Effects of Talinum paniculatum (Jacq.) Gaertn. leaf extract on general toxicity and pubertal development of rats.

Talinum paniculatum (Jacq.) Gaertn. (Talinaceae), popularly known as "major gomes," is a Brazilian Cerrado plant used in traditional medicine and as a food source. Recent studies have demonstrated its diuretic effects. However, no studies have been performed on its effects on the reproductive system. Therefore, we aimed to investigate the effects of the ethanol-soluble fraction of T. paniculatum leaves (ESTP) on general toxicity and on the pubertal development of male and female Wistar rats. For this purpose, the uterotrophic and the pubertal assays were performed. In the uterotrophic test, female immature rats were treated for three consecutive days with 30, 100, or 300 mg/kg of ESTP. Uterus without luminal fluid was weighed and the relative weight calculated. For the pubertal assay, male and female immature rats were submitted to 30-day treatment with 30 or 300 mg/kg of ESTP. Clinical signs of toxicity, biochemical, and histopathological parameters were evaluated. ESTP treatment did not promote estrogenic effects in female rats. In the pubertal test, no daily signs of toxicity or weight loss were observed. Moreover, ESTP did not affect the onset of vaginal opening and preputial separation and did not cause significant changes in biochemical parameters as well as in organ weight and histopathological analyses of animals.

Experimental cryptorchidism enhances testicular susceptibility to dibutyl phthalate or acrylamide in Sprague-Dawley rats.

Cryptorchidism (CPT), the most common male congenital abnormality, is variably associated with other male reproductive tract problems. We evaluated if cryptorchid rats develop enhanced testicular susceptibility to dibutyl phthalate (DBP) or acrylamide (AA) after extended exposure. Three studies with rats were performed: (1) in utero and postnatal exposure to DBP or AA; (2) establishment of CPT and orchiopexy; and (3) in utero and postnatal exposures to DBP or AA associated with CPT/orchiopexy. Seminiferous tubules were histologically scored according to the severity of lesions: (1) Rats exposed to DBP (score 1.5) or AA (score 1.1) presented mostly preserved spermatogenesis. Some seminiferous tubules showed vacuolated germinative epithelium, germ cell apoptosis, and a Sertoli cell-only (SCO) pattern. (2) CPT (score 3.3) resulted in decreased absolute testes weights, degenerated and SCO tubules, and spermatogenesis arrest that were reversed by orchiopexy (score 1.1). (3) Exposure to DBP or AA with CPT/orchiopexy led to atrophic testes, spermatogenesis arrest, germ cell exfoliation/multinucleation, and SCO tubules (both chemicals score 2.5). Exposure to chemicals such as DBP or AA prevented the recovery of cryptorchid testes by orchiopexy. The possible role of environmental contaminants should be considered when looking for factors that modulate human testicular disorders associated with CPT.

Domestic cat testicular aromatase activity as assessed by the tritiated water-release assay

Lower testicular tetosterone:17β-estradiol (T:E2) ratio was found in teratospermic domestic cats (<40% morphologically normal sperm). The aim of this study was to assess the reliability of the tritiated water-release assay (TWRA) to measure aromatase activity in domestic cat testes. Testicular T and E2 concentrations, measured by enzyme immunoassay, and sperm morphology were evaluated to verify the relationship between them. Aromatase activity was measured in microsomal fraction and in homogenates of cat testes. Rat ovaries and piglet testes were used for assay validation. Aromatase activity was not detected in cat testes microsomal fraction (n = 8), not even when the protein amount added to the assay was increased from 50 to 200 μg. In homogenates, however, it was detected (3.5 ± 0.5 pmol.g-1.h-1; n = 7), although in such low levels that no activity inhibition was detectedwhen homogenates were incubated with increasing fadrazole concentrations. Although none of the cats in this study were classified as teratospermic, some sperm defects were correlated with testicular T:E2 ratio (abnormal acrosome, r = -0.76) and with E2 concentration (proximal cytoplasmic droplet, r = 0.77). However, we did not find any correlation between aromatase activity and hormonalor sperm morphology data. To our knowledge this is the first demonstration of testicular aromatase activity in domestic cats. Despite that, due to the low aromatase activity measured and the lack of correlation with other reproductive data, we could not infer that TWRA is a reliable method to detect differences in testicular aromatase activity in normospermic cats. Perhaps this method could be used in teratospermic individuals that probably havean increased aromatase activity. As an alternative, we suggest that more sensitive techniques should be used to compare aromatase activity between normospermic and teratospermic cats.(AU)

Domestic cat testicular aromatase activity as assessed by the tritiated water-release assay

Lower testicular tetosterone:17β-estradiol (T:E2) ratio was found in teratospermic domestic cats (<40% morphologically normal sperm). The aim of this study was to assess the reliability of the tritiated water-release assay (TWRA) to measure aromatase activity in domestic cat testes. Testicular T and E2 concentrations, measured by enzyme immunoassay, and sperm morphology were evaluated to verify the relationship between them. Aromatase activity was measured in microsomal fraction and in homogenates of cat testes. Rat ovaries and piglet testes were used for assay validation. Aromatase activity was not detected in cat testes microsomal fraction (n = 8), not even when the protein amount added to the assay was increased from 50 to 200 μg. In homogenates, however, it was detected (3.5 ± 0.5 pmol.g-1.h-1; n = 7), although in such low levels that no activity inhibition was detectedwhen homogenates were incubated with increasing fadrazole concentrations. Although none of the cats in this study were classified as teratospermic, some sperm defects were correlated with testicular T:E2 ratio (abnormal acrosome, r = -0.76) and with E2 concentration (proximal cytoplasmic droplet, r = 0.77). However, we did not find any correlation between aromatase activity and hormonalor sperm morphology data. To our knowledge this is the first demonstration of testicular aromatase activity in domestic cats. Despite that, due to the low aromatase activity measured and the lack of correlation with other reproductive data, we could not infer that TWRA is a reliable method to detect differences in testicular aromatase activity in normospermic cats. Perhaps this method could be used in teratospermic individuals that probably havean increased aromatase activity. As an alternative, we suggest that more sensitive techniques should be used to compare aromatase activity between normospermic and teratospermic cats.

Influence of oily vehicles on fetal testis and lipid profile of rats exposed to di-butyl phthalate.

It has been hypothesized that oils containing high levels of omega-3 polyunsaturated fatty acids, such as canola and fish oil, could counteract some of the adverse effects induced by phthalates. In the present study, the influence of different oily vehicles on di-butyl phthalate (DBP)-induced testicular toxicity and lipid profile was investigated. Pregnant Wistar rats were treated by oral gavage from gestation days 13 to 20 with DBP (500 mg/kg/day) diluted in three different vehicles: corn, canola or fish oil. Male fetuses were analyzed on gestation day 20. DBP exposure lowered intratesticular testosterone levels and anogenital distance, regardless of the vehicle used. The percentage of seminiferous cords containing multinucleated gonocytes and cord diameter was increased in DBP-exposed groups, compared with vehicle controls, with no difference between the three DBP-exposed groups. Clustering of Leydig cells was seen in all DBP groups. Lipid profile indicated that administration of canola and fish oil can increase the content of omega-3 fatty acids in rat testis. However, content of omega-3 was diminished in DBP-treated groups. Overall, our results indicate that different oily vehicles did not alter fetal rat testicular toxicity induced by a high DBP dose.

Testicular testosterone: estradiol ratio in domestic cats and its relationship to spermatogenesis and epididymal sperm morphology.

The phenomenon of teratozoospermia in felids is not fully understood. In this study, we investigated the testicular androgen:estrogen balance in domestic cats and correlated these data with epididymal sperm morphology and the degree of spermatogenic activity. During spring and summer, testes and blood samples were obtained from 37 mixed-breed domestic cats (12 to 48 mo). The epididymal sperm were harvested and evaluated for sperm counts, motility, and morphology. Distal cytoplasmic droplets were not considered a defect, and samples were considered normozoospermic if they contained more than 60% normal sperm (N = 25) or teratozoospermic if they contained less than 45% normal sperm (N = 12). The testicular and serum concentrations of testosterone (T) and 17ß-estradiol (E2) were determined with an enzyme immunoassay. The gonadosomatic index and epididymal sperm numbers and motility did not differ between groups. The percentage of normal sperm was higher in normozoospermic (74.3 ± 2.0, mean ± SEM) than in teratozoospermic samples (43.1 ± 1.4). The most prevalent sperm defects in the teratozoospermic group were abnormal acrosomes (9.7 ± 2.0) and bent midpieces (12.2 ± 2.0) or tails (24.0 ± 2.7) with cytoplasmic droplets. Histomorphometric data were similar between groups, although there was a lower Leydig cell nuclear volume in teratozoospermic samples. Normozoospermic samples contained a higher percentage of haploid cells and had a higher index of total spermatogenic transformation than teratozoospermic samples. Serum concentrations of T (0.5 ± 0.1 vs. 0.8 ± 0.4 ng/mL) and E2 (9.5 ± 1.2 vs. 11.4 ± 2.3 pg/mL) and testicular T concentrations (471.6 ± 65.3 vs. 313.4 ± 57.6 ng/g) were similar between groups. However, compared with normozoospermic samples, teratozoospermic samples had higher testicular E2 concentrations (8.5 ± 3.6 vs. 5.4 ± 0.5 ng/g) and a lower T:E2 ratio (31.8 ± 4.1 vs. 87.2 ± 11.6). There were significant correlations between testicular E2 values and percentages of normal sperm (r = -0.55) as well as those with primary sperm defects (r = 0.58) or abnormal acrosomes (r = 0.64). The T:E2 ratio was also correlated with meiotic index (r = 0.45) and percentage of normal sperm (r = 0.58). In conclusion, a high testicular E2 concentration and a reduced T:E2 ratio were significantly associated with higher ratios of abnormal sperm types, suggesting that the balance between androgens and estrogens is an important endocrine component in the genesis of teratozoospermia in felids.