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Assessment of aging in the testis of Chinese hamster (Cricetrus griseus) by proton nuclear magnetic resonance spectroscopy.

By using nuclear magnetic resonance (NMR), the aged testis of Chinese hamster was studied by measuring relaxation times of tissue water protons. Spin-spin relaxation time (T2) was significantly prolonged compared to young testis when edema was apparent in the interstitial space of the aged testis. When edema was experimentally induced in the young testis by ligation of a subcapsular vein, T2 was prolonged to the similar extent as in the aged testis. Spin-lattice relaxation time (T1) and water content of the tissue were essentially the same in the testes of the young and aged animals. It is concluded that T2 can serve as a sensitive parameter for assessing the age-dependent histological changes in the testis of Chinese hamster.

Aging↗

Effects of exposure of lactating female rats to polychlorinated biphenyls (Pcbs) on testis weight, sperm production and sertoli cell numbers in the adult male offspring.

To investigate the effects of intermittent and continuous exposure of lactating rats to Aroclor 1242 (a PCB congener), testis weight, daily sperm production (DSP) and Sertoli cell number per testis were examined in the adult male offspring. Thyroxine (T4) was also measured because of the well-documented effects of polychlorinated biphenyls (PCBs) on this hormone. In experiment 1.3 groups of lactating female rats received daily subcutaneous injections of low (0.8 mg) or high (1.6 mg) doses of Aroclor 1242 in 0.1 ml corn oil from parturition to weaning of pups at 21 days. In experiment II, 3 groups of lactating rats received 2 subcutaneous injections per week of 0.8 or 1.6 mg Aroclor 1242, as in experiment 1. In both experiments, control rats received vehicle alone. Serum T4 was measured at 21 and 90 days of age, and testis weight, DSP and Sertoli cell numbers were examined at 90 days. In experiment I (continuous exposure), both the low (0.8 mg) and high (1.6 mg) doses suppressed T4 concentrations at 21 days of age. Testis weight was increased by 14.8% (LD) and 16.5% (HD) compared with controls. DSP was increased by 20.4% in the low dose and 25% in the high dose animals compared with controls. The number of Sertoli cells per testis was increased by 32.6 and 39.4% in low and high dose animals, respectively. A similar study in which the lactating females were only dosed twice per week (experiment 11) did not show any differences in these parameters. These results indicate that continuous exposure of lactating female rats to PCBs increases testis weight, sperm production and Sertoli cell numbers in the adult male offspring.

Aging↗

Developmental expression of p63 in the mouse testis.

p63 is a member of the p53 gene family and have structural similarities with p53. p63 encodes for multiple isotypes either with N-terminal transactivation domain (TAp63) or without it (DeltaNp63). In the mammalian testis, it has been shown that p53 plays important roles in the regulation of germ cell apoptosis and meiosis. However, little is known for the physiological function of p63 in the mammalian spermatogenesis. To investigate the potential roles of p63 in the developing mouse testis, we examined the expression pattern of p63 in the mouse testis from birth to adulthood. In addition to the TAp63 mRNA which was continuously expressed in the developing testis, transcripts encoding DeltaNp63 was detected at specific stages of testicular development by RT-PCR, from postnatal day 1 to day 7 and from 3 weeks to 4 weeks after birth. Western blot analysis of whole testis lysates with anti-p63 antibody revealed an approximately 68 kD band throughout development and a less abundant protein at 60 kD in the earlier period of postnatal development. Immunopositive reactions for p63 were observed as early as 10 days after birth and p63 protein was localized to the nuclei of spermatocytes and round spermatids. These findings strongly suggest that p63 might be involved in the regulation of proliferation and differentiation of spermatogenic cells in the developing mouse testis.

Animals↗

Alteration of trace element distribution and testis ACE activity in mice with high peritoneal aluminum.

The present investigation was conducted to assess the effects of subacute aluminum (Al) exposure on testicular zinc (Zn), copper (Cu), and iron (Fe) distribution in mice. Animals were intraperitoneally exposed to 0, 13, or 35 mg Al/kg body weight/d for a period of 14 d. Al concentrations in serum and testis in Al-treated animals were significantly higher than those of controls. The serum concentrations of Fe were lower, whereas serum Zn and Cu showed a pattern comparable to that of controls. The accumulation of testicular Fe and Cu remarkably increased in Al-exposed groups, whereas the Zn concentration in testis was significantly reduced only at the highest dose of Al exposure. The values of testicular thiobarbituric acid reactive substances (TBARS) were also increased after Al administration, indicating increased lipid peroxidation and oxidative stress. In addition, when the testicular Al was increased, the testis-specific angiotensin-converting enzyme (testis ACE) was noted. The results of this study indicated that part of the effect of Al intoxication on testis might contribute to abnormal metabolism of other minerals, such as Fe, Zn, and/or Cu. It was also suggested that reduced testis ACE activity presumably plays an important role in oxidative damage of Al-induced testicular toxicity.

Aluminum↗

Distributional map of the terminal and sub-terminal sugar residues of the glycoconjugates in the prepubertal and postpubertal testis of a subject affected by complete androgen insensitivity syndrome (Morris's syndrome): lectin histochemical study.

In the present research we have investigated the distribution of the sugar residues of the glycoconjugates in the prepubertal and postpubertal testes of a subject with Morris's syndrome (CAIS, Complete Androgen Insensitivity Syndrome). For this purpose a battery of six horseradish peroxidase-conjugated lectins was used (SBA, PNA, WGA, ConA, LTA and UEAI). We have obtained a complete distributional map of the terminal and sub-terminal oligosaccharides in the tunica albuginea, interstitial tissue, lamina propria of the seminiferous tubules, Leydig cells, Sertoli cells, spermatogonia, mastocytes and endothelial cells. Furthermore the present study has shown that a large amount of sugar residues were detectable in the prepubertal and postpubertal testes but that some differences exist with particular regard to the Sertoli cells. The Sertoli cells and the Leydig cells of the retained prepubertal testis of the patient affected by Morris's syndrome were characterized by the presence of alpha-L-fucose, which was absent in the retained prepubertal testis of the normal subjects. Comparing the results on the postpubertal testis with those obtained on the same aged testis of healthy subjects we have demonstrated that alpha-L-fucose in the Sertoli and Leydig cells and D-galactose-N-acetyl-D-galactosamine in the Leydig cells are a unique feature of the subject affected by Morris's syndrome. D-galactose (ss1,3)-N-acetyl-D-galactosamine and sialic acid, which are present in the Leydig cells of the normal testis were never observed in the same cells of the postpubertal testis of the CAIS patient.

Adolescent↗

Detection of anti-Mullerian hormone receptor II protein in the postnatal rat testis from birth to sexual maturity.

Anti-Mullerian hormone (AMH) produced by the immature Sertoli cells negatively regulates the postnatal Leydig cell (i.e. adult Leydig cells/ALC) differentiation, however, the mechanism is sparsely understood. AMH negatively regulates the steroidogenic function of fetal Leydig cells (FLC) and ALC. However, when this function is established in the ALC lineage and whether AMH has a function in FLC in the postnatal testis are not known. Therefore, the objectives of this study were to examine the presence of AMH receptor type II (AMHR-II) in FLC and cells in the ALC lineage in the postnatal mammalian testis using the rat model Male Sprague Dawley rats of days 1, 5, 7-21, 28, 40, 60 and 90 were used. AMHR-II in testicular interstitial cells was detected in testis tissue using immunocytochemistry. Findings showed that the mesenchymal and the progenitor cells of the ALC lineage, were negative for AMHR-II. The newly formed ALC were the first cell type of the ALC lineage to show positive labeling for AMHR-II, and the first detection was on postnatal day 13, although they were present in the testis from day 10. From days 13-28, labeling intensity for AMHR-II in the ALC was much weaker than those at days 40-90. FLC were also positive. The time lag between the first detection of the newly formed ALC in the testis and the first detection of AMHR-II in them suggests that the establishment of the negative regulatory role of AMH on ALC steroidogenesis does not take place immediately upon their differentiation; no change in cell size occurs during this period. The absence of AMHR-II in mesenchymal cells suggests that it is unlikely that the negative regulatory effect of AMH on ALC differentiation in the postnatal testis is achieved via a direct action of AMH on mesenchymal cells. The presence of AMHR-II in postnatal FLC suggests a possible role by AMH on FLC, which warrants future investigations.

Aging↗

Regulation of LH receptors and steroidogenesis in the foetal rat testis in vivo.

Foetal rats were injected with high doses of hCG or ovine LH (oLH) to determine if LH receptors were down-regulated and steroidogenesis impaired as is seen in the adult animal. Foetuses injected in utero with 52 micrograms/kg hCG had a prompt increase in intratesticular testosterone, demonstrating that the injected substance reached the foetal testis. Pregnant mothers were laparotomized and each foetus in one uterine horn injected with 52 micrograms/kg hCG. Twenty-four hours later, the foetal testes from both uterine horns were collected and free LH receptor content measured. In foetuses injected with hCG, free LH receptors were significantly reduced. LH receptors in foetal testes from the contralateral horn were not significantly different from those in controls. Foetuses injected with 52 micrograms/kg hCG at 17.5 days of gestation showed a delayed response in LH receptor up-regulation when compared with neonatal rats injected with comparable doses of hCG. Injection of foetal and neonatal rats with 400 micrograms/kg oLH showed no evidence of down-regulation, as is seen in the adult. Testes from foetuses injected with 52 micrograms/kg hCG 3 days earlier showed an increase in both basal and hCG-stimulated testosterone production during 3 h of incubation when compared with controls. These results show that the rat foetal testis in vivo does not demonstrate down-regulation of LH receptors or steroidogenic lesions of the testosterone synthetic pathway when exposed to high levels of hCG as does the adult rat testis. These results from the foetal testis in vivo are in agreement with results obtained from the foetal testis in vitro and the neonatal testis in vivo.

Animals↗

Endogenous circannual rhythms and photorefractoriness of testis activity, moult and prolactin concentrations in mink (Mustela vison).

Mink are seasonal photosensitive breeders; testis activity is triggered when days have less than 10 h light. Increasing and decreasing plasma concentrations of prolactin induce the spring and autumn moults. In a 5 year experiment, males were maintained under short days (8 h light:16 h dark) at 13 degrees C or long days (16 h light:8 h dark) at 21 degrees C, winter and summer conditions, respectively. Under winter and summer conditions, circannual cycles of prolactin secretion and moulting were observed at intervals of about 11 months. Recurrence of testis cycles was not evident. In a second experiment, males were maintained under an 8 h light:16 h dark cycle from the winter solstice or under 10 h light:14 h dark, 12 h light:12 h dark or 14 h light:10 h dark cycles from 10 February. Under 8 h light:16 h dark cycle, testis regression was slightly later than under natural conditions, indicating photorefractoriness. However, mink remained sensitive to light: the longer the photoperiod, the faster the testis regression. In a third experiment, males were transferred under 8 h light:16 h dark or 16 h light:8 h dark from 15 May (group 1), 12 June (group 2) or 4 July (group 3); males submitted to long days received melatonin capsules on the day of transfer. Increasing concentrations of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) and testis volume were shown by half the males in group 2 and nearly all the males in group 3; the constant release of melatonin from implants was more efficient than short days; but in the three groups, prolactin concentrations decreased in the few days after short-day or melatonin treatment. Overall, the results demonstrate endogenous circannual rhythms of prolactin secretion, body weight and moulting. Although a refractory period to short days was observed, the annual cycle of testis activity totally relies on the annual changes in daylength.

Animals↗

Increased testis growth and sperm production in adult rats following transient neonatal goitrogen treatment: optimization of the propylthiouracil dose and effects of methimazole.

Treatment of male rat pups with the reversible goitrogen 6-propyl-2-thiouracil (PTU), administered by adding 0.1% PTU to the mother's drinking water from birth to day 25, increases their testis size and daily sperm production (DSP) at 160 days of age by up to 80% and 140%, respectively. The purpose of this study was to examine the effectiveness of various concentrations of PTU and determine the PTU dose that would maximize testis growth while minimizing side effects such as decreased maternal water consumption and decreased pup growth. Whether this effect was specific to PTU was determined by evaluating the effects of another commonly used goitrogen, methimazole (MMI), in increasing adult testis size and sperm production. Dams were given PTU (0.1-0.0004% w/v) or 0.025% MMI (w/v) in their water from birth to day 25 post partum, then given no further treatment. Thyroxine concentrations were measured in all groups of pups at 25, 35 and 45 days, and testis weight and DSP were determined at 90 days of age. At 25 days of age, thyroxine concentrations were maximally decreased by PTU treatments of 0.0015% or greater; less severe decreases were produced by 0.0004% PTU or 0.025% MMI. Thyroxine concentrations increased in all treated groups at day 35 compared with day 25, and returned to normal by day 45. At 90 days of age, testis weight was increased by about 40% in rats whose mothers had been treated with doses of 0.006% PTU or greater, whereas testis weights in groups given 0.0015 and 0.0004% PTU or 0.025% MMI were increased 31, 15 and 18%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Morphometric and endocrine analyses of the effects of nutrition on the testis of mature Merino rams.

The effects of nutrition on the testis were investigated in groups of five mature Merino rams that were fed either a sub-maintenance (low) diet or a supra-maintenance (high) diet for 69 days. Testosterone, oestradiol and inhibin were measured in blood plasma sampled simultaneously from jugular and testicular veins after an i.v. injection of 200 ng ovine LH kg-1. Plasma concentrations of testosterone, inhibin and oestradiol were higher in testicular than in jugular vein plasma for both diets (P < 0.01). After the LH injection, jugular plasma testosterone increased more rapidly (P < 0.01) in rams fed the high diet than in rams fed the low diet. This was not seen in the testicular vein. Oestradiol concentrations were higher in rams on the high diet than in those on the low diet, in both the testicular (P < 0.0001) and the jugular vein (P < 0.02). Diet did not affect inhibin concentrations. Testes were surgically removed and processed for light microscopy. Testicular mass and seminiferous tubule length and diameter were higher with the high diet than the low diet (P < 0.01). The number of Sertoli cell nuclei per testis was also affected (high diet: 120 +/- 6 x 10(8); low diet: 77 +/- 7 x 10(8); P < 0.001), whereas the proportion of testis occupied by Sertoli cell nuclei was not affected. The number of Leydig cells per testis was not affected by diet, but Leydig cells occupied a greater volume of testis in rams on the high diet than in those on the low diet (P < 0.001). The effects of nutrition on Leydig and Sertoli cells are consistent with changes in the endocrine and exocrine functions of the testis. The finding that Sertoli cell population was altered in adult rams may be explained by the GnRH-independent effects of nutrition.

Analysis of Variance↗

Association of cytochrome b5 with 16-androstene steroid synthesis in the testis and accumulation in the fat of male pigs.

The 16-androstene steroids, one of the principal causes of boar taint, are synthesized in the testis by the andien-beta synthase enzyme system. This system has been shown in vitro to involve both cytochrome P450c17 and cytochrome b5. The objective of this work was to investigate the relationship between the levels of cytochrome b5 in the testis, in vitro steroidogenesis, and the accumulation of 16-androstene steroids in the fat of pubertal boars. We found that the in vitro rate of 16-androstene steroidogenesis in testis microsomes was correlated with 16-androstene steroid concentrations in fat (r = .66, P < .01). Western blots were used to determine the amounts of cytochrome b5 and cytochrome P450c17 protein in testis, and two immunoreactive cytochrome b5 proteins of approximately 12 and 16 kDa were found. Levels of cytochrome P450c17 or the high molecular weight cytochrome b5 in testis were not significantly correlated to levels of 16-androstene steroids in fat. However, levels of total cytochrome b5 immunoreactive protein and levels of the low molecular weight immunoreactive cytochrome b5 were correlated to fat 16-androstene steroid concentrations (r = .59, P < .001; r = .72, P = .0001, respectively). Levels of the low molecular weight immunoreactive cytochrome b5 were also correlated to 16-androstene steroid synthesis rates in vitro (r = .62, P < .05). These results indicate that increased levels of a low molecular weight immunoreactive cytochrome b5 protein, and not of cytochrome P450c17, are related to increased testicular 16-androstene steroid production and accumulation in fat. These results support the hypothesis that selection for reduced levels of this low molecular weight immunoreactive cytochrome b5 protein in the testis may result in decreased levels of 16-androstene steroids in fat and reduced boar taint in uncastrated male pigs.

Adipose Tissue↗

The autonomous innervation of the buffalo (Bubalus bubalis) testis. An immunohistochemical study.

The innervation pattern in the buffalo testis was determined by using histochemical and immunohistochemical methods. Nerves were concentrated in the tunica albuginea and septula testis, and did not show an uniform distribution. The tunica albuginea at the lateral and medial sides and at the free border of the testis is most densely innervated than at the epididymal border. At the cranial pole thick nerve bundles were observed between albugineal vessels and muscle bundles. Rare parenchymal nerves were found in perivascular position between seminiferous tubules and their occurrence is confined to lobules at the cranial and caudal testicular poles. An intense NPY immunoreactivity occurred in nerve bundles and in solitary varicose fibres. Nerves were concentrated in the tunica albuginea at the lateral and medial side and at the free border of the testis, and in the lobules at the cranial and caudal testicular poles. Sub P immunoreactivity was occasionally detected in some thicker nerve bundles and solitary fibers, in the tunica albuginea and in the wall of blood vessels, showing a similar distribution but less intensity and density than NPY immunoreactivity. TH immunoreactivity stained nerve fibers in the buffalo testis with a distribution pattern similar to that obtained with general neuronal markers. The histochemical reaction for AchE was negative, so cholinergic fibers cannot be detected in the buffalo testis. The histochemical NADPHd reaction stained rare nitrergic nerve bundles and solitary fibers. The majority of NADPHd activity was confined to the vascular endothelium, and rarely to the interstitial Leydig cells, whereas the Sertoli and germ cells did not show any reaction.

Acetylcholinesterase↗

[Studies on the existence of renin in the human testis].

Recent reports have shown the existence of renin in Leydig cells of the rat testis and immunoreactive renin activity in the human testis. But we could not rule out the possibility that the renin found in the testis is contamination of plasma components by endocytosis. In the present study we tried to demonstrate the renin production in the human testis by the following experiments. At first, we measured the plasma renin activity and testosterone in the internal spermatic vein simultaneously after treatment with hCG and obtained the results that suggested hCG-induced renin secretion in the human testis. Next, we showed the hCG-dependent renin production from the cultured human Leydig cell tumors in vitro. Thirdly, we demonstrated the existence of renin mRNA in the human testisusing reverse transcriptase-polymerase chain reaction (RT-PCR). Fourthly, we detected significant signals of human renin mRNA in the human testicular tissues by in situ hybridization. From the above results, it was demonstrated that the human renin gene was hCG-dependently expressed in the human testis.

Animals↗

[A case of testicular tumor with a small tumor focus in the testis].

The initial symptom of a testicular tumor is painless enlargement of the testis in most cases, but in some cases can manifest itself with metastatic lesions associated with a small focus. On the other hand, a occult testicular tumor accompanied by a minimal lesion in the testis, may be included in the category of extra gonadal germ-cell tumors. We have seen a 41-year-old male with a chief complaint of swelling of a cervical lymph node, who was found to have a tumor in the testis of normal size. A CT scan revealed a tumor in the retroperitoneal space. The serum levels of HCG and AFP were markedly elevated. Biopsy of the cervical lymph node established a diagnosis of embryonal cell carcinoma. The left testis, free from a palpable mass but tender, was excised. A gonadal cell tumor approximately 7 mm in size was just below the epididymis. A stage III-a testicular tumor was diagnosed in the patient. Three courses of VAB-6 yielded partial relief in the patient. We concluded that the testis should be carefully examined with echogram whenever a extra gonadal germ-cell tumor is suspected, since a tiny lesion can exist in the testis concomitantly in some cases.

Adult↗

[The vanishing testis: diagnosis and histological findings].

PURPOSE: Controversy exists on how to diagnose the vanishing testis and the degree of investigation required. In this series, we reviewed anatomical and histological findings in vanishing testes and investigated the effectiveness of diagnostic laparoscopy and imaging studies. MATERIALS AND METHODS: Between 1974 and March 1999, 107 boys with nonpalpable testis underwent surgery. Of the total, 52 had spermatic vessels, vas deferens, and/or nubbin, and as a result the diagnosis of vanishing testis was made. RESULTS: The affected side of vanishing testis was left 41, right 9 and bilateral 2.35 nubbins were found and the lengths of 24 nubbins were 5 mm or less. Histological examinations were performed in 43 cases including 27 nubbins. From that total, 31 had vas deferens and 11 had epididymis. Only two nubbins had seminiferous tubules but they included no germ cells. The two nubbins were greater than 5 mm long. Laparoscopic surgery was undertaken in 12 separate cases of the vanishing testis and as a result hypoplastic spermatic vessels were present in 7 of the 12 cases. CONCLUSION: The incidence of viable testicular tissue in vanishing testes was 4.7% in our series and it ranges from 0-16% in other series. We submit that one can diagnose the inguinal vanishing testis with preoperative imaging and laparoscopy, and that the nubbin seldom contains testicular tissue. Our results do not support the necessity to remove nubbins.

Cryptorchidism↗

Diagnostic findings from testis fine needle aspiration mapping in obstructed and nonobstructed azoospermic men.

PURPOSE: Although helpful for defining extratesticular obstruction, the testis biopsy offers limited information on nonobstructive azoospermic testes. Guided by diagnostic biopsies, testis sperm extraction procedures fail in 25% to 50% of patients with nonobstructive azoospermia, largely because it is clinically difficult to know where sperm are located. To provide a more complete assessment of spermatogenesis in nonobstructive azoospermic patients and to simplify the confirmation of sperm production in men with obstruction, we use a systematic, fine needle aspiration "mapping" procedure. We summarize the diagnostic findings in a series of azoospermic men. MATERIALS AND METHODS: From 118 azoospermic infertile men (22 with obstructed and 96 with nonobstructed azoospermia) fine needle aspiration data were used to generate location specific, sperm frequency maps for obstructed and nonobstructive azoospermic testes to determine if "sperm rich" locations existed. RESULTS: Fine needle aspiration map analysis revealed that all aspiration locations from obstructed cases showed sperm. In men with nonobstructive azoospermia, sperm was identified in the right testis in 134 of 652 (20.5%) and in the left testis in 151 of 716 (21.1%) separate aspirations. Rates of sperm detection among various intratesticular sites were not statistically different. In 27.1% of cases the fine needle aspiration map found sperm in men with sperm negative biopsies. The likelihood of heterogeneity in fine needle aspiration sperm findings was 25% within individual nonobstructive azoospermic testes and 19.2% between testis sides. At post-procedure followup of 88 patients (74%), no clinical or surgical complications were observed. CONCLUSIONS: Testis fine needle aspiration mapping is a simple, reliable and informative diagnostic tool in the evaluation of azoospermic infertile men.

Biopsy, Needle↗

Cysts and epithelial proliferations of the testicular collecting system (including rete testis).

Neoplastic and non-neoplastic proliferations of the testicular collecting system may be a source of diagnostic difficulty as they are rarely encountered and may have an appearance that leads to confusion with lesions arising in adjacent structures. The rete testis and epididymis are the 2 sites that are most likely to give rise to this unusual group of lesions. This article principally, although not exclusively, deals with those lesions that produce an intrascrotal mass. These include benign acquired or developmental cysts such as cystic dysgenesis of the rete testis or acquired cystic transformation of the rete testis, non-neoplastic epithelial proliferations such as adenomatous hyperplasia of the rete testis or cribriform hyperplasia of the epididymis, and benign or malignant neoplasms. Rete testis cystadenoma, including the sertoliform variant, and papillary cystadenoma of the epididymis are examples of benign tumors of the testicular collecting system that must be considered, the latter is the more likely of the 2 to be encountered. Primary carcinoma of either the rete testis or the epididymis is a rare occurrence; these entities are considered in some detail to allow for their recognition and distinction from other tumors that may occur in this region. Our own experience with 18 unpublished cases of lesions of the rete is also present.

Adult↗

[A case of traumatic rupture of the testis: usefulness of magnetic resonance imaging].

Traumatic rupture of the testis is rare because of the protection afforded by surrounding structures. Moreover, it is difficult to accurately diagnose preoperatively. A 17-year-old man was referred to our department with the complaint of painful swelling of the left testis after being hit by a basketball. Although ultrasonography and computed tomography did not reveal the rupture of the tunica albuginea, we preoperatively diagnosed the rupture of the left testis by magnetic resonance imaging (MRI). We repaired the tunica albuginea and preserved the left testis. In this report, the advantages of MRI for preoperative diagnosis of traumatic rupture of the testis are discussed. In addition, previous cases with traumatic rupture of the testis in the Japanese literature are also reviewed.

Adolescent↗