Unsuccessful Sperm Retrieval Research Articles

AI Model Developed Using Machine Learning for Predicting Sperm Retrieval in Micro-TESE for Nonobstructive Azoospermia Patients

Azoospermia is a severe problem that prevents couples from having their own children through natural pregnancy. In nonobstructive azoospermia (NOA), microdissection testicular sperm extraction (micro-TESE) is required to collect sperm and, at 40%–60%, the sperm retrieval success rate is not very high. Previous studies identified no single clinical finding or investigation that could accurately predict the outcome of sperm retrieval. It would be very valuable to have a factor for predicting the possibility of sperm retrieval in patients with NOA before performing micro-TESE. We retrospectively obtained data from the medical records of 430 patients who underwent micro-TESE from 2011 to 2020. Parameters extracted were age, height, body weight, body mass index, luteal hormone, follicle-stimulating hormone, PRL, total testosterone, E2, T/E2, sperm retrieval, G-band, AZF, medical history, Rt testis, and Lt testis. Prediction One, which does not require coding, was used to create the AI prediction model for sperm retrieval. Prediction One makes the best prediction model using an artificial neural network with internal cross-validation. Prediction One also evaluates the “importance of variables” using a method based on permutation feature importance. The AUC for the AI model was 0.7246, which is acceptable. In addition, among the variables, T/E2 ratios contributed most to predicting whether sperm retrieval was possible or not. However, the difference in T/E2 between successful and unsuccessful sperm retrieval was not statistically significant. In addition, our analysis of data from 20 patients who underwent micro-TESE in 2021 found that in 85%, the actual result matched the result predicted using our novel AI model. We created an AI model for predicting sperm retrieval in patients with NOA before undergoing micro-TESE. In addition, we found that T/E2 ratios contributed most to predicting possibility of sperm retrieval in NOA using machine learning.

MP30-02 A NOVEL PREDICTION MODEL FOR POSSIBILITY OF SPERM RETRIEVAL IN MICRO TESE FOR NON-OBSTRUCTIVE AZOOSPERMIA PATIENTS USING AUTOMATED ARTIFICIAL INTELLIGENCE

You have accessJournal of UrologyCME1 Apr 2023MP30-02 A NOVEL PREDICTION MODEL FOR POSSIBILITY OF SPERM RETRIEVAL IN MICRO TESE FOR NON-OBSTRUCTIVE AZOOSPERMIA PATIENTS USING AUTOMATED ARTIFICIAL INTELLIGENCE Hideyuki Kobayashi, Masato Uetani, Koichi Nakajima, and Koichi Nagao Hideyuki KobayashiHideyuki Kobayashi More articles by this author , Masato UetaniMasato Uetani More articles by this author , Koichi NakajimaKoichi Nakajima More articles by this author , and Koichi NagaoKoichi Nagao More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003258.02AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Azoospermia is a severe problem that prevents couples from having their own children through natural pregnancy. In non-obstructive azoospermia (NOA), a particularly severe type of male infertility, microdissection testicular sperm extraction (micro TESE) is required to collect sperm and, at 40-60%, the sperm retrieval success rate is not very high.Previous studies identified no single clinical finding or investigation that could accurately predict the outcome of sperm retrieval. In addition, although clinical and hormone parameters have been previously investigated as potential predictors of sperm retrieval, the evidence for them has been conflicting and a specific biochemical marker predicting the possibility of sperm retrieval has yet to be established. It would be very valuable to know the possibility of sperm retrieval for patients with NOA before performing micro TESE. Therefore, our objective was to make a model for predicting the possibility of sperm retrieval in patients with NOA before performing micro TESE, using machine learning. METHODS: We retrospectively obtained data from the medical records of 430 patients who underwent micro TESE from 2011 to 2020. Parameters extracted were age, height, body weight, body mass index, LH, FSH, FSH, PRL, total testosterone, E2, T/E2, sperm retrieval, G-band, AZF, medical history, Rt testis, and Lt testis. Prediction One, which does not require coding, was used to create an AI prediction model for sperm retrieval. Prediction One makes the best prediction model using an artificial neural network (ANN) with internal cross validation. RESULTS: The mean age of the 430 male patients who underwent micro TESE in this study was 36.78±7.58 years. Sperm was successfully retrieved in 151 (35.1%) patients.Statistically significant differences between successful and unsuccessful sperm retrieval were observed for age, LH, FSH, total testosterone, Rt testis, Lt testis and medical history. Medical history included none, Lt varicocele, cryptorchidism, inguinal hernia, torsion of spermatic cord, orchitis, cancer treatment, spinal injury, spina bifida, and anti-sperm antibody (+). The AUC for the AI model was 72.46%, which is acceptable. The values for Accuracy, Precision and Recall were 72.09%, 64.76% and 45.03%, respectively, when the F-value was 53.13%. CONCLUSIONS: We created an AI model for predicting sperm retrieval in patients with NOA before undergoing micro TESE. Source of Funding: none © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e391 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Hideyuki Kobayashi More articles by this author Masato Uetani More articles by this author Koichi Nakajima More articles by this author Koichi Nagao More articles by this author Expand All Advertisement PDF downloadLoading ...

P-011 Deep testicular tissue dissection is most often required to retrieve testicular sperm in patients with non-obstructive azoospermia undergoing microdissection testicular sperm extraction

Abstract Study question How successful is sperm retrieval from the testicular surface of patients with non-obstructive azoospermia (NOA) undergoing microdissection testicular sperm extraction (mTESE)? Summary answer Enough good quality sperm for ICSI could be retrieved from the testicular surface in only 33 out of 224 patients (14.7%) What is known already Due to the anatomic singularity of NOA, characterized by the heterogeneous distribution of histologically and functionally distinct seminiferous tubules, retrieving enough good quality sperm for ICSI may require a complete testicular dissection. While conventional TESE (cTESE) may reach only the seminiferous tubules located in the testis surface, mTESE allows the nearly complete exploration of the whole testicular parenchyma. This is the reason why mTESE provides better sperm retrieval rates compared to cTESE, according to two systematic reviews and one meta-analysis: the superiority of mTESE compared to cTESE, however, has been challenged by a recent meta-analysis. Study design, size, duration Retrospective cohort study on 224 patients, aged 37 (20-54) years, who underwent unilateral (71, 31.7%) or bilateral (153, 68.3%) mTESE from January 2018 through May 2021. 175 patients were naïve for mTESE, while 49 underwent a salvage mTESE following one or more previously failed conventional TESE. Participants/materials, setting, methods The testis was opened like a book following a transversal incision of the testis covering two-thirds to three-quarters of its circumference: the entire surface of the testicular parenchyma was explored first in search for dilated seminiferous tubules. If no/ not enough sperm were retrieved, the testis was opened till to be fully bivalve, and the deeper portion of the testicular parenchyma was explored both into hilum direction and orthogonally to the para-equatorial section plan. Main results and the role of chance Sperm was retrieved in 114 patients (50.8%), 67 (94.3%) undergoing unilateral and 47 (30.7%) undergoing bilateral mTESE: only 15/153 (9.8%) had their testicular sperm retrieved in the second testis following the unsuccessful sperm retrieval in the first testis, while in the remaining cases sperm had to be retrieved from both testes to obtain enough good quality sperm for ICSI. Histopathology demonstrated Sertoli-cell only syndrome in 65.6% of operated testes, while maturation arrest was found in 19.8%, hypospermatogenesis in 12.6% and hyalinosis in 1.5%. Sperm were obtained from the testis surface in 33 out of 114 patients (28.9%); in the remaining patients a deeper dissection was needed. No difference was seen among patients with sperm found in the surface or requiring deeper dissection in terms of age, testis volume, serum testosterone level, and histopathological subcategories; although serum FSH and LH level were significantly lower in patients with sperm retrieved from the testicular surface, on multivariate logistic regression they were not found to predict the chance of retrieving sperm from the surface (OR 0.96, 95% CI 0.92-1.007, p = 0.104 and OR 0.95, 95% CI 0.85-1.06, p = 0.405, respectively). Limitations, reasons for caution the amount of testicular tissue dissected was strictly dependent upon the chance of finding enough sperm for ICSI in the specimens handled by the biologist in the operating room: we cannot exclude that in some patients enough sperm could be obtained from the testis surface following further extensive search Wider implications of the findings mTESE is to be preferred to cTESE, since in most patients with NOA seminiferous tubules containing sperm may be found in the deeper portion of the testicular parenchyma. This may be particularly true for patients with unfavorable histopathological patterns, such as Sertoli cell only syndrome or maturation arrest. Trial registration number Not applicable

Whole-exome sequencing improves the diagnosis and care of men with non-obstructive azoospermia

Non-obstructive azoospermia (NOA) is a severe and frequent cause of male infertility, often treated by testicular sperm extraction followed by intracytoplasmic sperm injection. The aim of this study is to improve the genetic diagnosis of NOA, by identifying new genes involved in human NOA and to better assess the chances of successful sperm extraction according to the individual's genotype. Exome sequencing was performed on 96 NOA-affected individuals negative for routine genetic tests. Bioinformatics analysis was limited to a panel of 151 genes selected as known causal or candidate genes for NOA. Only highly deleterious homozygous or hemizygous variants were retained as candidates. A likely causal defect was identified in 16 genes in a total of 22 individuals (23%). Six genes had not been described in man (DDX25, HENMT1, MCMDC2, MSH5, REC8, TDRKH) and 10 were previously reported (C14orf39, DMC1, FANCM, GCNA, HFM1, MCM8, MEIOB, PDHA2, TDRD9, TERB1). Seven individuals had defects in genes from piwi or DNA repair pathways, three in genes involved in post-meiotic maturation, and 12 in meiotic processes. Interestingly, all individuals with defects in meiotic genes had an unsuccessful sperm retrieval, indicating that genetic diagnosis prior to TESE could help identify individuals with low or null chances of successful sperm retrieval and thus avoid unsuccessful surgeries.

Testicular microlithiasis defines a subgroup of azoospermic men with low rates of sperm retrieval.

To investigate the prevalence of testicular microlithiasis and its association with sperm retrieval rates and histopathology in men with non-obstructive azoospermia. A total of 120 men underwent scrotal ultrasonography prior to microsurgical testicular sperm extraction. Sperm retrieval rate, testicular histopathology, testicular size, reproductive hormones, karyotyping, Y chromosome microdeletion analyses, and presence of varicoceles and hydroceles were compared between men with and without testicular microlithiasis. The total sperm retrieval rate was 40%. Ten men with normal spermatogenesis were excluded. The remaining 110 men with non-obstructive azoospermia were analyzed and testicular microlithiasis was detected in 16 of them (14.5%). The sperm retrieval rate in that subgroup was only 6.2% (1/16) as opposed to 39.4% (37/94) in men with non-obstructive azoospermia and no evidence of microlithiasis (P = 0.009). The mean right and left testicular diameters were significantly lower in the microlithiasis group (P = 0.04). On multivariate logistic regression analysis, the presence of mictolithiasis (odds ratio 7.4, 95% confidence interval 2.3, 12.2; P = 0.01) was the only independent predictor of unsuccessful sperm retrieval. The 15 patients with microlithiasis and without successful sperm extraction were diagnosed by histopathology as having Sertoli cells only. The 16th patient with successful sperm retrieval had a histopathology of mixed atrophy and was diagnosed with Klinefelter syndrome. The presence of testicular microlithiasis is associated with low sperm retrieval rates among our cohort of men with non-obstructive azoospermia undergoing scrotal ultrasonography prior to microsurgical testicular sperm extraction. Larger, prospective studies should be conducted to confirm these findings.

P–076 Probability of sperm retrieval in azoospermic patients and mRNA expression profile of JMJD1A, TNP2 and PRM2 : in a subset of karachi population

Abstract Study question To access successfulness of sperm retrieval by evaluating the mRNA expression profile of JMJD1A, TNP1, TNP2, PRM1 and PRM2 in patients undergoing surgical sperm retrieval procedure. Summary answer Probability of sperm retrieval in azoospermia is decreased when mRNA expression profile of JMJD1A TNP2 and PRM2 in testicular tissue is decreased. What is known already Studies have been done on expression of JMJD1A in non-obstructive azoospermic patients in other part of the world with smaller sample size but this is the first study in Pakistan with larger number of patients. Study design, size, duration: Crossectional study, 100 azoospermic patients coming for purpose of sperm retrieval by TESE or micro-TESE in Australian Concept Infertility Medical Center, Karachi,from March, 2018 to December, 2019 Participants/materials, setting, methods All recruited azoospermic patients were evaluated by history, physical examination, and hormonal assessment. RNA was extracted by pureLink RNA Micro kit and mRNA expression of the JMJD1A, TNP1, TNP2, PRM1 and PRM2 genes was determined using innu-SCRIPT One Step RT_qPCR SyGreen kit. For quantitative variables independent t test and for categorical variables chi-square/ Fisher Exact test was used. Unadjusted and adjusted prevalence ratio were reported by using cox regression algorithm. Main results and the role of chance: The patients were categorized into (i) Group-I: Patients with successful sperm retrieval n = 42, (ii) Group-II: Patients with unsuccessful sperm retrieval n = 58. The patients were categorized into (i) Group-I: Patients with successful sperm retrieval n = 42, (ii) Group-II: Patients with unsuccessful sperm retrieval n = 58. Azoospermic men in the successful sperm retrieval group had significantly decreased expression of JMJD1A (P < 0.001), TNP2(P < 0.001), and PRM2 (P 0.008). In addition to this regarding hormonal parameters: FSH (P 0.004), LH(P < 0.001), TSH(P<.011) were significantly different in azoospermic men with successful and unsuccessful sperm retrieval. In multivariate analysis, after adjusting for the other covariates, a significant association was found between JMJD1A, TNP2, PRM2 and successful sperm retrieval (p-value <0.05). Limitations, reasons for caution It is unicentric and outcomes for fertilization were not assessed. Azoospermic patients from multi-centeres were difficult because of lack of facility of sperm retrieval procedures at these centers and it was difficult to follow the fertrilization outcome. Wider implications of the findings: This will be useful for making the decision in azoospermic men to proceed for ICSI or not. In addition to this, the repetition of unnecessary surgical procedures can be avoided, as the azoospermic men often undergo number of rounds of ICSI, with the hope of becoming biological father. Trial registration number non-clinical trials

Endocrine Tests and/or Testicular Volume are Not Predictive of Successful Sperm Retrieval by Conventional Multiple Testicular Sperm Extraction in Nonobstructive Azoospermia

Objectives: This study aimed to determine the role of presurgical markers in the prediction of sperm retrieval by conventional Multiple Testicular Sperm Extraction in infertile Vietnamese men with nonobstructive azoospermia (NOA). Patients and Methods: Retrospective descriptive analysis of 136 infertile men with azoospermia, examined from August 2014 to July 2018. Patients underwent stepwise surgical sperm retrieval via percutaneous epididymal sperm aspiration, testicular sperm aspiration, then conventional multiple testicular sperm extraction in up to three locations, and procedures stopped as soon as sperm were detected. Factors were analyzed to determine the prediction of the likelihood of successful sperm retrieval, in men with NOA. Results: The overall success rate of sperm retrieval in men with azoospermia was 49.3%, but it was only 18.4% in NOA group. The difference in testicular volume between men with successful sperm retrieval and unsuccessful sperm retrieval was not statistically significant in NOA group (5.68 ± 2.37 vs. 4.46 ± 2.83, p = 0.138). The differences in the endocrine tests between the two groups were also not significant in terms of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone (p [Formula: see text] 0.05). Multivariable analysis of predictive factors of sperm retrieval in NOA groups found no significant difference, except testicular density (p = 0.015). Conclusions: In infertile men with NOA, neither an endocrine test nor testicular volume should be used for predicting the results of surgical sperm retrieval by conventional multiple testicular sperm extraction.

Intronic variation of the SOHLH2 gene confers risk to male reproductive impairment

To evaluate whether SOHLH2 intronic variation contributes to the genetic predisposition to male infertility traits, including severe oligospermia (SO) and different nonobstructive azoospermia (NOA) clinical phenotypes. Genetic association study. Not applicable. Five hundred five cases (455 infertile patients diagnosed with NOA and 50 with SO) and 1,050 healthy controls from Spain and Portugal. None. Genomic DNA extraction from peripheral blood mononuclear cells, genotyping of the SOHLH2 polymorphisms rs1328626 and rs6563386 using the TaqMan allelic discrimination technology, case-control association analyses using logistic regression models, and exploration of functional annotations in publicly available databases. Evidence of association was observed for both rs6563386 with SO and rs1328626 with unsuccessful sperm retrieval after testicular sperm extraction (TESE-) in the context of NOA. A dominant effect of the minor alleles was suggested in both associations, either when the subset of patients with the manifestation were compared against the control group (rs6563386/SO: P=.021, odds ratio [OR] = 0.51; rs1328626/TESE-: P=.066, OR = 1.46) or against the group of patients without the manifestation (rs6563386/SO: P=.014, OR = 0.46; rs1328626/TESE-: P=.012, OR = 2.43). The haplotype tests suggested a combined effect of both polymorphisms. In silico analyses evidenced that this effect could be due to alteration of the isoform population. Our data suggest that intronic variation of SOHLH2 is associated with spermatogenic failure. The genetic effect is likely caused by different haplotypes of rs6563386 and rs1328626, which may predispose to SO or TESE- depending on the specific allelic combination.

Do partial AZFc deletions affect the sperm retrieval rate in non-mosaic Klinefelter patients undergoing microdissection testicular sperm extraction?

BackgroundThe purpose of this study is to evaluate the prognostic factors for sperm retrieval and determine if Y chromosome deletion is associated with deleterious effects on spermatogenesis in non-mosaic Klinefelter patients. Whether Y chromosome deletion determines the sperm retrieval rate in non-mosaic Klinefelter patients has not yet been addressed.MethodsWe retrospectively collected medical records of azoospermic patients from Sep 2009 to Dec 2018, and enrolled 66 non-mosaic 47, XXY patients who were receiving mTESE. The predictive values of patients age, serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone, prolactin, estradiol and Y chromosome deletion were assessed for successful sperm recovery.ResultsTesticular sperm recovery was successful in 24 (36.4%) of 66 men. The mean age (36.0 vs. 36.6 years), and levels of FSH (30.0 vs 36.9 IU/L), LH (17.7 vs 21.9 IU/L), testosterone (2.4 vs. 2.1 ng/ml), prolactin (9.1 vs. 8.8 ng/ml), and estradiol (19.4 vs. 22.3 pg/ml) did not show any significant difference when comparing patients with and without successful sperm retrieval. Partial deletion of azoospermic factor c (AZFc) was noted in 5 (20.8%) of 24 patients with successful sperm retrieval, including three b2/b3 and two gr/gr deletion cases, whereas 4 (9.5%) of 42 patients with unsuccessful sperm retrieval were noted to have AZFc partial deletion (one b2/b3, one sY1206 and two gr/gr deletion), though the difference was not statistically significant (p = 0.27).ConclusionAccording to present results, age and AZFc partial deletion status should not be a deterrent for azoospermic males with non-mosaic Klinefelter syndrome to undergo mTESE.

"Second-look" Micro Testicular Sperm Extraction (MicroTESE) in Patients with Non-obstructive Azoospermia Following Histopathological Analysis.

Introduction:Microdissection testicular sperm extraction (microTESE) is considered the gold standard method for surgical sperm retrieval among patients with non-obstructive azoospermia (NOA).Aim:This study aimed to evaluate the correlation between histopathological findings after failed microTESE procedure and outcomes of the „second-look“ procedure and to provide insight into the most common histopathological patterns after testicular biopsy within our population.Methods:The retrospective study included 33 selected patients with NOA, who had undergone unsuccessful sperm retrieval. The diagnosis of NOA was made after the assessment of the patient’s history data, a physical examination, semen analysis, the hormonal profile, and genetic studies. After negative sperm retrieval, histopathological report has been analyzed for „second-look“ microTESE attempt.Results:Five testicular histopathological patterns were found: hypospermatogenesis (9,1%), Sertoli cell-only syndrome (43%), germ cell maturation arrest (15%), seminiferous tubule hyalinization (15%), mixed pattern (21%). Y-microdeletions were detected in 5 patients, of which 3 patients showed AZFc region deletions. Only 3 patients (9,1%) underwent a „second-look“ procedure after the evaluation of histopathological reports. After the stimulation therapy and „second-look“ procedure, we had a positive outcome in a single patient (33,3%). Mean FSH value in patients with confirmed spermatogenesis was 17.26±3.11IU/l, while mean FSH value in patients without presence or germ cell statistically significantly exceeded and was 24.28±4.71IU/L (p=0.038).Conclusion:Histopathological reports following the microTESE procedure are obligatory for the proper selection of patients who are candidates for the „second-look“ microTESE attempt. Patients with Sertoli cell-only syndrome and hypospermatogenesis particularly can benefit from the “second-look” procedure.

445 Relevance of testicular histopathology on prediction of sperm retrieval rates after a conventional bilateral multiple TESE in case of non-obstructive azoospermia (NOA) and obstructive-azoospermia (OA)

To establish the relevance of testicular histopathology (TH) on sperm retrieval (SR) after testicular sperm extraction (TESE) in patients with non-obstructive azoospermia (NOA) and in obstructive azoospermia (OA) patients, already underwent a previous failure testicular fine needle aspiration (TEFNA). We evaluated a total of 82 azoospermic men, underwent TESE, referring to our Centre between January 2008 and March 2017. A general and genital physical examination, scrotal and trans-rectal ultrasound, semen analysis, hormone measurements, including follicle stimulating hormone (FSH), luteinizing hormone (LH) and total testosterone (TT) were collected. Successful sperm retrieval (SSR) was obtained in 36 men of total (43.9%).(Table 1) SSR was 29.5% in NOA patients, while OA men, underwent a previous failure TEFNA, had SR in 86% of cases. Mean LH was 6.55 IU/L, TT 4.70 ng/ml, testicular volume (TV) right 13.7 ml and TV left 13.6 ml. FSH mean was 13.45 IU/L in patients with negative SR and 8.18 IU/L in men with SSR. According to histology, 20.7% had Normal Spermatogenesis (NS), 35.3% Hypospermatogenesis (H), 35.3% Maturation Arrest (MA) and 8.5% Sertoli-Cell-Only-Syndrome (SCOS). SSR was 88.2% in patients with NS, 24.1% in the MA group and 48.27% in patients with Hypospermatogenesis, while negative SR was reported in SCOS patients. 2 patients with NS and unsuccessful SR, underwent a second TESE, of whom one had SR and cryopreservation.

Testicular piRNA profile comparison between successful and unsuccessful micro-TESE retrieval in NOA patients.

PIWI-interacting RNA (piRNA) is a sub-group of small RNAs about 30 nucleotides length which specifically expressed in mammalian germ cells. Although piRNAs play pivotal roles in spermatogenesis regulation, little is known in the testicular tissues of infertile men. To explore whether piRNA profile could serve as a biomarker for male infertility diagnosis in a clinic, in this study, we systematically investigated the expression profile of piRNAs in testicular tissues from the patients with non-obstructive azoospermia (NOA) between successful and unsuccessful sperm retrieval before micro-dissection testicular sperm extraction (micro-TESE). The differential expression levels of piRNAs were evaluated using small RNA-Seq method. Ontologic analyses were performed to determine the presence of enriched biological processes. A total of 18,324 Homo sapiens piRNAs were identified by small RNA-Seq from NOA patient testicular tissues; among them, 959 piRNAs were significantly altered between successful and unsuccessful sperm retrieval groups, of which 951 testicular piRNAs were significantly downregulated and 8 piRNAs were upregulated in NOA patients with unsuccessful sperm retrieval (USR) groups compared to those with successful sperm retrieval (SSR) groups, respectively. Unexpectedly, 553 testicular piRNAs were found completely absent in USR but showing abundant in SSR, which suggests that those piRNAs might serve as a biomarker for micro-TESE application. A total of 20 significantly differential piRNAs (hsa-piR-20830, hsa-piR-4731, hsa-piR-6254, hsa-piR-419, hsa-piR-7152, hsa-piR-7548, hsa-piR-14195, hsa-piR-5026, hsa-piR-11482, hsa-piR-17765, hsa-piR-17102, hsa-piR-4484, hsa-piR-17260, hsa-piR-17098, hsa-piR-20511, hsa-piR-5802, hsa-piR-19121, hsa-piR-2510, hsa-piR-4745, hsa-piR-11873) were selected to further validate the RNA-Seq data by quantitative real-time polymerase chain reaction. In addition, bioinformatic analyses revealed that those altered piRNAs were involved in many important biological pathways, including apoptosis, cell proliferation, and differentiation. Our results demonstrate that testicular tissues from NOA patients with successful and unsuccessful spermatozoa retrieval exhibit differential piRNA profiles. This study provides a useful resource to further elucidate the regulatory role of piRNAs in spermatogenesis and provides a profound clue to identify useful biomarkers for predicting residual spermatogenic loci in NOA patients during assisted reproductive treatment.

Sperm Retrieval in Patients with Klinefelter Syndrome: A Skewed Regression Model Analysis.

BackgroundThe most common chromosomal abnormality due to non-obstructive azoospermia (NOA) is Klinefelter syndrome (KS) which occurs in 1-1.72 out of 500-1000 male infants. The probability of retrieving sperm as the outcome could be asymmetrically different between patients with and without KS, therefore logistic regression analysis is not a well-qualified test for this type of data. This study has been designed to evaluate skewed regression model analysis for data collected from microsurgical testicular sperm extraction (micro-TESE) among azoospermic patients with and without non-mosaic KS syndrome.Materials and MethodsThis cohort study compared the micro-TESE outcome between 134 men with classic KS and 537 men with NOA and normal karyotype who were referred to Royan Institute between 2009 and 2011. In addition to our main outcome, which was sperm retrieval, we also used logistic and skewed regression analyses to compare the following demographic and hormonal factors: age, level of follicle stimulating hormone (FSH), luteinizing hormone (LH), and testosterone between the two groups.ResultsA comparison of the micro-TESE between the KS and control groups showed a success rate of 28.4% (38/134) for the KS group and 22.2% (119/537) for the control group. In the KS group, a significantly difference (P<0.001) existed between testosterone levels for the successful sperm retrieval group (3.4 ± 0.48 mg/mL) compared to the unsuccessful sperm retrieval group (2.33 ± 0.23 mg/mL). The index for quasi Akaike information criterion (QAIC) had a goodness of fit of 74 for the skewed model which was lower than logistic regression (QAIC=85).ConclusionAccording to the results, skewed regression is more efficient in estimating sperm retrieval success when the data from patients with KS are analyzed. This finding should be investigated by conducting additional studies with different data structures.

Cryopreservation of adult cervid testes

Several species of cervids are currently classified as threatened or endangered due to a rapid decline in their populations. Sperm cryopreservation, in association with assisted reproductive technologies, can find application for the conservation of endangered cervids. In cases of unsuccessful sperm retrieval through other means prior to the death of the animal, adult testis is the only source of sperm. Recovery of viable sperm from adult testes depends on the effective preservation of testicular tissues through optimization of cryopreservation protocols. The present study evaluated combinations of 10% dimethyl sulfoxide (DMSO) with 0% or 80% fetal bovine serum (FBS) and 20% DMSO with 0 or 20% FBS for the cryopreservation of testicular tissues of three adult cervids using uncontrolled slow freezing protocol. The cryopreserved testis was compared to chilled tissue without cryoprotectants. Results revealed that testicular tissues of barking deer cryopreserved in 20% DMSO (D20) had all the analyzed 7 parameters (number of TNP1-, PRM2 and acrosin-expressing cells/tubule and, the number of viable, morphologically normal, acrosome intact, Annexin V-negative sperm) comparable to the chilled testis. However, testicular tissues of sambhar and hog deer cryopreserved only in D20S20 had 5 of 7 parameters comparable to the chilled testis. In conclusion, D20 is acceptable for cryopreservation of barking deer and D20S20 for sambar and hog deer testes.

Prediction model for obtaining spermatozoa with testicular sperm extraction in men with non-obstructive azoospermia.

Can an externally validated model, based on biological variables, be developed to predict successful sperm retrieval with testicular sperm extraction (TESE) in men with non-obstructive azoospermia (NOA) using a large nationwide cohort? Our prediction model including six variables was able to make a good distinction between men with a good chance and men with a poor chance of obtaining spermatozoa with TESE. Using ICSI in combination with TESE even men suffering from NOA are able to father their own biological child. Only in approximately half of the patients with NOA can testicular sperm be retrieved successfully. The few models that have been developed to predict the chance of obtaining spermatozoa with TESE were based on small datasets and none of them have been validated externally. We performed a retrospective nationwide cohort study. Data from 1371 TESE procedures were collected between June 2007 and June 2015 in the two fertility centres. All men with NOA undergoing their first TESE procedure as part of a fertility treatment were included. The primary end-point was the presence of one or more spermatozoa (regardless of their motility) in the testicular biopsies.We constructed a model for the prediction of successful sperm retrieval, using univariable and multivariable binary logistic regression analysis and the dataset from one centre. This model was then validated using the dataset from the other centre. The area under the receiver-operating characteristic curve (AUC) was calculated and model calibration was assessed. There were 599 (43.7%) successful sperm retrievals after a first TESE procedure. The prediction model, built after multivariable logistic regression analysis, demonstrated that higher male age, higher levels of serum testosterone and lower levels of FSH and LH were predictive for successful sperm retrieval. Diagnosis of idiopathic NOA and the presence of an azoospermia factor c gene deletion were predictive for unsuccessful sperm retrieval. The AUC was 0.69 (95% confidence interval (CI): 0.66-0.72). The difference between the mean observed chance and the mean predicted chance was <2.0% in all groups, indicating good calibration. In validation, the model had moderate discriminative capacity (AUC 0.65, 95% CI: 0.62-0.72) and moderate calibration: the predicted probability never differed by more than 9.2% of the mean observed probability. The percentage of men with Klinefelter syndrome among men diagnosed with NOA is expected to be higher than in our study population, which is a potential selection bias. The ability of the sperm retrieved to fertilize an oocyte and produce a live birth was not tested. This model can help in clinical decision-making in men with NOA by reliably predicting the chance of obtaining spermatozoa with TESE. This study was partly supported by an unconditional grant from Merck Serono (to D.D.M.B. and K.F.) and by the Department of Obstetrics and Gynaecology of Radboud University Medical Center, Nijmegen, The Netherlands, the Department of Obstetrics and Gynaecology, Jeroen Bosch Hospital, Den Bosch, The Netherlands, and the Department of Obstetrics and Gynaecology, Academic Medical Center, Amsterdam, The Netherlands. Merck Serono had no influence in concept, design nor elaboration of this study. Not applicable.

Expression level of chromodomain Y (CDY): potential marker for prediction of sperm recovery in non-obstructive azoospermia

The availability of testis specific genes will be of help in choosing the most promising biomarkers for the detection of testicular sperm retrieval in patients with non-obstructive azoospermia (NOA). Testis specific chromodomain protein Y 1 (CDY1) is a histone acetyltransferase which concentrates in the round spermatid nucleus, where histone hyperacetylation occurs and causes the replacement of histones by the sperm-specific DNA packaging proteins, TNPs and PRMs. The aim was to evaluate CDY1 gene as a marker for predicting of successful sperm retrieval in NOA patients. This research was conducted on 29 patients with NOA who had undergone testicular sperm extraction (TESE) procedure. NOA patients were subdivided into patients with successful sperm retrieval (NOA+, n=12) and patients with unsuccessful sperm retrieval (NOA-, n=17). Relative expression of CDY1 gene and chromatin incorporation of CDY1 protein were measured by quantitative real-time polymerase chain reaction (qRT-PCR) and ELISA assay, respectively. Quantification of mRNA relative expression and incorporation of CDY1 protein in chromatin showed significant lower expressions and protein levels of CDY1 in testis tissues of NOA- in comparison to NOA+ group. The findings in this study demonstrated a correlation between the low levels of CDY1 function and unsuccessful sperm recovery in the testicular tissues of NOA- compared to NOA+ patients. Therefore, it can be reasonable to consider CDY1 as a potential biomarker for predicting the presence of spermatozoa, although the claim needs more samples to be confirmed.

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Although the study by Komori et al 1 Komori K. Tsujimura A. Miura H. et al. Serial follow-up study of serum testosterone and antisperm antibodies in patients with non-obstructive azoospermia after conventional or microdissection testicular sperm extraction. Int J Androl. 2004; 27: 32-36 Crossref PubMed Scopus (42) Google Scholar reported no change in testosterone after TESE, Ramasamy et al 2 Ramasamy R. Yagan N. Schlegel P.N. Structural and functional changes to the testis after conventional versus microdissection testicular sperm extraction. Urology. 2005; 65: 1190-1194 Abstract Full Text Full Text PDF PubMed Scopus (222) Google Scholar demonstrated a significant decrease in serum testosterone—20%—at 3-6 months. Ramasamy et al also reported that the initial decrease was followed by a return to 95% of the pre-TESE testosterone levels at the end of 18 months. Editorial CommentUrologyVol. 75Issue 3PreviewAlthough the authors did not show any pathologic findings of testis of nonobstuctive azoospermia patients, I guess that many of them might have Kleinfelter's syndrome whose total plasma testosterone levels are decreased in 50%-60% of the patients. Nevertheless, the mean baseline testosterone level 7.83 ng/mL in patients with unsuccessful sperm retrieval looks like to be much higher than that of nonobstructive azoospermia patients seen in the everyday practice. Full-Text PDF

Editorial Comment

Although the authors did not show any pathologic findings of testis of nonobstuctive azoospermia patients, I guess that many of them might have Kleinfelter's syndrome whose total plasma testosterone levels are decreased in 50%-60% of the patients. Nevertheless, the mean baseline testosterone level 7.83 ng/mL in patients with unsuccessful sperm retrieval looks like to be much higher than that of nonobstructive azoospermia patients seen in the everyday practice.

The value of repeat testicular sperm retrieval in azoospermic men

To determine the predictive value of a previous testicular biopsy to the chance of sperm retrieval in the next testicular sperm extraction (TESE) procedure, we retrospectively analyzed the outcome of past sperm collection procedures and histopathology diagnoses of patients with nonobstructive azoospermia. Repeated TESE ensured a high recovery rate (96%) when the first recovery procedure had been successful and when hypospermatogenesis was diagnosed (77%); when no spermatozoa were found on the first attempt, a repeat TESE procedure was successful in one-third of the patients.

Postoperative pain, complications, and satisfaction rates in patients who undergo surgical sperm retrieval

Objective To determine patients’ experiences with surgical sperm retrieval and its common complications. Design A questionnaire based survey using visual analogue scales (VAS) and closed questions to analyze complication, pain, and satisfaction rates. Setting Tertiary care university hospital. Patient(s) One hundred consecutive males undergoing surgical sperm retrieval by percutaneous epididymal sperm aspiration (PESA) or testicular sperm extraction (TESE). Intervention(s) None. Main outcome measure(s) We surveyed for pain perception complication rates and satisfaction scores. Result(s) Of the 85 patients who replied, 21 underwent retrieval for nonobstructive causes, 37 following failed reversal of vasectomy, and 27 for other obstructive causes. Retrieval was successful in 100% of obstructive causes and in 61% for nonobstructive azoospermia. Epididymal retrieval was successful in 23 patients, 30 patients underwent TESE after failed PESA, and 23 had TESE only. There were significant increases in pain perception scores and reported complications with TESE over PESA (31 vs. 16; and 21 out of 63 vs. 2 out of 22, respectively), but no difference in satisfaction rate. The cause of azoospermia did not affect pain perception or satisfaction in TESE. Complication rates were increased in larger testes (3 out of 22 vs. 24 out of 63). Unsuccessful sperm retrieval did not significantly affect patients’ pain perception or satisfaction. Surgical sperm retrieval was rated as significantly less painful than both vasectomy and reversal (21% vs. 42% vs. 57%, respectively) and was associated with significantly fewer days absent from work (3.0 vs. 8.5). Conclusion(s) Surgical sperm retrieval by PESA or TESE is a safe procedure with only minor complications that is tolerated well by patients.