Testicular Volume Research Articles

Spermatogenesis after gonadotoxic childhood treatment: follow-up of 12 patients.

What is the long-term impact of presumed gonadotoxic treatment during childhood on the patient's testicular function at adulthood? Although most patients showed low testicular volumes and some degree of reproductive hormone disruption 12.3 (2.3-21.0) years after gonadotoxic childhood therapy, active spermatogenesis was demonstrated in the semen sample of 8 out of the 12 patients. In recent decades, experimental testicular tissue banking programmes have been set up to safeguard the future fertility of young boys requiring chemo- and/or radiotherapy with significant gonadotoxicity. Although the risk of azoospermia following such therapies is estimated to be high, only limited long-term data are available on the reproductive potential at adulthood. This single-centre prospective cohort study was conducted between September 2020 and February 2023 and involved 12 adult patients. This study was carried out in a tertiary care centre and included 12 young adults (18.1-28.3 years old) who had been offered testicular tissue banking prior to gonadotoxic treatment during childhood. All patients had a consultation and physical examination with a fertility specialist, a scrotal ultrasound to measure the testicular volumes and evaluate the testicular parenchyma, a blood test for assessment of reproductive hormones, and a semen analysis. Testicular tissue was banked prior to the gonadotoxic treatment for 10 out of the 12 included patients. Testicular volumes were low for 9 patients, and 10 patients showed some degree of reproductive hormone disruption. Remarkably, ongoing spermatogenesis was demonstrated in 8 patients at a median 12.3 (range 2.3-21.0) years post-treatment. This study had a limited sample size, making additional research with a larger study population necessary to verify these preliminary findings. These findings highlight the need for multicentric research with a larger study population to establish universal inclusion criteria for immature testicular tissue banking. This study was conducted with financial support from the Research Programme of the Research Foundation-Flanders (G010918N), Kom Op Tegen Kanker, and Scientific Fund Willy Gepts (WFWG19-03). The authors declare no competing interests. NCT04202094; https://clinicaltrials.gov/ct2/show/NCT04202094?id=NCT04202094&draw=2&rank=1 This study was registered on 6 December 2019, and the first patient was enrolled on 8 September 2020.

Ovine fertility by artificial insemination in the breeding season could be affected by intraseasonal variations in ram sperm proteomic profile

It is important to note that seasonality could affect ram reproductive parameters, and therefore, fertility results after artificial insemination. In this work, 1) we assessed fertility rates after cervical artificial insemination of 11,805 ewes at the beginning (June 21st to July 20th) and at the end (November 20th to December 21st) of the reproductive season in the Assaf breed for the last four years, and 2) we aimed to identify male factors influencing the different reproductive success obtained depending on the time at the mating season in which ovine artificial insemination was performed. For this purpose, we evaluated certain ram reproductive and ultrasonographical parameters as well as we performed a multiparametric and proteomic sperm analysis of 6–19 rams at two very distant points in the mating season (July as Early Breeding Season –EBS– and November as Late Breeding Season –LBS–). Rutinary assessments carried out in the ovine reproduction centers (testicular volume, libido, sperm production and mass motility) showed non-significant differences (P ≥ 0.05) between both studied times, as well as the ram ultrasonographic evaluation (Resistive and Pulsatility Index as Doppler parameters; and pixels mean gray level, and hypoechoic areas percentage and density as echotexture parameters). However, at level of sperm functionality, although sperm quality appeared non-significantly lower (P ≥ 0.05) in the EBS, we identified a significantly different (P < 0.05) sperm proteomic profile between the seasonality points. The following proteins were identified with the lowest abundance in the EBS with a fold change > 4, a P = 2.40e-07, and a q = 2.23e-06: Fibrous Sheath-Interacting Protein 2, Disintegrin and Metalloproteinase Domain-Containing Protein 20-like, Phosphoinositide-Specific Phospholipase C, Tektin 5, Armadillo Repeat-Containing Protein 12 Isoform X3, Solute Carrier Family 9B1, Radial Spoke Head Protein 3 Homolog, Pro-Interleukin-16, NADH Dehydrogenase [Ubiquinone] 1 Alpha Subcomplex Subunit 8, Testis, Prostate and Placenta-Expressed Protein, and Acyl Carrier Protein Mitochondrial. In conclusion, while our basic analyses on male and sperm quality showed similar results between the beginning and the end of the breeding season, on a proteomic level we detected a lower expression of sperm proteins linked to the energy metabolism, sperm-oocyte interactions, and flagellum structure in the EBS. Probably, this different protein expression could be related to the lower fertility rate of Assaf ewes after cervical artificial insemination at this time. More importantly, sperm proteins can be used as highly effective molecular markers in predicting sperm fertilization ability related to intraseasonal variations.

Semen Analysis in "Urology-Naïve" Patients: A Chance of Uroandrological Screening in Young Males.

(1) Background: While females start their gynecological examinations during puberty, only few men decide to be visited by urologists in their youth. Given the participation in the EcoFoodFertility research project, our department had the opportunity to screen young males that were supposedly healthy. (2) Results: from January 2019 to July 2020, we evaluated 157 patients with sperm, blood analysis, and uroandrological examinations. The inclusion criteria were age 18-40 and absence of previous urological disease (urology-naïve). The primary endpoint of the study was to record uroandrological diseases that are occasionally discovered during examination in asymptomatic young men. The average age was 26.9 years (range 18-40); average testicular volume was 15.7 mL (range 12-22 mL); and 45.2% reported abnormal semen analysis: 62 cases of teratozoospermia, 27 asthenozoospermia, 18 oligozoospermia, and 2 azoospermia were discovered respectively; 4/157 patients were diagnosed with hypogonadism; 2 cases with suspicious testicular mass resulted in testicular cancer; and 31 suspected varicoceles and 8 patients with mild sexual dysfunctions were managed. (3) Conclusions: an uroandrological evaluation of young asymptomatic males allowed for the prompt diagnosis of different urological conditions, including cancerous ones, in our series. Despite being debatable, combining urological counselling with physical examination, semen analysis, and a laboratory profile could be useful and cost-effective in order to ameliorate male health.

Use of Doppler indices of vascular impedance for diagnosing testicular degeneration in stallions

Spectral Doppler ultrasonography is used to evaluate testicular blood flow in males. Further, the correlation between testicular function and values of Doppler indices, Resistive Index (RI), and Pulsatility Index (PI) were reported in men and several animal species (Velasco and Ruiz. Animals 2020;11:1-13). We have previously shown that stallions with oligospermia associated with medically induced testicular degeneration (TD) have higher values of Doppler indices of testicular blood flow than normal stallions (Pozor et al. JEVS 2014;34:38-9). The goal of this retrospective study was to validate Doppler indices of vascular impedance for the identification of stallions with TD. Medical records from five adult stallions with clinical signs of testicular degeneration (small, flaccid, hypoechoic testes; oligospermia or azoospermia, necrospermia; immature germ cells, detached sperm heads) were reviewed. Three of the five subfertile stallions were later castrated, and severe TD was confirmed histologically. Spectral Doppler ultrasound examination of the marginal aspect of the testicular artery was performed by the same operator, and values of RI and PI were determined for each testis. Doppler indices in the five subfertile stallions were compared with those of five normal fertile stallions, evaluated four times in different seasons, using Kruskal-Wallis One-Way ANOVA method. There were no significant differences betweensubfertile and normal stallions for RI (Mean: 0.66 vs. 0.63, SD: 0.08 vs. 0.11, respectively; P>0.5) or PI (Mean: 1.18 vs. 1.22, SD: 0.23 vs. 0.40, respectively; P>0.5). In addition, information from a pony stallion was obtained over four consecutive years. An initial testicular blood flow evaluation was performed on this stallion at 18 yrs. of age, when good fertility and normal semen parameters were confirmed. Over the four years, the pony stallion's testicular volume decreased from 95 cm3 to 41 cm3. The stallion's total sperm numbers fell from an average of 4.6 billion in Year 1 to 0 in Year 4, indicating progressive TD, which was later confirmed histologically. Values of Doppler indices of vascular impedance in the affected stallion were compared to a pony stallion of a similar size and age with normal testicular and seminal parameters. Doppler indices in the affected stallion did not change significantly over the 4 years evaluated (RI: 0.58 – 0.65; PI: 1.02 – 1.12). Further, the Doppler indices in the affected stallion were not different from the normal pony stallion. Based on data collected from this small population of stallions, Doppler indices of testicular blood flow may not be reliable for diagnosing testicular degeneration in stallions. We have no clear explanation for why the results presented here differ from our previous study. Perhaps different etiologies of TD, acute induced vs chronic progressive, contributed to this difference.

Blue LED light treatment of pregnant mares is not detrimental to gonadal development in their offspring

A light program transmitted via blue LED light directed at one eye from individual light masks (Equilume, Kildare, Ireland) to pregnant mares from the middle of December onwards advanced the onset of foaling with birth of mature and healthy foals, but they were slightly smaller than foals from untreated control pregnancies (Lutzer et al., Domestic Anim Endocrinol 2022;78:106675). Differences, however, equilibrated within the first year of life. In this study we followed the hypothesis that light treatment of pregnant mares is without detrimental effects on body size and gonadal development of offspring after puberty. A total of 47 warmblood horses born and raised on the same stud farm to blue LED light-treated (17 males and 9 females) or control mares (14 males and 9 females) were included. At 24 months of age, their body weight, height at withers and cannon bone circumference was determined. In males, the external genitalia were examined, testis size was measured by transcutaneous ultrasound. The total testicular volume (TTV) was calculated. In females, a transrectal ultrasound examination of the genital tract was performed, the presence of corpora lutea was detected via plasma progesterone concentration and the antral follicle count (AFC) was assessed. Statistical comparisons weremade by analysis of variance (general linear model) with group (blue light vs. control) and sex (male vs. female) as between subject factors (IBM SPSS 27). Results are given as mean ± SEM. There were no group or sex differences with regard to bodyweight or height at the withers. The cannon bone circumference was larger in male vs. female horses (p<0.001) but not affected by blue light treatment during pregnancy. Testes were classified as normal in size and location (Love et al., J Reprod Fertil Suppl 1991;44:99-105) in 13 of 17 blue light and 11 of 14 control colts (n.s., chi2 analysis). The TTV was 232±14cm3 in the blue light and 218±11cm3 in the control group (n.s.). Abnormal testicular development was diagnosed in 7 males with unequal testicular size in 3 blue light and 2 control animals, both testes too small (1 blue light) or unilaterally cryptorchid (1 control). In females, no abnormalities were present. A corpus luteum (progesterone >1ng/ml) was present in 8 / 9 females from the blue light and 5 / 8 from the control group. The AFC was 20.6±2.5 in blue light and 22.1±3.7 in control females (n.s.). Results demonstrate that blue LED light treatment of pregnant mares has no detrimental effects on body and genital tract development of their offspring at two years of age.

Predicting Micro-TESE among Heterogeneous Nonobstructive Azoospermic Patients: The Impact on Surgical Decision and ICSI

Background. Previously published predictive models for microdissection testicular sperm extraction (micro-TESE) were generally assumed patients with nonobstructive azoospermia (NOA) a homogenous population, i.e., the laboratory predictors were associated with sperm retrieval rate (SRR) in a similar way among different subpopulations. In addition, previous studies primarily regarded the success of sperm retrieval as the sole endpoint, although live birth is the ultimate goal for the couples. Objectives. The main objective is to develop and evaluate the clinical benefit of a model predicting the clinical outcome of micro-TESE in heterogeneous population with NOA. The outcome of pregnancy was taken into account via assessing the association between the predicted outcome of micro-TESE and pregnancy. Materials and Methods. A development cohort of 1,292 patients with NOA and an external validation cohort of 530 patients were included. Sperm retrieval was performed using micro-TESE. Clinical outcomes, including sperm retrieval, clinical pregnancy, and live birth, were collected. We developed a model using the machine learning method random forest and provided a web-based calculator. Results. The SRR was 38.1% (492/1,292) in the development cohort and 48.5% (257/530) in the validation cohort. The final model includes etiology, AMH, sperm retrieval surgical history, testicular volume, FSH, LH, and age as predictors (ordered by variable importance). The area under the curve of our model was 0.76 (0.74–0.79) in the development cohort and 0.75 (0.71–0.79) in the external validation cohort. The decision curve analysis showed that personalized model-based surgical decision provides additional clinical benefit. The clinical pregnancy rate (CPR) and cumulative live birth rate (CLBR) were 45.3% (405/895) and 57.6% (338/587), respectively, in the overall population. For patients of different SRR, the CPR and CLBR of whom had successful sperm retrieval were similar. Discussion and Conclusion. Our model predicting the SRR of micro-TESE was generalizable and easy to use. Predicted pregnancy outcomes like CPR and CLBR could also be derived from predicted SRR. A model-based surgical decision after personalized consultation would be beneficial to patients with NOA.

Timing of puberty in relation to semen characteristics, testicular volume, and reproductive hormones: a cohort study

To investigate if timing of puberty is associated with semen characteristics, testicular volume, and reproductive hormone levels. Cohort study. The Danish National Birth Cohort (DNBC) and its sub-cohort, the Fetal Programming of Semen Quality (FEPOS) cohort of 1,058 young men. Self-reported information on timing (younger, same age, older than peers) of the pubertal markers: voice break (primary exposure), pubic hair growth, regular shaving, and axillary hair growth. We estimated the relative differences with 95% confidence intervals (CI) for semen characteristics (semen volume, sperm concentration, total sperm count, sperm motility, percentage of morphologically normal spermatozoa), testicular volume, and reproductive hormones (follicle stimulating hormone [FSH], luteinizing hormone [LH], sex hormone-binding globulin [SHBG], testosterone, estradiol, free androgen index [FAI]) obtained at a median age of 19.2 years according to timing of pubertal development. Compared to men reporting voice break 'same age as peers', men reporting voice break 'older than peers' tended to have lower total sperm count (-12% [-25%, 4%]) and lower percent morphologically normal spermatozoa (-10% [-20%, 2%]), whereas men reporting voice break 'younger than peers' tended to have lower proportion of non-progressive and immotile spermatozoa (-6% [-13%, 1%]) and larger testicular volume (7% [1%, 13%]). The pattern was less consistent for the other pubertal markers.For reproductive hormones, voice break 'older than peers' tended to have higher FSH (24% [-1%, 55%]), higher SHBG (7% [0%, 15%]), lower estradiol (-14% [-23%, -5%]) and lower FAI (-8% [-14%, -1%]), whereas voice break 'younger than peers' tended to have higher LH (4% [-2%, 11%]), higher testosterone (5% [0%, 11%]), higher estradiol (17% [6%, 29%]) and higher FAI (4% [-2%, 11%]). When the categorical pubertal markers were analyzed as a linear term to assess for dose-dependency, older age at pubertal development were associated with higher FSH, higher SHBG, lower testosterone, lower estradiol and lower FAI for most pubertal markers. These results lend weak support to the hypothesis that older age at pubertal development is associated with markers of reduced male fecundity, especially reproductive hormone levels, although associations with semen characteristics and testicular volume were statistically insignificant.

The effect of scrotal versus inguinal orchiopexy on the testicular function of children with clinically palpable, inguinal undescended testis: a randomized controlled trial.

To compare the impact of the scrotal vs inguinal orchidopexy approach on the testicular function of infants with cryptorchidism, a randomized controlled trial was conducted involving boys who were 6-12 months old at surgery and were diagnosed with clinically palpable, inguinal undescended testis. Between June 2021 and December 2021, these boys at Fujian Maternity and Child Health Hospital (Fuzhou, China) and Fujian Children's Hospital (Fuzhou, China) were enrolled. Block randomization with a 1:1 allocation ratio was employed. The primary outcome was testicular function assessed by testicular volume, serum testosterone, anti-Müllerian hormone (AMH), and inhibin B (InhB) levels. Secondary outcomes included operative time, amount of intraoperative bleeding, and postoperative complications. Among 577 screened patients, 100 (17.3%) were considered eligible and enrolled in the study. Of the 100 children who completed the 1-year follow-up, 50 underwent scrotal orchidopexy and 50 underwent inguinal orchidopexy. The testicular volume, serum testosterone, AMH, and InhB levels in both groups increased markedly after surgery (all P < 0.05), but there were no apparent differences between groups at 6 months and 12 months after operation (all P > 0.05). No differences between the scrotal and inguinal groups were noted regarding the operative time ( P = 0.987) and amount of intraoperative bleeding ( P = 0.746). The overall complication rate (2.0%) of the scrotal group was slightly lower than that of the inguinal group (8.0%), although this difference was not statistically significant ( P > 0.05). Both scrotal and inguinal orchiopexy exerted protective effects on testicular function in children with cryptorchidism, with similar operative status and postoperative complications. Scrotal orchiopexy is an effective alternative to inguinal orchiopexy in children with cryptorchidism.

Clinical and genetic characteristics of 42 Chinese paediatric patients with X-linked adrenal hypoplasia congenita

BackgroundX-linked adrenal hypoplasia congenita (AHC) is a rare disorder characterized by primary adrenal insufficiency (PAI) and hypogonadotropic hypogonadism (HH), with limited clinical and genetic characterization.MethodsThe clinical, biochemical, genetic, therapeutic, and follow-up data of 42 patients diagnosed with X-linked AHC were retrospectively analysed.ResultsHyperpigmentation (38/42, 90%), vomiting/diarrhoea (20/42, 48%), failure to thrive (13/42, 31%), and convulsions (7/42, 17%) were the most common symptoms of X-linked AHC at onset. Increased adrenocorticotropic hormone (ACTH) (42/42, 100%) and decreased cortisol (37/42, 88%) were the most common laboratory findings, followed by hyponatremia (32/42, 76%) and hyperkalaemia (29/42, 69%). Thirty-one patients presented with PAI within the first year of life, and 11 presented after three years of age. Three of the thirteen patients over the age of 14 exhibited spontaneous pubertal development, and ten of them experienced delayed puberty due to HH. Six patients receiving human chorionic gonadotropin (hCG) therapy exhibited a slight increase in testicular size and had rising testosterone levels (both P < 0.05). The testicular volumes of the three patients with pulsatile gonadotropin-releasing hormone (GnRH) therapy were larger than those of the six patients undergoing hCG therapy (P < 0.05), and they also exhibited some growth in terms of luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone. Of the 42 patients, three had an Xp21 deletion, and 39 had an isolated DAX1 defect. Most patients (9/10) with entire DAX1 deletion accounting for 23.8% (10/42) of the total variants had early onset age of less than one year.ConclusionsThis study details the clinical features and genetic spectra of X-linked AHC. Patients with X-linked AHC show a bimodal distribution of the age of onset, with approximately 70% presenting within the first year of life. Pulsatile GnRH may be recommended for HH when hCG therapy is not satisfactory, although it is difficult to achieve normal testicular volume. The combination of clinical features and molecular tests provides information for an accurate diagnosis.

Phthalates exposure and pubertal development in a 15-year follow-up birth cohort study in Taiwan.

Phthalates are ubiquitous endocrine disruptors that can affect pubertal development in children. The association of fetal and childhood levels of phthalates with pubertal development were explored. We conduct a population-based birth cohort study to investigate the association between prenatal and childhood exposure to phthalates and pubertal development. Initially, a total of 445 children were recruited from 2000 to 2001, of which 90 children were followed for 15 years which measurements of urine and development assessed at 2, 5, 8, 11, and 14 years. We defined higher Tanner stage as the 14-year-old Tanner stage ≥ 4 and 5 for boys and girls, respectively. A logistic regression analysis was conducted to estimate the crude and adjusted odds ratio of a higher Tanner stage at 14 years old. The Pearson correlation coefficient and multiple linear regression were used to estimate the association of testicular volume, uterine volume, ovarian volume, and blood hormones at 14 years of age with the log-transformed concentration of phthalates at 2, 5, 8, 11, and 14 years. In boys, a significantly different geometric mean of mono-benzyl phthalate (MBzP) was observed in 11-year-olds; 6.82 and 2.96 in the lower Tanner stage group and higher Tanner stage group. In girls, a significant difference in the geometric mean of mono(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP) in 11-year-olds and mono-ethyl phthalate (MEP) in 2-year-olds was observed; MEHHP was 32.97 and 18.13 in the lower Tanner stage group and higher Tanner stage group, and MEP was 26.54 and 65.74 in the lower Tanner stage group and higher Tanner stage group, respectively. Uterine volume at 14 years old was negatively associated with several phthalate metabolites (MEHP at 8 years old, MnBP at 8 years old, MBzP at 14 years old, MMP prenatally, MMP at 8 years old, and MEP at 8 years old) after adjusting for covariates. However, no significant correlations were found between phthalate metabolites and ovarian or testicular volume. Phthalate exposure at certain time points may influence the reproductive development of children during puberty; however, further studies should be conducted to determine the causal nature of this association.

Identification and validation of CCL2 as a potential biomarker relevant to mast cell infiltration in the testicular immune microenvironment of spermatogenic dysfunction

BackgroundSpermatogenic dysfunction is an important cause of azoospermia. Numerous studies have focused on germ-cell-related genes that lead to spermatogenic impairment. However, based on the immune-privileged characteristics of the testis, the relationship of immune genes, immune cells or immune microenvironment with spermatogenic dysfunction has rarely been reported.ResultsUsing integrated methods including single-cell RNA-seq, microarray data, clinical data analyses and histological/pathological staining, we found that testicular mast cell infiltration levels were significantly negatively related to spermatogenic function. We next identified a functional testicular immune biomarker, CCL2, and externally validated that testicular CCL2 was significantly upregulated in spermatogenic dysfunctional testes and was negatively correlated with Johnsen scores (JS) and testicular volumes. We also demonstrated that CCL2 levels showed a significant positive correlation with testicular mast cell infiltration levels. Moreover, we showed myoid cells and Leydig cells were two of the important sources of testicular CCL2 in spermatogenic dysfunction. Mechanistically, we drew a potential “myoid/Leydig cells-CCL2-ACKR1-endothelial cells-SELE-CD44-mast cells” network of somatic cell–cell communications in the testicular microenvironment, which might play roles in spermatogenic dysfunction.ConclusionsThe present study revealed CCL2-relevant changes in the testicular immune microenvironment in spermatogenic dysfunction, providing new evidence for the role of immunological factors in azoospermia.

(255) Operating Characteristics of Sperm Concentration and FSH for Predicting Chromosomal Abnormalities

Abstract Introduction Microdeletions of the Y-chromosome (Yq) and karyotype abnormalities are frequent causes of male factor infertility. Current AUA/EUA guidelines recommend obtaining Yq microdeletion analysis, karyotype, and genetic counseling in men with non-obstructive azoospermia or severe oligospermia (&amp;lt;5 mil/mL). A recent retrospective cohort study suggests that Yq microdeletions are primarily found at a sperm concentration &amp;lt; 0.5 mil/mL and that by lowering the concentration threshold for genetic testing, specificity could be increased and relative financial cost decreased without adversely affecting the sensitivity. Studies have shown that FSH is the best predictor of NOA in combination with testicular volume as well as a significant predictor of microdeletion presence. Identification of Yq microdeletions and karyotype abnormalities have implications when counseling patients (and their partners) about the likelihood of retrieving sperm and need for further genetic counseling. Objective We hypothesized that the combination of sperm concentration and FSH would better predict the presence of chromosome abnormalities in infertile men. Methods A retrospective cohort study was conducted with subjects who were referred to a military tertiary Urology clinic between 2010 to 2020 and who underwent an infertility evaluation to include genetic testing (karyotype, Y-chromosome analysis), hormone profile (FSH), and at least one semen analysis. Sperm density and FSH were analyzed as independent predictors of chromosomal abnormalities using logistic regression. Receiver operating characteristics were generated for each independent variable that achieved significance in regression. Results A total of 356 patients met inclusion criteria. Over the ten-year period, the prevalence of Y-chromosome microdeletions and karyotype abnormalities was 3.3% (n=12) and 8.4% (n=30), respectively. The ability to predict chromosomal abnormality based on logistic regression was statistically significant for each independent variable. The area under the curve for sperm concentration and FSH was found to be statistically better in combination than concentration alone (0.71 vs 0.66, p=0.033). The best balance of sensitivity and specificity (i.e. for a diagnostic test) for concentration and FSH were found at cut-points of 8.345 mIU/mL, respectively. To optimize performance as a screening test and maintain sensitivity &amp;gt;90%, the cut points for concentration and FSH were 2.74 mIU/mL, respectively. There was no difference in sensitivity for concentration between 2-7 mil/mL and no abnormalities were found if concentration exceeded 7 mil/mL; thus a cutoff of 7 mil/mL would capture 100% of all chromosome abnormalities. Conclusions Based on receiver operating characteristics, lowering the threshold for chromosomal analysis to a sperm concentration of &amp;lt;1 mil/mL maintains acceptable performance as a screening test for genetic abnormality, with resulting cost savings expected. Incorporating FSH into this decision appears to improve both sensitivity and specificity in prediction of chromosomal abnormalities. Our military single-institution ten-year cohort, exemplifies the potential improvement in patient selection and results that can be achieved if the sperm concentration is lowered from the current cutoff Disclosure No

Predictors of successful salvage microdissection testicular sperm extraction (mTESE) after failed initial TESE in patients with non-obstructive azoospermia: A systematic review and meta-analysis.

There has been no systematic review and meta-analysis to analyze and summarize the predictive factors of successful sperm extraction in salvage microdissection testicular sperm extraction. We aimed to investigate the factors predicting the result of salvage microdissection testicular sperm extraction in patients with non-obstructive azoospermia who failed the initial microdissection testicular sperm extraction or conventional testicular sperm extraction. We conducted a systematic literature search in PubMed, Web of Science, EMBASE, and the Cochrane Library for literature that described the characteristics of patients with non-obstructive azoospermia who underwent salvage microdissection testicular sperm extraction after failing the initial microdissection testicular sperm extraction or conventional testicular sperm extraction published prior to June 2022. This meta-analysis included four retrospective studies with 332 patients with non-obstructive azoospermia who underwent a failed initial microdissection testicular sperm extraction and three retrospective studies with 177 non-obstructive azoospermia patients who underwent a failed conventional testicular sperm extraction. The results were as follows: among non-obstructive azoospermia patients whose first surgery was microdissection testicular sperm extraction, younger patients (standard mean difference: -0.28, 95% confidence interval [CI]: -0.55 to -0.01) and those with smaller bilateral testicular volume (standard mean difference: -0.55, 95% CI: -0.95 to -0.15), lower levels of follicle-stimulating hormone (standard mean difference: -0.86, 95% CI: -1.18 to -0.54) and luteinizing hormone (standard mean difference: -0.68, 95% CI: -1.16 to -0.19), and whose testicular histological type was hypospermatogenesis (odds ratio: 3.52, 95% CI: 1.30-9.53) were more likely to retrieve spermatozoa successfully, while patients with Sertoli-cell-only syndrome (odds ratio: 0.41, 95% CI: 0.24-0.73) were more likely to fail again in salvage microdissection testicular sperm extraction. Additionally, in patients who underwent salvage microdissection testicular sperm extraction after a failed initial conventional testicular sperm extraction, those with testicular histological type of hypospermatogenesis (odds ratio: 30.35, 95% CI: 8.27-111.34) were more likely to be successful, while those with maturation arrest (odds ratio: 0.39, 95% CI: 0.18-0.83) rarely benefited. We found that age, testicular volume, follicle-stimulating hormone, luteinizing hormone, hypospermatogenesis, Sertoli-cell-only syndrome, and maturation arrest were valuable predictors of salvage microdissection testicular sperm extraction, which will assist andrologists in clinical decision-making and minimize unnecessary injury to patients.

(145) Is LIM-15 an Important Marker in Patients Who Performed Second Micro–testicular Sperm Extraction after Failed First Micro–testicular Sperm Extraction in Men with Nonobstructive Azoospermia?

Abstract Introduction LIM15, also known as DNA meiotic recombinase 1 (DMC1), is a meiosis-specific recombinase. LIM15 is specifically expressed as a meiotic recombination protein in spermatogenesis. Objective Does the LIM15 gene expression value measured in the ejaculate predict the success of the second micro–testicular sperm extraction (mTESE) after the failed first mTESE? Methods Data from 111 men who failed the first mTESE and underwent the second mTESE between August 2018 and March 2022 were prospectively collected. LIM15 gene expression level was determined by real-time PCR in the materials from the ejaculate. Results Sperm was recovered successfully in 23 out of 111 (20.7%) men with second mTESE. The mean age of men for whom sperm could not be retrieved and men with successful sperm recovery were 34.6 ± 3.5 and 33.7 ± 5.2, respectively. The mean patient age, serum total testosterone and FSH levels, and testicular volume did not show any statistical difference between men with successful sperm recovery and men with failed sperm recovery (Table 1). According to the testicular histology, the sertoli cell-only, maturation arrest, and hypospermatogenesis in men with successful sperm recovery were 26.1%, 65.2%, and 8.7%, and in men for whom sperm could not be retrieved were 0%, 27.3%, and 72.7%. Comparing the groups in terms of testicular histology demonstrated meaningful correlation between sperm retrieval rate (SRR) and testis histology (p&amp;lt;0.001). The LIM15 gene expression level measured in ejaculate significantly correlated with the SRR in mTESE (p = 0.039). As shown in Figure 1, the maximal sensitivity (50%) and specificity (87%) were obtained for the LIM15 gene with a cutoff point of 4.61 and AUC of 0.65. Conclusions According to our results, LIM15 measured in the ejaculate before the second mTESE can provide information on sperm retrieval and reduce the potential complications and unnecessary costs associated with assisted reproductive techniques. Disclosure No

(524) Adolescent Varicocelectomy: Long-term Follow-up of Clinical Outcomes and Patient Satisfaction

Abstract Introduction Worldwide 10–15% of male adolescents have a varicocele, which is the most common surgically correctable cause of male subfertility. In young adult men, varicocele can be associated with smaller testis volume and abnormal semen parameters. However, there is still controversy in whether surgical repair has better long-term outcomes than just active surveillance. Objective To evaluate satisfaction, symptoms improvement, hypogonadism, and paternity in patients who underwent varicocelectomy when they were adolescents. Methods We performed a retrospective chart review of adolescents undergoing varicocelectomy from 2011-2021. Inclusion criteria were having primary varicocele, having 12-21 years at time of surgery, having had the varicocelectomy &amp;gt;1-year ago, ≥1 postoperative visits, and having answered the questionnaire. Exclusion criteria were having a diagnosis of secondary varicocele (renal malignancy or venous abnormality) and incomplete records. Data about clinical, surgical, and postoperative outcomes were gathered. Patients were contacted by phone and were asked about orchialgia, paternity, varicocele recurrence, diagnosis and treatment for hypogonadism, and satisfaction after surgery. Descriptive analysis and paired t-test were used, statistical significance was assessed at p&amp;lt;0.05. Results From a total of 50 patients meeting the inclusion criteria, we were able to contact 23 patients. The mean age at varicocele diagnosis was 17.43±2.52 years. Varicocele was grade 3 in 16 (69.6%) patients and left-sided in 17 (73.9%) patients. Eight (34.8%) patients had a preoperative testicular ultrasound, that in 2(25%) patients showed testicular volume discrepancy &amp;gt;20%. At the physical exam, 6(26.1%) patients were found to have asymmetric testicles. Overall, 2(8.7%) patients had a previous varicocele repair and 7(30.4%) had a preoperative abnormal semen analysis. Mean age at surgery was 18.78±2.02 years. Microscopic varicocelectomy was performed in 22 (95.7%) patients, mainly in the left-side (78.3%). Pre- and postoperative FSH and T levels were done in 10 patients, without any significant difference after surgery (p=0.074 and p=0.069, respectively). Pre- and postoperative semen analysis were performed in 12 patients and showed a significant improvement in motility after varicocelectomy (p=0.022). Mean follow-up after surgery was 1.42±1.40 months. The questionnaire was performed in a mean of 3.43±2.55 months after varicocelectomy and the mean age of the responders was 22.3±3.11 years. Orchialgia persisted in 7 patients (30.4%), predominately in the left side (71.4%). None of the patients had been diagnosed with varicocele recurrence nor had required a surgical reintervention. Twenty-one (86.9%) patients were not actively trying to achieve a natural conception nor had been involved in a pregnancy after varicocelectomy. Two patients (8.7%), with a preoperative abnormal semen analysis, were able to conceive. None of the patients referred to have hypogonadism nor being under testosterone therapy. The majority of patients (86.9%) referred to be satisfied with the postoperative results. Conclusions Our results suggested that long-term outcomes of varicocelectomy in adolescents would show high patient satisfaction, low rate of recurrence, no association with hypogonadism, and a significant persistence of testicular pain. Although male fertility after varicocelectomy demonstrated promising results, a longer follow-up assessment should be done when patients are actively trying to conceive, to have a more representative sample size. Disclosure Any of the authors act as a consultant, employee or shareholder of an industry for: Acerus, Boston Scientific, Endo Pharmaceuticals and Coloplast

Gonadal development and function after immature testicular tissue banking as part of high-risk gonadotoxic treatment.

Experimental fertility preservation programs have been started to safeguard the future fertility of prepubertal and pubertal males requiring high-risk gonadotoxic treatment protocols. However, long-term follow-up studies evaluating the effects on their gonadal development and function related to the testicular biopsy procedure are rather limited. This two-center follow-up study (between 2002 and 2020) evaluated the gonadal development and function of a cohort of 59 prepubertal and pubertal males who have been offered immature testicular tissue banking (TTB) prior to conventional high-risk chemo- and/or radiotherapy (HR-C/R) or conditioning therapy before hematopoietic stem cell transplantation (CT-HSCT). The aim is to investigate the long-term impact of the testicular biopsy procedure and the high-risk gonadotoxic treatment. Testicular growth and the reproductive hormones luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone (T), and inhibin B (INHB) were analyzed after treatment completion, and compared between males accepting TTB and those refusing TTB (control) as well as between HR-C/R and CT-HSCT treatment protocols. Of the 59 prepubertal and pubertal males included, 25 were treated by HR-C/R and 34 required CT-HSCT. TTB was accepted for 39 males and refused for 20 males. Most patients were prepubertal at diagnosis (85%), at TTB (79%), and at treatment completion (76%), and pubertal or postpubertal at their last follow-up visit (66%). After 5.0 (1.0-13.0) years post treatment, most patients show normal testicular volumes (83%) and normal LH (89%), FSH (87%), T (87%), and INHB (79%) serum levels. The testicular biopsy procedure did not have an effect on testicular growth, LH, FSH, T, and INHB. Significantly more small postpubertal testicular volumes (p=.0278) and low INHB serum levels (p=.0130) were recorded after CT-HSCT, especially after myeloablative conditioning. The clinical follow-up data demonstrate no effect related to the biopsy procedure, but a substantial risk for impaired gonadal development after high-risk gonadotoxic treatment, in particular myeloablative CT-HSCT. Longer follow-up studies with a larger study population are needed to confirm these preliminary findings.

Delayed Puberty.

Testicular volume ≥4ml and appearance of breast budding are the first signs of puberty. Delayed puberty is diagnosed in the absence of thelarche by 13 y or menarche by 15 y in girls and absence of testicular enlargement by 14 y in boys. Delayed puberty can be due to hypogonadotrophic hypogonadism, hypergonadotrophic hypogonadism or eugonadotrophic eugonadism characterised by low, elevated and normal gonadotrophin levels, respectively. Constitutional Delay of Growth and Puberty (CDGP) and systemic illness should be considered before pathological causes. Assessment of sexual maturity by Tanner's staging and anthropometric assessment on growth chart is pivotal. Lack of menarche in girls with thelarche suggests structural abnormalities of reproductive tract or disorders of sexual development. Measurement of bone age helps to interpret hormone measurements and decide on timing of pubertal induction. Ultrasound assessment of abdomen gives valuable clues to pubertal onset (in girls) and possible underlying etiology. Karyotyping is mandatory in all girls with delayed puberty and short stature, and delayed menarche and boys with hypergonadotrophic hypogonadism. Gonadotrophin releasing hormone analogue stimulation test may help distinguish hypogonadotrophic hypogonadism from CDGP. Pubertal induction is done with intramuscular testosterone and oral estradiol in boys and girls, respectively. Hormone replacement is begun at low doses and slowly escalated over 2 y to mimic a physiological puberty process. Short course of testosterone for 3 to 6 mo is helpful in adolescent boys with CDGP and psychological distress. Attainment of adult sexual maturity by 18 y is mandatory to rule out disorders of hypothalamic pituitary gonadal axis.

Food monotony compromises photoperiod induced responses in migratory redheaded bunting

Food is one of the key environmental factors affecting an organism’s behavior and physiology on daily and seasonal basis. Over the years, studies regarding human food habits and its subsequent impact on their behavior and physiology have been explored a lot but studies on same line are missing with reference to different animal taxa. Thus, in the present study, we investigated the impact of varied food availability over a monotonous food schedule on birds. We were interested in knowing how this treatment affects the birds’ behavior and physiology? The experiment was performed on migratory passerine finch; redheaded bunting ( Emberiza bruniceps), wherein two groups of bird ( N = 10 each) were taken. Group I (Kakuni group; KG) birds were fed only with seeds of Setaria italica (Kakuni) whereas group II (Mixed Diet group; MG) was provided with seeds of kakuni along with sesame seeds ( Sesamum indicum) and egg white, all three separately in different bowls. Results reveal that behaviorally, MG birds were more active with higher gross food intake when compared with KG. MG birds also had significant gain in body mass and testicular volume during the course of experiment, although, the accumulation of fat remained unchanged between the two groups. Thus, the study clearly demonstrates the motivation to eat and stay active besides being reproductively proficient in presence of varied food availability.

Zbtb40 Deficiency Leads to Morphological and Phenotypic Abnormalities of Spermatocytes and Spermatozoa and Causes Male Infertility.

Studies on the gene regulation of spermatogenesis are of unusual significance for maintaining male reproduction and treating male infertility. Here, we have demonstrated, for the first time, that a loss of ZBTB40 function leads to abnormalities in the morphological and phenotypic characteristics of mouse spermatocytes and spermatids as well as male infertility. We revealed that Zbtb40 was expressed in spermatocytes of mouse testes, and it was co-localized with γH2AX in mouse secondary spermatocytes. Interestingly, spermatocytes of Zbtb40 knockout mice had longer telomeres, compromised double-strand break (DSB) repair in the sex chromosome, and a higher apoptosis ratio compared to wild-type (WT) mice. The testis weight, testicular volume, and cauda epididymis body weight of the Zbtb40+/- male mice were significantly lower than in WT mice. Mating tests indicated that Zbtb40+/- male mice were able to mate normally, but they failed to produce any pups. Notably, sperm of Zbtb40+/- mice showed flagellum deformities and abnormal acrosome biogenesis. Furthermore, a ZBTB40 mutation was associated with non-obstructive azoospermia. Our results implicate that ZBTB40 deficiency leads to morphological and phenotypic abnormalities of spermatocytes and spermatids and causes male infertility. This study thus offers a new genetic mechanism regulating mammalian spermatogenesis and provides a novel target for gene therapy in male infertility.

Clinicopathological characteristics of Klinefelter syndrome: a testicular biopsy analysis of 87 cases

Objective: To investigate the clinicopathological characteristics of testicular biopsies from Klinefelter syndrome (KS) patients. Methods: The testicular biopsy specimens of 87 patients with KS (a total of 107 biopsy specimens) were collected from the Department of Pathology, Peking University Third Hospital, Beijing, China from January 2017 to July 2022. All patients were diagnosed as KS by peripheral blood karyotyping analysis. The testicular histopathologic features, testicular volume and hormone levels were evaluated retrospectively. The histopathologic analysis was used to assess the quantity and morphology of Leydig cells, the spermatogenic state of seminiferous tubules, the thickening of the basement membrane of seminiferous tubules and the changes of stroma. Results: Leydig cell proliferative nodules were seen in 95.3% (102/107) of KS testicular biopsy tissues. The eosinophilic inclusion bodies and lipofuscin in Leydig cells were found in 52.3% (56/107) and 57.9% (62/107) of specimens, respectively. The Sertoli cell only seminiferous tubules and the hyalinized tubules were found in 66.4% (71/107) and 76.6% (82/107) of the examined tissues, respectively. The tubules with complete spermatogenic arrest were found in 15.9% (17/107) of specimens, and 5.6% (6/107) of the specimens showed low spermatogenesis or incomplete spermatogenic arrest. In 85.0% (91/107) of the specimens, increased thick-walled small vessels with hyaline degeneration were identified. Conclusions: The most common features of KS testicular specimens are Leydig cell proliferative nodules, hyaline degeneration of seminiferous tubules and proliferation of thick-walled blood vessels. Testicular biopsy specimens of KS are rare. The pathologists can make a tentative diagnosis of KS based on the histological findings, combined with the ultrasound and laboratory results, which is helpful for further diagnosis and treatment of KS.