Achieving an optimal pregnancy outcome through the combined utilization of micro-TESE and ICSI in cryptorchidism associated with a non-canonical splicing variant in RXFP2.

PURPOSE: To identify the genetic cause of a cryptorchidism patient carrying a non-canonical splicing variant highlighted by SPCards platform in RXFP2 and to provide a comprehensive overview of RXFP2 variants with cryptorchidism correlation. METHODS: We identified a homozygous non-canonical splicing variant by whole-exome sequencing and Sanger sequencing in a case with cryptorchidism and non-obstructive azoospermia (NOA). As the pathogenicity of this non-canonical splicing variant remained unclear, we initially utilized the SPCards platform to predict its pathogenicity. Subsequently, we employed a minigene splicing assay to further evaluate the influence of the identified splicing variant. Microdissection testicular sperm extraction (micro-TESE) combined with intracytoplasmic sperm injection (ICSI) was performed. PubMed and Human Genome Variant Database (HGMD) were queried to search for RXFP2 variants. RESULTS: We identified a homozygous non-canonical splicing variant (NM_130806: c.1376-12A > G) in RXFP2, and confirmed this variant caused aberrant splicing of exons 15 and 16 of the RXFP2 gene: 11 bases were added in front of exon 16, leading to an abnormal transcript initiation and a frameshift. Fortunately, the patient successfully obtained his biological offspring through micro-TESE combined with ICSI. Four cryptorchidism-associated variants in RXFP2 from 90 patients with cryptorchidism were identified through a literature search in PubMed and HGMD, with different inheritance patterns. CONCLUSION: This is the first cryptorchidism case carrying a novel causative non-canonical splicing RXFP2 variant. The combined approach of micro-TESE and ICSI contributed to an optimal pregnancy outcome. Our literature review demonstrated that RXFP2 variants caused cryptorchidism in a recessive inheritance pattern, rather than a dominant pattern.

Clinical management of nonobstructive azoospermia: An update.

Approximately 1% of the general male population has azoospermia, and nonobstructive azoospermia accounts for the majority of cases. The causes vary widely, including chromosomal and genetic abnormalities, varicocele, drug-induced causes, and gonadotropin deficiency; however, the cause is often unknown. In azoospermia caused by hypogonadotropic hypogonadism, gonadotropin replacement therapy can be expected to produce sperm in the ejaculate. In some cases, upfront varicocelectomy for nonobstructive azoospermia with varicocele may result in the appearance of ejaculated spermatozoa; however, the appropriate indication should be selected. Each guideline recommends microdissection testicular sperm extraction for nonobstructive azoospermia in terms of successful sperm retrieval and avoidance of complications. Sperm retrieval rates generally ranged from 20% to 70% but vary depending on the causative disease. Various attempts have been made to predict sperm retrieval and improve sperm retrieval rates; however, the evidence is insufficient. Further evidence accumulation is needed for salvage treatment in cases of failed sperm retrieval. In Japan, there is inadequate provision on the right to know the origin of children born from artificial insemination of donated sperm and the rights of sperm donors, as well as information on unrelated family members, and the development of these systems is challenging. In the future, it is hoped that the pathogenesis of nonobstructive azoospermia with an unknown cause will be elucidated and that technology for omics technologies, human spermatogenesis using pluripotent cells, and organ culture methods will be developed.

A step closer to parenthood with non-obstructive azoospermia: Unveiling the impact of microdissection testicular sperm extraction in Australia's largest single-centre study.

BACKGROUND: Non-obstructive azoospermia (NOA) diagnosis poses challenges for couples seeking parenthood. Microdissection testicular sperm extraction (MD-TESE) excels in retrieving testicular sperm cells for NOA cases. However, limited live birth data in Australian NOA patients hinders accurate counselling. AIMS: This study aimed to determine the likelihood of infertile couples with a male partner diagnosed with NOA conceiving biological children using MD-TESE / intracytoplasmic sperm injection (ICSI). MATERIALS AND METHODS: A retrospective cohort study included 108 NOA men treated at a public fertility unit and a private fertility centre (May 2009-May 2022). PRIMARY OUTCOME: live birth rate (LBR); secondary outcomes: sperm retrieval rate, pregnancy rate, and neonatal outcomes. RESULTS: Among 108 patients undergoing MD-TESE, the positive sperm retrieval rate (PSRR) was 64.8% (70/108). Histology best predicted sperm retrieval success, with hypo-spermatogenesis yielding a 94.1% PSRR. Age, testicular volume, and hormonal parameters had no significant impact. Mean male age: 35.4 years; mean partner age: 32.7 years. Fertilisation rate: 50.7%. LBR per initiated cycle: 58.7% (37/63); per embryo transfer: 63.8% (37/58); per initially diagnosed NOA man: 34.3% (37/108). Cumulative LBR: 74.1% (43/58); twin rate: 10.8% (4/37). No neonatal deaths or defects were observed among 47 live offspring. CONCLUSION: This study provides valuable data for counselling NOA couples on the probability of conceiving biological offspring. MD-TESE and ICSI yielded favourable PSRR (64.8%) and LBR (63.8%). However, couples should be aware that once NOA is confirmed, the chance of taking home a baby is 34%.

Potential for artificial intelligence in medicine and its application to male infertility.

Background: The third AI boom, which began in 2010, has been characterized by the rapid evolution and diversification of AI and marked by the development of key technologies such as machine learning and deep learning. AI is revolutionizing the medical field, enhancing diagnostic accuracy, surgical outcomes, and drug production. Methods: This review includes explanations of digital transformation (DX), the history of AI, the difference between machine learning and deep learning, recent AI topics, medical AI, and AI research in male infertility. Main Findings Results: In research on male infertility, I established an AI-based prediction model for Johnsen scores and an AI predictive model for sperm retrieval in non-obstructive azoospermia, both by no-code AI. Conclusions: AI is making constant progress. It would be ideal for physicians to acquire a knowledge of AI and even create AI models. No-code AI tools have revolutionized model creation, allowing individuals to independently handle data preparation and model development. Previously a team effort, this shift empowers users to craft customized AI models solo, offering greater flexibility and control in the model creation process.

Simultaneous bioreduction of tellurite and selenite by Yarrowia lipolytica, Trichosporon cutaneum, and their co-culture along with characterization of biosynthesized Te-Se nanoparticles.

BACKGROUND: Natural and anthropogenic activities, such as weathering of rocks and industrial processes, result in the release of toxic oxyanions such as selenium (Se) and tellurium (Te) into the environment. Due to the high toxicity of these compounds, their removal from the environment is vital. RESULTS: In this study, two yeast strains, Yarrowia lipolytica and Trichosporon cutaneum, were selected as the superior strains for the bioremediation of tellurium and selenium. The reduction analyses showed that exposure to selenite induced more detrimental effects on the strains compared to tellurite. In addition, co-reduction of pollutants displayed almost the same results in selenite reduction and more than ~ 20% higher tellurite reduction in 50 h, which shows that selenite triggered higher tellurite reduction in both strains. The selenite and tellurite kinetics of removal were consistent with the first-order model because of their inhibitory behavior. The result of several characterization experiments, such as FE-SEM (Field emission scanning electron microscopy), dynamic light scattering (DLS), Fourier-transform infrared spectroscopy (FTIR), X-ray diffractometer (XRD), and dispersive X-ray (EDX) on Te-Se nanoparticles (NPs) revealed that the separated Te-Se NPs were needle-like, spherical, and amorphous, consisted of Te-Se NPs ranging from 25 to 171 nm in size, and their surface was covered with different biomolecules. CONCLUSIONS: Remarkably, this work shows, for the first time, the simultaneous bioreduction of tellurite and selenite and the production of Te-Se NPs using yeast strains, indicating their potential in this area, which may be applied to the nanotechnology industry and environmental remediation.

Micro-TESE strategy in patients with NOA caused by AZFc deletion: synchronous or asynchronous?

In the treatment of infertile patients with non-obstructive azoospermia (NOA) caused by the deletion of the azoospermia factor c region (AZFc) on the Y chromosome, synchronous and asynchronous surgical strategies are discussed. Clinical data from NOA patients with the AZFc deletion who underwent micro-TESE were analyzed retrospectively. The sperm retrieval rate (SRR) and sperm utilization rate of synchronous and asynchronous operation groups were followed up and compared. The fertilization rate, high-quality embryo rate, clinical pregnancy rate, abortion rate, and cumulative live birth rate of ICSI in patients with successful sperm retrieval were compared between the two groups. The two groups had sperm utilization rates of 98.9% (93/94) and 50.0% (14/28), respectively. The asynchronous group's sperm consumption rates were much lower than those of the synchronous operation group. Fertilization rate, high-quality embryo rate, clinical pregnancy rate of fresh transfer cycle, abortion rate, and cumulative live birth rate of patients in the synchronous operation group with fresh sperm, and the asynchronous operation group with thawed sperm, respectively, were 30.6% vs 33.8%, 33.8% vs 40.7%, 40.0% vs 12.5%, 30.4% vs 7.1%. Between the two groups, there was no significant difference. This suggests that individuals with NOA caused by the AZFc deletion have a high possibility of successfully acquiring sperm using micro-TESE and ICSI to conceive their own offspring. Synchronous micro-TESE is recommended to improve sperm utilization rate and the cumulative live birth rate.

Micro-TESE surgery combined with ICSI regimen in the treatment of non-obstructive azoospermia patients and its effect analysis.

This study aimed to analyze the clinical effects of microdissection testicular sperm extraction (micro-TESE) surgery combined with an intracytoplasmic sperm injection (ICSI) regimen in the treatment of non-obstructive azoospermia (NOA) patients with different etiologies. In total, 128 NOA patients participated in this study, in which they received infertility treatment by micro-TESE surgery combined with an ICSI regimen, and all patients were divided into three groups [the Klinefelter syndrome (KS), the idiopathic and the secondary NOA groups]. In addition, the sperm retrieval rate (SRR), fertilization rate, embryo development status and clinical treatment effects were analyzed. Among the 128 NOA patients, the SRR of KS NOA patients was 48.65%, those of idiopathic and the secondary patients were 33.82% and 73.91%, respectively. Regardless of etiologies, there was no correlation with age, hormone value or testicular volume. Further analysis showed that the SRR of the KS group was positively related with testosterone (T) values, and the SRR of the secondary group had a positive relationship with follicle-stimulating hormone or luteinizing hormone values. In the subsequent clinical treatment, the retrieved sperm was subjected to ICSI and achieved good treatment effects, especially in the secondary group, and the implantation rate (55.56%) and clinical pregnancy rate (68.42%) were both higher than those of the idiopathic group (28.75% and 40.00%) and KS group (22.05% and 30.77%). Micro-TESE surgery combined with ICSI insemination is the most effective treatment regimen for NOA patients. The SRR of NOA patients with different etiologies are related to certain specific factors, and micro-TESE surgery seems to be the ideal and only way to have biological children.

Telomere Length in Human Spermatogenic Cells as a New Potential Predictor of Clinical Outcomes in ART Treatment with Intracytoplasmic Injection of Testicular Spermatozoa.

Predicting the clinical outcomes of intracytoplasmic sperm injection (ICSI) cycles that use the testicular spermatozoa of azoospermic patients presents a challenge. Thus, the development of additional approaches to assessing the competence of a testicular-sperm-derived embryo without causing damage to gametes or the embryo is necessary. One of the key parameters in determining such developmental competence is telomere length (TL). We aimed to analyze TLs in spermatogenic cells from the testicular biopsy samples of azoospermic patients and determine how this parameter influences embryo competence for pre- and post-implantation development. Using Q-FISH, we studied the TL of the chromosomes in spermatogonia and spermatocytes I from the TESE biopsy samples of 30 azoospermic patients. An increase in TL was detected during the differentiation from spermatogonia to spermatocytes I. The patients' testicular spermatozoa were used in 37 ICSI cycles that resulted in 22 embryo transfers. Nine pregnancies resulted, of which, one was ectopic and eight ended in birth. The analysis of embryological outcomes revealed a dependence between embryo competence for development to the blastocyst stage and the TL in spermatogenic cells. The TLs in spermatogonia and spermatocytes I in the testicular biopsy samples were found to be higher in patients whose testicular sperm ICSI cycles resulted in a birth. Therefore, the length of telomeres in spermatogenic cells can be considered as a potential prognostic criterion in assessing the competence of testicular-sperm-derived embryos for pre- and post-implantation development. The results of this study provide the basis for the development of a laboratory test for the prediction of testicular sperm ICSI cycle outcomes.

Follicle-stimulating hormone and Inhibin B as predictors of successful sperm retrieval in men undergoing testicular sperm extraction: A review of 44 cases and meta-analysis.

The aim of the study and meta-analysis was to evaluate the predictive value of follicle-stimulating hormone (FSH) and inhibin B in sperm retrieval in men with non-obstructive azoospermia NOA. A total of 44 patients with a mean age of 36.1 years (SD=+/- 6.17) was included. We had 19 patients with successful sperm retrieval and 25 with failure. All patients had a normal karyotype. There was no difference between groups regarding patients mean age, prolactin or FSH. Patients with successful sperm retrieval had a significantly higher inhibin B level (134.62(+/-64,35) vs. 72.36(+/-67.78), p=0.006) and, paradoxically a higher body weight (92.38(+/-11.38) vs. 83.76(+/-11.90), p=0.027). The forest plots showed that a higher FSH level was significantly correlated with a negative success rate. Ahigher Inhibin B level was associated with a higher successful sperm retrieval (p=0.00001 respectively, p=0.0002). Surgical sperm retrieval could be, in some cases, the only chance to have a biological offspring.

Onco-TESE (Testicular Sperm Extraction): Insights from a Tertiary Center and Comprehensive Literature Analysis.

Background and Objectives: The peak of incidence of testicular cancer (TC) occurs among individuals in their reproductive age, emphasizing the importance of fertility preservation as an integral aspect of disease management. Sperm cryopreservation performed before orchiectomy is ineffective in azoospermic men, necessitating alternative approaches such as microdissection testicular sperm extraction (mTESE) at the time of orchiectomy (onco-mTESE) to obtain viable sperm. This study presents the findings from our institution's experience with onco-mTESE and critically discusses our results in light of the existing body of literature. Materials and Methods: This is a tertiary center retrospective analysis of onco-mTESE procedures performed at a single center between December 2011 and July 2022. The included patients were post-puberal men with testicular tumors requiring orchiectomy, along with concomitant severe oligozoospermia or azoospermia. Bilateral mTESE was performed in all cases. Surgical outcomes, sperm retrieval rates, the usage of preserved viable sperm, assistive reproductive techniques' results, and post-operative serum testosterone were recorded. Results: A total of nine patients were included, with a median age of 34 (IQR 29-36) years. All patients had germ cell tumors (GCTs), with seminomatous and non-seminomatous GCTs accounting for 44.4% (n = 4) and 55.6% (n = 5) of patients, respectively. Sperm retrieval occurred in three (33%) patients: one patient in the ipsilateral testis, one in the contralateral testis, and one in both testes. No complications were reported during the procedure, and no post-operative hypogonadism was observed. Among the three patients with successful sperm retrieval, an intracytoplasmic sperm injection (ICSI) was performed in two patients, resulting in two pregnancies, leading to one healthy live birth and one miscarriage. Conclusions: In the context of TC, it is essential to conduct a thorough evaluation of testicular function, including a semen analysis and cryopreservation. Onco-mTESE has proven its safety in preserving fertility in azoospermic cases while ensuring the efficacy of oncological treatment.

[Molecular markers for predicting the success of micro-TESE in non-obstructive azoospermia].

Non-obstract azoospermia (NOA) is a serious male infertility disease. At present, testicular sperm extraction (micro-TESE) is performed in combination with intracytoplasmic sperm injection (ICSI) technology, NOA patients can have their own consanguine offspring. However, due to the invasiveness and uncertainty of micro-TESE surgery, it is difficult for patients to accept it. Therefore, finding an accurate method to predict the possibility of micro-TESE successful sperm retrival would be beneficial to azoospermia patients. Many genes are transcribed and expressed during spermatogenesis, and molecular assays have irreplaceable sensitivity and specificity in predicting the success sperm retrivel of micro-TESE. This article reviews the methods to predict the success sperm retrivel of micro-TESE including mRNA, non-coding RNA (piRNA, microRNA, cirRNA, tFRNAs) and some protein so far, to provide certain reference value for clinical and subsequent research.

[Testicular and epididymal sperm extraction surgery]. / Chirurgies d'extraction de spermatozoïdes testiculaires et épididymaires.

BACKGROUND: Testicular and epididymal sperm extraction surgery is a frequent procedure. However, to date, there has been no French consensus on the technique to be preferred in the various situations of male infertility and on the best way to perform them. We therefore decided to proceed with a formalized expert consensus, using the method recommended by the French National Authority for Health. The aim was to provide the French urology community with practical advice on how best to perform these procedures. METHODS: Twenty-six international experts met online for the consensus. A research committee carried out a comprehensive literature review and prepared the 55 statements submitted to the rating group. After 2 rounds of scoring, 50 recommendations were validated in March 2023, having achieved a consensus of more than 85% among the experts. RESULTS: The consensus covered (1) preparation for sperm extraction surgery, (2) the different sperm extraction surgery techniques (Microsurgical epididymal sperm aspiration [MESA], Percutaneous epididymal sperm aspiration [PESA], Conventional Testicular Sperm Extraction [TESE], Microsurgical Testicular Sperm Extraction [micro-TESE]), (3) advice to be given to the patient concerning the postoperative period, before presenting the results of the sperm extraction surgeries. CONCLUSIONS: The expert consensus meeting on the performance of sperm extraction surgery presented a set of clinical guidelines based on the available literature and expert opinion. These guidelines should have a favourable effect on the development of this activity in France.

Non-invasive molecular biomarkers for predicting outcomes of micro-TESE in patients with idiopathic non-obstructive azoospermia.

Non-obstructive azoospermia (NOA), the most severe type of male infertility, affects approximately 1% of men worldwide. However, the aetiology of most NOA cases is not definite, that is defined as idiopathic NOA (INOA), posing a clinical conundrum worldwide. Most of these patients must receive donor sperm treatment until the emergence of microdissection testicular sperm extraction (micro-TESE). Although this procedure has recently become a promising treatment for INOA, the low sperm retrieval rate and testicular trauma have prompted us to explore appropriate non-invasive molecular biomarkers to predict the outcomes of sperm recovery preoperatively. Previous studies have identified a spectrum of biomarkers to address this challenging issue at various levels in different tissues, such as DNAs, RNAs, protein and steroid levels in the blood and seminal fluid. To better understand and assess the predictive values of diverse molecular biomarkers from different tissues on the outcome of sperm retrieval by micro-TESE in patients with INOA, we summarised recent findings and discussed the potential applications of these methods. The ultimate goal of this study was to provide references for further studies and clinical management.

Do inflammatory markers such as NLRP3 and IL1-ß have an effect on the prediction of testicular sperm extraction success in infertile men?

Infertility is defined as the inability of couples to have a baby without contraception after at least 1 year of regular sexual intercourse. Mechanisms to explain inflammation in male infertility of unknown causes are still being investigated. The inflammasome is a key regulator of innate immunity, which is involved in the inflammatory response to infections and various diseases through the activation of caspase-1 and the use of inflammatory cytokines. Although many factors are believed to affect the success of mTESE in infertile patients with non-obstructive azoospermia (NOA), the inflammation mechanisms in the environment have not been clearly explained in the aetiology of infertility. In this study, we aimed to investigate the effect of NLRP3 and similar inflammasome mechanisms on antioxidant mechanisms on the success of mTESE. A total of 24 NOA patients with micro-testicular sperm extraction (mTESE +) and no sperm found (mTESE -) participated in the study between January 2020 and January 2021. NLRP3, IL1-ß, TAS, TOS and OSI amounts in serum and seminal plasma parameters were compared statistically, and their effects on mTESE success were investigated. FSH, LH, estradiol and testosterone values did not differ significantly (p > 0.05) in the group with mTESE (-) and mTESE (+). Serum IL-1 Beta, NLRP3, TOS, TAS, and OSI values did not differ significantly (p > 0.05) in the group with mTESE (-) and mTESE (+). Seminal plasma TOS and OSI values were significantly lower in the group with mTESE (+) than the group with mTESE (-). Although inflammasomes such as NLRP3 and IL1-ß do not have a significant predictive value in the success of mTESE, we think the high seminal plasma values of infertile patients may be understandable with further studies. This study was conducted to determine how inflammasomes are involved in IL-1ß, pathway NLRP3, and sperm retrieval in micro testicular sperm extraction (mTESE) in infertile men.

Does testicular sperm retrieval adversely impact spermatogenesis over the long-term?

Testicular sperm retrieval (TSR) techniques are valuable in the context of severe idiopathic male factor infertility; however, there are few studies in the literature examining the long-term impact of TSR on testicular function. The objective was to determine whether testicular sperm aspiration (TESA) or microdissection testicular sperm extraction (micro-TESE) worsens the pre-existing spermatogenesis deficiency in men with either cryptozoospermia or severe oligozoospermia. The study population consisted of 145 men with either cryptozoospermia or severe oligozoospermia that underwent TESA or micro-TESE and had long-term post-operative semen analyses (SA). Patients with SA prior to and following TSR were included (n = 24). Amongst them, 16 men underwent TESA and 8 underwent micro-TESE. The follow-up SA was obtained at a mean of 3.0 ± 2.0 years following TSR (range: 0.3-8.3 years) amongst all participants. The post-operative semen parameters in the TESA group were similar to the pre-intervention parameters (p > 0.1). Similarly, the micro-TESE cohort did not demonstrate significant alterations in semen parameters post-intervention (p > 0.05). None of the men in the study became azoospermic following the TSR. Our study indicates TESA or micro-TESE do not appear to worsen the pre-existing spermatogenesis deficiencies in cryptozoospermic and oligozoospermic men over a long-term period. Larger studies are required to corroborate these findings.

Ten-year outcomes of patients who developed persistent azoospermia following chemotherapy associated with different oncological diagnoses: A retrospective cohort study from a different perspective.

BACKGROUND: This study evaluated the treatment procedures for chemotherapy (CT)-induced persistent azoospermia and their outcomes from a different perspective. METHODS: In 63 patients (mean age: 30.16 ± 4.91 years) who had undergone CT 11 ± 5 years earlier, the semen volume, gonadotropins level, FSH level, genetics, micro-testicular sperm extraction (m-TESE) result, sperm DNA fragmentation index (SDFI), semen reactive oxidative stress (ROS) rate, duration of embryonic development, and pregnancy and baby take-home rates were examined. The correlations between the ROS rates and the SDFIs, m-TESE results, sperm motility, pathology scores, time-lapses, and baby take-home rates were evaluated. RESULTS: The semen volumes were 3.5 ± 1.1/ml. The FSH level following CT was 17.87 ± 5.80 mIU/ml. A sperm rate of 34.9% was found from the m-TESE result. The mean SDFI and ROS rate were 4 (<15-30>) and 1.29 ± 0.51, respectively. The time-lapse was calculated as 5h. Pregnancy and live birth were achieved at 20.63% and 12.7%, respectively. In the patients with a low ROS (≤1.42) and SDFI (≤15), the m-TESE success rate was high, the FSH value was low, the pathological score and fertilization rate were elevated, the embryonic cleavage period was normal, and the pregnancy and baby take-home rates were high. DISCUSSION: The sperms may be detected using m-TESE in patients who develop persistent azoospermia associated with CT due to different oncological diagnoses. Our study revealed that a low FSH value and normal ejaculatory ROS rates are positive predictive factors of sperm detection before m-TESE. The motility of the sperms detected after m-TESE and normal SDFI rates were found to be positive predictive criteria of high fertilization, good embryonic cleavage, pregnancy, and live birth.

Translational aspects of novel findings in genetics of male infertility-status quo 2021.

INTRODUCTION: Male factor infertility concerns 7-10% of men and among these 40-60% remain unexplained. SOURCES OF DATA: This review is based on recent published literature regarding the genetic causes of male infertility. AREAS OF AGREEMENT: Screening for karyotype abnormalities, biallelic pathogenic variants in the CFTR gene and Y-chromosomal microdeletions have been routine in andrology practice for >20 years, explaining ~10% of infertility cases. Rare specific conditions, such as congenital hypogonadotropic hypogonadism, disorders of sex development and defects of sperm morphology and motility, are caused by pathogenic variants in recurrently affected genes, which facilitate high diagnostic yield (40-60%) of targeted gene panel-based testing. AREAS OF CONTROVERSY: Progress in mapping monogenic causes of quantitative spermatogenic failure, the major form of male infertility, has been slower. No 'recurrently' mutated key gene has been identified and worldwide, a few hundred patients in total have been assigned a possible monogenic cause. GROWING POINTS: Given the high genetic heterogeneity, an optimal approach to screen for heterogenous genetic causes of spermatogenic failure is sequencing exomes or in perspective, genomes. Clinical guidelines developed by multidisciplinary experts are needed for smooth integration of expanded molecular diagnostics in the routine management of infertile men. AREAS TIMELY FOR DEVELOPING RESEARCH: Di-/oligogenic causes, structural and common variants implicated in multifactorial inheritance may explain the 'hidden' genetic factors. It is also critical to understand how the recently identified diverse genetic factors of infertility link to general male health concerns across lifespan and how the clinical assessment could benefit from this knowledge.

Cumulative IVF outcomes after retrieval of testicular spermatozoa: should we use immotile spermatozoa for ICSI?

RESEARCH QUESTION: What are the cumulative clinical pregnancy rates (CPR) and live births rates (LBR) in intracytoplasmic sperm injection (ICSI) cycles using testicular motile compared with immotile spermatozoa, obtained from testicular sperm aspiration (TESA) or extraction (TESE)? DESIGN: A retrospective analysis of ICSI cycles using TESA or TESE over a period of 7 years. Cycles were divided into two groups according to the motility of the retrieved spermatozoa: Group A consisted of couples with motile spermatozoa; Group B of couples with immotile spermatozoa. Group B was subdivided into two groups: B1 consisted of couples with motile spermatozoa and B2 with immotile spermatozoa after the addition of pentoxifylline. RESULTS: No differences in CPR and LBR per transfer was found between the study groups after fresh embryo transfer. No pregnancies were achieved by vitrified-warmed embryo transfer in group B2. Fertilization rates decreased when using immotile spermatozoa (64.4%, 56%, 37.9%, for groups A, B1 and B2, respectively, P < 0.001). Top-quality embryo rates were higher in groups A and B1 compared with B2 (40.7% and 40.1% versus 19.1%, respectively, P = 0.015). Cumulative CPR (53%, 41.7%, 13.6% for groups A, B1 and B2, respectively, P = 0.005) and LBR (42.4%, 30%, 13.6% for groups A, B1 and B2, respectively P = 0.03) per oocyte retrieval was significantly higher when using motile spermatozoa compared with motile or immotile spermatozoa after adding pentoxifylline. CONCLUSIONS: Although fertilization, top-quality embryo rates, cumulative CPR and LBR decreased when using immotile spermatozoa, ICSI is still valid; therefore, it should be considered and offered to couples before embarking on a donor sperm insemination cycle, or cryopreserving oocytes for future additional testicular sperm retrieval.

TESE-ICSI outcomes per couple in vasectomized males are negatively affected by time since the intervention, but not other comorbidities.

RESEARCH QUESTION: Does time since vasectomy (as obstructive interval) and the presence of different male comorbidities adversely affect the likelihood of achieving a newborn for vasectomized males undergoing testicular sperm extraction (TESE) and intracytoplasmic sperm injection (ICSI)? DESIGN: This retrospective study included 364 couples with vasectomized males undergoing TESE-ICSI cycles with autologous oocytes at IVI Valencia. The main outcome was live birth rate (LBR). Subjects were divided according to the male risk factor evaluated into quartiles (obstructive interval, body mass index [BMI]) or groups (hypertension, diabetes mellitus, dyslipidaemia). The reproductive outcomes were calculated per embryo transfer, per ovarian stimulation completed, and per couple. RESULTS: The average obstructive interval was 11.3 years. The LBR was 34.4% (95% CI 30.1-38.6) per embryo transfer, 27.8% (95% CI 24.1-31.5) per ovarian stimulation and 46.2% (95% CI 41.8-51.3) per couple. When considering obstructive interval, a significantly lower LBR per couple (P = 0.04) was found in the group with the longest obstruction time: Q1 42.1% (95% CI 33.5-50.7), Q2 49.1% (95% CI 36.1-62.1), Q3 56.3% (95% CI 46.7-65.9) and Q4 37.2% (95% CI 26.5-47.9) but the cumulative live birth rate (CLBR) was not affected (P = 0.63). LBR per ovarian stimulation of males with hypertension was significantly lower (P = 0.04) than healthy males: 13.5% (95% CI 2.5-24.5) and 28.6% (95% CI 24.7-32.5), respectively. The group of diabetic vasectomized males had a significantly higher CLBR (P = 0.02). The remaining risk factors assessed (smoking, dyslipidaemia and a high BMI) did not affect LBR compared with their healthy counterparts. CONCLUSION: Time since vasectomy appears to negatively influence the LBR when assessed per couple. The CLBR was not affected by the obstructive interval or the presence of other male comorbidities apart from diabetes, which had a significant effect.

Prediction of the extent of germ cell loss utilising a noninvasive spectroscopy method in rat testicular damage model.

The aim of this study is to investigate the efficiency of elastic light single-scattering spectroscopy system, a noninvasive method, to acquire spectra during testicular biopsy from normal and damaged seminiferous tubules with various degrees of germ cell loss. Adult control rats and doxorubicin-injected rats to achieve seminiferous germ cell loss (for 10 days [10D], 20 days [D20], 30 days [D30], 40 days [D40], and 50 days [D50]) were used. Spectroscopic measurements were acquired utilising a single-fibre optical probe, and histopathology of the biopsied testicular tissue samples were compared. Time-dependent testicular damage comprising various degrees of seminiferous tubule degeneration after doxorubicin-administration was observed. In D30, D40 and D50 groups, where significant germ cell loss was identified, elastic light single-scattering spectroscopy system signals were well correlated with disturbed spermatogenesis where significant differences in spectral signals were obtained. Our findings indicate that the elastic light single-scattering spectroscopy system has the potential to enable instant imaging of spermatogenesis in rats and could also be useful in humans for clinical applications, such as to increase sperm recovery success during micro-TESE for men with nonobstructive azoospermia.