Sequential observation of 2,6-dimethylaniline-induced nasal lesions in a rat two-stage nasal carcinogenesis model after initiation with N-bis(2-hydroxypropyl) nitrosamine

Male F344 rats received diet containing 3,000 ppm 2,6-dimethylaniline (DMA) after initiation with a single subcutaneous injection of 2,400 mg/kg of N-bis(2-hydroxypropyl)nitrosamine (DHPN), and histological and electron microscopic examinations of the nasal cavity were performed at 4, 13, 26 and 52 weeks to examine sequential changes induced by DMA. Severe atrophy of Bowman's glands and epithelial disarrangement were apparent from week 4, followed by dilatation and/or proliferation of Bowman's glands, degeneration of epithelial cells, and proliferation of undifferentiated epithelial cells from week 13. Focal glandular hyperplasias, dysplastic foci, and adenomas were observed from week 26, and carcinomas at 52 week. These nasal lesions were mostly evident in the olfactory mucosa in the nasal cavity, and their severity and/or incidences, other than atrophy of Bowman's glands, increased with the treatment period. Electron microscopically, carcinoma cells demonstrated desmosomes, dense secretory granules identical to those in normal Bowman's glands, a basement membrane, and microvilli. These results suggest that Bowman's glands are the target of DMA, giving rise to nasal carcinomas after DHPN-initiation.     

The effects of different iodide availabilities on thyroid function during development in Japanese quail

Japanese quail (Coturnix japonica) were used to study the effects of different egg iodide (I) availabilities on thyroid function during development. Low (less than 50 micrograms 1/kg feed in the maternal diet) and high (1200 micrograms 1/kg feed) I availability were compared to control levels (800 micrograms 1/kg feed), a standard supplementation for game bird feed. We measured thyroid gland content of I, triiodothyronine (T3) and thyroxine (T4), plasma concentrations of T3 and T4, hepatic 5' monodeiodinase (5'-D) activity, and the response of the thyroid gland to thyrotrophin (TSH) stimulation. Embryos, on day 14 of the 16.5-17 day incubation period, and 1-day chicks were used for most studies but thyroid gland hormone content and plasma hormone concentrations were determined for more stages. With high I, thyroidal I content was elevated but thyroidal T4 and T3 were not different from controls. Plasma T3 and T4, the thyroid gland response to TSH stimulation, and hepatic 5'-D activity did not differ between control and high I. Reduced body weight occurred with high I. In general, thyroid gland weight was not altered, but some high I birds exhibited thyroid hypertrophy and altered thyroid gland function. With low I availability, thyroid gland contents of I and T4 were reduced but thyroidal T3 content was maintained. The thyroid gland response to TSH stimulation, plasma thyroid hormone concentrations, and the developmental patterns of plasma thyroid hormones, hepatic 5'-D activity, body weight and thyroid weight were not different between control and low I groups. Developing Japanese quail exhibit excellent ability to adjust thyroid function over a wide range of I availabilities. Regulation appears to occur at the level of thyroid hormone synthesis in the thyroid gland, which allows most aspects of thyroid dynamics to remain unchanged in the maintenance of circulating thyroid hormone concentrations.     

The effects of androgens and gonadotropins on testicular development in the prepubertal rat.

Treatment of male rat pups from five to 34 days of age with dihydrotestosterone or 5 alpha-androstane-3 alpha, 17 beta-diol, resulted in reduced testicular size at 35 days of age. This appeared to be due to decreased tubular diameters and reduced spermatocyte numbers, especially late pachytene cells in stages X to XIII. In rat pups treated with dihydrotestosterone or 5 alpha-androstane-3 alpha, 17 beta-diol, treatment with luteinizing hormone resulted in some restoration of tubular diameter and spermatocyte numbers. In 5 alpha-androstane-3 alpha, 17 beta-diol treated rat pups partial restoration of testes size resulted from follicle-stimulating hormone treatment, but tubular diameter and spermatocyte numbers were depressed. Single serum samples collected at 35 days of age were analysed for luteinizing and follicle-stimulating hormone concentrations. These data indicated that the effects of administered androgens were due primarily to depression of circulating gonadotropin concentrations but a direct inhibition at the level of the testis could not be ruled out.     

Maternal exposure to low doses of bisphenol a has no effects on development of female reproductive tract and uterine carcinogenesis in Donryu rats

Effects of maternal exposure to low doses of bisphenol A (BPA), including those comparable with human exposure levels, on growth and development of the female reproductive system and uterine carcinogenesis in Donryu rats were investigated. Dams were administered BPA (0, 0.006 and 6 mg/kg/day) daily by gavage from gestation day 2 up to the day before weaning (postnatal day 21 at offspring). The serum levels of BPA were significantly elevated in the dams receiving 6 mg/kg/day, however, BPA levels in the milk of dams, and those in the serum and liver of offspring were similar between control and treated groups. The treatment did not exert any influences on uterine development including weight, gland genesis and estrogen receptor alpha expression, vaginal opening and gonadotropin secretion in the female offspring up to puberty. After maturation, no effects were evident with regard to estrous cyclicity in female offspring treated with BPA. In addition, the treatment had no effects on age-related morphological changes of the reproductive and endocrine organs and uterine carcinogenesis until 15 months of age. The results demonstrate that maternal exposure to BPA at levels comparable to human exposure did not have any effects on the female reproductive system of offspring in rats. In addition, BPA was also found in the serum, milk and liver of control dams and pups, and low levels of BPA were detected in drinking water and pellet diet. The present study showed that the experimental animals were also exposed to environmental BPA in the animal room.     

Short term effects of carbon disulfide on the intracardial irritation development and transmission in the rat

Studies were conducted into effects of acute and subacute CS2 application on intracardiac irritability and conduction in rat. Aconitine-induced arrhythmia and the coronary occlusion method were used as pathophysiological models. Under physiological conditions, short-time exposure to CS2 caused deceleration of intracardiac impulse conduction, while under pathophysiological conditions, modified arrhythmia or reduced survival rate was the result.     

The neuroprotective effects of lidocaine and methylprednisolone in a rat model of retinal ischemia-reperfusion injury

Retinal ischemia is a common cause of visual impairment for humans and animals. The neuroprotective effects of lidocaine (LDC) and methylprednisolone (MP) upon retinal ischemic injury were investigated in a rat model. Sprague-Dawley rats were divided into 3 groups, the IR control, LDC and MP. A very high intraocular pressure (HIOP) and retinal ischemia were induced. In LDC group, LDC bolus (1.5 mg/kg) was i.v. injected 30 min before ischemia and then a constant rate infusion (CRI) with 2 mg/kg/hr was given until 60 min after reperfusion. In MP group, MP bolus (30 mg/kg) was i.v. administered twice at 2 min before and immediately after ischemia, respectively. The HIOP damage to retina was evaluated by electroretinogram (ERG) and morphometrical histology. The functional analysis of the retina by ERG revealed a 35.2% reduction of a-wave in the IR group, 49.7% reduction in the LDC group but no significant change in the MP group compared to normal controls. An 81.0% reduction of b-wave was observed in the IR group, 80.7% reduction in the LDC group and 17.6% reduction in the MP group. In the morphometrical histology, the retinal inner plexiform layer/outer nuclear layer (IPL/ONL) ratio was reduced to 48.8% in the IR group, 80.1% in the LDC group and 96.2% in MP group. In conclusion, the MP showed significantly good neuroprotective effects on retinal IR injury, and the LDC showed moderate neuroprotective effects demonstrated in retinal structure but not in retinal function.     

Hypofractionated radiation therapy for invasive thyroid carcinoma in dogs: a retrospective analysis of survival

Thirteen dogs with invasive thyroid carcinoma (WHO classification T2b or T3b) seen between January 1991 and October 1997 were treated by external beam irradiation. Four once-weekly fractions of 9 gray of 4 MeV X-rays were administered. Four of the dogs died of progression of the primary disease and four from metastatic spread. Of the remaining dogs, three died of unrelated problems, although two were still alive at the time of the censor. Kaplan-Meier analysis of the survival time from first dose to death from either primary or metastatic disease gave a median survival time of 96 weeks (mean 85 weeks, range six to 247 weeks). Radiographic evidence of pulmonary metastatic disease at presentation had no prognostic value whereas crude growth rate was a highly significant factor. The present series indicates that radiation therapy should be considered an important modality for the control of invasive thyroid carcinoma in the dog.     

Preventive effect of zinc against cadmium-induced oxidative stress in the rat testis

The aim of this study was to investigate the antioxidant role of zinc (Zn) in the Cd-exposed testes of Wistar rats. Subchronic exposure to Cd (CdCl(2), 40 mg/l, per os) for 30 days resulted in a significant reduction in growth rate (-11%) and relative weights of testes (-36%) and seminal vesicles (-80%). Treated rats displayed a decrease in testicular and plasma testosterone levels, respectively (-70%, P<0.05; -48%, P<0.05), epididymal sperm count (-22%, P<0.05), and spermatozoa motility (-35%, P<0.05). In contrast, Cd increased the malondialdehyde (+46%, P<0.05), metallothionein (+200%, P<0.05), and 8-oxodGuo concentrations (+71%, P<0.05) in the testis. In the gonad, Cd decreased the GPx (-30%, P<0.05), CAT (-32%, P<0.05), mitochondrial Mn-SOD (-34%, P<0.05), and cytosolic CuZn-SOD (-32%, P<0.05) activities. Zinc supplementation (ZnCl(2), 40 mg/l, per os) in the Cd-exposed rats restored the activities of GPx, CuZn-SOD, and Mn-SOD in the testes to the levels of the control group. Moreover, zinc administration was capable of reducing the elevated levels of malondialdehyde in the testis. Interestingly, zinc supplementation attenuated DNA oxidation induced by Cd in the gonad and restored the testosterone level and sperm count to the levels of the control group. Zinc administration minimized oxidative damage and reversed the impairment of spermatogenesis and testosterone production induced by Cd in the rat testis.     

中药复方促孕散对不孕大鼠模型卵巢、子宫发育的影响

为了探讨中药复方制剂促孕散对肾虚不孕大鼠模型(ASR)的卵巢、子宫发育的影响,试验取9日龄雌性SD大鼠于颈背部皮下注射丙酸睾丸酮制备肾虚不孕大鼠模型,将其分为模型组、中药治疗组、正常组,从第80日龄用药,28 d后断头处死,剖腹取出双侧卵巢、子宫,将卵巢、子宫剥离干净后,称重、记录,制作石蜡切片,观察其形态结构变化。结果表明:模型组卵巢重(0.057±0.009)g,子宫重(0.363±0.023)g,卵泡数(6.20±0.63)个,子宫内膜面积(2.71±0.27)mm2,明显小于正常组[卵巢重(0.129±0.019)g、子宫重(0.572±0.023)g、卵泡数(23.00±1.63)个、子宫内膜面积(20.74±2.21)mm2]和中药治疗组卵巢重[(0.115±0.035)g、子宫重(0.532±0.095)g、卵泡数(20.30±2.58)个、子宫内膜面积(10.03±0.95)mm2]。说明中药复方制剂促孕散能够有效治疗肾虚不孕大鼠,增加不孕大鼠卵巢、子宫重,促进其卵泡发育及成熟。     

Evaluation of the effects of clomipramine on canine thyroid function tests

To evaluate the effect of long-term clomipramine administration on the hypothalamic-pituitary-thyroid axis in healthy dogs, 14 healthy adult dogs were enrolled in a prospective study. Clomipramine (3 mg/kg PO q12h) was administered to all dogs beginning on day 0, and continued for 112 days. Serum total thyroxine (T4), free thyroxine (fT4), 3,5,3'-triiodothyronine (T3), 3,3',5'-triiodothyronine (reverse T3; rT3), and thyroid-stimulating hormone (TSH) were measured on days 0, 7, 28, 42, 56, and 112. Thyrotropin-releasing hormone (TRH) response tests were performed concurrently. Significant decreases were noted in serum T4, f4, and rT3 concentrations beginning on day 28 through the end of the study period. The lowest mean (+/-SEM) concentrations of T4 (26 +/- 1.2 to 17 +/- 0.5 nmol/L) and rT3 (1.21 +/- 0.13 to 0.83 +/- 0.08 nmol/L) occurred at day 112, whereas the lowest mean fT4 (29 +/- 2.4 to 18 +/- 1.7 pmol/L) was found on day 56 of clomipramine treatment. The effect of treatment over time on serum T3 concentration also was significant, but the deviation in T3 from baseline was variable. No significant effect of clomipramine treatment was noted on either pre- or post-TRH TSH concentrations. The 35 and 38% decreases in serum T4 and fT4 concentrations, respectively, during clomipramine administration may lead to a misdiagnosis of hypothyroidism. Although no evidence of hypothyroidism was noted in this study population, subclinical hypothyroidism may have occurred. A longer duration of treatment might further suppress thyroid function, and concurrent illness or other drug administration might exacerbate clomipramine's effects.     

The effects of thyroid hormones on the skin of beagle dogs

The effects of hypothyroidism on canine skin were determined by comparing morphologic, morphometric, and hair cycle differences in skin biopsy samples from 3 groups of age- and gender-matched Beagle dogs: (1) euthyroid dogs; (2) dogs made hypothyroid by administration of 131I; and (3) dogs made hypothyroid and maintained in a euthyroid state by treatment with synthetic thyroxine. After 10 months of observation, there was slower regrowth of hair 2 months after clipping in the untreated-hypothyroid dogs. Untreated-hypothyroid dogs had a greater number of follicles in telogen and fewer hair shafts (ie, a greater number of hairless telogen follicles) than did the control group. The control dogs had a greater number of telogen follicles but the same number of hair shafts as the treated-hypothyroid group. Treated-hypothyroid dogs had the greatest number of follicles in the growing stage of the hair cycle (anagen). This study suggests that, at least in Beagles, induced hypothyroidism does not affect the pelage as dramatically as has been described in naturally occurring disease. This is because normal Beagles retain hair shafts in follicles for long periods, and the alopecia of hypothyroidism appears to evolve slowly because of the prolongation of this haired telogen stage. The evaluation of thyroxine-treated hypothyroid dogs demonstrates that thyroid hormone supplementation of Beagle dogs with induced hypothyroidism stimulates hair growth.     

Effect of preservation methods on the performance of bovine pericardium graft in a rat model

This study investigates the effect of preservation methods on the performance of bovine parietal pericardium grafts in a rat model. Mid-ventral full thickness abdominal wall defects of 3 x 2.5 cm in size were created in 90 male Sprague-Dawley rats (300-400 g), which were divided into three groups of 30 rats each. The abdominal defects of group one and two were repaired with lyophilized and glycerolized bovine pericardium grafts, while the defects of group three were repaired with expanded polytetrafluoroethylene (ePTFE) Mycro Mesh as a positive control. Another group of 30 rats underwent sham operation and was used for comparison as negative control. Each group of rats (n = 30) was divided into five subgroups (n = 6) and killed at 1, 3, 6, 9 and 18 weeks post-surgery for gross and morphological evaluations. The rats tolerated the surgical procedure well with a total mortality of 0.05%. No serious post-operative clinical complications or signs of rejection were encountered. Adhesions between the grafts and the underlying visceral organs observed in the study were mostly results of post-surgical complications. Glycerol preservation delayed degradation and replacement of the grafts, whereas lyophilization caused early resorption and replacement of the grafts. The glycerolized grafts were replaced with thick dense fibrous tissue, and the lyophilized grafts were replaced with thin loose fibrous tissue. The healing characteristic of the bovine pericardium grafts was similar to those of the sham-operated group, and quite different from those of the ePTFE Mycro Mesh. The outcome of the present study confirmed the superiority of glycerolized bovine pericardium grafts over its lyophilized counter part.     

The effects of endometrial damage on placental and fetal development in a mare

A 13-year-old ex-polo pony maiden mare with an extensively damaged endometrium was initially used as an embryo donor in a research herd. Embryos recovered from the mare were of normal morphology and size. Subsequently, the mare was inseminated and left pregnant. Early conceptus development was normal and she foaled a small (22 kg) but healthy filly after a gestation of 360 days. Examination of the placenta showed that development of the microcotyledons accurately reflected the damage to the uterus. Plasma progestagen profiles throughout gestation also provided evidence for placental pathology and fetal stress.     

Immunohistochemical studies on the development of endocrine cells in the thyroid and parathyroid glands in the golden hamster

The development of endocrine cells in the thyroid and parathyroid glands in the golden hamster was studied immunohistochemically in relation to the formation of these glands. The thyroid was formed on day 9 of gestation by the ventral outpocketing of the foregut between the first and second branchial pouches. The thyroid epithelial cells were faintly thyroglobulin-immunoreactive on day 10.5 of gestation. This immunoreaction became intense thereafter, but was almost confined to the cytoplasm of epithelial cells until birth. It appeared in the follicular lumen in newborn animals. The ultimobranchial body was derived from the fifth pouch and fused with the thyroid on day 12 of gestation. Calcitonin-immunoreactive cells first appeared on day 14 of gestation in the dorsomedial part of the thyroid derived from the ultimobranchial body and increased in number and intensity thereafter. Somatostatin-immunoreactive cells also appeared in the dorsomedial part of the thyroid derived from the ultimobranchial body on day 13 of gestation, and increased in number in newborn animals, but decreased thereafter. The parathyroid was derived from the third pouch, situated on day 13 of gestation on the dorsolateral side of the thyroid, and surrounded by a common capsule with the thyroid. Parathormone-immunoreactive cells first appeared on day 15 of gestation in the parathyroid and increased in number and intensity after birth.     

Preventive treatment with vitamin E alleviates the poisoning effects of carbon tetrachloride in cattle

Fifteen yearling steers were used to study the preventive effect of vitamin E on the protection against free radicals produced by carbon tetrachloride (CCl4). The animals were randomly divided in three equal groups and treated as follows: group A--previously injected (i.m.) with 15 IU/kg BW on the 15th and second day before the trial and drenched with 0.05 ml/kg BW CCl4; group B--only drenched with the same dose of CCl4; group C--drenched with a placebo. Food intake was recorded and blood samples collected daily for 8 days after the CCl4 drenching to compare the activity of aspartate aminotransferase (AST), gamma-glutamyltransferase (gamma-GT) and the levels of erythrocyte reduced glutathione (GSH) and serum malonyldialdehyde (MDA). Food intake was reduced in group B for the first 3 days (P < 0.05); higher activities of AST and gamma-GT were observed in the poisoned groups, nevertheless the overall values were lower in the group A than B (P < 0.02); only the group A reached the basal values of AST at the seventh day; higher levels of GSH and MDA were recorded in the poisoned cattle indicating the generation of free radicals. It was concluded that the preventive use of Vitamin E lessened the damage in hepatic tissue caused by the free radicals and prevented the anorexia caused by CCl4.     

Anti-nutritional effects of a moderate dose of soybean agglutinin in the rat.

This study was conducted to investigate the effects of a moderate dose of purified soybean agglutinin on performance and nitrogen digestibility in rats as well as to determine its effects on the protein, DNA and RNA content of the small intestine and pancreas. Twenty-four Sprague-Dawley rats were randomly allotted into one of four groups for a 10-day nitrogen balance experiment. The four groups of rats were fed 7 g of a casein-cornstarch based diet or a similar diet supplemented with 0.1, 0.2 or 0.4 mg/g purified soybean agglutinin. All experimental diets were adjusted to an identical nutrient level. Dose of soybean agglutinin had no significant effect on rat performance. Incorporation of soybean agglutinin in the diet reduced apparent protein digestibility and the utilization of dietary protein by increasing nitrogen loss from the faeces and urine. Fresh pancreatic weight increased in rats fed soybean agglutinin at a level of 0.4 mg/g in the diet compared to the control, but the dry pancreatic weight and the protein content of the pancreas did not differ among the four groups. However the DNA and RNA content of the pancreas had a tendency to increase with a higher level of soybean agglutinin. The weight of the jejunum and its protein, DNA and RNA content were not significantly affected by soybean agglutinin, but the dry weight and the RNA of the jejunum tended to increase with higher levels of soybean agglutinin in the diet. In conclusion, purified soybean agglutinin, at moderate levels in the rats diet, had negative effects on digestive function, such as nitrogen digestibility, nitrogen retention and nitrogen balance. As the level of soybean agglutinin increased, the effects became more pronounced. Meanwhile, hypertrophy of the pancreas was observed with higher doses of soybean agglutinin incorporation in the diets.