Testes Of Patients Research Articles

Expression and characterization of androgen receptor coregulators, SRC-2 and HBO1, during human testis ontogenesis and in androgen signaling deficient patients

Androgen receptor (AR) is essential for testicular physiology and spermatogenesis. SRC-2 and HBO1 are two AR coregulators yet their expression and roles in human testis are unknown. For the first time, we studied by immunohistochemistry and RT-PCR, the expression and distribution of these two coregulators during human testicular ontogenesis, in patients with altered AR signaling (Androgen insensitivity syndrome, AIS) and evaluated the functional impact of SRC-2 and HBO1 on AR signaling in a Sertoli cell context. SRC-2 was present in Sertoli cells at all developmental stages. HBO1 was barely or focally detected in the fetal testis yet its expression, in Sertoli and germ cells, drastically increased postnatally from early infancy to adulthood. In transient co-transfection studies we showed that SRC-2 induced, while HBO1 inhibited AR-mediated transactivation of reporter constructs in murine Sertoli SMAT1 cells. HBO1, but not SRC-2, expression was reduced in testes of patients with AIS compared to normal testes.

The value of finding a closed internal ring on laparoscopy in unilateral nonpalpable testis

PurposeThere is an ongoing debate about whether initial laparoscopy is as efficient as initial extra-abdominal exploration for unilateral nonpalpable testis. The aim of this study was to clarify whether the finding of a closed internal ring on laparoscopy accurately predicts extra-abdominal findings in unilateral nonpalpable testis. MethodsBoth laparoscopy and subsequent extra-abdominal exploration were prospectively performed in consecutive patients with unilateral nonpalpable testis. Laparoscopic findings were reviewed with respect to the status of the vas deferens, spermatic vessels, and the appearance of the internal ring. Except for patients with an abdominal testis, all patients underwent subsequent open exploration regardless of the laparoscopic findings. Open explorations were primarily performed by scrotal incision. During open exploration, any testicular remnants were removed and evaluated histologically. ResultsA total of 100 patients with unilateral nonpalpable testis were evaluated, of whom 15 had an abdominal testis. Of the 85 patients without an abdominal testis, an open internal ring was recognized in only 8. Of the remaining 77 with a closed internal ring, subsequent open exploration revealed testicular nubbins in all patients (100%), regardless of the laparoscopic findings of the vessels and vas. Nine nubbins were associated with intra-abdominal vanishing vessels above the closed internal ring. All nubbins were confirmed by histological findings, and the incidence of viable germ cell elements was 7.4%. ConclusionsWhen laparoscopy demonstrates no abdominal testis in patients with unilateral nonpalpable testis, the finding of a closed internal ring on laparoscopy is always associated with the presence of an extra-abdominal nubbin, regardless of the status of the vessels and the vas at the internal ring. Subsequent extra-abdominal exploration can be obviated in this circumstance if nubbin excisions are skipped. Initial diagnostic laparoscopy for unilateral nonpalpable testis provides not only the intra-abdominal findings, but also exact prediction of the extra-abdominal findings in patients with a closed internal ring.

1043 Evaluation of Pediatric Testes After Surgery by Laser Doppler and White Light Spectroscopy

AimTo evaluate testes after unilateral orchiopexy Laser Doppler and white light spectroscopy and compare microcirculation with contralaterat testis and healthy controls.MethodsA combination of the laser Doppler (determination of blood flow)...

Developmental expression and function of DKKL1/Dkkl1 in humans and mice

BackgroundExperiments were designed to identify the developmental expression and function of the Dickkopf-Like1 (DKKL1/Dkkl1) gene in humans and mice.MethodsMouse testes cDNA samples were collected at multiple postnatal times (days 4, 9, 18, 35, and 54, as well as at 6 months) and hybridized to Affymetrix mouse whole genome Genechips. To further characterize the homologous gene DKKL1 in human beings, the expression profiles between human adult testis and foetal testis were compared using Affymetrix human Genechips. The characteristics of DKKL1/Dkkl1 were analysed using various cellular and molecular biotechnologies.ResultsThe expression of Dkkl1 was not detected in mouse testes on days 4 or 9, but was present on days 18, 35, and 54, as well as at 6 months, which was confirmed by RT-PCR and Western blot results. Examination of the tissue distribution of Dkkl1 demonstrated that while Dkkl1 mRNA was abundantly expressed in testes, little to no expression of Dkkl1 was observed in the epididymis or other tissues. In an in vitro fertilization assay, a Dkkl1 antibody was found to significantly reduce fertilization. Human Genechips results showed that the hybridization signal intensity of DKKL1 was 405.56-fold higher in adult testis than in foetal testis. RT-PCR analysis of multiple human tissues indicated that DKKL1 mRNA was exclusively expressed in the testis. Western blot analysis also demonstrated that DKKL1 was mainly expressed in human testis with a molecular weight of approximately 34 kDa. Additionally, immunohistochemical staining showed that the DKKL1 protein was predominantly located in spermatocytes and round spermatids in human testes. An examination of the expression levels of DKKL1 in infertile male patients revealed that while no DKKL1 appeared in the testes of patients with Sertoli cell only syndrome (SCOS) or cryptorchidism, DKKL1 was observed with variable expression in patients with spermatogenic arrest.ConclusionsThese results, together with previous studies, suggest that DKKL1/Dkkl1 may play an important role in testicular development and spermatogenesis and may be an important factor in male infertility.

Sonographic Patterns of Testicular Torsion

The objective of this study was to determine the sonographic patterns of the testes in patients with testicular torsion. Clinical data of 46 patients with testicular torsion confirmed by surgical exploration or manual detorsion were reviewed, and sonographic patterns of testicular torsion were categorized according to the sonographic characteristics of the testes, and their relation to the pathologic changes were analyzed. The sonographic appearances of testicular torsion were categorized into six patterns, reflecting pathologic changes of testicular torsion. Sonography of testicular torsion has different patterns, and recognition of these patterns may help the examiner establish a diagnosis and subsequently manage the patient.

Seminal molecular markers as a non-invasive diagnostic tool for the evaluation of spermatogenesis in non-obstructive azoospermia

Non-obstructive azoospermia (NOA) is currently evaluated by the use of conventional histopathological methods. In some cases, focal spermatogenesis is present in the testes of patients with NOA which may be almost undetectable by routine histopathological examinations. Application of molecular markers in semen to predict the spermatogenesis status in the testis will emphasize the probability of finding sperm in NOA testis through further search using TESE or mTESE. Detection of germ cell-specific transcripts in semen is a signal of germ cells present in the testis. In this study, we used molecular methods to evaluate spermatogenesis status in azoospermic men. Semen samples were collected from 203 men with azoospermia. Total RNA was extracted from the semen precipitates. First-strand complementary deoxyribonucleic acid (cDNA) was synthesized by reverse transcriptase then, (RT)-PCRs were carried out using primers for testis stage-specific genes (DAZ, AKAP4, PRM1, and PRM2). Testicular tissue biopsies were used for evaluating spermatogenesis status in testis. Histopathological examination and LH, FSH, and testosterone level measurements (chemiluminescence assay) were performed. The presence of DAZ and PRM2 transcripts in semen significantly indicated the presence of spermatogonia and spermatids in the testicular tissues. Absence of all four markers in semen confirmed the histopathological results corresponding to sertoli cell only syndrome (SCO). Although TESE should not be excluded solely on this criteria, using PRM1, PRM2, AKAP4, and DAZ transcripts in semen would provide a non-invasive molecular diagnostic tool to better counsel patients before undergoing TESE.

The Effects of Medical Castration on Testes in Patients With Advanced Prostate Cancer

Because of patients' fear of surgery, the psychological symbolism of the male character, and significant encouragement by urologists, the majority of patients with advanced prostate cancer prefer medical castration as a choice of hormonal treatment. We investigated how the preserved testes were affected by medical castration with luteinizing hormone-releasing hormone (LH-RH) agonist and/or antiandrogen therapy. Thirty-four consecutive patients with advanced prostate cancer who underwent a bilateral epididymal-sparing orchiectomy were enrolled in this study. Before surgery, 19 of them had received LH-RH agonists and/or antian-drogens, while the other 15 patients had not. Morphological measurements were made immediately after the operation, and a histological examination was performed by a uropathologist with confirmation by urologists. The axial length, weight, and histopathological findings of the testes and prostate-specific antigen (PSA)/tes-tosterone level were compared between the two groups. Histological changes and characteristics were recorded and analyzed. The median axial length and weight of the testes in the 19 patients with prior medical castration were 2.6 (2.0–3.6) cm and 8.0 (6.0–11.5) g, which significantly differed from those with primary surgical castration [4.0 (2.9–5.0) cm and 12.0 (9.0–17.0) g, both p< 0.001]. Moreover, profound atrophic and fibrosclerotic changes were noted in the seminiferous tubules in the group with prior medical castration. Specific findings, including basement membrane thickening and interstitial cell hyperplasia, were noted in specimens with prior medical castration. However, the postoperative serum PSA and testosterone levels did not significantly differ between the two groups. Medical castration in patients with advanced prostate cancer indeed caused a significant reduction in the volume and profound pathological change in the testes, which may compromise a patient's expectation of his self-image (symbol of sexual virility).

Increased apoptosis of germ cells in patients with AZFc deletions

AZFc deletions are associated with variable testicular histology ranging from the Sertoli cell only to spermatogenic arrest and hypospermatogenesis. Such variable phenotypes may be explained by progressive germ cell regression over time. Increased apoptosis is likely responsible for progressive regression of spermatogenic potential. This study evaluated germ cell apoptosis as a cause of the progressive decrease in the number of germ cells in patients with AZFc deletions. This study evaluated germ cell apoptosis in patients with AZFc deletions. A total of 151 patients who were diagnosed with either severe oligozoospermia or non-obstructive azoospermia were screened for Y chromosome microdeletions. Germ cell apoptosis was examined using terminal deoxy-nucleotidyl transferase-mediated digoxigenin-dUTP nick-end labeling (TUNEL) on formalin-fixed 5-microm sections of testicular specimens. Seven out of 117 (6.0%) patients with azoospermia and 4 of 34 (11.8%) patients with severe oligozoospermia had Y chromosome microdeletions. The percentage of apoptotic germ cells in the testes of patients with AZFc deletions were significantly increased compared to those of patients without AZFc deletions. These results suggest that increased apoptosis of germ cells is responsible for the progressive decline of spermatogenic potential in patients with AZFc deletions.

Laparoscopic-Assisted Management of Impalpable Testis in Patients Older than 10 Years

Cryptorchidism affects 1% of male births. The majority of patients with undescended testis are identified and treated in childhood, but a significant proportion of them especially in third-world countries are neglected and present late. Herein, we present our initial experience of managing impalpable testis in older children and adults with laparoscopic assistance. This study was conducted from 2003 through 2008 at LUMHS Jamshoro. Thirty-two patients with 40 impalpable testes were included in this study. Diagnostic laparoscopy was performed in 32 anesthetized patients. Laparoscopic-assisted orchiopexy or orchiectomy was performed in patients with intraabdominal testis. Testicular vessels and vas deferens were mobilized and after obtaining sufficient length were brought through the posterior wall of the inguinal canal by creating a neo-inguinal ring medial to the epigastric vessels after a small inguinal incision. Of 40 impalpable testis, ultrasound localized 16 (40%) of them, and on laparoscopy, 36 (90%) were localized, 30 (75%) as intraabdominal and in 6 (15%) cases vas and vessels were entering into the internal inguinal ring (intracanalicular). The remaining 4 patients were diagnosed as having vanishing testis (anorchia). Laparoscopic orchiectomy was performed in 14 (35%) of these testes, while single-stage laparoscopic-assisted orchiopexy using Prentiss' maneuver was performed in 16 (40%) testis. No major complications occurred. Seven testes were associated with ipsilateral hernias and were simultaneously repaired laparoscopically. Laparoscopic-assisted single-stage orchiopexy is a safe and successful procedure for intraabdominal testis in adolescents and older patients. The additional benefit of shortening the usual course of spermatic cord was beneficial to fix the testis in the scrotum without tension.

The lipid composition of human testis in patients with bilateral varicocele as cause of infertility.

The lipid composition of human testis in patients with bilateral varicocele as cause of infertility is presented. The total lipids formed 1.90 percent of the total wet weight of the tissue. Testicular cholesterol formed 26.50 percent, glycerides 28.50 percent and phospholipids 45 percent of the total lipids. The increase observed in total cholesterol, glycerides in the testis of patients with bilateral varicocele compared to the control group appears to suggest that non-utilization of cholesterol esters for androgen biosynthesis. Androgen deficiency would result in inhibition of spermatogenesis as shown by the histological examination of the bilateral testicular biopsy specimens. Testicular neutral lipids get altered whenever there is imparied steroidogenesis and spermatogenesis. Thus, testicular lipids may taken as one of the important indices of testicular function.

Further studies on testicular lipids and glycogen in human patients with unilateral varicocele.

In the present study the lipid and glycogen content of human testis in patients with unilateral varicocele is presented. Eight cases were taken for the present study. Testicular biopsy specimens taken from these patients were analyzed and the normal testis acted as control groups. There was marked increase in total lipids, ester cholesterol and glycerides in the testis of varicocele patients. The abnormal increase observed in estercholesterol may be due to non-utilization of cholesterol esters for androgen biosynthesis. The glycogen level was reduced significantly in the affected testis. Thus, varicocele seem to alter the availability of substrates of energy required for normal functioning of germinal and non-germinal cells which results in defective spermatogenesis and steroidogenesis.

Multifocality in Testicular Germ Cell Tumors

Standard treatment for testicular germ cell tumor is radical orchiectomy. Several groups have suggested an organ sparing approach in patients with bilateral tumors or tumor in a solitary testis. We determined the prevalence of multifocality in testicular germ cell tumor cases. Orchiectomy specimens from 145 consecutive patients treated for testicular germ cell tumor between 1995 and 2006 were included in the study. In the current series slides were reviewed by a single dedicated uropathologist. Multifocality was defined as 1 of 4 distinct pathological entities, including 1) distinct tumor focus conspicuously separable from the main tumor mass, 2) microinvasive tumor characterized by a single or small groups of malignant germ cells scattered within the normal interstitial parenchyma, 3) extra tumor vascular invasion and 4) rete testis invasion by pagetoid tumor spread. Multifocality was identified in 48 patients (33%), of whom 17 (12%) had an additional distinct tumor focus, 21 (14%) had microinvasive tumor, 17 had extra tumor vascular invasion and 2 had rete testis invasion by pagetoid tumor spread. Multifocality was more prevalent in men with smaller tumors and seminomatous histology (pure seminoma or as part of a mixed germ cell tumor). Multifocality was present in 63% of men with an index mass of 2 cm or less in diameter. Study limitations include potential pathological sampling errors resulting from the retrospective design. Multifocality is a frequent finding in testicular germ cell tumor cases that is associated with small mass size and seminomatous histology. Data suggest that additional invasive tumor outside the index mass may be present in up to 63% of men considered potential candidates for organ sparing surgery.

Histopathological Patterns of the Testes in Patients with Severe Burns Not Involving the Perineum (A Case Series Study)

Histopathological Patterns of the Testes in Patients with Severe Burns Not Involving the Perineum (A Case Series Study)

Dose reduction in computed tomography: the effect of eye and testicle shielding on radiation dose measured in patients with beryllium oxide-based optically stimulated luminescence dosimetry

The aim of this study was to assess the effect of eye and testicle shielding on radiation dose to the lens and the testes of patients undergoing CT examinations. Fifty-one male patients underwent CT twice with identical protocols initially without, the second time with protective garments. Doses to the testes and the lenses were recorded with beryllium oxide-based dosimeters. The dose to the testes and lenses from CT exposure was reduced by 96.2% +/- 1.7% and 28.2% +/- 18.5%, when testicle and eye shielding was used, respectively. The effect of the eye shielding on the eye lens dose was found to depend on the x-ray tube position when the eye is primarily exposed during the scan. The maximum eye lens dose reduction achieved was found to be 43.2% +/- 6.5% corresponding to the anterior position of the tube. A significant correlation between the patient's body mass index and dose exposure could not be found. Eye and testicle shields, apart from being inexpensive and easy to use, were proven to be effective in reducing eye lens and testicle radiation dose burden from CT exposures.

Sex reversal from functional disomy of Xp: Prenatal and post‐mortem findings

Translocations involving the short arms of the X and Y chromosomes are uncommon and are often associated with anomalies in gonadal development. Segmental duplications of the X chromosome interfere with the formation of the testis in patients with a 46,XY karyotype. The gene products located within the duplicated segment, when present in double dose, may affect on male sex development. We report on a fetus with karyotype 46,XY,der (14)t(X;14) (p10;p10)dn. Attached to chromosome 14 is the entire short arm of the X chromosome. Therefore, the fetus is affected with a disomy of Xp, resulting in complete male to female sex reversal, as well as other structural defects. To the best of our knowledge, this is the first description of an XY fetus with a pure duplication of the entire short arm of X chromosome.

RGS22, A Novel Testis-Specific Regulator of G-protein Signaling Involved in Human and Mouse Spermiogenesis along with GNA12/13 Subunits1

The heterotrimeric G-protein pathway controls numerous cellular processes, including proliferation, differentiation, migration, membrane trafficking, and embryonic development. Regulator of G-protein signaling (RGS) proteins are known to function at the G-protein level. Here, the functional role of a novel RGS protein, regulator of G-protein signaling 22 (RGS22), in the testis was investigated at the mRNA and protein levels. Our results demonstrate that RGS22 is a testis-specific gene. However, significantly decreased expression of RGS22 was found in the testes of patients with azoospermia. RGS22 was translated or posttranslationally modified into multiple proteins of different molecular sizes in prokaryocytes as well as in the testes. Its protein (NP_056483) was localized in spermatogenic cells and Leydig cells and could interact with guanine nucleotide binding protein, alpha 12, 13, and 11 (GNA12, GNA13, and GNA11). Fragmental GFP-fusion protein tracking revealed that the N-terminal of RGS22 was localized in the nucleus. RGS22 and GNA13 were localized in the nucleus from the elongated spermatid stage onward. Indirect immunofluorescence studies revealed defective expression of GNA13 in macrocephalic and global nucleus spermatozoa. These findings suggest that their functions in this subcellular compartment are likely related to the postmeiotic developmental phase, spermiogenesis. RGS22 may also play a role in GNA13 translocation from the cytoplasm to the nucleus during spermiogenesis.

Differential expression of androgen and estrogen receptor of appendix testis in patients with descended and undescended testes

The incidence of appendix testis has been shown to be 76% in descended and 24% in undescended testis in our previous intraoperative survey. To determine the possible role of the appendix testis in the process of testicular migration, we compared the androgen and estrogen receptor status of appendix testis in descended and undescended testes. Thirty-seven appendix testes were collected intraoperatively and the expression of androgen and estrogen receptors were examined with immunostaining and immunofluorescence labeling. Based on the diagnosis, the specimens were divided into three groups. Group H (groin hernia, n = 11, as a group of descended testis), Group AU (acquired undescended testis, n = 14), and Group CU (congenital undescended testis, n = 12). The testicular appendages were found to express both androgen and estrogen receptors in Group H and Group AU, but specimens in Group CU were only estrogen receptor positive, whereas androgen receptors were not present. The presence of the androgen receptor in the appendix testis of the descended testes and acquired undescended testes and its absence in patients with congenital undescended testis suggests that the appendix testis might play a role in the process of testicular descent.

Primary melanoma of testis

Primary melanoma of testis is extremely rare and even the existence of such an entity is questioned. We present the case of a 60-year-old man with primary malignant melanoma in the testis. We report this case to emphasize the need for awareness of the possibility of the testis being the primary site in the patient with a melanoma and to underline the necessity of meticulous investigation of suspicious lesions of the testis in patients with or without a past history of malignant melanoma.

Quantification of DDX3Y, RBMY1, DAZ and TSPY mRNAs in testes of patients with severe impairment of spermatogenesis

Y chromosome microdeletion is the most important genetic cause of impairment of spermatogenesis. Nevertheless, a significant proportion of patients with spermatogenic failure do not have this condition. This study investigated the expression level of AZF genes, DDX3Y (DBY), RBMY1, DAZ and TSPY in testicular tissues of 42 subjects with impaired spermatogenesis compared with 33 with normal spermatogenesis. Histopathological evaluation was performed in all subjects and tissues were classified according to Johnsen Score. Transcript amounts were determined by quantitative-competitive RT-PCR. Patients with complete Sertoli cell-only syndrome (SCOS) did not exhibit RBMY1, DAZ and TSPY gene expression, however, we detected very low expression of DDX3Y transcript. Tissue samples with focal SCOS showed significantly decreased expression of all genes (P < 0.001). Maturation arrest and hypospermatogenesis tissues expressed significantly low levels of DDX3Y testicular transcript (P < 0.001), while the mRNA levels of the other genes were similar to that in tissues from the normal spermatogenesis group. Negative or diminished gene expression of DDX3Y, RBMY1, DAZ and TSPY in tissues samples with SCOS or focal SCOS reflects the absence or the lower number of germ cells, respectively. The finding that the testicular transcript of DDX3Y is significantly decreased in patients with severe spermatogenenic failure, especially in those presenting maturation arrest, suggests an important role of DDX3Y during spermatogenesis.

Expression and Possible Functions of a Novel Gene SPATA12 in Human Testis

In our previous study, a novel human testis-specific gene, SPATA12, was identified using the digital differential display program. In the current study, both SPATA12 mRNA and protein levels in the developmental stage of the testis were detected by SYBR real-time reverse transcriptase polymerase chain reaction and Western blot. A high level of SPATA12 gene expression was observed in normal adult testis but was completely absent in fetal testis. Both in situ hybridization and immunohistochemical analysis showed that SPATA12 was expressed in seminiferous tubules of adult testis-more precisely in spermatocytes, spermatids, and spermatozoa-but there was no expression in Sertoli and Leydig cells. These results showed that SPATA12 is a stage-specific and germ cell-specific gene, which suggests that it must be involved in the development of testicular maturation. It was also found that the expression level of SPATA12 mRNA in the testes of infertile men was associated with the amount and density of germ cells. With a decrease in the number of germ cells, the expression of SPATA12 mRNA was lower. In addition, the signal in the testes of patients with cryptorchidism or Sertoli cell only syndrome was not detected. Flow cytometry analysis of SPATA12 in human HeLa cells and mouse GC-1 spg germ cells indicated that the expression of the SPATA12 gene may delay the progression of G(1) to S in the cell cycle. SPATA12 was also shown to be lost in testicular germ cell tumors both at the level of transcription and translation. We hypothesized that the putative function of SPATA12 is to maintain the cell in a differentiated state and/or to suppress cell proliferation.