Seminal transforming growth factor-beta in normal and infertile men.
Transforming growth factor-beta (TGFbeta) is a cytokine with autocrine and paracrine action in the testis and potent immunoregulatory and anti-inflammatory activities. In the present study, we examined the concentration of latent (acid-activatable) and free (active) TGFbeta in seminal plasma from normal subjects (n = 23) and infertile (n = 40) patients, by using a TGFbeta specific immunoenzymological assay, and a bioassay (CCL64 cell line growth inhibition) detecting any form of TGFbeta. Free TGFbeta1 was present in normal subjects at a concentration (1.82 +/- 1.06 ng/ml) close to that known to give maximal stimulation in vitro. In pathological groups, the mean concentrations were not significantly different from the normal ones. Latent TGFbeta1 was present in normal seminal plasma at a high concentration (92.4 +/- 29.2 ng/ml). In subjects with pathologies of both testis and genital apparatus, or with epididymal occlusion, mean latent TGFbeta1 concentrations were normal, whereas transferrin concentrations were lower. The concentrations found in the epididymal occlusion group indicate that TGFbeta1 is, for a large part, secreted by the genital tract. In the testicular pathology group, TGFbeta1 concentrations were 130.7 +/- 61.2 ng/ml, a mean not statistically different from normal, although higher. No differences were found between patients with high and normal blood plasma follicle stimulating hormone, and this is consistent with the notion that most TGFbeta1 in seminal plasma is not of testicular origin. The TGFbeta bioassay ensured that immunologically detected TGFbeta was present in a bioactive or bioactivatable form. Furthermore, the values found in normal and pathological seminal plasmas were usually higher than those detected by the immunoassay, suggesting that other forms of TGFbeta might be present. Together, the present data show that very large amounts of TGFbeta are present in human seminal plasma. The TGFbeta ligand assay in the seminal plasma appears to indicate no differences between normal and infertile subjects. (+info)
Absence of testicular DAZ gene expression in idiopathic severe testiculopathies.
Deletions of the DAZ (deleted in azoospermia) gene family are frequently responsible for male infertility and are generally assessed by analyses of genomic DNA extracted from peripheral leukocytes. The multicopy nature of this gene prevents the distinction of intragenic deletions or deletions not involving the whole DAZ gene cluster. Thus it is still unclear whether each DAZ copy is effectively expressed in the testis. We analysed, by reverse transcription-polymerase chain reaction (RT-PCR), the expression of DAZ, RBM and SRY genes, in testicular cells from infertile men affected by idiopathic severe hypospermatogenesis, obstructive azoospermia and Sertoli cell-only syndrome. Normal mRNA for DAZ, RBM and SRY were observed in obstructive azoospermia, whereas only SRY transcripts were detected when only Sertoli cells were present. Nine out of 10 patients affected by idiopathic severe hypospermatogenesis had normal expression of SRY, RBM and DAZ, while in one patient no DAZ transcript was detected, suggesting that his testiculopathy was related to the absence of DAZ expression. The lack of DAZ mRNA in testicular cells with an apparently normal DAZ gene constitution on DNA extracted from leukocytes may be explained by different hypotheses: (i) not all the copies of the DAZ gene cluster are transcribed in the germ cells and the reported patient had a small deletion involving only the active ones; (ii) the patient may be mosaic for the DAZ gene having a normal constitution in leukocytes and be deleted for DAZ gene in the testis; (iii) abnormalities of DAZ transcription may exist. These findings highlight the intrinsic interpretative difficulties of normal PCR analysis for DAZ and RBM on leukocytes and suggest caution in the use of germ cells for assisted reproductive techniques in these cases to avoid transmission of genetic abnormalities to male offspring. (+info)
Testicular adrenal rests: evidence for luteinizing hormone receptors and for distinct types of testicular nodules differing for their autonomization.
We report one patient with 21-hydroxylase deficiency and associated bilateral macro-orchidism caused by nodular hyperplasia of testicular adrenal rests (TAR). The boy, referred to us when 10 years old, was born with bilateral cryptorchidism that was treated unsuccessfully with i.m. injections of human chorionic gonadotropin (hCG) and later on with orchidopexy. He was treated with oral dexamethasone (0.625 mg per day) for the following 13 years. After one year, there was a marked reduction in steroid hormone levels (17-hydroxyprogesterone (17-OH P) from 27.2 to 1.2 nmol/l, testosterone from >104 to 4.8 nmol/l, estradiol (E(2)) from 481 to 33 pmol/l). After the same period of time, both testicular volume and nodularity decreased: from 45 to 18 ml and from numerous to four nodules in the right testis, and from 40 to 13 ml and from numerous to three nodules in the left testis. At the third year, there were transient increases in serum gonadotropins, testicular volume (right testis = 25 ml, left testis = 20 ml) and steroid hormones, including cortisol (serum ACTH and dehydroepiandrosterone sulfate remained suppressed). At the fourth year of follow-up, there were still four nodules in the right testis and three in the left testis. The LH-dependency (which implies possession of LH/hCG receptors) of these nodules was also substantiated by their steroidogenic response to an acute i.m. hCG test. An exogenous ACTH stimulation test increased serum 17-OH P and cortisol. Since these nodules, unlike the majority of those present initially, were not suppressed by the corticosteroid therapy and since they were not detected when the patient returned for control at 23 years of age, they had partial autonomy from ACTH. At 23 years of age, the patient had a single nodule in the right testis (right testis volume = 13 ml, left testis volume = 10 ml), which should have accounted for the consistent difference in size between the two gonads. Serum LH was about 7 mU/l and FSH about 23 mU/l. The responsiveness of plasma steroid hormones to hCG had changed quantitatively and qualitatively. Secretion of cortisol was absent, secretion of 17-OH P and testosterone was reduced, and secretion of E(2) was much increased. The ACTH stimulation test showed that serum cortisol did not respond, while the other steroids responded in the order of 17-OH P>E(2)> testosterone. We conclude that there were three different groups of TAR when the patient was already 10 years old: (i) ACTH-sensitive (the majority), (ii) partially ACTH-insensitive but LH/hCG-sensitive (three nodules in the left testis and three in the right testis), (iii) almost entirely ACTH-insensitive and partially hCG-insensitive (a single nodule in the right testis). Probably, the never suppressed gonadotropin levels (presumably due to the bilateral testicular damage subsequent to the cryptorchid state) and the hCG therapy were major etiological factors for the appearance of the second and third population of TAR. (+info)
Unusual sonographic appearance of an epidermoid cyst of the testis.
Epidermoid cyst of the testis is a rare benign testicular tumor with varied sonographic appearances. We present a case in which two specific ultrasonographic patterns were seen: (1) an "onion ring" configuration of alternating hyperechoic and hypoechoic regions, described previously as being characteristic of this lesion, and (2) a heterogeneous region with multiple punctate hyperechoic foci. (+info)
Specific expression of heat shock protein HspA2 in human male germ cells.
In the mouse, the heat shock protein 70-2 (Hsp70-2) has been found to play a critical role in spermatogenesis. The HspA2 gene is the human homologue of the murine Hsp70-2 gene with 91.7% identity in the nucleotide coding sequence. We examined the expression of HspA2 in human tissues. To detect HspA2 expression, antiserum 2A that was raised against mouse Hsp70-2 and that cross-reacted with human HspA2 protein expressed in Escherichia coli was used. The results of Western blotting indicate that significant HspA2 expression occurs in testes with normal spermatogenesis, whereas only a low amount of HspA2 was expressed in testis with Sertoli cell-only syndrome. Only a small amount of HspA2 was detected in breast, stomach, prostate, colon, liver, ovary, and epididymis. Immunoreactivity to HspA2 was present in spermatocytes and spermatids in the testes with normal spermatogenesis, while immunoreactivity to HspA2 in testis with Sertoli cell-only syndrome was remarkably decreased or inconspicuous over the entire cell. These results demonstrate that the HspA2 protein is highly expressed in human male specific germ cells, suggesting that HspA2 protein may play a specific role during meiosis in human testes as found in the murine model. (+info)
Highly sensitive quantitative telomerase assay of diagnostic testicular biopsy material predicts the presence of haploid spermatogenic cells in therapeutic testicular biopsy in men with Sertoli cell-only syndrome.
The role of a telomerase assay in the recognition of Sertoli cell-only syndrome with testicular foci of haploid cells was evaluated. Men with Sertoli cell-only syndrome (n = 23) were given a new diagnostic testicular biopsy. Part of the biopsy was stained and the remainder was processed for the quantitative telomerase assay. After 3-13 months, a therapeutic testicular biopsy was performed. This material was minced and then examined using confocal laser scanning microscopy and fluorescent in-situ hybridization. Histology of diagnostic testicular biopsy material confirmed the diagnosis of Sertoli cell-only syndrome in all the participants. All seven men with a telomerase assay value in their diagnostic testicular biopsy of >42 total product generated (TPG) U/microg protein had haploid cells (i.e. spermatozoa and/or spermatids) in their therapeutic testicular biopsy. Among participants with telomerase assay values <42 TPG U/microg protein, only one man had haploid cells in his therapeutic testicular biopsy. Thus, telomerase assay values >42 TPG U/microg protein in the diagnostic biopsy identified 87.5% of the Sertoli cell-only syndrome men with haploid cells in their therapeutic testicular biopsy. Significantly higher values of the telomerase assay were found in men with testicular foci of haploid cells than in men without these foci. The use of a quantitative telomerase assay biopsy appears to be important for identifying those men with Sertoli cell-only syndrome who have foci of haploid cells and can be candidates for assisted reproduction techniques. (+info)
Ultrasonography of intratesticular lesions: its role in clinical management.
Ultrasound is the primary imaging modality in the investigation of patients with symptoms related to the scrotum, and is pivotal to the diagnosis of suspected testicular malignancy. This retrospective study analysed the results of testicular ultrasound at a large teaching hospital over a five year period. We wished to examine the clinical consequences for patients in whom ultrasound findings were suspicious of testicular cancer, and the accuracy of the ultrasound diagnosis. Real time ultrasound examinations were performed, providing multiplanar imaging of the testis and para testicular tissues. Over a five year period 661 examinations were carried out. An intratesticular lesion was identified in 44 patients; nineteen of these patients were shown to have testicular malignancy following tissue diagnosis. When ultrasound was used to identify testicular malignancy in those patients with an intratesticular lesion, it had a sensitivity of 94.7% and a specificity of 59.1%. A tissue diagnosis was obtained in 93% of those patients thought likely to have a testicular malignancy on sonographic assessment, and in 40% of those in whom a diagnosis of malignancy was possible, but less likely. Our study shows that this modality can be used to aid the clinician in deciding which patients should undergo orchidectomy, invasive biopsy or clinical surveillance. (+info)
Halicephalobus gingivalis (H. deletrix) infection in two horses in southern California.
Two horses, a 16-year-old male Holsteiner and a 5-year-old male miniature horse, were diagnosed with halicephalobiasis at the California Veterinary Diagnostic Laboratory System, San Bernardino Branch, in April and June of 1998. Over a period of 4 weeks, the Holsteiner horse developed renal dysfunction, blepharospasm, and blindness in the right eye. A 15-cm-diameter mass was detected on ultrasound examination in the right kidney. Terminally, the animal developed seizures and was euthanized. The miniature horse had a 6-week-long illness characterized by testicular enlargement and uveitis. This animal developed ataxia and died. Necropsy examination revealed bilateral enlargement of the kidneys in both horses, petechial hemorrhages of the optic nerve (Holsteiner), and a diffusely firm and enlarged left testicle (miniature horse). Microscopic evaluation of tissues revealed granulomatous nephritis, optic neuritis, retinitis, and encephalitis in both horses and orchitis in only the miniature horse with intralesional rhabditiform nematodes. Halicephalobus gingivalis was found in the urine sediment of both animals and in semen of the Holsteiner horse. (+info)