Akira Sugenoya

Proliferative activity of human thyroid tumors evaluated by proliferating cell nuclear antigen/cyclin immunohistochemical studies

Background. To clarify the proliferative activity of human thyroid tumors, the labeling index of various thyroid tumors was evaluated by proliferating cell nuclear antigen (PCNA)/cyclin immunostaining.

Association of HLA-DPB1 ∗0501 with early-onset graves' disease in Japanese

To investigate the association between HLA antigens and Graves disease among Japanese, serologic typing and DPB1 genotyping using the PCR-RFLP method have been performed. HLA alleles of 106 patients with Graves disease were determined, and the frequency of HLA-B46 was found to be significantly increased. Furthermore, the frequencies of HLA antigens were compared between two age groups: early-onset and late-onset patients (under and over 20 years, respectively). It was found that the frequency of DPB1*0501 (88.9%) was significantly increased (pc < 0.004) in the early-onset group as compared with the healthy controls (55.0%) but not in the late-onset group (60.7%). On the other hand, a significant increase of HLA-B46 was observed in the late-onset patients (pc < 0.0004). These results suggest that the genetic background of Japanese patients with early-onset Graves disease is different from late-onset patients. Namely, the HLA-DP allele (DPB1*0501) and the HLA-B allele (B46) are primarily involved in the pathogenesis of early-onset and late-onset Graves disease in Japanese, respectively.

The outcome of patients with thyroid carcinoma and graves' disease

A total 847 consecutive patients with Graves disease who underwent thyroidectomy between 1965 and 1990 were found to have a 4.3% incidence of coincident carcinoma of the thyroid. In 68.2% of these cases the tumors were less than 1 cm in diameter and were detected in only 7 patients (19.4%) prior to surgery. Dissection of the regional lymph nodes was necessary in only 11 patients. A review of the patients in 1990 revealed no carcinoma-related deaths with metastases occurring most commonly in the patients with larger tumors and in those whose cancer had been detected prior to thyroidectomy. Although the detection of potential tumors occurring with Graves disease using preoperative echogram may be of interest, our results suggest that occult micro-carcinoma occurring coincidentally with Graves disease is not clinically significant as it did not cause any recurrence in this study. Moreover, there was no evidence to suggest that Graves disease was associated with the increased or rapid growth of these tumors.

Clinical and pathologie properties of small differentiated carcinomas of the thyroid gland

Ninety-one small differentiated thyroid carcinomas less than 10 mm in size treated during the last 33 years were subjected to clinical study. In 44 of the 91 cases, tumor was found incidentally at surgery, while in the remaining 47 a thyroid mass was palpable before operation. A preoperative diagnosis of carcinoma could not be made in nodules less than 5 mm in size. Ultrasonographic examination is now extending the limits of exact diagnosis. Metastasis was found in 16 of 33 patients who had lymph nodes dissected. Metastatic lymph nodes were distributed in the same sites as those with thyroid carcinomas larger than 10 mm. Pathologic examination was carried out in 108 small carcinomas including cases from an affiliated hospital. Histologically, all of them were papillary or follicular carcinomas. The growth pattern of the tumors was divided into 5 groups: 23 were nonencapsulated sclerosing type, 29 encapsulated type without capsule invasion, 48 encapsulated type with capsule invasion, 4 the infiltrative type, and 4 the round type. Lymph node metastasis and intraglandular dissemination were found most frequently in the infiltrative type, followed by the encapsulated type with invasion. The results suggest the necessity of lymph node dissection and lobectomy for patients with these histological types of small carcinomas who had only local removal of the tumor.RésuméQuatre-vingt un cancers de la thyroïde, de moins de 10 mm de diamètre, traités ces 33 dernières années ont été analysés. Dans 44 des 91 cas, la tumeur a été découverte fortuitement lors de la cervicotomie alors que dans 47 cas, une masse était cliniquement palpable avant lintervention. Le diagnostic préopératoire de cancer nétait pas possible lorsque le diamètre de la tumeur était inférieur à 5 mm. Léchographie élargit actuellement les possibilités du diagnostic. Parmi les 33 patients qui ont eu un curage ganglionnaire, 16 avaient des métastases. La distribution des métastases était similaire à celle quon observait lorsque le diamètre de la tumeur est supérieur à 10 mm. Lexamen anatomopathologique de 108 cas de cancer de petite taille a démontré que la plupart de ces tumeurs étaient du type papillaire ou folliculaire. Selon leur type de développement, on distinque 5 groupes: 23 faisaient partie du type non encapsulé sclérosant, 29 étaient du type encapsulant sans invasion de la capsule, 48 étaient du type encapsulé avec invasion de la capsule, 4 étaient du type infiltrant, and 4, du type circonférentiel. Les métastases lymphatiques et latteinte disséminée de la glande étaient retrouvées le plus souvent dans le type infiltrant, ensuite dans les cancers encapsulés avec invasion. Ces résultats suggèrent quil faut pratiquer une lobectqmie avec curage ganglionnaire pour les “petits” cancers de la thyroïde, surtout lorsque la première intervention a consisté seulement en une résection locale.ResumenEl sujeto del presente estudio clínico estuvo constituido por noventa y un pacientes con carcinomas pequeños de la glándula tiroides tratados en los pasados 33 años. En 44 de los 91 casos el tumor fue hallado en forma incidental en el curso de la cirugía y en los 47 restantes la masa tiroidea aparece palpable antes de la operación. El diagnóstico preoperatorio de carcinoma no pudo ser establecido en los nódulos menores de 5 mm. El examen ultrasonográfico ha logrado ampliar las posibilidades de un diagnśtico exacto. Se encontraron metástasis en 16 de 33 pacientes en quienes se disecaron los ganglios linfáticos. La distribución de los ganglios metastásicos fue la misma que en los casos de carcinoma tiroideo mayor de 10 mm. El examen patológico fue efectuado en 108 carcinomas pequeños, incluyendo casos de un hospital afiliado; histológicamente todos fueron papilares o foliculares. El patrón de crecimiento de los tumores fue clasificado en 5 grupos: 23 del tipo esclerosante no encapsulado, 29 del tipo encapsulado sin invasión de la capsula, 48 del tipo encapsulado con invasión de la capsula, 4 del tipo infiltrante y 4 del tipo redondo. Las metástasis ganglionares y la diseminación intraglandular fueron halladas más frecuentement en el tipo infiltrante, seguido del tipo encapsulado con invasión. Los resultados sugieren la necesidad de efectuar disección ganglionar y lobectomía en pacientes con estos tipos histológicos de carcinomas pequenos que hayan sido sometidos a la remoción local del tumor.

Therapeutic efficacy of the angiogenesis inhibitor O- (Chloroacetyl- carbamoyl) fumagillol (TNP-470; AGM-1470) for human anaplastic thyroid carcinoma in nude mice

TNP-470 (AGM-1470), a synthetic analog of fumagillin (6-chloroacetyl-carbamoyloxy-4-(1,2-epoxy-1,5-dimethyl- 4-hexenyl)-5-methoxy-1-oxaspiro [2,5] octane 1, has been reported to reduce the supply of nutrients to experimental tumors by inhibiting angiogenesis. In this study, we investigated anti-tumor activity of TNP-470 against human thyroid anaplastic carcinoma with a view to developing a new treatment for this thyroid tumor. A transplantable tumor was established from thyroid anaplastic carcinoma of a 78-year-old woman, as a xenograft in nude mice (BALB/c, nu/nu, male). This transplantable tumor, with chromosomal abnormality was shown to be non-functional in excretory hormones and to preserve morphological characteristics of the original anaplastic tumor tissue. TNP-470 was given at a dose of 50 mg/kg b.w. to nude mice transplanted with human thyroid anaplastic carcinoma by different routes of administration: intratumoral, peritumoral, subcutaneous and intraperitoneal. Intratumoral and peritumoral administration were effective, and especially the TNP-470 administered by the former route completely inhibited tumor growth. Immunohistochemical analysis using anti-factor VIII antibody revealed the density of microvessels to be significantly decreased by local administration of TNP-470 (intratumoral administration, 7.8 +/- 2.9/mm2, control, 27.0 +/- 6.3/mm2; peritumoral administration, 9.7 +/- 2.6/mm2, control, 21.1 +/- 5.1/mm2). Our findings suggested the possibility of clinical application of TNP-470 to control the growth of human anaplastic thyroid carcinoma.

Epidermal growth factor receptor on human thyroid neoplasms.

AbstractMorphological observation of epidermal growth factor receptor (EGF-R) was attempted for human thyroid neoplasms, and the results were evaluated compared with the various histological types.nThirty-four malignant tumors, 24 benign tumors, 7 adenomatous goiters, and 7 normal thyroid tissues obtained at surgery were subjected to this study. They were fixed in 10% formalin solution, embedded in paraffin. After sectioning at 4μm, immunohistochemical staining by avidin-biotin-peroxidase complex technique was performed employing anti-human EGF-R monoclonal antibody.nBoth normal thyroid tissues and benign adenomas showed no apparent immunoreactive staining. The majority of the malignant tumors, however, demonstrated dark brown reaction products indicating location of EGF-R on the cell surface, cytoplasm, and nuclear envelope. Medullary and anaplastic carcinomas were stained more intensively than papillary and follicular carcinomas. In adenomatous goiter, the immunoreactive products were occasionally observed on the follicular cells of some limited areas, whereas its staining pattern was different from the malignant neoplasms.nThe correlative study between UICC classification of thyroid tumors and EGF-R staining in the papillary carcinoma revealed a higher frequency of positive staining in cases of multifocal metastasis.RésuméDans cette étude, on a mis en évidence le récepteur du facteur de croissance épithélial (R-FCE) dans les tumeurs de la thyroïde. Les caratéristiques immunohistologiques ont été étudiés dans les différentes variétés histologiques.On a donc étudié 34 tumeurs malignes, 24 tumeurs bénignes, 7 goitres adénomateux, et 7 pièces comportant du tissu normal, prélevés lors de la chirurgie de la thyroïde. Tous les tissus ont été fixés au formol à 10% et inclus dans de la paraffine. Après des coupes de 4μm, on a effectué une coloration immunohistochimique par la technique du complexe avidine-biotineperoxydase en utilisant des anticorps R-FCE humains monoclonaux.La coloration des tissus normaux et des adénomes bénins était apparemment immunonégative. La plupart des tumeurs malignes cependant ont montré des substances marron foncé, indiquant la présence de R-FCE sur la surface cellulaire, dans le cytoplasme et sur la membrane du noyau. Les cancers anaplasiques et médullaires ont présenté une coloration plus intense que celle des cancers papillaires et folliculaires. Dans le goitre adénomateux, on a mis en évidence, par endoit, des substances immuno-réactives dans quelques cellules folliculaires, mais les résultats de coloration différait de ceux obtenus dans les tumeurs malignes.Une étude de corrélation entre la classification UICC des tumeurs thyroïdiennes et la coloration R-FCE des cancers papillaires, a démontré une fréquence accrue de coloration positive en cas de métastases multifocales.ResumenEl factor de crecimiento epidermal (FCE) es un subgrupo de diversos polipéptidos de crecimiento involucrados en la regulación del crecimiento y diferenciación celulares. El FCE se liga a receptores específicos (FCE-R) de la membrana celular de sus órganos blanco (target organs); es considerado como un oncogen por su homología con el oncogen transformador v-erb B del virus de la eritroblastosis de las aves. Se ha demostrado la presencia de FCE-R en algunos tumores del esófago, estómago, seno, vejiga, pulmón, cerebro, ovario, y útero. La identificación del FCE-R es esencial en la yaloración del comportamiento biológico de los tumores sólidos. Este trabajo demuestra la reactividad de un anticuerpo monoclonal anti FCE-R con células de neoplasmas tiroideas así como la ubicación de FCE-R.El estudio tuvo como propósito la observación morfológica de FCE-R en neoplasmas tiroideos y su comparación con los diversos tipos histológicos.Treinta y cuatro tumores malignos, 24 tumores benignos, 7 bocios adenomatosos, y 7 tejidos normales obtenidos durante la cirugía fueron sometidos a estudio. Los especímenes fueron fijados en solutión de formol al 10% incluídos en parafina. Una vez seccionados a un espesor de 4μm, se realizó la coloración inmunohistoquímica con la técnica del complejo de avidinabiotina-peroxidasa empleando un anticuerpo monoclonal murino contra FCE-R humano.Tanto los tejidos tiroideos normales como los adenomas benignos exhibieron ausencia de coloración inmunorreactiva. Sin embargo, la mayoría de los tumores malignos demostraron la presencia de productos con una reacción marrón oscura indicativa de la ubicación de FCE-R en la superficie celular, el citoplasma, y la envoltura nuclear. Los carcinomas medulares y anaplásicos aparecieron con coloraciones más intensas que los carcinomas papilares y foliculares. En el bocio adenomatoso los productos inmunorreactivos fueron ocasionalmente observados en las células foliculares de áreas limitadas, al tiempo que su patrón de coloración fue diferente del de los neoplasmas malignos.El estudio correlativo entre la clasificación de la UICC (Union International Contra el Cáncer) de los tumores tiroideos y la coloración de FCE-R en el carcinoma papilar demostró una mayor frecuencia de coloración positiva en casos con metastasis multifocales.De los resultados del estudio se puede concluír que: (a) el carcinoma tiroideo puede poseer mayor cantidad de FCE-R y mayor afinidad de ligazón que el tumor benigno, (b) la presencia de FCE-R puede correlacionarse con los diversos grados de malignidad en el carcinoma tiroideo, (c) el bocio adenomatoso puede poseer algún potencial de malignidad.

Calcium regulating hormones and bone mineral content in patients after subtotal gastrectomy

Twenty-nine men who had undergone Billroth I gastrectomy and 19 men who had undergone Billroth II gastrectomy were studied to examine the changes in their calcium regulating hormones and bone mineral content following surgery. The serum calcium and phosphate concentrations in the patients with Billroth I and Billroth II were normal. The Billroth II group had an elevated level of serum alkaline phosphatase and reduced bone mineral content. The 24,25(OH)2D concentration was reduced (P<0.01) and 25(OH)D and 1,25(OH)2D concentrations were increased (P<0.01,P<0.05, respectively) in the Billroth II group. It was suggested by our study that the Billroth II patients had a reduced bone mineral content and an elevated 1,25(OH)2D concentration. Therefore, the pathophysiology of postgastrectomy bone metabolic disease is not due to vitamin D deficiency, but may instead be due to reduced calcium absorption in the intestine.

The relationship between multiple intrathyroidal involvement in papillary thyroid carcinoma and chronic non-specific thyroiditis

The relationship between multiple intrathyroidal involvement (MII) and chronic non-specific thyroiditis (CT) was investigated in 69 papillary thyroid carcinoma patients who received a subtotal or total thyroidectomy. The overall incidence of MII in patients with CT and the incidence of MII in the affected lobe of the patients with CT were significantly higher than that without CT (P = 0.0012 and 0.0425, respectively). Because Hashimotos thyroiditis is believed not to carry the increased risk of associated thyroid malignancy, the high incidence of MII in the affected lobe in the case with CT is postulated to be caused by intraglandular metastases.

Squamous Cell Carcinoma Arising from Thyroglossal Duct Remnants: Report of a Case and Results of Immunohistochemical Studies

We report on the case of a 57-year-old male found to have squamous cell carcinoma (SCC) arising from thyroglossal duct remnants. The patient presented with an asymptomatic tumor in his anterior neck which was immovable on palpation. Aspiration biopsy cytology revealed class V malignancy with many atypical clusters and marked keratinization. After preoperative radiation, a radical operation employing Sistrunks procedure with bilateral neck dissection was performed. Histopathological examination confirmed a diagnosis of moderately differentiated SCC, but revealed ciliated columnar epithelium in the walls of the cyst without a normal layer of squamous cells. Furthermore, immunohistochemical studies demonstrated the tumor to be negative for thyroglobulin staining, but positive for cytokeratin and carcinoembryonic antigen. These histopathological findings proved attributable to squamous metaplasia occurring in the ciliated columnar epithelium of the thyroglossal duct. Thus, SCC might originate in the metaplastic portion of the thyroglossal duct remnants. Although the prognosis associated with SCC in the thyroglossal duct is not as optimistic as that associated with papillary carcinoma, no evidence of recurrence has been observed in this patient in the 7 years since his operation. This suggests the effectiveness of our therapeutic approach for this unusual disease.

Expression of basic fibroblast growth factor in thyroid disorders

Morphologic and biologic studies were undertaken to clarify the biologic significance of basic fibroblast growth factor (bFGF) in human thyroid neoplasms. A total of 71 malignant tumors (50 papillary carcinomas, 14 follicular carcinomas, 7 anaplastic carcinomas), 11 follicular adenomas, 6 adenomatous goiters, and 6 Graves disease tissues were examined employing immunohistochemical methods (avidin-biotin-peroxidase complex technique). An affinity-purified polyclonal rabbit antiserum to human bFGF was used as a primary antibody. The eluate of malignant thyroid tumor tissues from the heparin-Sepharose column was examined by Western blot analysis to elucidate the molecular weight form. With immunohistochemical staining, bFGF was frequently detected in the cytoplasm of malignant thyroid tumors compared to tissues of the benign diseases and normal controls. With anaplastic carcinoma, immunoreactivity of the tumor cells was particularly strong. In the correlative analyses between UICC TNM classification and bFGF staining in papillary carcinoma, there were significant differences when relating positive staining to the grade of nodal metastases. By Western blot analysis, the bFGF immunoreactivity was specifically detected in the two forms, with molecular weights of 18 and 33 kDa. The high-molecular-weight form was detected in only anaplastic carcinoma. The present investigations demonstrated a close correlation between the expression of bFGF and the degree of malignancy. bFGF might play an important role in promoting lymph node metastases. Moreover, the high-molecular-weight form of bFGF might have an intense influence on tumor growth.RésuméUne étude morphologique et biologique a été effectuée pour clarifier la signification du facteur de croissance des fibroblastes de base (bFGF) dans les tumeurs de la thyroïde chez lhomme. On a étudié 71 tumeurs de la thyroïde (50 cancers papillaires, 14 cancers folliculaires, 6 adénomes solitaires, et 7 cancers anaplasiques, 11 adénomes folliculaires, 6 goitres adénomateux et 6 Maladies de Basedow) en utilisant des méthodes immunohistochimiques et notamment la technique du complexe avidinebiotine-peroxydase. On a employé un anticorps primaire fabriqué à partir dun antisérum polyclonal de lapin purifié dirigé contre le bFGF humain. Léluat des tissus thyroïdiens malins provenant de la colonne héparine-sepharose a été examiné selon la technique Western Blot pour déterminer son poids moléculaire. Le bFGF a été détecté plus fréquemment dans le cytoplasme des tumeurs thyroïdiennes malignes que dans les maladies bénignes et les contrôles. Dans le cancer anaplasique, limmunoréactivité des cellules cancéreuses a été particulièrement forte. En ce qui concerne la corrélation entre la classification TNM de lUnion Internationale contre le Cancer et la coloration bFGF des cancers papillaires, il y avait des différences significatives correspondant au degré denvahissement des ganglions lymphatiques. Dans lanalyse selon la technique du Western blot, limmunoréactivité bFGF a été détectée spécifiquement sous les deux formes de bFGF ayant des poids moléculaires de 18 et de 33 K, respectivement. Le poids moléculaire de 33K na été détecté que dans le cancer anaplasique. Cette étude démontre la corrélation étroite entre lexpression bFGF et le degré de malignité. Le bFGF peut probablement jouer un rôle important dans la survenue de métastase lymphatique. Le type à poids moléculaire élevé de bFGF pourrait influencer dadvantage la croissance tumorale.ResumenSe emprendió un estudio morfológico y biológico, con el propósito de clarificar la significación biológica del factor básico de crecimiento de fibroblastos (FBCF) en los neoplasmas de la glándula tiroides humana.Setenta y un tumores malignos (50 carcinomas papilares, 14 carcinomas foliculares y 7 carcinomas anaplásico), 11 adenomas foliculares, 6 bocios adenomatosos y 6 tejidos de glándula con enfermedad de Graves fueron examinados mediante métodos inmunohistoquímicos (técnica del complejo avidina-biotinaperoxidasa); se utilizó un antisuero policlonal purificado contra el FBCF humano, como anticuerpo primario. Además, se utilizó el análisis de Western blot para elucidar la forma de peso molecular.Con la coloración inmunohistoquímica, el FBCF fue detectado con frecuencia en el citoplasma de los tumores malignos de la tiroides, en comparación con lo observado en la enfermedad benigna y en los controles normales. La inmunorreactividad tumoral fue particulamente fuerte en el carcinoma anaplásico. En el análisis correlativo entre la clasificiación TNM UICC y la coloración del FBCF en el carcinoma papilar, se hallaron diferencias significativas relativas a la coloración positiva y al grado de la metástasis ganglionares.En el análisis Western blot, la inmunorreactividad FBCF fue específicamente detectada en las dos formas diferentes con pesos moleculares de 18 K y 33 K. La forma de alto peso molecular fue detectada sólamante en el carcinoma anaplásico.La presente investigación demuestra una estrecha correlación entre la expresión de FBCF y el grado de malignidad. El FBCF puede jugar un papel importante en promover las metástasis ganglionares. Además, la forma de alto peso molecular del FBCF puede tener una influencia más intensa sobre el crecimiento del tumor.