Expression of D2-40 in adjunct diagnosis of papillary thyroid carcinoma

The clinical pathologic criteria for nuclear features of papillary thyroid carcinoma are subjective and sometimes cannot distinguish carcinoma from adenomatous goiter and follicular neoplasms. No single antibody has demonstrated high sensitivity or specificity in making these distinctions. Using quantitative analysis of immunohistochemical staining with D2-40, a recently available monoclonal antibody used as a lymphatic endothelial marker, we examined 72 cases of papillary carcinoma. Controls included 36 follicular adenomas, 36 follicular carcinomas, and 20 adenomatous goiters with papillary hyperplasia. Cytoplasmic D2-40 immunoreactivity was present in 60 of 72 papillary carcinomas, 2 cases of follicular adenoma and 2 cases of follicular carcinoma, whereas no adenomatous goiter or normal thyroid glands contained positive epithelial cells. Overexpression of D2-40 in papillary thyroid carcinomas thus has potential diagnostic utility in differentiating these tumors from their potential histologic mimics.     

Dipeptidyl aminopeptidase IV staining of cytologic preparations to distinguish benign from malignant thyroid diseases.

Staining for dipeptidyl aminopeptidase IV (DAP IV [EC: 3.4.14.5]) activity was applied to aspiration biopsy specimens or imprint preparations of surgical biopsy specimens from thyroid tumors. Material was obtained from 55 patients with histologically proven thyroid diseases: 9 with papillary carcinoma, 5 with follicular carcinoma, 11 with follicular adenoma, 13 with adenomatous goiter, 8 with Hashimoto's thyroiditis, and 9 with other benign conditions. Most tumor cells, follicular lumina in cell clusters, and intranuclear inclusions were strongly positive for DAP IV in all examples of papillary or follicular carcinoma. In contrast, only a few epithelial cells were labeled for DAP IV in follicular adenoma and adenomatous goiter. Some Hürthle cells in Hashimoto's thyroiditis also were positive for DAP IV. When a DAP IV scoring system based on the percentage of positive cells and staining intensity was used, all benign tissues except one (from a follicular adenoma) were found to have extremely low scores. These results indicate that staining for DAP IV activity is a simple but useful tool to aid in distinguishing benign from malignant thyroid neoplasms.     

Thyroid pathologic findings in patients with Cowden disease

We describe the histologic findings in thyroid glands from six female and five male patients with Cowden disease. The patients were aged 9 to 43 years (mean age, 26 years). The salient thyroid lesions in this syndrome were multicentric follicular adenomas and adenomatous (parenchymatous, hyperplastic) nodules showing a wide range of nonspecific cytoarchitectural patterns. Multiple tiny cellular foci, so-called microadenomas, were also a feature. Specific lesions composed of oxyphil or clear cells, a tumor with features of hyalinizing trabecular adenoma, and an adenolipoma also occurred. Two cases showed a follicular carcinoma in addition to multiple benign follicular cell proliferations. The follicular carcinomas occurred at an older age and were larger in size than the clinically significant benign nodular lesions, suggesting tumor progression. All tumors showed thyroglobulin immunoreactivity and were negative for calcitonin. The histologic findings of a multiple adenomatous goiter or multiple follicular adenomas, particularly in children and young adults, should alert the pathologist and physician to the possibility of an inherited trait, such as Cowden disease, with its implications for family screening. The tumors are usually benign and well demarcated, but, because of multicentricity and increased risk of recurrence or progression to carcinoma, total thyroidectomy should be advocated.     

An immunohistochemical study of calcitonin-containing cells in benign and malignant thyroid lesions

Parafollicular cells (C-cells) in benign and malignant thyroid lesions were studied immunohistochemically with a polyclonal anti-calcitonin (CT) antibody. The C-cells were seen most frequently in the middle third of the lateral lobes in the thyroid gland of normal individuals and patients with Graves' disease and chronic thyroiditis, although in the latter the number of such cells was significantly decreased (p less than 0.05). In adenomatous goiter, C-cells were present in nodular lesions from an early stage of nodule development (frequency about 19%), whereas in the later stage these cells were rarely observed inside type 1 nodules, which were generally characterized by an admixture of follicles with considerably different sizes. However, C-cells were not observed inside type 2 nodules, which were composed of similar-sized follicles, or in the parenchyma of 56 cases of benign and malignant thyroid tumors. These findings suggest that since C-cells are present in nodular lesions, the histogenesis of adenomatous goiter is quite different from that of follicular adenoma; thyroid neoplasms generally contain no C-cells in the parenchyma.     

Papillary carcinoma of the thyroid. A histochemical, immunohistochemical and ultrastructural study with special reference to the follicular variant

Twenty-one papillary thyroid carcinomas (PTCs), grouped into predominantly papillary (14 cases), predominantly follicular (5 cases), and extremely follicular, i.e., follicular variant (2 cases) types, were studied in comparison with three cases each of follicular lesions including follicular carcinoma, follicular adenoma, adenomatous goiter and Graves' disease. Histochemical, immunoperoxidase, and electron microscopic analyses demonstrated no remarkable differences between the predominantly papillary and predominantly follicular PTCs, but the presence of common characteristics distinct from those of the follicular lesions. These two types of PTCs showed less glycogen, more mucoid material, more epidermal keratin, less thyroid hormone with relative predominance of T3 over T4, and more interdigitating reticulum cells (IDCs) than most of the follicular lesions. Ultrastructurally, the tumor cells of these PTCs had markedly irregular nuclei, a vesicular chromatin pattern, and small basally located lysosomes, in contrast with the cells in the follicular lesions which had smooth round nuclei, more heterochromatin, and apical or dispersed lysosomes of various sizes. The follicular variant PTCs showed some mixed features, such as glycogen in the follicular portion and mucoid material in metastatic papillary foci, positive keratin and IDCs but greater amounts of thyroid hormone, and a rather intermediate type of ultrastructure with only mildly irregular but vesicular nuclei and large apical as well as small basal lysosomes. These findings cytologically support the WHO definition of papillary carcinoma that includes tumors with variable mixtures of papillary and follicular patterns. However, separate consideration may be necessary with regard to the follicular variant.     

Histochemical and biochemical study on adenylate cyclase and 5'-nucleotidase activity in thyroid glands with normal and various thyroid diseases

The adenylate cyclase and 5'-nucleotidase activity was measured biochemically in the thyroid glands from patients with various thyroid diseases in comparison with normal thyroid. The basal adenylate cyclase activity in normal thyroid was 159.3 p-moles cAMP/min./g tissue. The activity was elevated to 230% of basal with 20 mM NaF and 190% of basal with 100 mU/ml TSH. These values in chronic thyroiditis and Graves' disease were not significantly different from the values of normal thyroid. In adenomatous goiter, adenoma and carcinoma, the basal adenylate cyclase activity was significantly higher than that of normal thyroid. Parallel to the biochemical determination of both enzyme activities, the distribution of histochemically demonstrable adenylate cyclase and 5'-nucleotidase activity was described in the follicular cells with normal and various thyroid diseases. The reaction product of adenylate cyclase and 5'-nucleotidase activity was restricted to the plasma membrane of the follicular cells. However, the distribution and intensity of the adenylate cyclase reaction varied in each thyroid disease, except for the absence of reaction product in the basal plasma membrane. The lack of demonstrable adenylate cyclase activity in the basal plasma membrane suggests the possibility that the basal plasma membrane may not play an important role of TSH-reception.     

Occurrence and significance of vascular invasion in multinodular adenomatous goiter

Solitary follicular neoplasms of the thyroid gland are usually classified as adenomas or carcinomas according to two main criteria: vascular invasion and capsular penetration. No information is available on the occurrence of vascular invasion in multinodular goiter lesions, except for the case of a follicular carcinoma within a goiter. One thousand consecutive cases of multinodular adenomatous goiter were reviewed. After screening all H&E-stained slides, 5 patients with histological features typical of adenomatous goiter but displaying foci of vascular invasion at the periphery of the nodes were selected. A single vessel (2 patients) and 2–4 vessels (3 patients) at the periphery of different nodules were involved, with clusters of follicular cells lined by endothelium and partly filling the lumen. Clinical information was obtained from all patients: No recurrences or progressive disease were reported 14 to 16 years after operation. These findings indicate that presence of minimal vascular invasion, although a valuable criterion of differentiation in solitary follicular lesions of the thyroid, has little diagnostic importance in the setting of multinodular adenomatous goiter. It does not appear to justify a diagnosis of malignancy and does not indicate a need for further therapy.     

HISTOCHEMICAL AND BIOCHEMICAL STUDY ON ADENYLATE CYCLASE AND 5'-NUCLEOTIDASE ACTIVITY IN THYROID GLANDS WITH NORMAL AND VARIOUS THYROID DISEASES

The adenylate cyclase and 5'-nucleotidase activity was measured biochemically in the thyroid glands from patients with various thyroid diseases in comparison with normal thyroid. The basal adenylate cyclase activity in normal thyroid was 159.3 p-moles cAMP/min./g tissue. The activity was elevated to 230% of basal with 20 mM NaF and 190% of basal with 100 mU/ml TSH. These values in chronic thyroiditis and Graves'disease were not significantly different from the values of normal thyroid. In adenomatous goiter, adenoma and carcinoma, the basal adenylate cyclase activity was significantly higher than that of normal thyroid. Parallel to the biochemical determination of both enzyme activities, the distribution of histochemically demonstrable adenylate cyclase and 5'-nucleotidase activity was described in the follicular cells with normal and various thyroid diseases. The reaction product of adenylate cyclase and 5'-nucleotidase activity was restricted to the plasma membrane of the follicular cells. However, the distribution and intensity of the adenylate cyclase reaction varied in each thyroid disease, except for the absence of reaction product in the basal plasma membrane. The lack of demonstrable adenylate cyclase activity in the basal plasma membrane suggests the possibility that the basal plasma membrane may not play an important role of TSH-reception.     

Histologic changes in Graves' thyroid gland after 131I therapy for hyperthyroidism.

The effect of 131I therapy was examined in 13 thyroid glands affected by Graves' disease 3 to 29 years after irradiation for hyperthyroidism. All of the patients had clinically palpable thyroid nodules. Two patients were in a latent hypothyroid, 6 in a euthyroid and 5 in a hyperthyroid state. The microscopic changes in the thyroids showed a pattern of multiple adenomatous nodules with cystic changes, marked oxyphilic cell changes with nuclear atypism and various degrees of chronic thyroiditis. Immunohistochemical staining for TG and T4 was negative to mildly positive in these oxyphilic cells and entirely negative for EGF and CEA. The DNA ploidy pattern was diploid pattern in 6 cases. One papillary-type microcarcinoma occurred, but there was no evidence of a relationship between the tumor and the irradiation. The pathologic findings in Graves' thyroid gland after 131I therapy are not specific, but pathologists should differentiate this lesion from adenomatous goiter, which occurs with no apparent cause, or from thyroid carcinoma because of the marked nuclear atypism of this lesion.     

Thyroid carcinoma in benign thyroid diseases. An analysis from minute carcinoma

The relative frequency of thyroid carcinoma in benign thyroid diseases such as toxic diffuse goiter (toxic goiter), adenomatous goiter (goiter), adenoma, and chronic thyroiditis (thyroiditis) was studied using 3,219 surgically removed thyroid glands. Coexistence of carcinoma and benign diseases was found in 257 glands. Among them, 157 glands had minute carcinoma (diameter of cancer nodule is smaller than 10 mm) and 100 glands had advanced carcinoma (larger than 10.1 mm in diameter). The incidence of carcinoma including minute carcinoma was 29.4% in goiter (98/333), 21.0% in thyroiditis (11/53), 8.6% in adenoma (55/638), and 5.3% in toxic goiter (98/1852). Chi-square test also revealed that the rate of carcinoma not only advanced carcinoma but also minute carcinoma was higher in goiter than in the other diseases (P less than 0.01, respectively). Among the patients aged under 39, the incidence of advanced carcinoma and minute carcinoma in thyroiditis were the highest, respectively (83% and 100% in thyroiditis, 9.0% and 11.4% in goiter, 2. 6% and 1.9% in adenoma, and 0.7% and 3.4% in toxic goiter), however, among the patients aged over 40, they were secondary lower and the lowest, respectively (19% and 16.8% in goiter, 5.2% and 7.5% in adenoma, 4.2% and 6.7% in thyroiditis, and 1.4% and 7.5% in toxic goiter). We concluded that adenomatous goiter accompanies carcinoma more frequently than other benign thyroid diseases and the high incidence of carcinoma in chronic thyroiditis is probably due to a preoperative selection of the patients.     

The diagnostic value of flow cytometric DNA measurements in selected disorders of the human thyroid

A flow cytometric (FCM) analysis of biopsies from 19 patients with thyroid lesions (Hashimoto's thyroiditis, hyperplastic goiter, colloid and adenomatous nodular goiter, follicular adenoma, follicular carcinoma, anaplastic carcinoma) was performed to evaluate the relevance of such results for the individual diagnosis and prognosis and prognosis of such diseases. All cases of Hashimoto's thyroiditis, the colloid nodular goiters, and the hyperplastic goiter had a unimodal diploid pattern, while the anaplastic carcinomas were aneuploid. These lesions are, however, easily diagnosed on morphological grounds and their FCM classification would only be redundant. Follicular adenomas and follicular carcinomas may be either unimodal diploid or aneuploid, and our single case of adenomatous colloid goiter showed a bimodal DNA distribution. This means that a diploid modal DNA content in a follicular tumor does not rule out malignancy, while aneuploidy is not an unequivocal sign on malignancy. The evaluation of the percentage of cells in S-phase did not seem to contribute valuable information for the individual diagnosis of malignancy within the group of thyroid lesions studied. Our study indicates a rather limited clinical value of FCM measurements of DNA in the individual case of thyroid disease.     

THYROID CARCINOMA IN BENIGN THYROID DISEASES An Analysis from Minute Carcinoma

The relative frequency of thyroid carcinoma in benign thyroid diseases such as toxic diffuse goiter (toxic goiter), adenomatous goiter (goiter), adenoma, and chronic thyroiditis (thyroiditis) was studied using 3,219 surgically removed thyroid glands. Coexistence of carcinoma and benign diseases was found in 257 glands. Among them, 157 glands had minute carcinoma (diameter of cancer nodule is smaller than 10 mm) and 100 glands had advanced carcinoma (larger than 10.1 mm in diameter). The incidence of carcinoma including minute carcinoma was 29.4% in goiter (98/333), 21.0% in thyroiditis (11/53), 8.6% in adenoma (55/638), and 5.3% in toxic goiter (98/1852). Chi-square test also revealed that the rate of carcinoma not only advanced carcinoma but also minute carcinoma was higher in goiter than in the other diseases (P<0.01, respectively). Among the patients aged under 39, the incidence of advanced carcinoma and minute carcinoma in thyroiditis were the highest, respectively (83% and 100% in thyroiditis, 9.0% and 11.4% in goiter, 2. 6% and 1.9% in adenoma, and 0.7% and 3.4% in toxic goiter), however, among the patients aged over 40, they were secondary lower and the lowest, respectively (19% and 16.8% in goiter, 5.2% and 7.5% in adenoma, 4.2% and 6.7% in thyroiditis, and 1.4% and 7.5% in toxic goiter). We concluded that adenomatous goiter accompanies carcinoma more frequently than other benign thyroid diseases and the high incidence of carcinoma in chronic thyroiditis is probably due to a preoperative selection of the patients. ACTA PATHOL. JPN. 35: 781–788, 1985.     

Enhanced B-Raf protein expression is independent of V600E mutant status in thyroid carcinomas

BRAF (7q24) encodes a serine/threonine protein kinase, and its expression level varies in different tissues. Although a high prevalence of BRAF mutation has been suggested as an important event in thyroid tumorigenesis, little is known about the expression pattern of B-Raf in the thyroid. Thus, we examined the expression of B-Raf in various neoplastic and nonneoplastic thyroid tissues and compared it with BRAF mutational status. Normal and hyperplastic thyroid tissues showed focal and faint immunoreactivity for B-Raf, especially in cuboidal follicular cells of small follicles. In contrast, diffuse expression of B-Raf was observed in follicular adenomas and well-differentiated carcinomas. The missense point mutation BRAF(V600E) was identified in 42% (13/31 cases) of papillary carcinomas and 33% (5/15 cases) of undifferentiated carcinomas but not in normal thyroid tissues, nodular hyperplasia, follicular adenomas, or follicular carcinomas. The immunohistochemical expression of B-Raf did not correlate with BRAF mutational status. Moreover, Western blotting revealed that B-Raf expression in thyroid carcinoma cell lines was also independent of BRAF mutation. Serum or fibroblast growth factor-1 stimulation further activates ERK1/2 in heterozygous BRAF(V600E)-positive carcinoma cells as well as BRAF(V600E) mutation-negative carcinoma cells. In conclusion, heterogeneous focal expression of wild-type B-Raf in nonneoplastic tissues may play a role in the growth or functional activity of thyroid follicular cells. In contrast, diffuse expression of wild-type and/or mutant B-Raf may be involved in the tumorigenic process resulting in activation of the mitogen-activated protein kinase signaling pathway in cooperation with other genetic abnormalities and activation of ligand-receptor signaling pathways.     

胃动素及其前体mRNA在正常人甲状腺和甲状腺肿瘤组织中表达的研究

目的 探讨胃动素和胃动素前体mRNA基因在正常人甲状腺的表达,比较人甲状腺胃动素前体mRNA基因序列与人小肠胃动素前体mRNA基因序列的异同;观察胃动素活性肽和胃动素前体mRNA在甲状腺肿瘤中的表达,并比较它们在良、恶性甲状腺肿瘤表达的异同及临床意义.方法 采用RT-PCR、Southern杂交和分子克隆等技术,克隆并测定人甲状腺和人小肠黏膜内胃动素前体mRNA基因序列;采用荧光免疫组织化学双染技术、Western印迹和即时荧光定量PCR(real-time PCR),观察胃动素和胃动素前体mRNA在正常甲状腺和甲状腺良、恶性肿瘤组织中表达的异同.结果 (1)正常人甲状腺组织有胃动素和胃动素前体mRNA的表达,且胃动素和降钙素共表达于同一细胞,即甲状腺C细胞;(2)人甲状腺组织内胃动素前体mRNA基因序列与基因库报道的人小肠胃动素前体mRNA基因序列(BC112314,NCBI,美国)完全相同;(3)免疫荧光组织化学、Western印迹以及real-time PCR结果均显示,正常人甲状腺和甲状腺肿瘤组织内均有胃动素和胃动素前体mRNA的表达,其中甲状腺髓样癌和嗜酸性腺瘤胃动素及其前体mRNA的表达高于正常甲状腺组织(均P＜0.05);但甲状腺乳头状癌和滤泡癌胃动素和胃动素前体mRNA的表达则明显降低(均P＜0.05);而结节性甲状腺肿与正常甲状腺组织相比胃动素和胃动素前体mRNA的表达差异均无统计学意义(P＞0.05).结论 人甲状腺组织有胃动素和胃动素前体mRNA的表达,且胃动素主要表达于甲状腺C细胞;人甲状腺组织胃动素前体mRNA基因序列与人小肠胃动素前体mRNA基因序列完全相同;甲状腺髓样癌和嗜酸性腺瘤内胃动素及其前体mRNA的表达明显增高,而甲状腺乳头状癌和滤泡癌内胃动素及其前体mRNA的表达明显降低.提示胃动素可能通过影响甲状腺滤泡旁细胞的分泌活动参与其生理活动的调节,胃动素可能与临床甲状腺髓样癌和甲状腺嗜酸性腺瘤疾病的发生和发展有关.     

Patterns of basement membrane laminin distribution in nonneoplastic and neoplastic thyroid tissue.

Laminin, a major basement membrane component, is typically absent or partially lost around the epithelial elements of most invasive carcinomas. To evaluate the distribution of laminin in both primary and metastatic thyroid tumors, we studied 14 benign thyroid lesions (eight adenomas, two Graves' disease, two Hashimoto's thyroiditis, one adenomatous hyperplasia, one nodular goiter), 20 carcinomas (seven papillary, six tall cell variant, four follicular, three Hürthle), and eight metastases (five tall cell variant, three follicular) utilizing a polyclonal antibody against highly purified, nidogen-free laminin. All benign lesions showed positive, linear immunostaining along basement membranes. Partial loss or absence of laminin was seen in the solid areas of all types of thyroid carcinomas examined; well-differentiated papillary and follicular tumors, as well as papillary and follicular areas of more poorly differentiated neoplasms, maintained linear laminin immunostaining in the papillary cores beneath the epithelial cells and around follicles. A similar correlation between laminin deposition and architectural organization was seen in metastatic lesions. Hürthle cell carcinomas had a unique fragmented, pericellular immunostaining pattern around individual tumor cells, suggesting uncontrolled laminin synthesis. Our findings suggest that preservation of laminin production in thyroid tumors reflects their degree of differentiation and that absence of laminin correlates with lack of structural organization rather than reflecting invasive and metastatic potential.     

Morphometric evaluation of histological sections of the thyroid gland in benign and malignant follicular lesions

A morphometric analysis was made on histological sections of thyroid tissue in various aspects of follicular cell pathologies (goiter, adenoma, and carcinoma) and in normal follicular cells. Three cases of each aspect and 150 cells of each case were measured. The results obtained demonstrate the importance of the nuclear area in the differential diagnosis of normal thyroid, goiter, adenoma, and carcinoma. The other parameters considered, i.e., perimeter, maximum diameter, and nuclear form factor, proved to be able to exclusively distinguish between benign lesions (normal thyroid, goiter and, adenoma) and malignant ones (carcinoma).     

Zur Ultrastrukturpathologie der vorbehandelten Basedow-Struma

Summary This electron microscopic study is based on 12 cases of toxic goiter treated preoperatively with iodide, thionamides and lithium. Essential features of Graves' disease include hypertrophy of the follicular epithelium, papillary follicular infoldings and frequently, lymphocytic infiltration and fibrosis. After preoperative therapy most thyroid glands show involutional changes of varying degree with reaccumulation of colloid and flattening of the follicular epithelium. In a small number of histologically examined cases (n=84) colloid goitre (13%) and nodular goitre (8%) were noted. Lymphocytic infiltration and interstitial fibrosis do not show any correlation with preoperative treatment. Ultrastructurally, different stages of cellular involution are seen. The hypertrophic follicular cell shows an increased cell surface with numerous long microvilli and some pseudopodia at the apical border and an occasionally thickened, basal lamina. The cytoplasm contains a well developed system of organelles which synthesizes (rough endoplasmic reticulum, Golgi apparatus, cytoplasmic vesicles) and degrades (colloid droplets, lysosomes) thyroglobulin. With increasing cellular atrophy we found cells to be reduced in size but to contain well developed thyroglobulin-synthesizing organelles. Finally, we observed completely atrophic flat cells with only a few organelles.There are at date no electron microscopic communications on lithiumpretreated thyroid glands.After lithium therapy characteristic changes are observed at the membranes of the cytocavitary system, which are often found to be thin and occasionally to be damaged. The rough endoplasmic reticulum is virtually deprived of ribosomes and the Golgi apparatus seems to be underdeveloped. The number of apical microvilli is reduced. The morphological findings following iodide-, thionamide- and lithium pretreatment in Graves' disease are discussed in light of recent biochemical results.