Role of Ozone Therapy in Preventing Testicular Damage in an Experimental Cryptorchid Rat Model.

BACKGROUND Cryptorchidism is the most common developmental abnormality of the male reproductive system. If left untreated, it results with infertility and testicular cancer. According to current evidence, surgery is the mainstay of treatment, and hormonal therapy approaches are still under investigation. For the protection of testicular functions, antioxidants have emerged as novel options. This study aimed to evaluate the protective properties of ozone, a strong antioxidant, on testicular tissue. MATERIAL AND METHODS Thirty-five male Wistar-albino rats, 1-month-old, were used for the study. Groups were formed as follows: 1) control, 2) sham surgery (cryptorchidism), 3) cryptorchidism plus ozone, 4) cryptorchidism plus human chorionic gonadotropin (hCG), and 5) ozone plus hCG. Surgical procedures were performed on all rats except the control group. All rats except the control group were used to create an experimental cryptorchidism model, and left testes of animals were surgically placed into the abdomen. After 1 month of surgery, groups 3, 4, and 5 were given corresponding treatments intraperitoneally for 4 weeks. At the end of the study period, testicular atrophy index (TAI) and testicular sperm motility (TSM) were assessed and biochemical, histopathological, and immunohistochemical tests were performed. RESULTS TAI and TSM were higher in the ozone, hCG, and ozone plus hCG groups than in the sham surgery group (p=0.001). TSM in the ozone group was significantly higher than in the hCG and ozone plus hCG groups. In biochemical analyses, the parameters of oxidative stress (GPx1, MDA, CAT, GSH, SOD) indicated increased oxidative activity in cryptorchidism, which was resolved by applying ozone and hCG (p=0.001). In addition, apoptotic markers, Caspase 3 and bcl-2 were significantly decreased by applying ozone and hCG (p=0.001). CONCLUSIONS Results of this study suggest that ozone therapy, either as a single agent or in combination with hCG, is a promising approach for protection of testicular functions.

Study of the protective effect of vitamin C on testicular tissue following experimental unilateral cryptorchidism in rats.

This study investigated the role of vitamin C as an antioxidant in protecting the testis against damage in experimental unilateral cryptorchidism. Forty-five male Wistar Albino rats were divided into three groups. The control group had intact rats, the cryptorchid group had unilateral cryptorchid rats and the treatment group had unilateral cryptorchid rats that it received vitamin C at a dose of 50 mg kg(-1) body weight intraperitoneal, once a day, during experimental period. Histopathological samples were obtained from five cases of 15 animals of each group at 15, 30 and 60 days after induction of cryptorchidism. The results showed histopathological parameters of the cryptorchid (left) testes in the cryptorchid group significantly decreased compared with the control group (P < 0.05), and treatment with vitamin C after 60 days significantly improved all parameters of these testes compared with the cryptorchid group (P < 0.05). In addition, the left testes on unilateral cryptorchid rats had noticeable adverse effects on the scrotal (right) testes (P < 0.05). Treatment with vitamin C after 60 days significantly improved all parameters of these testes compared with the cryptorchid group (P < 0.05). It can be concluded that treatment with vitamin C significantly improved histopathological parameters in scrotal testes on unilateral cryptorchid rats.

Protective effects of N-acetylcysteine on experimentally undescended testis.

PURPOSE: We evaluated the efficacy of N-acetylcysteine for testicular damage induced by undescended testes in rats. MATERIALS AND METHODS: Flutamide was injected in the abdomen of pregnant rats daily from days 14 to 20 of gestation. Male offspring with cryptorchidism were randomly divided into 2 groups. Healthy male rats without undescended testes comprised the control group (group 1). Group 2 (undescended testes without N-acetylcysteine) received no treatment. Group 3 (undescended testes plus N-acetylcysteine) received intraperitoneal N-acetylcysteine daily. At 70 days after experiment initiation the testes were removed for histopathological and biochemical analysis. RESULTS: Mean malonyl dialdehyde values were lowest in group 1 and highest in group 2. In group 3 malonyl dialdehyde levels were significantly lower than in group 2 (p <0.001). Conversely, mean glutathione peroxidase was highest in group 1 and lowest in group 2. Glutathione peroxidase levels in group 3 were significantly higher than in group 2 (p <0.001). Histopathological differences between groups 1 and 3 in the modified Johnsen score were not significant (p = 0.041). However, the differences between these groups and group 2 were significant (p <0.001). The median apoptotic cell count did not differ between groups 1 and 3 but it was significantly higher in group 2 than in the other groups (p = 0.03 and <0.001, respectively). CONCLUSIONS: N-acetylcysteine may alleviate undescended testis induced damage to testes through its antioxidant effects. The underlying mechanism of these effects merits further investigation. Long-term studies are also needed as well as comparative animal and human studies.

Protective effect of α-tocopherol on damage to rat testes by experimental cryptorchidism.

It is thought that the degeneration of germ cells associated with an increase in the temperature due to cryptorchidism involves oxidative stress. α-Tocopherol is a powerful antioxidant that prevents oxidation of polyunsaturated fats found in membranes and stabilizes peroxyl radicals. For this reason we were interested in determining the role of α-Tocopherol using experimental cryptorchidism, followed by orchidopexia in neonatal rats. Eighty-four, 10-day-postpartum (dpp) male rats (Wistar strain) were used and divided into 7 groups: healthy control, sham with α-Tocopherol treated with 30 or 100 mg/kg doses, sham vehicle, cryptorchidism treated with α-Tocopherol at 30 or 100 mg/kg doses and cryptorchidism vehicle. Cryptorchidism was surgically induced at 10 dpp. At 25 dpp the animals were treated with α-Tocopherol and the vitamin vehicle. Lipoperoxidation and testicular morphology was determined in half of the animals at 40 dpp (short term). The remaining animals underwent orchidopexia and fertility was determined at 90 dpp. Testicular morphology was determined at 120 dpp (long term) in these animals. A significant reduction of lipoperoxidation was observed in the cryptorchid group treated with α-Tocopherol compared to the untreated cryptorchid group, in addition to short-term histological alterations. At long term, we observed an increase in the area and maturation of the seminiferous epithelium, a decrease in apoptosis and histological alterations and an increase in fertility from α-Tocopherol treatment. α-Tocopherol treatment decreased lipoperoxidation, possibly stabilizing free radicals produced during cryptorchidism, reducing morphological testicular alterations and favoring fertility.

Inguinal hernias and genital abnormalities in young Jordanian males.

This study was carried out to document the prevalence of inguinal hernia, hypospadias, undescended testis and varicocele in 3057 male applicants to the military wing of Mu'ta University in the south of Jordan. Age range was 17-20 years. A total of 250 men had one the 4 conditions: 93 (3.0%) had inguinal hernia; 15 (0.5%) had undescended testis (26.7% bilateral); 59 (1.9%) had hypospadias; 83 (2.7%) had varicocele (98.79% on the left side). Prevalence of inguinal hernia and undescended testis were comparable with international prevalence rates, while the rate for hypospadias was higher and that for varicocele lower. A birth defects registration system would help in planning preventive and treatment strategies.

The use of a genetic-counselling program by Dutch breeders for four hereditary health problems in boxer dogs.

Our group developed a genetic-counselling program for boxer-dog breeders in The Netherlands, using data for cryptorchidism (uni- and/or bilateral), epilepsy, knee-problems (including ligament rupture, fractured or ruptured meniscus, severe osteo-arthrosis of the knee, or a combination of these disorders), and schisis (including cheiloschisis, palatoschisis, or cheilopalatoschisis). We transformed the estimated breeding values (EBVs) into odds ratios (ORs), to enable the breeder to compare the risk for each of the traits for a certain dam-sire combination with the average population risk (set at 1). The goal of the study was to evaluate the use of our genetic-counselling program by Dutch breeders of boxer dogs. We asked breeders of the Dutch Boxer Club to send in an application form for genetic-counselling from June 1 to December 1, 2000. Breeders indicated on this application form three desirable sires for their dam (sire 1, sire 2, sire 3) in random order. On the basis of this information, a counselling report was produced which included ORs for the four diseases in litters of the dam-sire combinations indicated on the application form. Together with the counselling report, the breeders received an evaluation form. We received 129 application forms from 70 breeders, and collected 125 evaluations. Of these evaluations, 96 were informative about the influence of the counselling report on sire choice. The most-important criteria used by breeders to select sires were: the exterior characteristics (60%) and known progeny (52%). Although it was the first time breeders could make use of genetic-counselling, 32% of the breeders indicated that the genetic-counselling played a major role in their sire selection. Breeders expressed little difference in importance for the four genetic traits, but there was a tendency to consider epilepsy more than the others. Breeders hesitated to put long-term population interest above short-term personal interest. Nevertheless, the general conclusion of this study is that breeders acknowledge the value of counselling, especially to reduce the frequency of a threatening disease such as epilepsy.

Criptorquidia en el Servicio de Urología del Hospital II IPSS Tarapoto, San Martín / Cryptorchidia a t the urology service of Tarapoto IPSS Hospital

Se revisaron los datos de 352 operaciones urológicas rfealizadas entre mayo de 1990 y diciembre de 1995, en el Hospital IPSS de Tarapotol. Se encontró que se había ubicado 36 operaciones correctivas de criptorquidia, en la mayoría de las cuales usaron la técnica de Bevan. Los resultados incluyen la presentación de la frecuencia de patologías en las 352 operaciones ujrológicas realizadas en el período de estudio. También se describen los tipos y localización de las criptorquidias, así como su distribvución por edad, la incidencia de hernia inmguinal la frecuencia de la posición testicular operatoria y el diagnóstico anatómico del tamaño testicular. Se concluye que la orquidopexia es la tercera en frecuencia en las operaciones urológica revisadas y qaue la criptorquidia unilateral derecha es la más frecuente. Finalmente, se recomienda la realización de cursos de educación médica conrtinua acerca de criptorquidias, dirigida a médicos generales y pediatras de esta región geográfica. Asimismo que se debe dar el mayor énfasis al problema en el examen de todo infante en el Programa de Control del Niño Sano, para evitar futuras complicaciones

Prevention of seminiferous tubular atrophy in a naturally cryptorchid rat model by early surgical intervention.

In an attempt to determine whether the seminiferous tubular atrophy of the cryptorchid testis is preventable by early surgical correction of the cryptorchid state, aberrantly developed gubernacula destined to result in a cryptorchid testis in the Long-Evans cryptorchid (LE/ORL) rat were surgically reimplanted to the bottom of the scrotum on day 10 to 12 of age. Testis descent was monitored and the changes in testicular histology and in the volumes of the seminiferous tubules and Leydig cells were examined at day 60. As expected, normal testis descent occurred on or about day 25. Compared to untreated undescended testes at day 60, relative seminiferous tubular volumes (volume: % +/- SEM) were significantly increased by early surgical reimplantation of the gubemacula (89 +/- 1 vs. 66 +/- 3; P < 0.01). Absolute seminiferous tubular volumes (microliter +/- SEM) were also significantly increased by early surgical intervention when compared to undescended nontreated testes (893 +/- 27 vs. 170 +/- 12; P < 0.01). The testes of the surgically corrected cryptorchid animals were similar in all respects to those found in the descended testes of the sham-operated controls. Relative Leydig cell volume (% +/- SEM) was increased in the untreated cryptorchid testes compared to the surgically corrected testes (5.2 +/- 0.6 vs. 1.2 +/- 1.0; P < 0.05). Relative Leydig cell volumes in the surgically corrected testes were not significantly different from those found in the sham-operated descended controls. A modest but significant (P < 0.05) increase in absolute Leydig cell volume was also noted in the cryptorchid testes when compared both to normal controls or surgically corrected cryptorchid testes. From these observations, we conclude that early gubernaculopexy reverses the histologic changes normally seen in the cryptorchid rat testis to a relatively normal histologic architecture. These data provide experimental evidence to support the value of orchiopexy in the treatment of cryptorchidism.

Epidermal growth factor reverses antiandrogen induced cryptorchidism and epididymal development.

Epidermal growth factor, an androgen responsive paracrine factor, administered to pregnant mice has been reported to result in persistent wolffian ducts in female offspring. This fact led us to investigate whether epidermal growth factor can reverse the undescended testes and epididymal abnormalities associated with time specific flutamide administration. Timed pregnant Sprague-Dawley rats were treated with flutamide (undescended testes 74% and epididymal anomalies 53%) or flutamide plus epidermal growth factor (undescended testes 24% and epididymal anomalies 9%). The decrease in undescended testes and epididymal abnormalities following epidermal growth factor treatment was significant at p < 0.01. We performed immunohistological studies to evaluate whether flutamide alters epidermal growth factor expression in the paratesticular tissues during the time of maximal androgenic activity. These investigations revealed that antiandrogens did not alter epidermal growth factor expression in the fetal testes or epididymides. This finding suggests that epidermal growth factor does not reverse epididymal abnormalities or undescended testes by direct stimulation of the wolffian ducts or fetal testis.

Alterations in maternal epidermal growth factor (EGF) effect testicular descent and epididymal development.

OBJECTIVE: (1) To determine the ability of epidermal growth factor (EGF) to reverse antiandrogen-induced cryptorchidism and epididymal abnormalities; (2) to evaluate whether alterations in maternal EGF would result in abnormal testicular descent or mal development of the epididymis. METHODS: Experiment 1: Timed pregnant ICR mice were treated with either flutamide, flutamide plus EGF, or vehicle alone on gestational days 11 through birth. Experiment 2: Maternal EGF was abolished by removing the submandibular glands. Following timed mating, dams were treated with either flutamide, anti-EGF, DHT, or vehicle alone on gestational days 11 through birth. RESULTS: Experiment 1: Treatment with flutamide resulted in a 36 percent (26/72) incidence of undescended testes (UDT), and a 43 percent (31/72) incidence of abnormal epididymides. Rats treated simultaneously with flutamide plus EGF had a reduced incidence of UDT (14%, 6/42) and epididymal anomalies (19%, 8/42); p < 0.01. Experiment 2: The absence of maternal EGF resulted in a significant incidence of cryptorchidism in 11/50 (22%) testes, and epididymal anomalies in 19/50 (38%); p < 0.01. CONCLUSIONS: Our findings suggest that EGF stabilizes the wolffian duct system and partially mediates testicular descent.

Medical audit of community screening for undescended testes in Halton District.

OBJECTIVES: To evaluate the quality of screening in the community for undescended testis and to determine the percentage of boys referred for surgery before their second birthday. DESIGN: Population-based retrospective study of hospital and community clinic records of boys resident in Halton Health District who underwent surgery for undescended testicle during 1987-89. SETTING: Community Child Health Clinics. Medical Records Department of referral hospitals surrounding Halton district. RESULTS: During 1987-89, 76 boys from Halton district underwent surgery for undescended testis in the neighbouring hospitals: 64 (84%) of these boys had attended at least one screening examination in the community clinics; 12 (16%) did not attend any. Among the 64 who attended 18 (28%) were not considered to have undescended testis. Of the 30 boys diagnosed to have undescended testis after school entry, six were diagnosed incidentally in hospital and 24 were detected by general practitioners and School Medical Service. The age range for surgical referral varied from six months to 12.5 years. The percentage of boys referred for surgery before their second birthday was only 13%. CONCLUSION: Many cryptorchid boys were not picked up in community screening and many of these did not receive early surgery. This audit report suggested the following recommendations: (a) increased awareness of the condition amongst doctors and health visitors performing screening examination; (b) early surgical referral. These recommendations have been implemented in the district. Our further plan is to repeat the audit in three years to assess changes.

Screening for undescended testes.

There is a general consensus among surgeons and paediatricians that cryptorchidism should be detected and treated by the age of 5 years, if not earlier. In reality, incomplete descent is often detected well beyond the age of 5. The effectiveness of local screening was assessed by reviewing the screening history and subsequent management of boys treated for undescended testes between 1983 and 1986. The detection rates for undescended testes in the boys under 5 years were low both for doctors and health visitors. In contrast, the school entry medical examination for 5 and 6 year olds had a higher screening yield. Over 40% of cases came to light as a result of parents recognising the condition and seeking medical opinion. Late detection had also occurred in some boys who had had previous genital surgery or examination. Methods to improve early detection and ways of monitoring proposed changes are discussed.

[The place of the screening method in the early discovery and treatment of cryptorchism]. / Miasto na skriningoviia metod za rannoto otkrivane i lechenie na kriptorkhizma.

According to modern concepts, only early operative treatment of cryptorchidism may guarantee good results. This has been proven by the results obtained by the authors, who for five years have on mass prophylactic examination (1984-1988) searched for cryptorchidism in children attending kindergartens in the town of Tolbukhin. In this way the authors succeeded in realising highest operative activity in the age from 3 to 4 years.