Thyroid Dysplasia in Wistar Hannover GALAS Rats

Thyroid dysplasia was recognized in WistarHan GALAS rats and confirmed as a heritable congenital disorder. The gene or genes involved were not identified, but homozygous animals with thyroid dysplasia also exhibited stunted growth, had reduced pituitary gland growth hormone (GH) and were hypothyroid. Heterozygous animals exhibited thyroid dysplasia with normal thyroid hormonal homeostasis and no difference in the incidence of preneoplastic or neoplastic lesions in oncogenicity studies.     

灰雁甲状腺的组织学研究

为研究灰雁与鹅的甲状腺结构的异同,采用不同方法,观察灰雁的甲状腺组织结构,在H-E染色的组织切片中,甲状腺实质由滤泡组成,滤泡由单层上皮细胞组成;在PAS反应中,甲状腺滤泡腔内的胶质深红染,显示胶质为糖类物质;在银染切片中,C细胞胞质内含有黑色的特异性嗜银颗粒,与太湖鹅、四季鹅不同。     

Thyroid follicular adenocarcinoma in a ferret

A 5-year-old male castrated ferret was presented to the Washington State University College of Veterinary Medicine for evaluation of progressive hair loss and a large, rapidly growing ventral neck mass. The patient had been diagnosed previously with an insulinoma, which was managed medically. Fine-needle aspirates of the neck mass were performed. The cytologic results were most consistent with epithelial neoplasia, likely a carcinoma; thyroid origin was considered likely based on tumor location and cell morphology. The tumor grew rapidly, and the owners elected euthanasia 1 week after examination. At necropsy, a circumscribed, ovoid mass disrupted the right cervical musculature next to the right lobe of the thyroid gland. Histopathologic evaluation revealed an infiltrative mass consisting of cuboidal cells arranged in solid sheets and irregular follicles enclosing colloid. The cells were large, with prominent nucleoli, and had a high mitotic rate. The histopathologic diagnosis was consistent with thyroid follicular adenocarcinoma. Immunochemical findings confirmed thyroglobulin production by neoplastic cells, but to a lesser extent than in normal ferret thyroid tissue. To our knowledge, this is the first case of thyroid follicular adenocarcinoma to be reported in a ferret, with only 1 other case of thyroid carcinoma, a C-cell carcinoma, described previously.     

Thyroid histological changes in lambs from iodine-supplemented and untreated ewes

AIM: To identify the histological changes in the thyroid associated with subclinical goitre. METHOD: Thyroids were histologically examined from lambs that died within 24 hours of birth in a trial to investigate the effect of iodine supplementation to Merino ewes on perinatal mortality. RESULTS: Thyroids with follicles depleted of colloid, infolded and lined by columnar epithelium are indicative of hypothyroidism. These findings can occur in thyroids considered to be in the normal weight range of less than 1.3 g. CONCLUSION: Thyroid histological changes are a useful aid in diagnosing subclinical goitre.     

Congenital diffuse hyperplastic goiter associated with perinatal mortality in 11 captive-born bottlenose dolphins (Tursiops truncatus).

Diffuse hyperplastic goiter was diagnosed by histopathology in 11 perinatal bottlenose dolphins (Tursiops truncatus) that died at four separate zoos and aquaria. Thyroid morphology of these animals was compared with the histologically normal thyroids of two stranded wild bottlenose dolphin calves, a neonate and a 2-mo-old calf. Histologic changes included reduced follicular luminal diameter, markedly reduced or absent luminal colloid, hypertrophy of follicular epithelium, and follicular dysplasia. The etiology of the thyroid gland lesion was not identified. Cause of death was not determined for most of these animals, but they were presumed to have died from metabolic derangements associated with the thyroid lesion, drowning, or dystocia.     

二狼山白山羊遗传性甲状腺肿发病机理及诊断研究——Ⅱ.甲状腺组织中TG分布、TG带谱分析及血清TG含量检测

应用BA-免疫组化结合图像分析测定了遗传性甲状腺肿病山羊(以下简称IHO)甲状腺组织中甲状腺球蛋白(TG)的含量和分布,用梯度-PAGE结合免疫印迹法分析了甲状腺组织中异常TG相关抗原蛋白的带谱及分子量;用BA-ELISA法测定了血清中TG含量.结果表明:(1)IHG纯合子病山羊甲状腺组织中TG含量明显低于对照组山羊(P<0.01),且TG在胶质中多为弱阳性,在增生上皮细胞内几乎为阴性;(2)IHO纯合子病山羊甲状腺组织中异常TG相关抗原蛋白为331000、110000,而IHG杂合子病山羊异常TG相关抗原蛋白为110000;(3)IHG杂合子病山羊血清中TG含量明显低于对照组(P<0.05).提示:IHG病山羊甲状腺激素合成不足是由于TG合成障碍,且其具体发病环节可能是TG亚单位(110000、331000相关TG抗原蛋白)进一步聚合为1200000的TG过程缺陷的结果;异常TG相关抗原蛋白110000,可作为净化IHG杂合子病山羊的特异性生化指标,其血清中TG含量降低,亦可作为剔除IHG杂合子山羊的生化指标之一.     

Histomorphometry and quantification of nucleolar organizer regions in bovine thyroid containing methylthiouracil residues

In order to study the morphology and morphometry and to characterize and quantify the nucleolar organizer regions (NORs) of bovine thyroids containing methylthiouracil (MTU) residues, five animals were orally treated with a suspension of MTU (5 g/animal/day) for 20 days (group A). This treatment protocol was interrupted 5 days before the the animals were slaughtered. Six animals receiving placebos composed group B. A third group (group C) was composed of normal thyroids obtained from a slaughterhouse. All glands were previously assessed for detection of antithyroid residues by chromatography, and only those glands from MTU-treated animals were positive. Follicles of glands from group A showed wide variation in size and shape. There was a predominance of small follicles covered by multiple layers of columnar cells, sometimes forming papillary projections into the lumen, characterizing severe interfollicular and intrafollicular adenomatosis. Many follicles had vacuolated cells with nuclei showing karyolysis or pyknosis and reduced amounts of a low-density and very excavated colloid. They also showed higher follicular epithelia and larger proportions of their structural components when compared with glands of groups B and C. In the thyroids from group A, the argyrophilic nucleolar organizer region-associated proteins (AgNORs) were greater in number, with small ones scattered all over the nucleus. Although the size of AgNORs in thyroids from groups B and C was variable, these AgNORs were fewer and larger than were those in glands from group A. In conclusion, the MTU induces proliferation and regressive changes in follicular cells, and the AgNOR technique is efficient to distinguish different degrees of thyroid hyperplasia.     

猪不同生长阶段甲状腺结构的形态计量学研究

为观察猪不同生长发育阶段甲状腺的形态学变化,本研究选用大约克夏猪,分别于初生、体质量为20、40、80和100kg时屠宰,测定甲状腺质量、长度、宽度、增殖细胞数量以及血清中甲状腺素的表达,并对猪的甲状腺石蜡切片进行组织学观察,应用Nikon NIS-Elements Documentation图像分析系统对甲状腺滤泡腔以及滤泡上皮细胞进行形态计量学指标的测量,并在不同生长阶段进行比较分析。结果表明,随着猪的生长发育,甲状腺增殖细胞的数量、甲状腺素的表达以及滤泡腔数密度、上皮细胞高度均逐渐降低;滤泡腔截面积及其等效直径、滤泡腔截面积密度逐步升高;而上皮细胞核截面积及其等效直径、上皮细胞核数密度变化规律不明显。相关分析表明,血清中FT3的表达与甲状腺宽度存在显著的负相关、与上皮细胞高度呈显著正相关;FT4、TT4的表达与体质量、甲状腺质量、甲状腺长度、甲状腺宽度、滤泡腔截面积、滤泡腔截面积等效直径存在显著负相关,与上皮细胞高度以及上皮细胞核数密度存在显著正相关。上述形态学测量结果为深入研究猪甲状腺在不同生长发育阶段所发生的形态学变化提供了准确的定量资料。     

Pathological characteristics of thyroid glands from Japanese Black Cattle living in the restricted area of the Fukushima Daiichi Nuclear Power Plant accident

To study the effect of ionizing radiation on thyroid glands, 66 Japanese Black cattle residing in the restricted area of the Fukushima Daiichi Nuclear Power Plant in 2013–2017 were pathologically examined. There were no clinical symptoms of thyroid disease in these cattle. Three cases of goiter and seven of atrophy were found in two among the four farms examined. Cases of goiter exhibited normal morphological structure without mass or nodule formation in thyroid glands. Cellular atypia or capsular invasion of the follicular epithelium was absent. The estimated integrated dose of external radiation in goiter cases ranged from maximum 797 mSv to minimum 24 mSv. All lobules in the seven atrophic thyroid glands were affected, but pathological findings, such as inflammatory cell infiltration or stromal fibrosis, were not observed. The estimated integrated dose of external radiation in atrophic thyroids ranged from maximum 589 mSv to minimum 8 mSv. Immunohistochemical analysis of anti‐nitroguanosine and the TUNEL method in goiter and atrophic thyroid glands did not reveal any positive findings. The present study indicates that there was no significant relationship between a radiation effect and pathological findings in any thyroid glands.     

Uterine Carcinosarcoma in a 2-year-old Female Wistar Hannover GALAS Rat

Carcinosarcomas are rare tumors in humans as well as rats and most commonly occur in the uterus. Recently, we observed a case of incidental carcinosarcoma of the uterus in a female Wistar Hannover GALAS [BrlHan:WIST@ Jcl (GALAS)] rat at 2 years of age. Histopathologically, the tumor was characterized by an admixture of malignant epithelial and nonepithelial elements. The carcinomatous components represented a type of endometrial carcinoma, consisting of glandular and solid proliferation of large-sized tumor cells. Prominent mitoses and tumor cell invasion were observed. The sarcomatous components were characterized by multifocal proliferation of severe atypical cells with cartilage matrix and were diagnosed as chondrosarcoma. Transitions between carcinomatous and sarcomatous components were observed, and many tumor cells in the solid lesion showed immunohistochemical reactivity with both cytokeratin and vimentin. Based on these findings, this tumor was diagnosed as a uterine carcinosarcoma. This is the first report of uterine carcinosarcoma in Wistar Hannover GALAS [BrlHan:WIST@Jcl (GALAS)] rats.     

Overexpression of c-Ras in hyperplasia and adenomas of the feline thyroid gland: an immunohistochemical analysis of 34 cases

Formalin-fixed, paraffin-embedded thyroid glands from 18 cats diagnosed with hyperthyroidism were evaluated immunohistochemically for overexpression of the products of oncogenes c-ras and bcl2 and the tumor suppressor gene p53. Fourteen thyroid glands from euthyroid cats without histologically detectable thyroid lesions were examined similarly as controls. Results from these investigations showed that all cases of nodular follicular hyperplasia/adenomas stained positively for overexpression of c-Ras protein using a mouse monoclonal anti-human pan-Ras antibody. The most intensely positively staining regions were in luminal cells surrounding abortive follicles. Subjacent thyroid and parathyroid glands from euthyroid cats did not stain immunohistochemically for pan-Ras. There was no detectable staining for either Bc12 or p53 in any of the cats. These results indicated that overexpression of c-ras was highly associated with areas of nodular follicular hyperplasia/adenomas of feline thyroid glands, and mutations in this oncogene may play a role in the etiopathogenesis of hyperthyroidism in cats.     

绵羊甲状腺细胞原代培养及鉴定

试验旨在建立一套完整的绵羊甲状腺细胞的原代培养方法,为绵羊甲状腺功能的体外研究奠定基础。通过常规消化分离得到甲状腺滤泡上皮细胞,光镜下对不同培养时间的细胞进行形态学观察,并结合RT-PCR、细胞免疫荧光和ELISA,对培养的绵羊甲状腺细胞从形态,甲状腺球蛋白(TG)、甲状腺过氧化物酶(TPO)、促甲状腺激素受体(TSHR)特异基因mRNA表达,TG特异抗原蛋白表达和甲状腺激素T3、T4的分泌功能等方面进行鉴定。结果显示,绵羊甲状腺细胞在体外呈贴壁式生长,具有上皮样细胞特点;TG染色呈胞浆阳性,具有特异TPO、TG、TSHR mRNA表达及分泌T3、T4的功能,但T3、T4的分泌量随培养时间的延长呈递减趋势。本研究成功地建立了简便可行的绵羊甲状腺细胞的原代培养方法,为深入研究绵羊甲状腺的功能提供了试验平台。     

Thyroid hyperactivity induced by methimazole, spironolactone and phenobarbital in marmosets (Callithrix jacchus): histopathology, plasma thyroid hormone levels and hepatic T4 metabolism

To determine drug-induced hyperfunction of marmoset thyroids due to inhibition of synthesis or enhancement of metabolic elimination of thyroid hormones, males were orally administered 10 and 30 mg/kg/day methimazole (MMI), 30 and 100 mg/kg/day spironolactone (SPL), or 50 mg/kg/day phenobarbital (PB) for 4 weeks. MMI caused marked hypertrophy of follicular epithelial cells in accordance with a significant decrease in the plasma thyroxin (T4) level. Hypertrophied epithelial cells were filled with dilated rough endoplasmic reticulum and reabsorbed intracellular colloids, and the luminal surface was covered with abundant microvilli. The colloid included vacuoles positive to anti T4 immuno-staining. SPL and PB also caused similar histomorphological changes, although they were less severe than those due to MMI and were not clearly associated with decrease in the plasma T4 levels. Hepatic T4 UDPGT activities tended to increase due to SPL and PB treatment, however, which were not so significant as increases in microsomal cytochrome P-450 contents. Some animals treated with SPL and PB showed marked increases in thyroid weights due to inactive dilated follicles. In conclusion, hyperactivity of thyroid follicles was induced in marmosets not only due to inhibition of T4 synthesis produced by MMI but also because of enhancement of hepatic T4 elimination produced by SPL and PB. However, hypertrophic effects of SPL and PB were less severe than MMI, because plasma T4 levels were maintained at almost pretreatment or control levels after SPL or PB treatment.     

Expression of class II beta-tubulin by proliferative myoepithelial cells in canine mammary mixed tumors

Benign mammary mixed tumors in dogs resemble human salivary pleomorphic adenomas with regard to their histogenesis, including the occurrence of cartilaginous or bony metaplasia as well as the expression pattern of cytoskeletal proteins in proliferative myoepithelial cells. Recently, a monoclonal antibody specific for class II beta-tubulin has been developed. The epitope it recognizes was determined to be the heptapeptide Glu-Glu-Glu-Glu-Gly-Glu-Asp, which is the common sequence found among the canine, rat, mouse, and human class II beta-tubulin-specific regions. We carried out immunohistochemical studies on mammary mixed tumors obtained from three female dogs using this the monoclonal antibody. The antibody to class II beta-tubulin reacted intensely with proliferative myoepithelial cells in canine mammary mixed tumors, whereas staining was barely detectable in normal myoepithelial cells surrounding alveoli and alveolar ducts within the tumor and adjacent normal tissue. Proliferative myoepithelial cells also expressed vimentin, but alpha-smooth muscle actin (alphaSMA) staining was barely detectable. Immunoblot analysis showed that class II beta-tubulin and vimentin were expressed in myoepithelial cell lines prepared from the three mammary mixed tumors. On the other hand, only one cell line, which was negative for alphaSMA, produced cartilage-specific type II collagen. These results suggest that class II beta-tubulin could be a new molecular marker of proliferating myoepithelial cells in canine mammary mixed tumors and that differential expression of cytoskeletal components is associated with cartilaginous metaplasia of proliferative myoepithelial cells in mixed mammary tumors.     

Background Data on Developmental Parameters During the Gestation Period in Rats

Abstract: Background data during the gestation period were obtained from 128 Wistar Hannover GALAS rats and 26 Crl:CD(SD) pregnant rats in the control groups of our previous toxicity studies. The body weights of dams in the Wistar Hannover GALAS rats were significantly lower throughout the gestation period than those in the Crl:CD(SD) rats. In contrast, the time-dependent change in the body weight gain (%) of dams showed very similar trends in both strains. The mean number of live embryos/fetuses in the Wistar Hannover GALAS rats was 12.0, and was lower than that (14.5) in the Crl:CD(SD) rats. The placental weights gradually increased with pregnancy progression and reached a plateau on gestation day (GD) 19, although the embryo/fetal weights rapidly increased from GD 17 to GD 21. The embryo/fetal weights in the Wistar Hannover GALAS rats were significantly lower on only GD 21 than those in the Crl:CD(SD) rats. It is considered that this fetal weight difference between the strains develops during the fetal period, but not during the organogenesis period. In contrast, there were no differences in the placental weights between the two strains. Microscopically, the thickness of the labyrinth zone in the Wistar Hannover GALAS rats was thicker throughout the gestation period than that in the Crl:CD(SD) rats.     

Pancreatic Ductal Adenocarcinoma in a Wistar Hannover GALAS Rat

There are no reported spontaneous cases of pancreatic ductal adenocarcinoma (PDAC), and there are few reports about chemically-induced PDAC in rats. We encountered a PDAC in a Wistar Hannover GALAS rat that had been subjected to a medium-term multiorgan carcinogenicity bioassay. This article describes the histological and histochemical findings of the tumor. The tumor was located in the pancreatic tissue and had not invaded the liver parenchyma or the mucosal layer of the alimentary tract. The tumor cells were atypical and were mainly arranged in small tubules. In addition, abundant stroma and mucus production were observed in the tumor. In an immunohistochemical examination, the tumor cells were positive for cytokeratin, Sox9 and pancreas duodenum homeobox 1 and negative for amylase 2A and insulin. Therefore, the tumor was diagnosed as a PDAC based on its histological and histochemical findings. We considered that the tumor was caused by the carcinogens administered during the abovementioned bioassay.     

Dysregulation of beta-catenin is common in canine sporadic colorectal tumors.

Human colorectal tumorigenesis is often initiated by APC (adenomatous polyposis coli) or beta-catenin (CTNNB1) mutations, which result in dysregulation of beta-catenin expression, followed by alterations in E-cadherin and/or p53. We examined 32 canine intestinal tumors for expression and intracellular distribution of beta-catenin, E-cadherin, and p53 using immunohistochemistry. beta-Catenin in normal mucosal epithelial cells was restricted to lateral cell membranes, but 13/13 (100%) colorectal adenomas had intense cytoplasmic and/or nuclear reactivity. Three of six (50%) colorectal carcinomas, 2/13 (15%) small intestinal carcinomas, and dysplastic cells in 1/2 focal hyperplastic lesions in the small intestine had a similar pattern of staining; remaining tumors had normal membranous beta-catenin reactivity. There was a correlation (P = 0.007) between abnormal beta-catenin and E-cadherin staining with 11/13 (85%) colorectal adenomas, 3/6 (50%) colorectal carcinomas, and 3/13 (23%) small intestinal carcinomas showing decreased membranous reactivity compared with normal mucosal epithelium. E-cadherin staining was reduced more often in adenomas than in carcinomas (P = 0.04). There were two patterns of nuclear p53 staining: > 60% of nuclei in 2/26 (8%) carcinomas (one colorectal, one small intestinal) were strongly labeled, whereas three colorectal adenomas and one small intestinal carcinoma had fainter staining in 10-20% of cells. Dysregulation of beta-catenin appears to be as important in canine colorectal tumorigenesis as it is in the human disease and could be due to analogous mutations. Malignant progression in canine intestinal tumors does not appear to be dependent on loss of E-cadherin or beta-catenin expression or strongly associated with overexpression of nuclear CMI antibody-reactivity p53.     

Hepatic microsomal enzyme induction and thyroid function in rats treated with high doses of phenobarbital or chlorpromazine.

Rats were treated daily with high doses of phenobarbital or chlorpromazine for 6 weeks. The animals were sacrificed after 1, 2, 4 and 6 weeks of treatment. At each occasion, plasma levels of ASAT, ALAT, LDH, AP, total T4 and T3, free T4 and T3, and TSH were determined. The thyroids and liver weights were recorded, and the microscopical examination was performed on the liver, thyroids and pituitary gland. The results showed hepatic morphological changes indicative of enzyme induction on weeks 1, 2 and 4. During the first 4 weeks, plasma T4 and T3 were decreased in many individuals and increased in few others, and after 6 weeks of treatment the values for T4 and T3 returned to near normal. An increase in ASAT and ALAT was found at the same occasions. Follicular cell hypertrophy in the thyroids was observed during the first 4 weeks of treatment, in both treated groups, and after 6 weeks the thyroid glands returned to normal, except for a few individuals from the phenobarbital group. The present study has shown that the effect of phenobarbital and chlorpromazine on the thyroid gland function is predominantly due to their effect on the peripheral hormone disposition, as a result of the hepatic microsomal enzyme induction.     

Induction of Thyroid Proliferative Changes in Rats Treated with Antithyroid Compound

This study was designed to investigate the histologic changes in goitrogen-induced thyroid growth of the rat. The animals were orally treated by 1% potassium perchlorate except the controls and were sacrificed in intervals ranging from 1 to 12 months. The thyroid weight increased progressively along the treatment and after 2 months showed a diffuse homogenous hypertrophy and hyperplasia of follicular cells, decreased amount of colloid and increased vascularity. After sixth month of treatment true nodules appeared with complex morphology.