstructive azoospermia had elevated serum FSH concentrations whereas only 6.7% (1/15) men with obstructive azoospermia had high FSH levels, and this was significantly different (P&lt;0.01). Conclusions: FSH concentrations increase in men between 20-50 years, and clinical interpretation of serum FSH results in men must be made by using age-based reference intervals.

An investigation into the use of the ParaDNA® body fluid identification system in forensic examinations

Examinations carried out in forensic laboratories currently utilise chemical screening tests and microscopic analysis to locate and identify body fluids. However, there are currently no reliable tests to positively identify vaginal material, to differentiate between venous and menstrual blood and to confirm the presence of azoospermic semen. Furthermore, for cases involving allegations of oral intercourse, the attribution of a positive amylase reaction to saliva (as opposed to vaginal secretions) can be problematic.LGC has developed a system that uses the ParaDNA system together with one-tube mRNA detection chemistry for body fluid identification. This can be utilized at the laboratory examination stage, prior to DNA testing, to positively identify body fluids. A series of mRNA markers were selected to positively identify saliva, vaginal secretions, peripheral blood, menstrual blood, seminal fluid and sperm cells. We have carried out an assessment of this system in an active casework environment using mock-case samples. To investigate the sensitivity and specificity of the mRNA markers, the system was tested both by direct sampling of the body fluid stain (using the ParaDNA sampling device), and using aliquots of the retained aqueous supernatant following cell-harvesting.This paper summarises the findings obtained from the initial experiments carried out, together with the results of a further investigation into the effect of mixed body fluid samples on the detection of the individual body fluids.

An easy, reproducible and cost-effective method for andrologists to improve the laboratory diagnosis of nonobstructive azoospermia: a novel microcentrifugation technique.

ABSTRACTThis study describes a new method of microcentrifugation as an improved, viable, cost-effective option to the classical Cytospin apparatus to confirm azoospermia. Azoospermic semen samples were evaluated for cryptozoospermia by a centrifugation method similar to that of World Health Organization guidelines (2010; entire specimen centrifuged at 3000g for 15 min, and aliquots of the pellet examined). Then, if no sperm were detected, the pellet from that procedure was resuspended in culture medium, centrifuged (2000g for 15 min), and the entire pellet spread on a 4 X 6mm area of a slide and stained using the Christmas tree method (Nuclear-Fast solution and picric acid). The entire stained area was examined for the presence or absence of sperm. A total of 148 azoospermic samples (after standard WHO diagnosis) were included in the study and 21 samples (14.2%) were identified as sperm-positive. In all microcentrifugation slides, intact spermatozoa could be easily visualized against a clear background, with no cellular debris. This novel microcentrifugation technique is clearly a simple and effective method, with lower cost, increasing both sensitivity and specificity in confirming the absence or presence of spermatozoa in the ejaculate. It may represent a step forward of prognostic value to be introduced by andrology laboratories in the routine evaluation of patients with azoospermia in the initial semen analysis.

Evaluation of minerals in the seminal plasma of azoospermic semen

Background: Several studies have reported that mineral elements are involved in the performance and maintenance of male fertility.Aim: The aim of present study was to evaluate the mineral elements such as sodium, potassium, calcium, magnesium, phosphorus, chlorine and iron found in the seminal plasma of men was examined at the Pasteur Institute of Ce DIvoire to determine new biochemical markers in azoospermia.Material and Methods: Analysis of seminal fluid was carried out with 30 samples of normozoospermic semen and 30 samples of azoospermic semen. The assay of calcium, phosphorus, magnesium and Iron was done using the automated Hitachi Roche Cobas c 311. The evaluation of the concentration level of chlorine, sodium and potassium in the seminal plasma was performed using the automated 9180 Roche Electrolyte Analyzer. The data collected were analyzed using the software Graph Pad Prism 5. P values less than 0.05 was considered the level of significance.Results: The results of this study showed that the correlation test performed between concentrations level of minerals in the seminal plasma of normozoospermic and azoospermic semen showed no significant difference statistically with P > 0.05.Conclusion: The evaluation of the concentration level of mineral elements such as sodium, potassium, calcium, magnesium, phosphorus, chlorine and iron in the seminal plasma of azoospermic and normozoospermic semen showed that the concentration level of these mineral elements has no diagnostic value in the male reproductive system disorders, including azoospermia.

Influencia de los soportes, tiempo, origen e interferentes en la observación de fluidos biológicos con luces forenses

&lt;p&gt;&lt;em&gt;Introducción&lt;/em&gt;: al laboratorio llegan diferentes soportes para ser analizados, los cuales pueden contener fluidos como sangre, semen, saliva u orina. Una herramienta de apoyo en la identificación de manchas biológicas no visibles es la observación con luces forenses. En la actualidad, se cuenta con avances investigativos en los que se hace referencia a la longitud de onda y a la combinación con diferentes filtros para la observación de fluidos biológicos. &lt;em&gt;Metodología&lt;/em&gt;: en esta investigación, se utilizó el equipo de luces alternas Polilight&lt;sup&gt;®&lt;/sup&gt; Flare con luz azul, junto con filtro y gafas naranjas, para la observación de manchas de fluidos biológicos depositados en distintos tipos de textiles y otras superficies que se reciben con frecuencia en el laboratorio. &lt;em&gt;Resultados&lt;/em&gt;: el principal resultado de este estudio fue determinar la confiabilidad del equipo en términos de reproducibilidad y repetitividad de los resultados, al igual que la influencia de variables tales como el sustrato, la edad de la mancha, el origen del fluido y posibles interferentes en la observación de fluorescencia de fluidos biológicos. &lt;em&gt;Conclusiones&lt;/em&gt;: se encontró que las manchas biológicas que emitieron la fluorescencia más fuerte fueron las de semen, seguidas de las manchas de orina y saliva. Las manchas de sangre no emitieron fluorescencia y fueron denominadas como no-fluorescentes. La permeabilidad de los sustratos hizo que el halo de fluorescencia fuera mayor sobre todo en los de colores claros. De otro lado, no se observaron diferencias estadísticamente significativas en la emisión de fluorescencia del semen azoospérmico con respecto al normospérmico.&lt;/p&gt;

Cysteine-rich secretory protein 1 in seminal plasma: potential biomarker for the distinction between obstructive and nonobstructive azoospermia

To investigate the presence of cysteine-rich secretory protein 1 (CRISP1) in seminal plasma as a means of distinguishing between obstructive azoospermia (OA) and nonobstructive azoospermia (NOA). Seminal plasma from normospermic donors (n = 45) and azoospermic donors (n = 80) was examined to determine CRISP1 levels. Neutral alpha-glucosidase (NAG) enzymatic activity was measured for comparison with CRISP1 levels. Research unit of an academic medical center. Normospermic and azoospermic donors from the clinical andrology laboratory of the centre hospitalier universitaire de Québec and from Mount Sinai Hospital. Seminal CRISP1 measurement by Western blot analysis. Neutral alpha-glucosidase activity was evaluated by a photometric method. Seminal plasma CRISP1 levels, NAG activity, cutoff value, sensitivity, and specificity. All seminal plasma samples from normospermic and nonobstructive azoospermic donors were CRISP1 positive, whereas CRISP1 was absent or present at low levels in samples from patients with OA. A significant correlation between seminal CRISP1 levels and NAG activity was found in azoospermic semen samples. The cutoff point to distinguish between donors with NOA or OA was established at 0.655 (relative intensity). At this threshold, specificity was 85% and sensitivity was 92%. Seminal CRISP1 combined with NAG activity can potentially distinguish between OA and NOA.

Study into the Iron Content of Seminal Plasma in Normal and Infertile Subjects

The iron content in seminal plasma of normal (n19), oligozoospermic (n11), azoospermic (n12), oligoasthenozoospermic (n19), and asthenozoospermic (n17) subjects was estimated by using atomic absorption spectrophotometer. The concentration of iron in normal seminal plasma varied from 265 to 365 mg%. The source of iron in seminal plasma seems to be the adnexal glands and not spermatozoa, as azoospermic semen also contained it. A statistically highly significant difference was seen when normal was compared with azoospermia and with asthenozoospermia. The necessary average wastage of iron through semen is calculated as 2.52 mg/day. This value is highly variable according to the seminal volume and frequency of ejaculation.

Demonstration of the DNA Integrity in Semen Samples with Normospermia, Oligozoospermia and Azoospermia by Comet Method and Immunohistochemical Determination of EGF-R and TGF-a

Objective: The aim of this study was to compare the rate of sperm cells with DNA damage by the comet assay, and to determine EGF-R and TGF-α localization by using immunohistochemical methods in normospermic, oligozoospermic and azoospermic events. Methods: Semen samples were obtained from 30 men attending the Infertility Clinic of Baskent University Hospital. We used the comet assay to determine the DNA damage of sperm cells. Indirect immunohistochemical methods were used with EGF-R and TGF-α polyclonal antibodies. Results: To determine the DNA damage of sperm cells, the comet assay was used in normospermic, oligozoospermic and azoospermic events and a statistically significant difference was found among the treatment groups (p=0.038). The differences between normospermic and azoospermic events is greater than normospermic and oligozoospermic events (p=0.05). Furthermore, EGF-R and TGF-α immunoreaction on the sperm cell surface of normospermic and oligozoospermic events were stronger than azoospermic sperm cells. Reactivity of these growth factors was related with sperm morphology. Conclusion: In azoospermic semen samples, DNA damage was higher, and also the reactions for EGF-R and TGF-α were weaker when compared to normal semen samples. (Gazi Med J 2011; 22: 33-40)

The use of fluorescence in situ hybridisation and laser microdissection to identify and isolate male cells in an azoospermic sexual assault case

In cases of sexual assault where sperm are not present, preferential lysis fails to yield the DNA profile of the assailant. The Forensic Science Service ® has developed a technique combining fluorescence in situ hybridisation and laser microdissection to enable the identification and isolation of male cells that may be present in azoospermic semen on vaginal swabs from victims of sexual assault. This technique has been used successfully by The Forensic Science Service ® in a sexual assault case providing evidence for the assertion that the suspect had vaginal intercourse with the victim rather than he had not.

Quick recovery and characterization of cell-free DNA in seminal plasma of normozoospermia and azoospermia: implications for non-invasive genetic utilities

We established a quick and reliable method for recovering cell-free seminal DNA (cfsDNA), by using the binding-washing-elution procedure on the DNA purification column. Low variations (below 15%) among the triplicate values of cfsDNA quantity verified the reproducibility of our cfsDNA recovery method. Similar cfsDNA yield and size distribution between seminal plasma acquired by filtration and centrifugation confirmed the presence of cfsDNA. To investigate the general characterization of cfsDNA, the quantitation and size distribution of cfsDNA from normozoospermic and azoospermic semen were analyzed by real-time PCR and electrophoresis, respectively. CfsDNA concentration in semen with normozoospermia (n = 11) was 1.34 +/- 0.65 microg mL(-1), whereas a higher cfsDNA concentration was observed in azoospermia (2.56 +/- 1.43 microg mL(-1), n = 9). The continuous distribution of DNA fragments ranging from approximately 1 kb to 15 kb and a spectrum of multiples of 180-bp fragments were observed in each normozoospermic and azoospermic sample. Distinct characteristic DNA ladder fragmentations in some azoospermic samples implicated that cfsDNA originate partly from apoptotic cells. CfsDNAs of 36 selected azoospermic patients with known information of Y chromosome microdeletion were subjected to the same microdeletion analysis by multiplex PCR and PCR amplification of sY114 (1450 bp). All multiplex PCR reactions with cfsDNA amplified successfully and provided the same result as leukocyte DNA. PCR amplification of sY114 gave a 1450-bp amplicon as expected. Our data suggested the potential use of cfsDNA in search of biomarker or diagnostic procedures.

Editorial Commentary

Editorial Commentary

1657: Single Azoospermic Semen Analysis at Three Months Post Vasectomy is Adequate to Determine the Patient Azoospermic

1657: Single Azoospermic Semen Analysis at Three Months Post Vasectomy is Adequate to Determine the Patient Azoospermic

Identification and purification of an aspartic proteinase from human semen

To purify and evaluate the molecular changes associated with an aspartic protease (Cathepsin D) in human semen from infertile subjects. Cathepsin D was purified from normo-, oligo- and azoospermic semen, by a procedure involving detergent solubilisation, affinity chromatography and gel filtration chromatography. The enzyme from normo-, oligo- and azoospermic samples was purified 86, 60 and 44 fold respectively. The purified enzyme appeared as a single band on SDS as well as on native PAGE irrespective of the pathological conditions. The molecular weight of Cathepsin D from oligospermic and normospermic samples was 40 kDa while that of azoospermic sample was found to be 43 kDa. The enzyme was inhibited by pepstatin while other proteinase inhibitors and metal ions did not have any effect. Purified Cathepsin D from azoospermic sample differs from normospermia and oligospermia.

Termination of a randomized controlled trial of two vasectomy techniques

Family Health International (FHI) and EngenderHealth conducted a multicenter, randomized controlled trial to evaluate a fascial interposition (FI) component of a vas occlusion procedure for male sterilization. A data monitoring committee (DMC) was established to provide independent review of the interim report. The DMC met to review the interim report after enrollment of 552 men. As planned, an estimate of the hazard ratio (HR) for successful vasectomy for the FI group versus the no-FI group controlling for age of the participant and level of experience of the surgeon was obtained for the interim report using Cox's proportional hazards regression. The main analysis comparing time to vasectomy success, defined as two consecutive azoospermic semen samples, was highly significant and met the prespecified level for stopping enrollment (HR=1.54, p<0.01). However, a troublesome age by vasectomy technique interaction became apparent in the analysis. FI was clearly beneficial for younger males. However, the trend for older men was not as clear. The DMC recommended continuing the trial to better study this interaction and obtain more data to clarify the effect of FI for older men. After consulting further with the DMC, the study statisticians presented the interim report to senior management at FHI. A meeting between FHI senior management and the DMC followed. After much deliberation, trial enrollment was terminated. Follow-up of participants already enrolled in the study continued as planned. This paper presents the issues involved in the decision to terminate the study.

DNA Profiling of Azoospermic Semen Samples from Vasectomized Males by Using Y-PLEX™6 Amplification Kit

Post-vasectomized azoospermic semen samples (N = 6) were analyzed for short tandem repeats (STR) on the Y-chromosome by using Y-PLEX 6 and the 310 Genetic Analyzer. We have observed a wide variation in the yield of extracted DNA from 12.5-1,000 ng. This variation was attributed to the number of epithelial and/or white blood cells that are present in these azoospermic samples. DNA profiles of these vasectomized males were obtained for all six Y-STR loci, namely DYS393, DYS 19, DYS389II, DYS390, DYS391, and DYS385 amplified by using the Y-PLEX 6.