Adenylate cyclase activity as a predictor of thyroid tumor aggressiveness

Prior studies in our laboratory have shown that human thyroid neoplasms have a greater adenylate cyclase activity in response to thyroid stimulating hormone (TSH) than does the adjacent histologically normal thyroid tissue. However, there is little information relating activity of the TSH receptor-adenylate cyclase system to the type of thyroid neoplasm. Thyroid tissue from 67 patients was divided by clinical and histological criteria into 6 categories: normal (59), benign tumors (20), stage 1 carcinoma—intrathyroidal involvement only (25), stage 2 carcinomaregional lymph node involvement (6), stage 3 and 4 carcinoma—tissue invasion or distant metastasis (11), and medullary carcinoma (3). Adenylate cyclase activity in an 8,000 x g thyroid membrane preparation was determined in the basal state and when maximally stimulated with 300 mU/ml TSH. The cyclase responsiveness was the ratio of TSH stimulated adenylate cyclase activity compared to basal adenylate cyclase activity. The cyclase responsiveness by category is: normal, 2.8±0.2 (mean ± SEM); benign, 17.9±2.4; stage 1 carcinoma, 9.2±1.9; stage 2 carcinoma, 4.0±1.0; stage 3 and 4 carcinoma, 1.6±0.4; and medullary carcinoma, 1.05±0.04 (for the neoplasms,p <0.02 by ANOVA). Tumor stage was the only correlate with this trend as other prognostic risk factors (age, sex, a history of neck irradiation, or papillary versus follicular histology) showed no difference in cyclase responsiveness. These studies demonstrate a consistent inverse correlation between adenylate cyclase responsiveness and tumor stage or aggressiveness. Cyclase responsiveness appears to have clinical application for predicting which thyroid tumors will behave aggressively.

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Resumen Estudios previos en nuestro laboratorio han demostrado que los neoplasmas tiroideos humanos poseen una mayor actividad de adenilato ciclasa en respuesta a la administración de hormona estimulante de la tiroides (TSH) que el tejido tiroideo histológicamente normal adyacente. Sin embargo, existe muy poca información sobre la relation de la actividad del sistema receptor de TSH-adenilato ciclasa y el tipo del neoplasma tiroideo. Tejido tiroideo proveniente de 67 pacientes fue dividido mediante criterios chlínicos e histológicos en 6 categorias: normal (59), tumores benignos (20), extensión intratiroidea solamente en estado 1 (25), carcinoma-extensión ganglionar regional en estado 2 (6), carcinoma-invasión tisular o metástasis distantes en estados 3 y 4 (11), y carcinoma medular (3). La actividad de la adenilato ciclasa en una preparación de membrana tiroidea de 8,000 × g fue determinada en el estado basai y en estado de maxima estimulación con 300 mU/ml TSH. El grado de respuesta de la ciclasa fue la tasa de actividad de la adenilato ciclasa estimulada por TSH comparada con la actividad basai de la adenilato ciclasa. El grado de respuesta por categorías fue: normal, 2.8±0.2; tumor benigno, 17.9±2.4; carcinoma estado 1, 9.2±1.9; carcinoma estado 2, 4.0±1.0; carcinoma estados 3 y 4, 1.6±0.4; y carcinoma medular, 1.05±0.04 (para los neoplasmas,p < 0.02 por ANOVA). El estado del tumor apareció como el único factor de correlatión con esta gradación, ya que otros factures de pronóstico (edad, sexo, historia de irradiación cervical, histología papilar versus folicular) no demostraron diferencia en cuanto al grado de respuesta de la ciclasa. Estos estudios demuestran una consistente relación inversa entre el grado de respuesta de la adenilato ciclasa y el estado o agresividad tumoral. El grado de respuesta de la ciclasa parece tener aplicación clínica para predecir qué tumores tiroideos se habrán de comportar en forma agresiva.

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Résumé Les études antérieures provenant de nos laboratoires ont démontré une augmentation de l'activité enzymatique de l'adénylate-cyclase en réponse à la thyroïd stimulating hormone (TSH) dans le tissu thyroïdien humain tumoral, par rapport au tissu thyroïdien adjacent normal. Cependant il existe peu de données concernant l'activité du système récepteur TSH/adénylatecyclase par rapport au type de tumeur de la thyroïde. Les tissus thyroïdiens provenant de 67 patients différents ont été repartis en 6 groupes selon des critères cliniques et histologiques: normal (59), tumeur bénigne (20), cancer stade 1 (intrathyroïdien uniquement) (25), cancer stade 2 (envahissement ganglionnaire régional) (6), cancer stade 3 et 4 (envahissement tissulaire avoisinant ou métastases à distance (11), et cancer médullaire (3). A partir d'une préparation de membrane thyroïdienne centrifugée à 8,000 × g, l'activité de l'adénylate-cyclase a été déterminée en l'état basai et après stimulation maximale par 300 mU/ml de TSH. La réponse enzymatique a été mesurée comme étant le rapport de l'activité stimulée par la TSH/activité basale. Les résultats selon les 6 groupes étaient (moyen±ET): tissu normal, 2.8±0.2; tumeur bénigne, 17.9±2.4; cancer stade 1, 9.2±1.9; cancer stade 2, 4.0±1.0; cancers stade 3 et 4, 1.6±0.4; et cancer médullaire, 1.05±0.04 (p < 0.02 par l'analyse de variance pour les néoplasies). Le stade tumoral était la seule variable corrélée avec l'activité enzymatique. L'activité enzymatique n'était pas corrélée avec l'âge, le sexe, les antécédents d'irradiation cervicale antérieure ou l'histologie (papillaire vs. folliculaire). Ces études montrent un rapport inversement proportionnel entre l'activité d'adénylate-cyclase et le degré d'agressivité tumorale ou le stade. La réponse d l'adénylate-cyclase paraît avoir une application clinique: prévoir quelles tumeurs thyroïdiennes auront une évolution agressive.

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Presented at the International Association of Endocrine Surgeons in Sydney, Australia, September, 1987.

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Supported in part by the Medical Research Service of the Veterans Administration.     

用荧光探剂DPH研究家蝇线粒体膜脂流动性

本文探讨了用ＤＰＨ作为荧光探剂，研究家蝇飞翔肌线粒体膜脂流动性的方法。实验表明：ＤＰＨ确是用来研究家蝇生物膜流动性的有效探剂。同时，初步观察了不同杀虫剂对ＤＰＨ与线粒体膜结合后荧光强度的影响。其中氯菊酯和倍硫磷、马拉硫磷、敌百虫、氧化乐果，可使ＤＰＨ探剂与线粒体膜结合后的荧光相对强度增加；而氰戊菊酯、氯氰菊酯，溴氰菊酯及辛硫磷、乙酰甲胺磷，则可使荧光强度减弱。     

二氧化硅活化巨噬细胞中早期生长反应因子-1及其信号转导通路的研究

目的 探讨早期生长反应因子(Egr-1)及其信号转导在矽肺发生发展中的作用。方法用细胞免疫荧光、原位杂交方法检测二氧化硅(SiO2)刺激后Egr-1的表达定位,用报道质粒及EMSA检测其活性改变;用激酶活性分析法检测si0：刺激巨噬细胞后ERK1／2活性改变,进一步用激酶抑制剂初步探讨SiO2活化Egr-1的信号转导通路。结果SiO2刺激RAW264．7细胞短时间Egr-1核蛋白表达及转录因子明显增加;且在处理后30～60min,Egr-1核蛋白结合活性明显升高(为未处理组的20倍);在刺激后15min ERK1／2活性开始升高,30min达高峰(活性为对照组的29倍)而后渐降至基础水平;进一步用激酶阻断发现,Egr-1 mRNA及蛋白表达均减少。结论SiO2能激活巨噬细胞中Egr-1,且此过程可能由ERK1／2、p38介导,提示SiO2-ERK1／2、p38-Egr-1通路可能在矽肺发生发展过程中起重要作用。     

Concomitant translocation of Puralpha with its binding proteins (PurBPs) from nuclei to cytoplasm during neuronal development

Two Puralpha-binding proteins (PurBPs) were found in nuclear extract from mouse brain during P4-P10 by the overlay assay. At P14, they were decreased significantly in nuclear extract and increased in the S3 fraction, indicating their dynamic translocation during development. Western blot analysis also demonstrated concomitant translocation of Puralpha with the PurBPs during P7-P14, when neuronal circuit proceeds. Immunocytochemical study with cultured hippocampal neurons from rat E18 confirmed that nuclear Puralpha was translocated to cytoplasm after plating for 7-14 days. These results suggest that spatiotemporal translocation of Puralpha with the PurBPs from nuclei to cytoplasm has a crucial role in neuronal development.     

脑血流断层显像Bullseye显示

为了探讨用Bullseye显示脑血流灌注显像数据的可能性,对8例典型脑梗塞病例及15例脑血流断层显像常规分析可疑病例的脑血流断层显像的横断面数据进行Bullscye显示,结果8例典型病人病灶用普通Bullseye即显示良好;常规分析可疑病例病变用普通Bullseye 8例显示良好,变黑Bullseye 13例显示出病灶;结果提示利用Bullseye显示脑血流灌注显像数据的可能。     

小鼠甲胎蛋白与结核杆菌热休克蛋白70基因融合载体的构建及体外表达

目的 构建小鼠甲胎蛋白(α-Fetoprotein，AFP)与结核杆菌热休克蛋白70(Mycobacterium tuberculosis heat shock protein 70，Mt．HSP70)基因融合载体．并研究其在真核细胞中的表达情况。方法 以pcDNA3.1为基本单位构建AFP及HSP70的融合表达载体，融合基因以G-S-G-G-S连接子连接；用脂质体将系列载体导入COS-7细胞．48h后以免疫化学方法检测其表达。结果 构建的AFP和HSP70的单独及融合表达载体导入COS-7细胞48h后经RT-PCR检测可扩增出相应片段，细胞免疫化学染色为阳性。结论 AFP和HSP70的单独及融合表达载体构建成功，并能在真核细胞中表达。     

Translocation of Enterococcus faecalis strains across a monolayer of polarized human enterocyte-like T84 cells

We used a two-chamber system to study transcytosis of Enterococcus faecalis across monolayers of human colon carcinoma-derived T84 cells, which show structural resemblance to the native intestine. Among 16 E. faecalis isolates from different sources, the well-characterized strain OG1RF and 8 other isolates (2 endocarditis isolates, 1 urine isolate, and all 5 fecal isolates) showed translocation in this assay, while 6 clinical isolates (3 endocarditis and 3 urine isolates), the recipient strain JH2-2, and the control, Escherichia coli DH5alpha, had no detectable translocation. Of two OG1RF mutants involving the previously studied epa (enterococcal polysaccharide antigen) gene cluster, known to be needed for virulence and resistance to killing by polymorphonuclear leukocytes, one epa mutant (TX5179) was unable to translocate, while TX5180, with an epa disruption farther downstream, showed a moderate decrease in translocation relative to that of the wild-type strain OG1RF (P < 0.01), indicating that the epa gene cluster is important for translocation across a T84 monolayer. This observation was confirmed by complementation of the epa mutant (TX5179) with epa genes and restoration of its translocation ability. In conclusion, we have demonstrated translocation of at least some strains of E. faecalis across T84 monolayers, although strains differ considerably in this ability, and we have demonstrated that epa mutations can cause marked changes in successful translocation. These results suggest that this model may be a useful in vitro system for studying the process of translocation from the intestinal tract.