Undescended Testes Group Research Articles

AZIN2 is associated with apoptosis of germ cells in undescended testis

Undescended testis (UDT), known as cryptorchidism (CRY), is a common congenital disorder in which one or both testicles do not descend normally into the scrotum. A unilateral UDT model was established by inducing UDT in mice through surgery. The results showed that the testis in the UDT model group was abnormal; the lumen of the seminiferous tubule was atrophic; apoptosis, necrosis and shedding were observed in many of the germ cells; the level of sex hormones was abnormal; and mature sperm was reduced. Subsequently, transcriptome sequencing was conducted on the testicular tissue of UDT model mice. Through analysis and verification of differential genes, AZIN2 was identified as playing a key role in the decline in male fertility caused by cryptorchidism. AZIN2 expression and spermine content was down-regulated in the testis of the UDT group. We then used a combination of hypoxanthine and xanthine to create a GC-1 cell damage model. In this model, AZIN2 expression and spermine content was down-regulated. When si-Azin2 transfected GC-1 cells, cell viability and proliferation were decreased. However, in the GC-1 cell damage model transfected with Azin2 over-expressed plasmid, AZIN2 expression and spermine content was up-regulated, reversing the cell damage caused by hypoxanthine and xanthine, and restoring the proliferation ability of GC-1 cells. These results indicate that in UDT, down-regulated AZIN2 expression is a factor in testicular damage. This discussion of the connection between AZIN2 and germ cells has important clinical significance as it provides an important reference for the diagnosis and treatment of cryptorchidism.

Diagnostic role of gray-scale and shear-wave elastography in pediatric patients with undescended testes: a prospective controlled study.

Ultrasound elastography is a simple non-invasive method for measuring tissue elasticity in relation to tissue fibrosis. The aim of this study was to compare echogenicity, volume and shear wave velocities of undescended vs normally descended testes. Sixty-six boys with undescended testes were included in this study. The median age range was 35.5 (10-118) months old. The cases included in this prospective study consisted of 66 patients with non-operated undescended testes, with 51 of them being affected unilaterally and 15 affected bilaterally, as diagnosed by physical examination. The control group consisted of 31 healthy boys without any particular health problems. This prospective study was performed by gray-scale ultrasonography and shear wave elastography in boys with undescended testes and healthy testes. The testicular volumes were established by ultrasound measurement, the echogenicity and shear wave elastography values were measured in boys with unilateral and bilateral undescended testes, and the results were compared with healthy boys' testes and their contralateral testes. The stiffness values were recorded for speed (m/s) and elasticity (kPa), and the stiffness values of undescended testes were compared with the healthy control group. Echogenicity values were lower in the bilateral undescended testes group than in the healthy group, and the healthy group's echogenicity was normal (p <0.001). The ROC curve was used to identify a cut-off shear wave elastography value for predicting decreased testicular echogenicity by using average shear wave elastography values. The area under the curve for the undescended testes was 0.78 (95% CI: 0.70-0.85, sensitivity 83.7%, specificity 68.7%, p <0.001), with an average shear wave elastography value of 2.32 (m/s) for above the cut-off point indicates. This was found to be significantly associated with reduced echogenicity on gray-scale ultrasonography, suggesting that it may be correlated with fibrosis developing in patients with undescended testes. The study provides interesting findings in that it proposes an alternative non-invasive method for the assessment of testicular tissue in undescended testes. We used shear wave elastography to compare the stiffness of normal testes in both heathy patients and in the contralateral healthy testes of boys with undescended testes, with the values obtained for the undescended testes reflecting the level of fibrosis of the parenchyma. Another outcome of this study was observed in patients with unilateral undescended testes, where the normally descended testes showed increased shear wave elastography values, which could be an early indication of parenchymal change.

Oxidative DNA Damage and NOX4 Levels in Children with Undescended Testes.

Undescended testis (UDT) is a common urological disorder. Patients with UDT have a risk of malignancy and infertility. The development of these conditions may be due to oxidative stress mediated by reactive oxygen species. The aim of this study was to investigate the relationship between these parameters by detecting oxidative DNA damage (8-hydroxy 2 deoxyguanosine/106 deoxyguanosine), ischemia-modified albumin (IMA), malondialdehyde (MDA), and nicotinamide adenine dinucleotide phosphate oxidase 4 (NOX4) levels in children with UDT and healthy control group. The blood samples were obtained from 30 patients with UDT and 40 healthy male subjects. The levels of oxidative DNA damage were detected by high-pressure liquid chromatography method. We used commercially available kits that use enzyme-linked immunosorbent assay method to measure IMA, MDA, and NOX4 levels. The levels of MDA, IMA, NOX4, and oxidative DNA damage in children with UDT were statistically significantly higher than control group. In addition, we found that the levels of NOX4, IMA, and oxidative DNA damage after 12 months of age was significantly higher than before 12 months of age. We identified increased lipid peroxidation, oxidative DNA damage, IMA, and NOX4 levels in children with UDT. Delay in the treatment of UDT may cause oxidative damage. That is why, according to us the antioxidant treatment may be beneficial in children with UDT.

A history of undescended testes in young men with Klinefelter syndrome does not reduce the chances for successful microsurgical testicular sperm extraction

Klinefelter syndrome (KS) and undescended testes (UDT) are known etiologies for non-obstructive azoospermia (NOA), and coexistence of both etiologies is not uncommon. Patients with both KS and a history of UDT are therefore considered to have extremely reduced chances for paternity. We aimed to analyze the impact of previous surgically corrected unilateral or bilateral UDT on sperm retrieval rates (SRRs) by microsurgical testicular sperm extraction (mTESE) in azoospermic men with KS. Age, testicular volumes, and hypothalamo-pituitary-gonadal axis function were investigated in relation to SRRs in 29 non-mosaic KS patients (47,XXY) with a history of UDT (group 1) who underwent mTESE between 2008 and 2016 in our center and compared to the data of age- and serum testosterone-matched non-mosaic KS controls with eutopic testes at birth (group 2), and to those of 51 men with NOA and a normal male karyotype (46,XY), but previous UDT (group 3). SRRs in KS patients with surgically corrected UDT during childhood were comparable to SRRs of KS patients with eutopic testes at birth: 31% (35% in unilateral and 22% in bilateral UDT) vs. 38% (p=0.581). SRRs and Leydig cell function in group 1 were negatively correlated with age. Significantly higher SRRs (66%) were found in euploid azoospermic men with surgically corrected UDT (p<0.001). A history of UDT does not preclude chances for future fatherhood in young azoospermic males with KS. In one of three men with previous unilateral UDT and in one of 4-5 in those with previous bilateral UDT, spermatozoa can be harvested by mTESE during late adolescence or young adulthood for immediate or future use in assisted reproduction.

Detection of insulin-like growth factor receptor-1 in the human cremaster muscle and its role in the etiology of the undescended testis

Previous studies of undescended testis (UT) has focused on insulin-like hormone 3 (INSL3), the genitofemoral nerve, and androgens in the testicular descent. Leydig cells, which are under the control of insulin-like growth factor-1 (IGF1), produce both androgens and INSL3. We aimed to investigate whether insulin-like growth factor receptor-1(IGFR1) exists in the cremaster muscle (CM) complex and is associated with normally descended testis as well as UT cases in humans. We studied 30CM from 15 patients who comprised the UT group (UTG), and 15 patients with unilateral testicular torsion (Control group; CG). Muscles, nerves, and vessels within the CM specimen were examined to determine the presence of IGFR1. The mean staining score (MSS) of IGFR1 in CM and its nerves were higher in the CG than in the UTG. These results were statistically significant (p=0.01 and p=0.02). Although the MSS of IGF1R was higher in the vessels of CM in the CG than the UTG, this was not statistically significant (p=0.48). IGFR1 with heterotetrameric receptor via IGF1, IGF2, insulin, and probably androgen, contribute to the remodeling and development of CM as well as the testis descent. In the current study, the presence of the IGFR1 in the CM was shown. Additionally, the IGFR1 density of the CM was lower in the UT cases than in the CG cases. Further evaluation of IGFR1 and other etiological factors can elucidate how they interact.

Impact of Preoperative Ultrasonographic Evaluation for Detection of a Viable Testis in Patients With a Unilateral Nonpalpable Testis.

To investigate the impact of preoperative ultrasonography (US) for detecting a viable testis in patients with a unilateral nonpalpable testis. Patients with a unilateral nonpalpable testis or unilateral palpable undescended testis who underwent preoperative US were enrolled. Patients were divided into 3 groups as follows: nonpalpable testis/no testis (n = 27), which included patients who had a unilateral nonpalpable testis with no viable testis detected at surgery; nonpalpable testis/viable testis (n = 10), which included patients who had a unilateral nonpalpable testis with a viable testis identified at surgery; and palpable undescended testis (n = 63), which included patients who had a unilateral palpable undescended testis. Preoperative US findings were compared among each group. The testicular volume on the contralateral descended side in the nonpalpable testis/no testis group was significantly greater than that in the nonpalpable testis/viable testis and palpable undescended testis groups. When a testicular volume of 0.54 mL was used as the cutoff value, the sensitivity, specificity, positive predictive value, and negative predictive value for the presence of the affected testis were 75.3%, 100%, 100%, and 60.0%, respectively. The testis on the affected side was detected in none of the nonpalpable testis/no testis group, 7 of the nonpalpable testis/viable testis group, and all of the palpable undescended testis group. When a visible testis on the affected side and a testicular volume of 0.54 mL or less were defined as positive, all patients in the nonpalpable testis/viable testis and palpable undescended testis groups had positive findings versus none in the nonpalpable testis/no testis group. Preoperative US provides valuable information for predicting the presence of a viable testis in patients with a unilateral nonpalpable testis by estimating both the unaffected testis and the affected side.

Testicular volume and elasticity changes in young children with undescended testes.

To evaluate the differences and changes of testicular volume and elasticity in normal (NL) testes and undescendedtestes (UDTs) of children using shear wave elastography (SWE). Testicular ultrasound images fromchildren younger than 60 months old were retrospectively reviewed. Testicular volumes and elasticities were compared betweenthe UDT group and NL group. In patients with unilateral UDT (uUDT), we also compared the values between uUDTand contralateral grossly normal (CGN) testis groups. There were 25 UDTs including 4 bilateral in the UDT groupand 54 normal testes in the NL group. While testicular volume was significantly smaller in UDT (vs. NL) and uUDT (vs. CGN)groups, the elasticity was not different. Testicular volume was positively correlated with age in both NL (r=0.474) and CGN(r=0.729) groups (p<0.001), while there was no correlation in the UDT group. Testicular stiffness showed negative correlationwith age in the NL group (r=-0.390, p=0.004) and positive correlation in the UDT group (r=0.426, p=0.034). Instead of increasing volume and decreasing stiffness of normal testes during development of the early 60 months, UDTsexhibited smaller volume and increasing stiffness. The CGN testes of uUDT patients showed increasing volume without stiffnesschange.

Malondialdehyde and CA II autoantibody levels are elevated in children with undescended testes

In the pathogenesis of sub-fertility/infertility and testicular cancer related to undescended testes, oxidative stress, inflammation and autoimmunity are important factors. Therefore, the present study was designed to determine serum oxidative stress markers and carbonic anhydrase (CA) II autoantibodies in boys with undescended testes (UDT), and to investigate the relationship between these parameters. Serum CA II autoantibody titers, malondialdehyde (MDA), ischemia modified albumin (IMA), protein carbonyl content and soluble CD40 ligand (sCD40L) levels were measured in 59 boys with UDT and 30 healthy subjects. MDA levels were significantly higher in the UDT group compared with the control group (p = 0.003). There was no significant difference between serum IMA, sCD40L or protein carbonyl levels. CA II autoantibody titers in the UDT group were significantly higher compared with those of the control group (p = 0.048). A weak positive correlation was determined between anti-CA II antibody titers and MDA and IMA levels (p = 0.041, p = 0.005, respectively). MDA is the most reliable and decisive biochemical marker displaying oxidative damage in undescended testes, and an autoimmune response may be triggered by oxidative stress against CA II during the UDT process.

Measurement of hydroxyproline and nitric oxide, and comparison of sac fluid acidity in different inguinal pathologies

PurposeTo compare hydroxyproline and nitric oxide levels, and fluid acidity between sac tissues from inguinal hernia, hydrocele and undescended testis patients. Material and methodThirty patients were in the inguinal hernia group, 11 patients in the hydrocele group and 5 patients in the undescended testis with inguinal hernia group. Intrasaccular fluid acidity, and nitric oxide and hydroxyproline levels in sac tissues were measured and analyzed statistically between groups. ResultsIntrasaccular fluid could be obtained from 24 inguinal hernia and 6 hydrocele patients. Acidity level (pH) was 7.5 ± 0.74 in the hernia group and 7.86 ± 0.63 in the hydrocele group (p < 0.05). All patients in the three groups were evaluated for nitric oxide and hydroxyproline. Results for hydroxyproline (mg/g dry tissue)/nitric oxide (nmol/g wet tissue) were 3.53 ± 0.54/2.02 ± 0.39 for inguinal hernia, 2.89 ± 0.9/1.92 ± 1.05 for hydrocele and 3.02 ± 1.23/2.86 ± 1.09 for undescended testis group (p > 0.05). ConclusionHydroxyproline and nitric oxide pathways do not seem to be responsible for the different characteristics of inguinal diseases. However, differences in the acidity of sac fluid may be evidence of the different mechanisms of hernia and hydrocele formation.

Oxidative, inflammatory and immunologic status in children with undescended testes

In order to better understand the pathogenesis of risk of future sub-/infertility in children with undescended testes (UDT), we designed this prospective study to examine the oxidative stress, inflammatory response and autoimmunity in children with UDT. We examined the concentrations of malondialdehyde (MDA), interleukin-6 (IL-6) and antisperm antibodies (ASA) in children with UDT and healthy controls. The UDT group consisted of 88 boys (aged 1-14 years, unilateral in 67 and bilateral in 21 cases), and 44 boys with normal descended testes served as a control group. Clinical evaluation revealed no testicular or other system abnormalities. MDA was used as lipid peroxidation index. IL-6 levels were measured using a commercial enzyme-linked immunosorbent assay kit. ASA was determined with an anti-human spermatozoa immunoglobulin G test. Mean age values ± SD were 4.6 ± 3.2 in the UDT group and 4.7 ± 3.4 in the control group (P= 0.872). MDA and IL-6 results for the UDT and control groups were significantly different (P= 0.003 and P= 0.019, respectively), but those for ASA were not (P= 0.473). The mean MDA and IL-6 values were significantly higher in bilateral cases than the respective values in the unilateral cases (MDA: 4.03 ± 3.68 vs 3.49 ± 5.22, P= 0.015; IL-6: 7.70 ± 6.86 vs 3.48 ± 6.50, P= 0.001) (P= 0.015). The results indicate that children with UDT are exposed to high levels of oxidative stress and inflammatory reaction. This could negatively affect the future fertility in these children.

Different surgical findings in congenital and acquired undescended testes

To compare surgical findings in congenital and acquired undescended testes (UDT). A review of 139 boys with 158 congenital UDT and 69 boys with 84 acquired UDT was performed. The most caudal testicular position preoperatively, testis position at surgery, patency of the processus vaginalis and epididymal anomalies were prospectively recorded. In the congenital group, orchiopexy had been performed at median age (range) 4.9 (1.5-14.6) years, while the median age (range) in the acquired group was 11.9 (3.8-23.3) years. Preoperatively, only congenital UDT were found not palpable or emergent inguinal, while only acquired UDT could be manipulated in an unstable scrotal position. In comparison with congenital UDT, acquired UDT were significantly more often located at the scrotal entrance, 27/158 vs 32/84 respectively (P < 0.001). At surgery anorchia, vanished testis or testes lying intra-abdominally were only registered in the congenital UDT group. Also 37/158 congenital UDT were located in the superficial inguinal pouch vs 52/84 of the acquired UDT (P = 0.04). In congenital UDT the processus vaginalis was wide open in 74/158, while in acquired UDT the processus vaginalis was closed in 46/84 (P < 0.001) and small open in 26/84 (P = 0.04). Epididymal anomalies were more often seen in the congenital UDT group (37%) than in the acquired group (11%). The most caudal position of congenital UDT after manipulation before surgery was at the scrotal entrance. These testes were frequently associated with epididymal anomalies and wide open processus vaginalis. This was in contrast to acquired UDT, which can often be pushed down well below the scrotal entrance and are more likely to be situated in the superficial inguinal pouch, with a normal epididymis and closed processus vaginalis.

Attenuation of Spermatogonial Stem Cell Activity in Cryptorchid Testes

To elucidate the mechanism of infertility caused by cryptorchidism we focused on early stage spermatogenesis and spermatogonial stem cell activity in undifferentiated spermatogonia in cryptorchid testes. Histological findings and expression patterns of the stem cell marker undifferentiated embryonic cell transcription factor 1 were examined in a unilateral cryptorchid rat model. We removed unilateral descended testis and contralateral descended testis from cryptorchid and normal rats (control), respectively, 18 days postcoitum to 144 days postpartum. In descended testes gonocyte differentiation into early A spermatogonia occurred at 9 days postpartum. However, this transformation was altered in undescended testes. Furthermore, the undifferentiated embryonic cell transcription factor 1 negative early A spermatogonia-to-positive early A spermatogonia ratio was significantly higher in the undescended testis group (mean ± SD 0.69 ± 0.04) than in the control (0.46 ± 0.10, p = 0.037) and descended testis (0.44 ± 0.05, p = 0.022) groups, indicating decreased early A spermatogonia with spermatogonial stem cell activity in cryptorchid testes. In cryptorchid testes the differentiation from gonocytes into early A spermatogonia and the stem cell activity of early A spermatogonia were altered during the early stage of spermatogenesis, suggesting that the loss of spermatogonial stem cell activity in cryptorchid rats resulted in altered spermatogenesis, thus interfering with fertility.

Perioperative surgical findings in congenital and acquired undescended testis

Purpose Perioperative surgical findings in congenital and acquired undescended testis (UDT) were prospectively assessed. Methods We included all boys with congenital or acquired UDT who underwent orchidopexy at our hospital between January 2006 and August 2009. Perioperatively, we scored the position and volume of the testis, the insertion of the gubernaculum, the patency of the processus vaginalis, and the obtained position. Results We included 69 boys (aged 0.9-14.6 years) with 76 congenital UDT and 28 boys (aged 2.2-18.5 years) with 30 acquired UDT. In the congenital group, the testis was in intracanalicular position in 55 cases (72%), whereas in the acquired UDT group, this was in 11 cases (37%; P < .001). The insertion of the gubernaculum was at the bottom of the scrotum in 13 cases (17%) of the congenital UDT group and in 12 cases (40%) of the acquired UDT group ( P < .05). The processus vaginalis was open in 63 cases (83%) of the congenital and in 9 cases (30%) of the acquired UDT group ( P < .001). Conclusion Compared to congenital UDT, acquired UDT are more likely to be situated in the superficial inguinal pouch, to have a normal insertion of the gubernaculums, and to have a closed processus vaginalis.

Prospective Evaluation of Human Chorionic Gonadotropin in the Differentiation of Undescended Testes From Retractile Testes

We prospectively evaluated the efficacy of human chorionic gonadotropin (HCG) in the treatment of undescended testis and sought to determine whether HCG assists in the differentiation of undescended testis from retractile testis. Patients with undescended testes were offered HCG. Testis position, laterality and the presence or absence of a hypoplastic scrotum were noted. The same physician (G. W. K.) recorded physical findings prospectively and stated clinical impression of descent. A total of 67 patients with 90 undescended or retractile testes were treated and evaluated with HCG. Of the 64 undescended testes 13 (20%) descended with HCG therapy, with none requiring subsequent surgery. Of the 26 retractile testes 15 (58%) descended with HCG (p <0.001). Based on physical examination, 100% of retractile testes descended if the testis was in the high scrotal position but only 40% descended if the testis was in the superficial pouch or inguinal area. In the undescended testes group no ectopic or nonpalpable testis descended with HCG. Evaluation of HCG with age demonstrated minimal response (15%) to HCG at less than 24 months, and a peak response between ages 2 and 6 years (75%) with response decreasing thereafter. HCG may have a limited role in the evaluation of undescended testis in patients younger than 2 years. HCG can serve as an adjunct in the clinical diagnosis of retractile testis in older children.

Is there a correlation between organochlorine compounds and undescended testes?

Some pesticides and synthetic chemicals are known to act as hormonal modulators, often possessing oestrogenic activity (xenooestrogens). They are persistent and accumulate in fatty tissue. Aim of our study is to address the question, whether a selection of such compounds is to be found in the fatty tissue of children undergoing surgical procedures and whether there are differences in values obtained from patients with or without undescended testes. Fat samples of 48 patients, 18 of whom had undescended testes, were examined by high-resolution gas chromatography and mass spectrometry for DDT and metabolites, polychlorinated biphenyls (PCB), toxaphenes, hexachlorocyclohexane (HCH), chlorinated cyclodienes and chlorinated benzenes. We were able to find accumulation of all substances in every patient. Statistical analysis revealed a highly significant difference between patients from the control group and those from the undescended testes group for two compounds, namely heptachloroepoxide (HCE) and hexachlorobenzene (HCB), increased values being found in the patients with undescended testes. Since the aetiology of this entity is unknown in most of the cases, prenatal exposure to exogenous oestrogens is an attractive and plausible hypothesis. In order to confirm this, some questions will have to be answered in further studies: effect of exposure to xenooestrogens during a specific period of development, probable role of other substances with proven or suspected hormonal activity, potential synergism of such compounds and differences in individual susceptibility.

THE INCIDENCE OF INTERSEXUALITY IN CHILDREN WITH CRYPTORCHIDISM AND HYPOSPADIAS: STRATIFICATION BASED ON GONADAL PALPABILITY AND MEATAL POSITION

Purpose The combined findings of cryptorchidism and hypospadias often indicate the existence of an intersex state. Testicular maldescent and incomplete tubularization of the urethral plate occur in a spectrum with the severity of the 2 processes likely dependent on the degree of pathophysiology in the androgenic hormonal axis. The incidence of intersexuality in children with undescended testes, hypospadias and otherwise nonambiguous male genitalia has been reported to be 27%. Although the likelihood of genotypic or gonadal ambiguity has previously been associated with meatal position in this population, to our knowledge our study is the first to evaluate the incidence of intersexuality relative to whether the undescended testis is palpable or nonpalpable. Materials and Methods The database at our hospital was searched for all cases of undescended testes (2,105) and hypospadias (1,057) between 1982 and 1996. Radiographic, histological and karyotypic data were compiled for all patients presenting with both diagnoses. Gonadal palpability (palpable versus nonpalpable) and meatal position (anterior versus mid versus posterior) were recorded and correlated with the likelihood of identifying an intersex condition. Ten boys with a diagnosis of undescended testes subsequent to inguinal hernial repair were excluded from study. Patients with congenital adrenal hyperplasia or complete testicular feminization were also excluded from study due to the clearly female appearance of the external genitalia. Statistical significance was assessed using Fisher's exact test. Results We identified 79 patients presenting with undescended testes, hypospadias and a phallus that was believed to be a possible penis. Intersex conditions were identified with nearly equal frequency in the 44 cases of unilateral (30%) and 35 of bilateral (32%) cryptorchidism. In the unilateral undescended testes group patients with a nonpalpable testis were at least 3-fold more likely to have an intersex condition than those with a palpable undescended testis (50 versus 15%, p = 0.02). In the bilateral group patients with 1 or more nonpalpable testes were also nearly 3-fold as likely to have an intersex condition than those with bilateral palpable undescended gonads (47 versus 16%, p = 0.07). Meatal position was graded as anterior in 33% of cases, mid in 25% and posterior in 41% with the more posterior location conferring a significantly greater likelihood of an intersex condition (anterior 2 of 26, mid 1 of 20 and posterior 21 of 33). Conclusions Gonadal palpability is an important predictor of an intersex state in unilateral and bilateral cases of cryptorchidism with hypospadias. Patients with an undescended testis that cannot be palpated are significantly more likely to have an intersex condition than those in whom the undescended testis may be palpated on physical examination. The severity of hypospadias likewise has a strong positive correlation with an intersex state.