Abstracts

1 Molecular Endocrinology Group, Division of Medicine & Medical Research Council Clinical Sciences Centre, Imperial College London, Hammersmith Hospital, London, United Kingdom 2 Queen Mary University of London, Oral Growth and Development, Institute of Dentistry, Bart's and The London School of Medicine, London, United Kingdom 3 Division of Restorative Dental Sciences, University College London, Eastman Dental Institute, London, United Kingdom 4 Department of Civil and Environmental Engineering, University College London, London, United Kingdom 5 Departments of Physiology and Medicine, Dartmouth Medical School, Lebanon, NH Oral 1 Optimal Bone Mineralization and Strength Requires the Type 2 Iodothyronine Deiodinase in Osteoblasts

Thyroid Hormone Metabolism and Regulation Thursday Oral Basic 11:15 AM

Both hypothyroidism and thyrotoxicosis are associated with increased fracture risk. T4 is the predominant circulating thyroid hormone, but target cell responses are determined by local availability of the active hormone T3. The D1 and D2 iodothyronine deiodinases convert T4 to T3 in peripheral tissues; in bone conversion is restricted to osteoblasts. To investigate the role of locally produced T3 in bone, we characterized mice harboring deletion of Dio2 (D2KO) or both Dio1 and Dio2 (D1/D2KO) in which circulating T3 levels are normal.

Mice were examined during growth and in adulthood between postnatal days 1 and 112. Skeletal samples were analyzed by histology, dynamic histomorphometry, digital X-ray analysis and backscattered electron scanning electron microscopy. Biomechanical characteristics were determined by destructive three point bend tests.

Adult D2KO mice had increased bone mineralization and brittle bones that underwent abrupt high energy fracture without normal plastic deformation following mechanical loading. These biomechanical abnormalities were accompanied only by reduced osteoblastic bone formation whereas skeletal development, linear growth and osteoclastic bone resorption were normal, indicating that D2KO mice have a discrete defect of osteoblast function. Additional deletion of D1 in D1/D2KO mutants did not modify the D2KO skeletal phenotype.

These data identify a novel and essential role for D2 in osteoblasts. The restricted expression of D2 in osteoblasts suggests that their T3 content is enhanced relative to other bone cells and that increased T3 availability is essential for normal osteoblast function. As in other tissues, D2 activity in osteoblasts is increased in hypothyroidism but reduced in hyperthyroidism, thus providing a physiological mechanism to buffer the detrimental effects of altered thyroid status on the skeleton. We propose that optimal bone mineralization and strength are maintained over the physiological range of systemic thyroid hormone concentrations by the regulated activity of D2 in osteoblasts.

1 Thyroid Cancer Research Laboratory, Endocrine Surgery, Massachusetts General Hospital, Harvard Medical School, Boston, MA 2 Institute for Genome Sciences and Policy, Duke University, Durham, NC 3 Division of Cancer Biology and Angiogenesis, Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 4 Department of Pathology, University of Michigan Medical School, Ann Arbor, MI 5 Department of Medical Oncology, Dana Farber Cancer Institute, Harvard Medical School, Boston, MA 6 Department of Pathology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 7 Endocrinology Unit, Department of Medicine, University of Perugia, Perugia, Italy 8 Endocrinology Unit, Internal Medicine Department, Catholic University, Rome, Italy 9 Endocrine Pathology, Department of Pathology, Brigham and Women's Hospital, Harvard Medical School, Boston, MA Oral 5 A Robust B-RAFV600E Genome-Wide Signature in Papillary Thyroid Cancer

Thyroid Cancer Thursday Oral Basic 12:15 PM

B-RafV600E is the most frequent genetic alteration (29-83%) in papillary thyroid cancer (PTC); transforms normal thyroid cells and does not require Ras function to induce cell-growth. However, mechanisms by which B-RafV600E induces tumor aggressiveness are not fully understood. We aimed to identify and validate a B-RafV600E genome-wide signature associated with tumor progression in PTCs.

We applied a new genome-wide microarray analysis to identify gene sets with the same function associated with PTCs-B-RafV600E vs. PTCs-wild type (wt) B-Raf, and normal thyroid (NT) tissue samples. Gene validation was performed with real time-PCR, and for some genes using short-hairpin-RNAs (sh), three dimensional cultures (3D), and a novel orthotopic mouse model using the human thyroid cancer cell line 8505c (B-RafV600E+/+). Seven of 17 (41.1%) up-regulated gene sets were mainly enriched in functionally interconnected genes that belong to extracellular matrix (ECM) remodeling: cell-adhesion molecules (integrins, CD44 and fibronectin-1), or key-regulators of ECM (TSP-1 and TGF-beta1), or proteases (cathepsins). PTCs-B-RafV600E showed higher mRNA expression levels of these genes vs PTCs-wt-B-Raf and NT. Importantly, the only down-regulated gene set included genes involved in the establishement of cell-polarity in epithelial cells. In vitro knockdown of B-RafV600E by sh in the 8505c cells resulted in a dramatic reduction of cell adhesion, and migration/invasion in 3D assays. In addition, B-RafV600E over-expression in normal human cells conferred invasive properties. Finally, sh-B-RafV600E 8505c cells were implanted orthotopically into the thyroid of immunocompromised mice. Five weeks post-tumor implantation, animals with orthotopic tumor implants using B-RafV600E-sh 8505c cells have significantly smaller tumor volume and less lymph-node and lung micro-metastases than those injected with sh-control 8505c cells. B-RafV600E's role in aggressive PTCs may be linked to the expression of ECM genes, which may represent new molecular markers and therapeutic targets in this type of cancer.

1 Medicine, Obstetrics & Gynecology, Touro University College of Medicine, Hackensack, NJ 2 Division of Endocrinology, V. Fazzi Hospital, Lecce, Italy 3 Medical Education, University of Illinois, Chicago, Chicago, IL 4 Obstetrics & Gynecology, Casa di Cura Salus, Brandisi, Italy 5 Neonatalogy & ICU, V. Fassi Hospital, Lecce, Italy 6 Obstetrics and Gynecology, V. Fazzi Hospital, Lecce, Italy Oral 6 Universal Screening vs. Case Finding for Detection and Treatment of Thyroid Hormonal Dysfunction During Pregnancy

Thyroid Diseases Thursday Oral Clinical 11:15 AM

Whether or not all women should be screened for thyroid disease during pregnancy is controversial. The goal of the present study is to determine if treatment of thyroid disease during pregnancy decreases the incidence of adverse outcomes, and to compare the impact of universal screening (US) versus case-finding (CF).

Women were recruited from two ambulatory clinics in Southern Italy. Women in the first trimester were randomly assigned to the US group or CF group. Women in both groups were stratified as high-risk or low-risk based on risk factors for thyroid disease. All women in the US group, and high-risk women in the CF group, were immediately tested for fT4, TSH and TPO antibody Low-risk women in the CF group had their sera tested postpartum. Out of 4657 women, 95 were excluded on the basis of known thyroid disease and 46 women dropped out. Women were randomly assigned by computer program to the US (n = 2257) or CF group (n = 2259). Intervention consisted of levothyroxine in women with a TSH above 2.5 mIU/l in TPO-Ab+women and antithyroid medication in women with a suppressed TSH and elevated fT4. The primary outcome measure was the total number of adverse outcomes during pregnancy or in the immediate neonatal period to the mother or fetus, controlling for clustering of outcomes within women.

3590 women (78%) were classified as low risk (1808 in the case-finding group, 1782 in the screening group). Low-risk women in the screening (treated) group were less likely to have adverse outcomes than those in the case-finding (untreated) group (OR = 0.48, 95% CI = [0.33,0.71]). The number needed to screen to detect one hypo- or hyperthyroid woman in the low-risk group was 36 [28,49]; the number needed to treat to prevent one hypo- or hyperthyroid woman from experiencing adverse outcomes was 1.8 [1.4,2.6]. Overall, screening low-risk women prevented 2.48% of adverse events that would have otherwise been expected (p = .012). Treatment of thyroid hormonal abnormalities during pregnancy results in a significant decrease in adverse outcomes. All high-risk women should be screened for thyroid disease in the first trimester and serious consideration should be given to universal screening.

1 Neuroscience and Mental Health Program, Research Institute, The Hospital for Sick Children, Toronto, Canada 2 Psychology, University of Toronto, Toronto, Canada Oral 9 Grey Matter Reductions in Children with Congenital Hypothyroidism: A Voxel-Based Morphometry (VBM) Neuroimaging Study

Thyroid and Development Thursday Oral Clinical 12:00 PM

Extensive animal research has shown low pre- and postnatal thyroid hormone (TH) levels affect vision and memory and lead to brain abnormalities. Behavioral findings are supported by human studies of children with congenital hypothyroidism (CH) or hypothyroxinemia of prematurity or offspring of women with hypothyroidism in pregnancy. While our recent studies show hippocampal atrophy in CH, we still do not know if the other conditions are similarly affected and where effects are localized within brain systems dedicated to vision and memory abilities.

Fifteen children with CH and 21 normal controls (C, 10–16 yrs) underwent neuroimaging (1.5T scanner). T1-weighted images were analyzed using voxel-based morphometry (VBM), an advanced neuroimaging tool parcellating brain into grey and white matter and CSF tissue subcomponents to identify significant clusters differentiating groups.

CH showed 37 regions of reduced grey matter and one region of increased grey matter relative to C. For C vs CH, VBM identified two very large clusters of >30,000 voxels in each frontal lobe as well as more grey matter in bilateral hippocampus (near entrorhinal cortex and subiculum), multiple regions belonging to visual pathways (e.g., left calcarine sulcus, bilateral occipital/parietal sulcus, right cuneus, right fusiform gyrus, right MT), and bilateral amygdala and putamen. For CH vs C, CH showed more grey matter only in a region of unknown function.

OPEN IN VIEWER

Statistical parametric map showing regions of reduced grey matter volumes in CH compared with Controls.

In CH, neurogenesis is reduced in brain regions subserving cognitive abilities on which these children consistently show difficulty. This effect likely reflects their lack of TH from onset of disease until euthyroidism is achieved following early treatment. Although children with CH do not typically manifest executive function difficulties, which normally reflect frontal lobe abnormalities, the large frontal clusters observed presently may signify disrupted functioning of larger systems (e.g., frontal/parietal, frontal/hippocampal circuits) versus localized frontal deficits. Additionally, these system deficits may be contributing to observed visuospatial and memory deficits.

1 Internal Medicine, Federal University of Parana, Curitiba, Brazil 2 Internal Medicine, Odense University Hospital, Copenhagen, Denmark 3 Internal Medicine, University of Siena, Siena, Italy 4 Internal Medicine, University of Pisa, Pisa, Italy 5 Internal Medicine, Boston University, Boston, MA 6 Internal Medicine, Federal University of Rio de Janeiro, Rio de Janeiro, Brazil 7 Internal Medicine, University of Wuerzburg, Wuerzburg, Germany 8 Internal Medicine, CHRU de Lille-Clinique Marc Linquette, Lille, France 9 Internal Medicine, Catharina Hospital, Eindhoven, The Netherlands 10 Internal Medicine, Hamilton General Hospital, Hamilton, Canada 11 Internal Medicine, London Health Sciences Centre, London, Canada 12 Endocrinology, Genzyme Corp, Cambridge, MA Oral 10 Modified-Release Recombinant Human TSH (MRRHTSH) Augments the Effect of 131I in Moderately-Sized Benign Multinodular Goiters: Results from a Global, Randomized, Placebo Controlled Study

Thyroid Nodules and Goiter Thursday Oral Clinical 12:15 PM

The efficacy and safety of an intramuscular (i.m.) injection of modified-release recombinant human TSH (MRrhTSH) 24 h before oral 131I to achieve shrinkage of symptomatic benign multinodular goiters (MNG) was evaluated in a global, randomized, single-blinded trial.

One hundred forty one goiter patients were screened to randomize 96 patients, of whom 95 received either a single i.m. injection of placebo (n = 32), 0.01 mg MRrhTSH (n = 30) or 0.03 mg MRrhTSH (n = 33), followed the next day by 131I therapy calculated to deliver 100 uCi/mL based on the thyroid 131I uptake prior to the i.m. injection. The primary endpoint was goiter shrinkage by CT at 6 months compared to baseline goiter volume. The mean patient age was 57 yrs (85% females, 83% Caucasians), with baseline goiter volumes of about 100 mL (range 32–242 mL). The average 131I dose was 36 mCi.

For the placebo and the 0.01 and 0.03 mg MRrhTSH groups, the mean goiter shrinkages at 6 months were 23%, 23% and 33%, respectively (placebo and 0.01 mg MRrhTSH vs. 0.03 mg, P < 0.05). A responder was predefined as a patient who achieved ≥28% goiter shrinkage at 6 months, a number determined by literature review. The percentages of responders in the three groups were 25%, 37% and 64%, respectively. There were no related serious adverse events.Transient hyperthyroidism occurred in 6%, 27% and 30% of the patients in the groups, respectively.

MRrhTSH at the dose of 0.03 mg i.m. plus oral 131I achieved significantly more goiter shrinkage at 6 months than did placebo or the 0.01 mg dose. Although transient hyperthyroidism occurred in some patients, MRrhTSH was well tolerated.

1 School of Medicine & Dentistry, University of Rochester, Rochester, NY 2 University of Rochester Eye Institute, University of Rochester, Rochester, NY 3 Environmental Medicine, University of Rochester, Rochester, NY 4 Biostatistics, University of Rochester, Rochester, NY Poster 12 Orbital Fibroblasts from Thyroid Eye Disease Patients Differ in Proliferative and Adipogenic Responses Depending on Disease Sub-Type

Autoimmunity Thursday Poster Basic

Thyroid Eye Disease (TED) is an autoimmune process primarily associated with Graves' Disease. Imaging is used to classify patients into 2 groups: Type I with expansion of the fat compartment and Type II with enlargement of the extraocular muscles. Some patients have both components. We tested the hypothesis that orbital fibroblasts (OFs) from Type I TED were prone to adipogenesis, whereas OFs from Type II TED were prone to proliferation.

CT scans and tissue samples were obtained from TED and non-TED patients undergoing orbital surgery. OFs from 3 Type I, 3 Type II, and 2 non-TED patients were treated with transforming growth factor beta-1 (TGFβ1) and/or insulin-like growth factor-1 (IGF1), 10% fetal bovine serum (FBS), co-culture with autologous T-cells, pioglitazone, or 15d-prostaglandin J₂ (15d-PGJ₂). Proliferation and adipogenesis were measured using the tritiated thymidine assay after 72 hours and the AdipoRed assay after 8 days, respectively. A mixed effects model was used to compare proliferation and adipogenesis of OFs from the three groups.

There was a trend toward more proliferation in Type II OFs compared to Type I OFs with autologous T-cell co-culture (P = 0.0633). Type II OFs proliferated more with 5 ng/ml TGFβ1 (P < 0.0001) and 10% FBS (P < 0.0001) than Type I OFs. Type I OFs proliferated more than Type II OFs with combined 5 ng/ml TGFβ1 and 5 ng/ml IGF1 (P = 0.044) treatment. However, proliferation was lower with TGFβ1 and IGF1 in combination than with TGFβ1 alone for all groups. Type I OFs had more adipogenesis than Type II OFs with 5 μM 15d-PGJ₂ (P = 0.0259) and 10 μM pioglitazone (P = 0.006) treatments.

Type II OFs were driven to proliferate more than Type I OFs, while Type I OFs were driven to adipogenesis more than Type II OFs. Further studies are needed to understand IGF1's role in modulating TGFβ1's effects on OFs. This study demonstrates the heterogeneity of OFs from Type I and Type II TED patients. Further investigation is required to determine if factors that influence the fate of TED OFs are genetic, environmental, or a combination of the two. Hopefully, this line of research will provide a basis for more targeted therapies based on the type of TED. Research supported by: RO1 EY11708, TL1 RR024135.

1 Dept. of Medicine I, Gutenberg University, Mainz, Germany 2 Dept. of ENT, Gutenberg University, Mainz, Germany 3 Dept. of Ophthalmology, Gutenberg University, Mainz, Germany 4 Dept. of Medical Statistics, Gutenberg University, Mainz, Germany 5 Dept. of Neuroradiology, Gutenberg University, Mainz, Germany Poster 13 Novel Combined Endoscopic and Transpalpebral Decompression Surgery in 150 Consecutive Patients with Severe Graves Orbitopathy

Autoimmunity Thursday Poster Clinical

Disfiguring proptosis and dysthyroid optic neuropathy (DON) lead to cosmetic impairment and loss of vision in severe Graves orbitopathy (GO). Surgical decompression of the crowded orbit offers a valid therapeutic option.

150 consecutive patients (median age 54 years, 113 female, 96 smokers, 280 orbits) with severe GO had a novel endonasal bone and transpalpebral fat decompression surgery at a university joint thyroid-eye-clinic. A complete multidisciplinary endocrine and ophthalmic assessment was performed pre, three and 12 months post-operatively.

Cosmetic reasons (196 orbits, 73%), DON (67, 24%), and one corneal ulceration (0.4%) were the indications for surgery. The medial orbital wall and the palpebral adipose tissue (median 8 ml, range 3–21 ml) were removed. Preoperatively, relative afferent pupillary defects and vision field defects were noted in 16% and in 43% of DON-patients but in 8% only after 12 months (p < 0.001). Proptosis decreased from 23 mm (13–32) to 20 (10–30) and 19 mm (11–30) at 3 and 12 months, respectively (p < 0.001). Lid fissure width decreased by 2 mm, p = 0.0001. Upgaze intraocular pressure dropped from 23 mmHg (10–44) by 4 mmHg at 12 months (p < 0.001). Median severity score (NOSPECS) declined from 7 points (2.5–13) to 4 (1–11) and 3.5 points (1–7) at 3 and 12 months (p < 0.001). Subjective eye signs significantly decreased in 89% of the patients and satisfaction rate was >90%. NOSPECS and intraocular pressure decreased less in GO smokers vs. non-smokers (both p < 0.001). Two patients (1.3%) only complained about new-onset constant diplopia three months after decompression, while 22/150 (15%) patients reported de-novo inconstant or intermittent diplopia. Prisms and/or subsequent squint surgery corrected all cases of diplopia. Preoperative coexistent sinusitis (n = 97, 65%) was successfully treated by decompression surgery in all cases. No major side-effects were registered.

The combined endonasal bone and transpalpebral fat orbital decompression is a safe and efficient therapy for severe GO.

1 UOC di Medicina Generale, PO S. Giovanni Bosco, Naples, Italy 2 UOC di Patologia Generale, PO S. Giovanni Bosco, Naples, Italy Poster 15 Helicobacter Pylori and Graves' Disease: The Autoimmune Disease is an Important Cause of the HP Infection Recurrence

Autoimmunity Thursday Poster Clinical

Aim: The Helicobacter Pylori (HP) infection is worldwide diffused. In the civilized countries is present in nearly 50% of the population. In many studies, the rate of HP recurrence, after the drug eradication, is about 3% for one year. Recently, we have demonstrated the high rate of prevalence of the HP infection in patients with the ongoing Graves' Disease. The aim of our work was to investigate if the autoimmune disease increases the rate of HP recurrence after a successful drug regimen of eradication.

Materials and Methods: The HP presence was investigated on a fresh stool sample using an immunoassay amplification technology test (IDEIA Hp StAR, DakoCytomation). In an HP-positive group of hyperthyroid GD patients at first diagnosis and control subjects we performed the standard eradication therapy with amoxicillin 1 gr, clarythromicin 0,5 gr and esomeprazole 20 mg b.i.d. for one week. After five weeks we tested the obtained HP eradication on the stool sample. We selected 15 HP-eradicated GD patients (medium age 38 ± 2 SD years) and 19 HP-eradicated subjects as control (medium age 40 ± 2.1 years). After at least six months (13 ± 3.2) we tested newly both the GD patients, now euthyroid under methimazole treatment, and the controls for the presence of HP. We analyzed the obtained results using statistical test such as the Fisher Test.

Results: Seven of 15 GD patients resulted newly infected by HP versus 1 of 19 in the control population. The rate of recurrence of the infection was significantly different between the patients affected by GD (46,6%) versus the respective controls (5,2%, P = < 0.001).

Conclusions: The GD is an important predisposing factor, probably independent by the hormonal status, for the HP infection and its recurrence.

1 Health Sciences Group, Exponent, Inc. , Menlo Park, CA 2 Section of Endocrinology, Diabetes, and Nutrition, Boston University Medical Center, Boston, MA Poster 17 Is the Iodine Content of Cat Food Responsible for “Toxic Nodular Goiter” in Older Cats?

Iodine Uptake and Metabolism Thursday Poster Basic

Three investigators of this abstract had cats that developed hyperthyroidism, similar to human toxic nodular goiters (TNG), and were treated with I-131 or thyroidectomy. The etiology of TNG in cats remains unknown although it is considered multifactorial. Since there are no reports of RAIU values in thyrotoxic cats, we speculated that iodine excess (Jod-Basedow) might be one cause.

Iodine content was measured in 71 randomly-selected canned, pouched, and dry cat foods from three areas of the U.S. to determine whether iodine concentrations in current commercial cat foods meet recommendations for supplementation, whether regional differences in iodine concentrations exist, and whether seafood ingredients are associated with iodine concentrations.

Canned foods' iodine concentrations ranged from 0.9 to 67.7 mg/kg dry weight, nearly an 80-fold variation, while pouched and dry foods each varied over 3-fold between minimum and maximum values. Iodine concentrations in cat foods from Northern California, Florida, and Indiana did not differ overall, with mean± standard deviation values of 8.5 ± 15.7, 8.1 ± 15.3, and 7.5 ±14.7 mg/kg dry weight, respectively. However, iodine concentrations for specific foods varied across the three locations, with a 13-fold variation in one brand of dry food (0.2–2.6 mg/kg dry weight). The highest iodine concentrations were associated with “fish”, salmon, and salmon broth as the first five ingredients, though the lowest concentration among cans had salmon as the fourth ingredient.

A previous review of iodine supplementation for cat foods revealed that recommendations for feline iodine intake changed in the 1970s, when this disease was first reported. Recommendations were revised again in 2006: under these, a 4-kg cat would consume 87.5 mcg iodine/day based on a 350-mcg iodine/1000-kcal metabolizable energy diet. The dramatic variation here among canned foods (resulting in possible ingestion of approximately 62–4643 mcg iodine/day) suggests that the disparity in iodine concentrations may lead to development of nodular hyperplasia and later clinical hyperthyroidism, if cats consume diets that are at first iodine deficient and later contain excessive iodine, as the human experience has shown.

1 Pathology, University of Verona, Verona, Italy 2 Morphological and Biomedical Sciences, University of Verona, Verona, Italy 3 Internal Medicine, University of Pisa, Pisa, Italy 4 Internal Medicine, University of Roma La Sapienza, Rome, Italy Poster 18 Claudins-1,3,4,5,7 and PAX8 During Human Thyroid Development: An Immunohistochemical Evaluation

Thyroid and Development Thursday Poster Basic

thyroid follicles develop from aggregates of unpolarized precursor cells. Pax8 belongs to a family of developmental control genes that encode transcription factors. It is expressed at the most immature stage of the thyroid anlage and plays a fundamental role in thyroid cell differentiation, maintenance of the differentiated state and cell proliferation. Tight junctions (TJs) are dynamic structure, that at different stages of epithelial tissue development play an essential role in maintaining integrity and physiological function of thyroid follicles. Claudins (CLDNs) are the major components of TJs. Tissues are characterized by distinctive CLDNs patterns changing during differentiation and tumor formation. CLDNs expression during thyroid ontogenesis is unknown. The aim of this study was to examine CDLNs-1,3,4,5,7 and Pax8 immunohistochemical staining pattern in human foetal thyroid glands.

18 thyroid glands were obtained from human foetuses (gestational age range: 15–22 weeks). Immunostaining was performed using a panel of polyclonal (CLDNs-1 and 3) and monoclonal (Pax8 and CLDNs-4,5,7) antibodies.

CLDN7 was constantly expressed showing strong, diffuse and linear basolateral positivity. CLDN4 and 5 staining was similar to CLDN7 but weaker. CLDN1 exhibited a weak, focal membranous staining at the gland periphery. CLDN3 immunoreactivity was negative. Pax8 was diffusely observed in a nuclear pattern in all samples.

CLDN7 and 4 are constitutively expressed in thyroid epithelium during ontogenesis at a similar level from foetal up to adult thyroid tissue, thus suggesting an essential role in architectural stability of follicular cells. CLDN1 was expressed at the border of the foetal gland where the first follicles, containing colloid, are localized. Conversely, CLDN1 was absent in adult normal tissue while up-regulated in thyroid cancer, thus emerging as an oncofetal antigen and a potential marker of thyroid cancer. Our study demonstrates CLDN5 expression in thyroid gland, suggesting its possible role in normal and neoplastic adult thyroid tissue. We also confirm that Pax8 is expressed at an early stage of thyroid cell differentiation playing a key role in embryogenesis.

1 Neuroscience and Mental Health Program, Research Institute, The Hospital for Sick Children, Toronto, Canada 2 Psychology, University of Toronto, Toronto, Canada Poster 19 Hippocampal Morphometry in Offspring of Hypothyroid and Hyperthyroid Women and Children with Congenital Hypothyroidism

Thyroid and Development Thursday Poster Clinical

Adequate thyroid hormone (TH) is needed in gestation and postnatally for normal hippocampal development. Animal research shows type or location of hippocampal abnormality is linked to timing of TH insufficiency. Children of women with hypothyroidism (HYPO) or hyperthyroidism (HYPER) in pregnancy or having congenital hypothyroidism (CH) present distinct human models of timing of TH insufficiency: HYPO has earliest TH loss since these children experienced TH insufficiency in early gestation before onset of fetal thyroid function; HYPER has later pregnancy TH insufficiency because although exposed to normal maternal TH levels, thyroid functioning was blocked in late gestation by maternal anti-thyroid medication; CH, still later TH deficiency in late gestation and postnatally until replacement TH took effect. We queried whether these groups differ in hippocampal structure (size, shape), chemical constitution, integrity, and function. This study describes our findings on hippocampal volumes.

Participants are 45 children (15 per group) and 15 controls (C) who have been followed longitudinally since birth. At 10–12 years, they receive extensive memory testing and structural and functional magnetic resonance imaging. Hippocampal volumes are determined with Analyze 7.0 by tracing according to set landmarks <80 right and left hippocampi per child.

Preliminary findings for 43 cases (9 HYPO, 5 HYPER, 14 CH, and 15 C) indicate smaller volumes in HYPER and CH (3068 and 3070 mm³) than HYPO and Control (3463, 3266 mm³). However, one HYPO whose mother had severe hypothyroidism in second and third trimesters (TSH > 25) had a very small hippocampus (2547 mm³).

TH insufficiency during late gestation and early infancy is associated with reduced hippocampal size in late childhood. The implications of these findings on memory functioning and in terms of hippocampal subcomponents specifically affected will also be evaluated.

1 Endocrinology & Metabolism, USC/Keck School of Medicine, Los Angeles, CA 2 Pediatric Endocrinology, USC/Keck School of Medicine, Los Angeles, CA Poster 20 A Unique Case of Congenital Hypothyroidism (CH) and Myxedema Coma

Thyroid and Development Thursday Poster Clinical

CH is screened for at birth and has largely been eradicated in the developed world. However, CH remains a major health risk in developing countries. If CH is unrecognized and untreated, devastating neurological and developmental consequences result. A unique case of untreated CH and myxedema coma is presented.

A 36 year-old female with untreated CH, who emigrated from Mexico 5 years ago, presented in myxedema coma secondary to a URI. The patient was 81 cm in height and weighed 16.5 kg corresponding to the 5th percentile in height and the 95th percentile in weight for a 3-year old child. Baseline functional development corresponded to that of a 10-month old child. Physical exam on presentation revealed a BP of 123/72, HR ranging between 60–70 BPM (normal SBP and HR for a 3 year-old child are 86–111 mmHg and 110–115 BPM, respectively), and T of 97.80F. Initial TFT's included a TSH of 208, a FT4 <0.4 and a FT3 <1.1. Work-up of CH consisted of a thyroid U/S, which showed no thyroid tissue, and a TG of 4.2.

Besides receiving supportive care, the patient was loaded with 150 mcg of LT4 IV followed by maintenance dose of 25 mcg LT4 daily. Over the ensuing 72 hours after LT4, SBP decreased from the 120's to the 100's, HR increased to the 90's to 100's (consistent with those expected of a three year old) and T increased to 98–990 F. At her 6 month follow-up visit the patient has grown 13 cm and lost 2.7 kg. She is now more active and able to walk, has grown hair after initial hair loss, and eruption of secondary teeth has occurred. Her most recent TFT's include a TSH of 56.53 and a FT4 of 1.08

1. CH was due to agenesis of the thyroid, 2. Untreated CH produces severe neurological and physical impairment, 3. This is a unique presentation of a largely extinct disease and the oldest patient on record with untreated CH. It gives us the exceptional opportunity to study and observe changes in response to L-T4 therapy and perhaps better understand the role of thyroid hormone in development.

1 Medicine, Endocrine Division and Otolaryngology, Head & Neck Surgery, Mount Sinai Hospital and University of Toronto Medical School, Toronto, Canada 2 Otolaryngology, Head & Neck Surgery, Mount Sinai Hospital and University of Toronto Medical School, Toronto, Canada Poster 22 Secretome Analyses Unravels Pathway-Based Candidate Biomarkers for Thyroid Carcinoma

Thyroid Cancer Thursday Poster Basic

Identification of molecular signatures of aggressive cancers will facilitate the discovery of specific biomarkers for predicting their anticipated clinical course and the development of non-invasive blood-based biomarkers. Herein, we have explored the conditioned serum-free media of several thyroid carcinoma cell lines which may be enriched by secreted/shed proteins or the adjacent disease-relevant cells of human thyroid thyroid carcinomas. Studies on such surrogate proteins secreted by these cells could identify biomarkers which could be further evaluated in biological fluids.

Conditioned serum-free media was obtained for three thyroid cancer cell lines (WRO, MRO and TPC-1) for secretome analysis. Using liquid chromatography-tandem mass spectrometry, the secreted proteins for the two aggressive thyroid cancer cell lines (WRO and MRO) were compared with those secreted by the non-aggressive cell line (TPC-1).

From these analyses, we observed that there were 77 non-redundant proteins identified. Among these 77 proteins, there were 7 that were identified in all the cell lines, while 9 were unique for the aggressive carcinoma cell lines and 35 proteins were detected only in the non-aggressive cell line TPC-1. Integration of these proteins into a cellular context using Ingenuity Pathway Analysis software revealed that secretome proteins are interconnected with several intracellular canonical pathways known to be associated with cancer development. The detected pathways included: glycolysis, gluconeogenesis, ERK/MAPK, PI3K/Akt, Oncostatin M and IL6 signaling pathways.

Using liquid chromatography-tandem mass spectrometry for secretome analyses has demonstrated in cultured human carcinoma cell lines: (1) the feasibility of detecting protein biomarkers which can distinguish between aggressive and non-aggressive thyroid carcinomas, (2) unravel novel pathways and cross-talks that can lead to rational biomarker discovery through linkage with cancer relevant pathways, and (3) facilitate the development of a selective non-invasive blood-based platform of biomarkers for predicting the clinical course and responses to treatment in thyroid carcinomas.

1 Surgery, Boston University, Boston, MA 2 Medicine, Boston University, Boston, MA 3 Surgery, Johns Hopkins Medical Institutes, Baltimore, MD Poster 23 Microrna 222 Affects the Invasive Potential in Papillary Thyroid Cancer Cells via E-Cadherin without Altering Proliferative Potential

Thyroid Cancer Thursday Poster Basic

MicroRNAs (miRNAs) are small noncoding RNAs that may act to modulate gene expression at the posttranscriptional level and act as factors contributing to cancer progression. mir222 has been shown to affect the proliferation potential of prostate carcinoma cell lines and regulate p27kip1 in human thyroid papillary carcinomas; NPA, a human papillary thyroid carcinoma cell line, has been shown to overexpress mir222. The aim of this study was to evaluate mir222 inhibition on NPA cells and its' implication for the proliferation and invasive potential.

NPA cells were transfected with antimiR 222 using the AMAXA nucleofector; cell growth was assessed using the MTT assay and invasiveness was investigated using a Boyden chamber assay. Comparison was made to an antimiR scramble control. Gene expression was evaluated by qRTPCR using primers for mir222 and mir221 normalized to rnu44 and E-cadherin normalized to GAPDH.

NPA cells transfected with antimir222 showed a 13-fold reduction in mir222 expression and a 10-fold reduction in mir221 expression, while mRNA levels for E-cadherin rose 5-fold by 96 hours. There was no reduction in cell proliferation but a 46% and 38% reduction in invasiveness at 72 and 96 hours respectively. Inhibition of mir222 expression in NPA cells caused no alteration in cell proliferation but a marked reduction in cell invasiveness.

Our data might suggesting a oncogenic function of mir222 and a potential therapeutic target for use in patients with papillary thyroid carcinoma.

1 Endocrinology, VA Greater Los Angeles Healthcare System, Los Angeles, CA 2 Medicine, UCLA School of Medicine, Los Angeles, CA Poster 24 Stability of Recombinant Human TSH (Thyrogen) Potency Based on Bioassay in FRTL-5 Cells

Thyroid Cancer Thursday Poster

Recombinant human TSH (Thyrogen^®, Genzyme), 4–12 IU/mg, is approved for use in a 0.9 mg dose/day for 2 consecutive days for diagnosis and treatment of differentiated thyroid cancer. It is recommended that it be injected IM immediately after reconstitution in the distilled water diluent supplied. However, Thyrogen has been used off-label in doses less than the standard 0.9 mg dose for stimulation of radioiodine uptake in the treatment of multinodular goiter. In order to determine whether the biologic activity of Thyrogen can be preserved after dilution, we designed experiments to assess the biologic stability of Thyrogen under different durations and temperatures of storage. Proving the stability of reconstituted Thyrogen would allow practitioners to store aliquots for future use, thus lowering the cost per use.

Thyrogen was diluted in 1% BSA in PBS to a concentration of 0.9 mg/ml and further diluted to 0.1mg/ml. Aliquots of 0.5 ml were stored at room temperature, 4°C, −11°C, and −60°C for various lengths of time. In addition, Thyrogen aliquots were subjected to several more extreme temperature conditions: incubation for 1 hour at 50°C and, in another experiment, 10 cycles of freezing in dry ice alternating with thawing at 37°C. Bioassays were performed in FRTL5 cells that were down-regulated in media lacking TSH for 5 to 7 days. Thyrogen was added to the media at a final concentration of either 5 ng/ml or 20 ng/ml and the cells were then incubated for 48 hr. Potency was assessed by measurement of 125I-iodide uptake in comparison to cells also treated with perchlorate to block iodide uptake.

Samples stored at 4°C and −60°C retained activity after storage periods up to 59 days. Samples subjected to the freeze-thaw cycle retained activity similar to those of samples that were not frozen. Samples heated to 50°C for 1 hour also stimulated iodide uptake more than that of non-stimulated cells. Thyrogen kept at 4°C, −11°C, and −60°C maintained good biologic potency for 59 days of storage.

The biologic activity of Thyrogen is very stable. The data provide reassurance that storage in the cold for at least one month does not result in loss of Thyrogen's biologic activity.

1 Dept of Endocrinology, University of Pisa, Pisa, Italy 2 Dept of Surgery, Section of Pathology, University of Pisa, Pisa, Italy 3 Dept of Oncology, Division of Pathology, University of Pisa, Pisa, Italy 4 Dept of Statistics and Applied Mathematics to Economy, University of Pisa, Pisa, Italy 5 Departments of Internal Medicine, Endocrinology and Metabolism, and Biochemistry, University of Siena, Siena, Italy Poster 25 Analysis of Clinical, Epidemiological and Pathological Features in a Series of 4187 Differentiated Thyroid Cancer Diagnosed Between 1969 and 2004: Did they Really Change in the Last Decades?

Thyroid Cancer Thursday Poster Clinical

In the last two decades, a marked increase in the prevalence of differentiated thyroid cancer (DTC) has been observed worldwide. The aim of the present study was to evaluate the changing features of DTC referred to our institution between 1969 and 2004.

Epidemiological, clinical, pathological features and prognostic factors were analyzed in 4187 DTC patients, subdivided into two groups: Group1 (n = 1215) and Group2 (n = 2972) diagnosed before and after 1990, respectively. The series was characterized by a considerable homogeneity in treatment and follow-up strategies, a very low percentage of patients lost at follow-up (<6%) and an almost unique long-term follow-up, with more than 500 patients followed for more than 20 years.

Group2 was characterized by an increased proportion of papillary carcinoma, especially of small size (<1 cm) with a concomitant decrease of follicular carcinoma. Male percentage was greater in Group2, while age at diagnosis was unchanged. DTC of Group2 were more frequently associated with multinodular goiter or autoimmune thyroiditis (AT), many were unexpected findings and only a few patients had a history of neck irradiation. Typical features of aggressiveness were much less frequent in Group2. The survival rate of Group2 (98.7%) was significantly greater than in Group1 (91.4%) (P < 0.0001). Gender, age, histotype, tumor size, extrathyroidal macro-invasion, lymph node and/or distant metastases and clinical class were found to be prognostic factors in both groups using univariate analysis. With multivariate analysis, we found that only age (>40 years), advanced stage (Class III/IV) and the time of diagnosis (before 1990) were independently correlated with a lower survival both in Group1 and Group2.

DTC patients diagnosed after 1990 were more likely to have small tumors, less advanced stage and a better prognosis than those diagnosed before 1990. Interestingly, a higher prevalence of DTC associated with AT was observed in Group2. However, despite these sharped differences, an advanced age and/or an advanced stage at diagnosis are still the most poor prognostic factors for survival as well as they were in the past.

1 Hospital Pathology, Seoul St. Mary's Hospital, The Catholic University of Korea, Seoul, South Korea 2 Surgery, Seoul St. Mary's Hospital, The Catholic University of Korea, Seoul, South Korea 3 Internal Medicine, Seoul St. Mary's Hospital, The Catholic University of Korea, Seoul, South Korea Poster 26 Tumor Growth Patterns Identified by Cyclin D1 Staining are Related with the Risk of Lymph Node Metastasis in Patients with Papillary Thyroid Carcinoma

Thyroid Cancer Thursday Poster Clinical

Many attempts have been made to predict the occurrence of lymph node metastases from papillary thyroid carcinoma. Despite these efforts, there is at present no reliable means to accurately predict cervical nodal metastasis. We present a new prediction system, based on the histological and immunohistochemical features.

We analyzed the clinicopathological factors in 123 consecutive patients who underwent total thyroidectomy and central neck dissection for papillary carcinoma and also evaluated the cyclin D1 expression and exon 15 BRAF mutation in tumor tissue.

Seventy-one (57.7%) of all patients had lymph node metastasis. Cyclin D1 was constantly expressed at invasive tumor front and revealed well-defined isolated glands of tumor cells in extra-tumoral region and laterally spreading tubular growth along the fibrous septa around the invasive front of the tumor. On univariate analysis, age (p = 0.014), tumor size (p = 0.011), isolated gland (p < 0.001), tubular growth (p < 0.001), and tumor multiplicity (p = 0.027) predicted lymph node metastasis, whereas patient gender, extrathyroidal extension, and BRAF mutation did not. On multivariate analysis, age (p = 0.035), isolated gland (p < 0.001) and tubular growth (p < 0.001) proved to be strong independent predictors of lymph node metastasis.

The cyclin D1 staining of papillary thyroid carcinoma is very useful for identifying the intrathyroidal spreading of the tumor. Tumor growth patterns verified by cyclin D1 staining can be used in identifying papillary thyroid carcinomas with metastatic potential.

1 Surgery, St. Joseph Mercy Hospital, Ann Arbor, MI 2 Surgery, University of Michigan, Ann Arbor, MI Poster 27 Routine Central Lymph Node Dissection (CLND) for Papillary Thyroid Cancer (PTC)

Thyroid Cancer Thursday Poster Clinical

Routine CLND in PTC treatment is controversial, because of possible increased operative morbidity and uncertain clinical impact.

Retrospective cohort study comparing total thyroidectomy (TT) to TT plus CLND. Inclusion criteria: initial thyroid operations for clinically node-negative PTC ≥1 cm. Endpoints: complications, staging, and post-treatment thyroglobulin (TG) levels.

From 5/02–12/07, of 270 patients operated for thyroid cancer, 103 patients met inclusion criteria. Surgeon and patient preference dictated that 52 underwent TT and 51 TT+CLND. Tumor size and MACIS (metastases/age/completeness-of-resection/invasion/size) scores were equivalent. Temporary hypocalcemia was more frequent with CLND (40.4% vs. 19.2%; p = 0.02). Three TT+CLND patients with normal PTH levels continue calcium/vitamin-D supplements >6 months after surgery (5.8%) versus none after TT (p = 0.11). Permanent nerve injury rates were equivalent (TT:1/52 vs. TT+CLND:0/51). 59% of TT+CLND patients had positive nodes (30/51). Node involvement was equivalent between those with suspicious nodes (58%) and non-suspicious nodes (59%). Node pathology results upstaged 11/22 TT+CLND patients over 45 y (TT: 3/22 stage III vs. TT+CLND: 12/22 stage III; p < 0.01). Mean serum TG levels were equivalent after radioiodine ablation (TT: 4.7 ng/mL vs. TT+CLND: 2.0 ng/mL; p = 0.51) as was the fraction of patients with undetectable TG levels (77% vs. 73%). Recurrences affected two TT (one central neck) and one TT+CLND patient.

Routine CLND during TT does not change the permanent complication rate for primary thyroid cancer surgery. Intraoperative clinical node assessment was inaccurate. Routine dissection upstaged 50% of patients over 45 y. To date, disease outcome is similar and excellent.

1 Department of Internal Medicine, University of Pisa, Pisa, Italy 2 Department of Surgery, University of Pisa, Pisa, Italy Poster 28 Ppargamma Agonists and Antiblastics Exert an Additive Antiproliferative Activity in Primary Cultured Human Papillary Dedifferentiated Thyroid Cancer Cells

Thyroid Cancer Thursday Poster Clinical

Dedifferentiated papillary thyroid carcinoma (DPTC) is characterized by aggressive growth, metastatic spread and loss of iodide uptake ability, making the tumor resistant to radioiodine. Agonists (TZD) of peroxisome proliferator-activated receptor-gamma (PPARg) induce apoptosis and exert antiproliferative effects on continuous thyroid carcinoma cell lines. Until now, no study has evaluated the possible antiproliferative effect of TZD in “primary cultured human DPTC cells” (DPTC-cells), that could be very useful to identify patients responsive to therapy, and no comparison has been reported with the antiproliferative effect of antiblastics, or about a possible additive antiproliferative effect.

The proliferation of DPTC-cells was evaluated by: 1) cell counting; 2) a proliferation assay (WST-1). DPTC-cells were incubated with increasing concentrations of two different PPARg agonists (rosiglitazone, pioglitazone), or four different antiblastics (bleomycin, cisplatin, gemcitabine and etoposide), or the combination of PPARg and antiblastics.

The results of WST-1 assay in DPTC-cells, mean of 7 different experiments, showed a reduction of proliferation by TZD with respect to the control both with rosiglitazone (20 and 30 mcM; respectively, −23% and −29%; p < 0.001) or with pioglitazone (20 and 30 mcM; respectively, −22 and −26%; p < 0.001). Bleomycin, cisplatin, etoposide and gemcitabine significantly (>40%) inhibited DPTC-cells proliferations. An additive antiproliferative effect (p < 0.05) was observed with rosiglitazone 30 mcM in combination with etoposide, and with pioglitazone 30 mcM in combination with bleomycin. Cell counting confirmed the above mentioned results.

TZD exert an antiproliferative effect in DPTC-cells in vitro; an antiproliferative effect of antiblastics was observed, too; TZD in combination with antiblastics have additive antiproliferative effect in human DPTC-cells.

1 Surgery, Buddhist Tzu Chi General Hospital , Taipei Branch, Taipei, Taiwan 2 Surgery and Traumatology, National Taiwan University Hospital, Taipei, Taiwan Poster 29 Serum Vascular Endothelial Growth Factor D Levels Correlate with Cervical Lymph Node Metastases in Papillary Thyroid Carcinoma

Thyroid Cancer Thursday Poster Clinical

Vascular endothelial growth factor D (VEGF-D), being one of the most potent directly acting lymphangiogenic factors, induces lymphatic endothelial proliferation and facilitates nodal metastases. Although tissues VEGF-D have been shown to contribute to tumor metastases in PTC, the clinical relevance of serum VEGF-D remains unknown. The aim of this multicenter study is to evaluate the clinical relevance of serum vascular endothelial growth factor D (VEGF-D) in papillary thyroid carcinoma (PTC)

This prospective study consisted of 77 primary PTC and 15 benign thyroid nodules patients, treated during the years 2008–2009. Preoperative serum samples were collected and serum levels of VEGF-D were compared with the clinicopathologic features and lymph node metastases.

There is no difference of serum VEGF-D levels between PTC and benign thyroid nodules group (223.17 vs 238.84 pg/ml respectively, P = 0.286). In PTC group, serum VEGF-D levels was significantly increased in patients with lymph node metastases (246.65 vs 212.62 pg/ml respectively, P = 0.008). Age, gender, and tumor size had no impact on the serum VEGF-D expression.

Serum VEGR-D concentration correlated significantly with cervical lymph node metastases in PTC patients. Further study is necessary to evaluate the clinical application.

1 Endocrinology, Hospital Churruca Visca, Buenos Aires, Argentina 2 Endocrinology, Hospital de Clínicas-University of Buenos Aires, Buenos Aires, Argentina Poster 31 Finding the Correct Recombinant Human TSH (RHTSH) Dose in Patients with Thyroid Carcinoma and End-Stage Renal Disease

Thyroid Cancer Thursday Poster Clinical

We have previously reported the TSH levels in patients with end-stage renal disease and thyroid cancer after different doses of rhTSH (Pitoia et al. Endocr Pract 2008:961–6) We are now completing this previous report with a new rhTSH dosage.

We administered five rhTSH different schemes of dosage in four hemodialysed patients with renal failure. Three received rhTSH to administer a radioiodine dose (100 mCi, 60 mCi and 70 mCi 131I). Two patient received rhTSH for the assessment of the disease status.

The first patient, who received two rhTSH 0.9 mg injections, administered in day 1 and 3, presented persistent elevated serum TSH levels during 17 days. The peak serum TSH measured at day 5 after the first injection was 644 mUI/L. Only a self-limited diarrhea was observed. Patients 2,3 and 4 who received 0.45 mg of rhTSH in two consecutive days also had a persistent elevated serum TSH level during 15 days. The serum TSH surge reached a peak level around 400 to 500 mUI/L at day 3 after the first rhTSH injection. The fourth patient was again evaluated with the administration of two 0.225 mg rhTSH injections. Peak TSH levels at day three was 178 mUI/L, similar to that obtained in subjects with normal function. No adverse events were observed in these patients.

Two rhTSH doses of 0.225 mg might probably be the optimal dosage in hemodialized patients with end-stage renal disease.

1 Surgery, Tulane University School of Medicine, New Orleans, LA 2 Otolaryngology-Head and Neck, Johns Hopkins School of Medicine, Baltimore, MD Poster 32 Comparing the Efficacy and Safety of “No Tie” Thyroidectomy with the Harmonic Focus Dissector to Standard Open Conventional Thyroidectomy for Well Differentiated Thyroid Cancer as It Relates to “Completeness” of Resection

Thyroid Cancer Thursday Poster Clinical

Harmonic technology (Ethicon endosurgery) using the CS 14 endoshears for the control of thyroid vessels during thyroidectomy was proved to be safe, and to shorten the operative time. However, to our knowledge, no previous study has evaluated the effect of the new Harmonic-Focus hemostat dissection technique without supplementary ligation at the ligament of berry in a homogenous group of patients with papillary thyroid cancer undergoing total thyroidectomy +/− central lymph node dissection to evaluate the resulting radioiodine uptake (I-123) postoperatively.

This is a nonrandomized, prospective analysis of a series of papillary thyroid cancer patients undergoing total thyroidectomy +/− central lymph node dissection at a teaching institution. Twenty nine consecutive patients underwent total thyroidectomy +/− central lymph node dissection by the senior surgeon. Demographics, pathological characteristics, thyroid size, operative time, intra operative and postoperative complications and (I-123) uptake following surgery were collected.

Twelve patients out of twenty nine patients underwent surgery with the Harmonic Focus Dissector “no tie” technique. The 2 groups were similar regarding age, sex, thyroid size and primary tumor size. Mean ± SD operative time (minutes) was shorter in the harmonic focus group compared with the conventional technique group (210 ± 49 vs. 246 ± 97, p = 0.20). There was no difference between the 2 techniques regarding the intraoperative or postoperative complication rate. I-123 percentage uptakes following surgery were similar between the Harmonic Focus Dissector Group and the conventional technique group. (0.22 ± 0.25, versus 0.43 ± 0.48, p = 0.15).

The use of the Harmonic Focus dissector without supplementary ligation is safe, and significantly reduced the operative time compared to the use of the conventional hemostatic techniques in patients with papillary thyroid cancer undergoing total thyroidectomy+/−central lymph node dissection. Moreover, postoperative percent radioactive iodine uptake is similar to that seen with conventional hemostatic techniques.

1 Surgery, Tulane University, New Orleans, LA 2 Medicine, Tulane University, New Orleans, LA 3 Pathology, Tulane University, New Orleans, LA Poster 34 Black Thyroid Associated with Thyroid Carcinoma

Thyroid Cancer Thursday Poster Clinical

Black thyroid is a rare pigmented change seen mainly in patients on minocycline ingestion. Black thyroid is previously thought to be a benign disease. There are 61 reported cases of black thyroid and 13 cases in association with thyroid carcinoma. To date, no study has reported the incidence of black thyroid and the incidence of associated thyroid cancer. The aim of the current study is to analyze retrospectively the incidence of black thyroid pigmentation in thyroid surgery specimens and examine the relation to thyroid carcinoma.

This is a retrospective study of 560 Consecutive patients, all of who underwent thyroid resection surgery at our institution. Data on patients' demographics, preoperative fine needle aspiration (FNA), clinical and pathology reports were collected. Patients with black thyroids and thyroid cancers were identified.

Forty-one patients (7.3%) had evidence of black thyroid pigmentation in their surgical specimens. The mean age (±SD) was 52 (±25) years. All patients were clinically and biochemically euthyroid. Black pigmentation was never diagnosed in preoperative FNA, and only one patient had a preoperative diagnosis of papillary thyroid carcinoma. Other patients underwent surgery for various reasons. Twenty-four patients (58.5%) had incidental thyroid carcinoma associated with black pigmentation of the thyroid. Thirteen of these patients (54.2%) had papillary carcinomas, and multifocal disease represented 61.5% of cases.

FNA does not diagnose black thyroid. Black thyroid pigmentation is not always a benign disease. Black thyroid is associated with different types of thyroid cancers, mainly papillary thyroid carcinomas. There is increase incidence of multifocal disease with black thyroid pigmentation. Thyroid glands with black pigmentation deserve through pathologic examination, which should include increase in the number of blocks sampled from each specimen to rule out multifocal disease. Further studies are necessary to confirm these data.

1 NIDDK/Clinical Endocrinology Branch, National Institutes of Health, Bethesda, MD 2 NICHD, National Institutes of Health, Bethesda, MD 3 NCI, National Institutes of Health, Bethesda, MD Poster 38 Primary Thyroid Lymphoma in the Absence of Thyroid Autoimmunity (Case Report)

Thyroid Cancer Thursday Poster Clinical

Primary thyroid lymphoma constitutes <2% of thyroid neoplasia. It is most frequently observed in long-standing Hashimoto's thyroiditis and in the presence of anti-thyroid peroxidase (TPO) antibodies. We report a case of rapidly-progressive primary thyroid lymphoma in the absence of thyroid autoantibodies.

N/A

Case: A 59 year old female with an 8 year history of hypothyroidism presented for evaluation of an incidental thyroid nodule and increased L-T4 requirement. Anti-TPO and anti-thyroglobulin antibodies were negative. Thyroid ultrasound revealed a poorly differentiated 2.3x1.7 cm mixed lesion in the context of an inhomogeneous gland. Fine needle aspiration (FNA) showed lymphocytes with atypia of likely reactive origin and neutrophils in the setting of proteinaceous and necrotic debris. Grocott Methanamine Silver and Fite stains ruled out fungal and acid fast bacilli infections. 8 weeks later, she noticed a diffuse enlargement of the nodule. Ultrasound showed a heterogeneous mass anterior to the trachea, with loss of overall thyroid echotexture. A large superficial lymph node (LN) was noted in the lateral cervical area. FNA of the mass and lymph node were thus performed. Cytology showed large atypical lymphoid cells with enlarged irregular nuclei and scant basophilic cytoplasm with lymphoglandular bodies in the background. Flow cytometry revealed a population of large abnormal B cells expressing CD19, CD20, CD22 and CD45, and negative for surface immunoglobulin; findings consistent with a B-cell lymphoma. Excision biopsy of the superficial LN confirmed the diagnosis of diffuse large B-cell lymphoma. Imaging (CT of the neck, chest, abdomen, and pelvis) confirmed the thyroid as the primary site, excluding extra-cervical disease. The patient was started on Rituximab+CHOP chemotherapy.

Thyroid lymphoma is a rare and aggressive neoplasm of the thyroid. The occurrence of thyroid lymphoma in the absence of TPO antibodies is a particularly uncommon event. Nonetheless a high index of suspicion should be kept for this condition in the presence of a rapidly growing thyroid mass. Aggressive early medical intervention represents the cornerstone in the management of thyroid lymphoma.

1 Surgery, St. Martin De Porre Hospital, Chia-Yi, Taiwan 2 Biomedical Information Science, National Chiao-Tung University, Hsien-Chu, Taiwan Poster 39 Gastric Adenocarcinoma Metastatic to the Lung and the Thyroid Gland

Thyroid Cancer Thursday Poster Clinical

Clinical diagnosis of cancer metastatic to the thyroid is extremely rare, and more commonly diagnosed at autopsy. The more common primary sites for metastasis to the thyroid are kidney, breast, lung, colon, esophagus and uterus. Thyroid metastasis from stomach (including carcinoma and sarcoma) were only reported in 3 cases in English literature.

Herein we report a 71 year old man with the presentation of growing neck mass, tarry stool and progressive dyspnea for 2 months. Bilateral thyroid multinodular lesions and four tiny (0.3 to 0.5 cm) lesions were noted on computed tomography. A 3.5 cm ulcerated mass lesion was noted at the body of stomach by fiberoptic esophgogastroscopy.

Bilateral near-total thyroidectomy was performed due to airway compression with related symptoms. Wedge resection on one of these pulmonary nodules was performed through video assisted thoracoscopic surgery (VATS) for diagnosis, and the lesion on the stomach was diagnosed by trans-endoscopic biopsy. Poorly differentiated adenocarcinomas with the same histological profiles were noted on these three sites. The immunohistochemical staining for thyroglobulin of them were also negative. The I-131 imaging of the residual lung nodules and stomach was also negative. Patient underwent subsequent chemotherapy and the condition was stable.

For any patients with other sites of malignancy, appearance of synchronous or metachronous thyroid nodule cannot exclude the possibility of a metastatic lesion. Aggressive surgical and medical therapy might be beneficial for some of these patients.

1 Department of Clinical Biochemistry, Royal Free Hospital, London, United Kingdom 2 Centre for Hepatology, Royal Free Hospital, London, United Kingdom 3 Pharmacy, Royal Free Hospital, London, United Kingdom 4 Infectious Diseases Unit, Royal Free Hospital, London, United Kingdom 5 Department of Endocrinology, Royal Free Hospital, London, United Kingdom Poster 41 Thyroid Dysfunction in a Hepatitis C Population Treated with Interferon and Ribavirin Combination Therapy in a Uk Cohort

Thyroid Diseases Thursday Poster Clinical

Thyroid dysfunction (TD) is a recognised side effect of Interferon-α (IFNα) therapy in the treatment of hepatitis C (HCV) infection, with incidence reported between 5 and 33% in different studies worldwide. No data exist for a UK cohort. This retrospective study has determined the incidence of TD in a UK cohort of patients with HCV treated with IFNα and Ribavirin combination therapy (CT).

A retrospective analysis of 275 patients treated at the Royal Free Hospital London in a two year period from January 2006. Thyroid function, viral load, HCV genotypes and liver function were assessed during a 6 or 12 month course of CT. Pre-treatment serum TSH measurements were available for 62% of the cohort. Liaison occurred between the Hepatology Specialist Nurse team and the Department of Endocrinology regarding those patients who developed TD. All those with an undetectable serum TSH had a diagnostic thyroid isotope scan.

20% (n = 56; 15% of men, 32% of women) developed TD within 1–11 months of commencing CT. The mean time of presentation of TD was (mean ± standard deviation) 3.8 ± 2.7 months. In the total cohort 9% (n = 24) developed an initially suppressed serum TSH (1% Graves', 8% transient thyroiditis) and 12% (n = 32) developed an initially elevated serum TSH (2% permanent hypothyroidism, 10% transient hypothyroidism). The relative risk of developing TD was 2.14 (95% confidence interval between 1.35 and 3.36; p = 0.002) for women. No statistically significant differences in risk were observed between ethnic groups. HCV genotype status, abnormalities of liver enzymes and success of CT as determined by sustained viral response were not associated with an increased risk of progression to TD. The mean pre-CT serum TSH was 1.65 ± 1.31 mU/L for those remaining euthyroid and 2.89 ± 1.36 mU/L for those developing hypothyroidism (p = 0.0003). ROC curve analysis (see figure) determined a serum TSH of 2 mU/L as the optimal cutoff for the prediction of hypothyroidism (Sensitivity = 79.2%, Specificity = 78.1%, PPV = 37.3%, NPV = 95.8%).

Although there was a relatively high incidence of TD in this UK cohort, only 3% required long-term treatment for TD at the completion of CT. A pre-CT serum TSH ≥2 mU/L predicted the development of hypothyroidism.

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1 Section of Endocrinology, Boston University School of Medicine, Boston, MA 2 Division of Cardiology, Brigham and Women's Hospital, Boston, MA Poster 44 Race and Sex are Predictors of Weight Gain Following Radioactive Iodine Treatment for Hyperthyroidism

Thyroid Diseases Thursday Poster Clinical

Weight gain is a common complaint in patients being treated for hyperthyroidism.

We conducted a retrospective chart review of 111 patients who underwent radioactive iodine treatment (RAI) for hyperthyroidism during 2002–2005. We collected data on age, sex, race, and etiology of hyperthyroidism. Weights (kg) and TSH values (uIU/L) were obtained at diagnosis, at time of RAI, and at 0–4 months, 4–8 months, 12 months, and 24 months post treatment. Absolute and percent weight increase were calculated. We conducted subgroup analyses by sex, race, and etiology of disease. We also compared TSH values at each time point.

Mean age of our cohort was 49 with 86 females and 25 males. Of these, 38% were black, 48% white, and 14% Hispanic. 72% of the patients had a diagnosis of Graves disease. Mean weight was 74.4+/−16 kg at time of RAI (median TSH 0.03). There was a statistically significant absolute weight increase of 1.57+/−0.3 kg 0–4 months after RAI (median TSH 1.43), 2.72+/−0.4 kg 4–8 months after RAI (median TSH 1.72), 3.19+/−0.5 kg 8–12 mo after RAI (median TSH 2.48), and 3.78+/−0.7 kg two years after RAI (median TSH 1.25). Percent weight increase was statistically significant at each time point (p < 0.01). Subgroup analyses showed a significant difference in absolute and percent weight gain by sex, with males gaining more weight than females. Blacks gained significantly more weight than whites. We did not find a difference in absolute and percent weight gain when comparing Graves vs. other etiologies for hyperthyroidism. We did not find a correlation between degree of TSH elevation and observed weight gained.

Our data support prior studies suggesting weight gain following RAI for hyperthyroidism. In addition, we make the new observation that male patients and blacks are at higher risk for weight gain following RAI. The etiology for the patient's hyperthyroidism does not seem to play a role in weight gain. Likewise, the degree of hypothyroidism immediately post treatment does not seem to affect the eventual weight gain.

1 Dept. of Medicine and Endocrinology, Aarhus University Hospital, Aarhus, Denmark 2 Dept. of Breast and Endocrine Surgery, Aarhus University Hospital, Aarhus C, Denmark Poster 46 Sternotomy in Thyroid Operations: Retrospective Study of 48 Operations

Thyroid Diseases Thursday Poster Clinical

Surgical treatment of substernal goiter occasionally involves sternotomy. Classification and handling of these operations are widely discussed. Objective: To review surgical results after thyroid operations including median sternotomy.

A retrospective review of all thyroid operations performed from 01.01.95 to 31.12.08. In 48 of 1573 thyroid operations (3%) median sternotomy was performed. Indications for sternotomy were substernal malignant lymph nodes in 2 cases (4%) and signs of mechanical compression from substernal goiter in 46 (96%).

Pathologic examinations discovered malignant disease in 5 patients (10%) and multinodular goiter in 43 patients (90%). Mean operation time was 4 hours and 8 minutes (n = 43). Mean estimated blood loss was 433 ml (n = 42). Blood transfusion was given in 5 operations (10%). Median duration of postoperative hospitalization were 7 days (range 4 to 27 days). Radical resection was not possible in one patient (male, 73 years) with an anaplastic carcinoma and severe tracheal compression. This patient died postoperatively due to pneumonia and septicemia. Pulmonary complications occurred in 11 patients (23%): pneumonia or atelectasis (6), pneumothorax (3) and pleural effusion (2). Three patients (6%) had postoperative hypocalcaemia (permanent in 2 patients (4%)). Vocal cord paresis was not observed in this series of patients.

Thyroid operations with sternotomy are complicated procedures accompanied with considerable pulmonary complications, but in experienced hands the risk of the specific complications hypoparathyroidism or recurrent laryngeal nerve injury is not increased.

1 Endocrinology, Georgetown University Hospital/Washington Hospital Center, Washington, DC 2 Endocrinology, Washington Hospital Center, Washington, DC Poster 49 A Very Unusual Presentation of Graves' Disease

Thyroid Diseases Thursday Poster

Graves' disease is a common cause of hyperthyroidism. Uncommon manifestations include thyrotoxic periodic paralysis, thymic hyperplasia and hypercalcemia. We describe a case with all the above manifesting concomitantly, as an initial presentation of Graves' disease.

A 27 year old African-Amercian male patient presented with acute onset bilateral lower extremity weakness, tachycardia, hypertension, thyrotoxic orbitopathy and enlarged thyroid gland. Laboratory findings revealed hypokalemia, hypomagnesemia, hypercalcemia, hyperthyroidism and high thyroid stimulating immunoglobulin levels. Chest CT scan showed a homogeneous thymic mass >6 cm in dimension. Lower extremity weakness resolved after normalization of potassium. It was likely that his Graves' hyperthyroidism had precipitated the paralysis, thymic hyperplasia and hypercalcemia. Hence, he was prescribed methimazole and propranolol. One month later, he was euthyroid and normocalcemic. 15 weeks later, CT scan of the chest showed a significant decrease in the thymic mass to less than 2 cm confirming the initial diagnosis.

Thyrotoxic periodic paralysis (ThPP) is a rare complication of hyperthyroidism characterized by various degrees of muscle weakness and hypokalemia due to a massive intracellular shift of potassium. It has a predilection for young Asian males. Treatment consists of potassium, non-selective beta-blockers and anti-thyroid drugs. Thymic hyperplasia is a benign enlargement of thymic tissue, thought to result from a combination of hyperthyroidism and autoimmunity in Graves' disease, and typically regressing with its treatment. Hypercalcemia in hyperthyroidism is usually due to activation of osteoclastic bone resorption by excess thyroid hormone and is not thought to be PTH mediated.

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Thymic hyperplasia Image 1 (Before treatment of Graves' disease).

To our knowledge, this is the first report of Graves' disease complicated by the simultaneous manifestation of ThPP, thymic hyperplasia and hypercalcemia. African-American males have only rarely been described to suffer from ThPP. Clinicians should consider Graves' disease in their differential when they see occurrence of ThPP, thymic hyperplasia or hypercalcemia either in isolation or in combination.

1 Endocrinology, University of Vermont, Burlington, VT 2 ENT, University of Vermont, Burlington, VT 3 Pathology, University of Vermont, Burlington, VT Poster 50 Imaging and Histological Features of a Woman with an Unusual Case of New Onset Hypothyroidism and Thyroiditis

Thyroid Diseases Thursday Poster Clinical

A 47-year old woman was sent to the Endocrinology clinic by her primary care because of abnormal thyroid functions, thyroid enlargement and weight loss. The patient complained of difficulty swallowing for 4 months and had lost ten pounds. Her TSH was 69, and she was started on 25 mcg/d of L-thyroxine by her primary care. Four weeks later, this was increased to 50 mcg/d. A thyroid scan and uptake was done by her primary care which revealed a low uptake of 0.5%. An ultrasound showed the gland to be diffusely enlarged at 56 × 23 × 29 mm on the right and 51 × 21 × 14 mm on the left. The gland had a heterogenous echotexture with two nodules on the right, both measuring 15 mm. The patient had no previous history for thyroid problems, no family history of thyroid disease and no exposure to radiation.

Blood tests were ordered for thyroid autoantibodies and TSH. The patient underwent a fine-needle aspirate of the right-sided thyroid nodules, and then due to non-diagnostic findings, a core biopsy.

Blood testing revealed partially-corrected hypothyroidism with a TSH of 26 (normal = 0.35–5.5) and a free T4 of 0.9 (normal =0.8–1.8). Thyroid antibodies were very elevated: Anti-thyroglobulin >9000 IU/mL (normal: 0–40 IU/ml) and Thyroperoxidase Ab 2560 IU/mL (normal: <35 IU/ml). The FNA was non-diagnostic, showing only fibrous tissue and blood; no follicular cells seen. The core biopsy showed predominantly fibrotic tissue with entrapped atrophic follicles, lymphocytic infiltrate, and squamous metaplasia. Staining for amyloid was negative.

This patient presented with goiter, hypothyroidism and autoimmune markers consistent with Hashimoto's thyroiditis, but her weight loss and the hard texture of the gland, only apparent during the fine needle aspiration were unusual features. The core biopsy revealed fibrotic tissue and atrophic follicles, which in this clinical setting is consistent with Riedel's thyroiditis. The patient was placed on steroids, and experienced almost immediate clinical improvement in terms of being able to eat.

1 Pharmaceutical Research Institute, Albany College of Pharmacy and Health Sciences, Albany, NY 2 Physiology, Uludag University, Gorukle, Turkey 3 Ordway Research Institute, Albany, NY 4 Medicine, Albany Medical College, Albany, NY Poster 53 Tetraiodothyroacetic Acid (TETRAC) and its Nanoparticle Formulation Inhibit Pancreatic Cancer Proliferation, Tumor Growth and Its Associated Angiogenesis

Thyroid Hormone Action Thursday Poster Basic

Tetrac (tetraiodothyroacetic acid) is a deaminated analogue of L-thyroxine (T₄) that inhibits the actions of T₄ and 3, 5, 3′-triiodo-L-thyronine at the cell surface receptor for thyroid hormone on integrin αvβ3. The integrin is expressed on cancer cells and on endothelial cells and vascular smooth muscle cells and tetrac blocks the proliferative effects of thyroid hormone on tumor cells and the pro-angiogenic actions of the hormone on tumor-relevant vasculature. In the absence of thyroid hormone, tetrac also blocks angiogenesis induced by vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and other pro-angiogenesis factors. Tetrac as a nanoparticle (tetrac NP), covalently linked to poly (lactide-co-glycolide), does not gain access to the cell interior and acts exclusively at the integrin receptor. Human pancreatic cancer is an aggressive, progressive tumor that is poorly responsive to current treatment modalities.

In the present studies, the activity of tetrac and tetrac NP against human pancreatic cancer cells (PANC-1) was studied in the nude mouse xenograft model. Subcutaneous implantation of 10⁷ PANC-1 cells in the left and right flanks of each animal was carried out in control animals and tetrac-treated and tetrac NP-treated mice (n = 6 each for group). Tumor volume was 200–250 mm³ by 15 d, at which time treatment was introduced for 15 additional d.

Tetrac or tetrac NP resulted in significant suppression (P < 0.01) of tumor growth and of tumor angiogenesis. Tumor volume in treated mice was reduced by 15 d to less than the implantation volume and decrease in tumor size was apparent and significant at 5 d. Tetrac NP was at least 10-fold more potent than tetrac. Tumor vasculature was estimated by tumor hemoglobin content and was reduced by treatment by 60%. Animal weight gain was comparable in the control and treatment groups.

These observations document the anti-tumor and anti-angiogenic effectiveness of tetrac NP against human pancreatic cancer xenografts.

1 Pharmaceutical Sciences, College of Health Sciences, Kuwait, Kuwait 2 General Surgery, School of Medicine, University of Miami Miller, Miami, FL 3 Nuclear Medicine, Faculty of Medicine, University of Kuwait, Kuwait, Kuwait Poster 56 Radionuclide Evaluation of the Effect of Hyperthermia on the Thyroid Gland Function

Thyroid Imaging Thursday Poster Basic

Hyperthermia may be a consequence of environmental conditions, bacterial or viral infections and/or thyroid storm. The aim of this study is to investigate the acute effect of body temperature elevation on thyroid function and on its radionuclide evaluation.

Thyroid scintigraphy studies were performed on fifteen New Zealand White rabbits weighing approximately 3–3.5 kg. Each rabbit was injected with 115 MBq (3.1 mCi) technetium-99m pertechnetate (99mTc pertechnetate). Static images were acquired using Gamma camera. Two days later the same protocol was repeated for the same rabbit after increasing the body temperature by 2°C. The experiment was repeated again after a 2 day interval at 3°C, and then after another 2 day interval at 4°C. Plasma Free thyroxine (FT4), free triiodothyronine (FT3) and thyroid stimulating hormone (TSH) were measured at control and at different hyperthermic temperatures (+2, 3, 4°C). Isometric tension of isolated rabbit thyroid artery strips were also recorded in organ baths during stepwise temperature elevation.

During hyperthermia the significant decrease in thyroid function and thyroid scintigraphy studies were proportional to body temperature. The reduction was nearly 50%. The recording of isometric tension in rabbit thyroid artery strips in organ baths, showed vasoconstriction during hyperthermia which is significantly proportional to the heating temperature, (n = 10; *p < 0.05). Plasma FT4 and FT3 level were decreased while TSH levels were not affected by acute fever.

Our results indicate that acute hyperthermia causes a transient decrease in thyroid gland function and scintigraphic patterns on radionuclide studies. Thus, body temperatures must be measured before radionuclide studies in order to ensure that interpretation of data is not influenced by hyperthermia.

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1 Surgery, Tulane University School of Medicine, New Orleans, LA 2 Medicine, Tulane University School of Medicine, New Orleans, LA Poster 57 A Comparison of Ultrasound-Guided Thyroid Biopsy Techniques with Respect to Adequacy of Cytological Material

Thyroid Imaging Thursday Poster Clinical

Ultrasound-guided (US) fine-needle aspiration biopsy (FNAB) sampling of the thyroid represents a standard diagnostic procedure in the evaluation of thyroid nodules. The biopsy specimen is usually acquired using two different techniques. In the first technique, short axis is used with observation only of the tip of the needle whilst in the nodule. In the second technique a biopsy needle guides using the long axis to observe the entire needle. Either technique is usually used according to the operator's preference. We performed a retrospective review to compare these two techniques with regard to specimen adequacy.

We performed sonography-guided FNA in 105 thyroid nodules between May 2008-May 2009. The biopsy specimen was acquired using the two different methods after localization by one single operator. The type of technique was correlated with diagnostic accuracy.

63 out of 105 thyroid nodules were biopsied using long axis technique. The rates of inadequate material for long axis and short axis were significantly different (p < 0.05). Interestingly, the difference was more for lesions deeper than 3 cm from the skin (p < 0.05).

This is the first study to compare long and short axes techniques with regard to specimen adequacy for neck lesions. Long axis technique decreased the rate of inadequate material; number of required needle sticks and provides more accurate cytologic evaluation. We consider long axis technique using a biopsy needle guide easier and superior to traditional short axis technique.

1 Endocrinology Unit, Catholic University School of Medicine, Rome, Italy 2 Endocrinology Unit, Bambino Gesù Children Hospital, IRCCS, Rome, Italy 3 Department of Internal Medicine, AUSL Modena, Modena, Italy Poster 59 Nodular Recurrence After Hemithyroidectomy: Role of Levothyroxine (LT4) Therapy

Thyroid Nodules and Goiter Thursday Poster Clinical

Hemithyroidectomy is the surgical treatment of choice in patients with benign monolobar nodular disease. Nodular recurrence after lobectomy is a frequently observed event especially in iodine deficient areas. Although patients affected by diffuse non-toxic goitre often experience a reduction of the thyroid volume when treated with LT4, the role of LT4 in the management of nodular thyroid disease and prevention of nodular recurrence after lobectomy is still controversial. The aim of this retrospective study was to investigate whether post-operative treatment with LT4 reduces the incidence of nodular recurrences in patients who had undergone hemithyroidectomy for benign monolobar nodular diseases.

233 patients (38 males, 195 females; age 49.9 ± 13.1 years) with no post-surgical evidence of nodular disease in the remnant, were followed-up yearly with serum TSH and ultrasound (US). Nodular recurrence was defined as a lesion of at least 5 mm at US. Patients were divided in two groups: Group 1 (45 patients) who did not receive any LT4, and Group 2 (188 patients) treated with LT4. Group 2 was further subdivided in Group 2a (123 patients) receiving LT4 substitutive therapy (TSH ≥ 0.5 and ≤3 mUI/l) and Group 2b (65 patients) receiving LT4 at TSH-suppressive dose (TSH < 0.5 mUI/l). Mean observation period was 5.8 ± 4.7 years.

71 out of 233 (30.5%) patients experienced recurrence of thyroid nodular disease: 29 patients (64.4%) in Group 1, 24 (19.5%) patients in Group 2a, and 18 (27.7%) patients in Group 2b. The recurrence rate was significantly lower (p < 0.001) in Group 2 compared with Group 1, but no significant difference was observed between Groups 2a and 2b. TG levels showed an inverse relationship with recurrence. Indeed, the within-group difference was statistically significant in Group 1 and Group 2b, but not with 2a, which had the lowest overall recurrence rate among all three groups (64.4%, 27.7% and 19.5%, respectively).

In patients who have undergone hemithyroidectomy for benign monolobar nodular disease, LT4 therapy may prevent recurrence of nodular disease. TSH suppression may not be required for prevention of nodular recurrence in the remnant thyroid tissue.

1 Radiology, University of North Carolina, Chapel Hill, NC 2 Endocrinology, University of North Carolina, Chapel Hill, NC Poster 60 Effectiveness of Ultrasound-Guided Thyroid FNA with On-Site Pathology

Thyroid Nodules and Goiter Thursday Poster Clinical

Fine needle aspiration of thyroid nodules is a common procedure performed at UNC by the Departments of Endocrinology and Radiology. We aim to document the added value of on-site pathology for ultrasound guided thyroid fine needle aspiration biopsies. This will be measured in terms of specimen adequacy and number of needle biopsy passes required in two groups: biopsies performed by radiology which use pathology on-site and biopsies performed by endocrinology (without pathology). Both sets of biopsies are reviewed by the same pathologists, allowing for a well-controlled study.

This is a retrospective chart review study of all ultrasound guided adult thyroid biopsies performed at UNC by staff of the Departments of Radiology and Endocrinology. 100 cases performed by radiology and 100 cases performed by endocrinology have been reviewed for specimen adequacy and number of passes required. All cases were performed under ultrasound guidance with a 23G biopsy needle. All pathology samples were reviewed by the UNC Department of Pathology.

The primary data analysis compares specimen adequacy (measured as yes or no) within the endocrinology data subset and radiology data subset. The number of passes required in each group is also assessed. Analysis demonstrates a 6% inadequacy rate with onsite pathology versus a 14% inadequacy rate without onsite pathology. A student t-test analysis demonstrates that this is a statistically significant difference (P < 0.05). Our analysis indicates that there would be an approximate 57% reduction in the number of callback or repeat biopsies by having onsite pathology.

Success of ultrasound guided biopsy in acquiring an adequate tissue sample is significantly improved when performed under the supervision of a radiologist with on-site pathology. Clinical implications include a potential marked reduction in number of repeat biopsies.

1 Department of Surgery, Massachusetts General Hospital, Boston, MA 2 Department of Pathology, Massachusetts General Hospital, Boston, MA 3 Surgical Research Lab, Tufts Medical Center, Boston, MA 4 Department of Medicine, Massachusetts General Hospital, Boston, MA Poster 61 The Management of Thyroid Nodules with Atypical Cytology on Fine Needle Aspiration Biopsy

Thyroid Nodules and Goiter Thursday Poster Clinical

Fine needle aspiration biopsy (FNAB) of thyroid nodules can yield atypical cytology results in some patients. In this study we aimed to identify different variables affecting the management of patients with atypical FNAB to determine an algorithm of management, including timing of repeat FNA.

An IRB approved retrospective search of the MGH Cytology clinical database identified 254 patients with “atypical” FNAB between 1/2005–12/2007. Clinical, cytological and imaging variables were reviewed, and data was analyzed using SPSS statistical software.

Of the 254 patients with atypical FNAB, 203 patients had data available for analysis, 80% females and 20% males. Further subclassification included: 19% predominantly microfollicular lesions, 21% macrofollicular, 30% mixed cellularity and 30% other. 62% of patients went directly to surgery (group I), 25% had a repeat FNAB (group II) and 13% were followed up (group III). Significant differences between these 3 groups included: younger patients were more likely to go directly to surgery; 2.4% of males had follow-up only vs. 16% of the women; 76% with predominantly microfollicular cellularity went directly to surgery and only 2.6% were followed-up vs. 46.5% and 32% of those with macrofollicular cytology, respectively; 32% of nodules with cystic component were followed-up vs. 9% without. No influence on management was associated with age, history of radiation, Hashimoto's thyroiditis and other US characteristics. Analysis of repeat FNAB timing revealed that 59% of patients had repeat FNAB in <3 months and 41% ≥3 months, with the only variable significantly different between those groups being history of radiation- all those patients had repeat biopsy within 3 months. Repeat FNAB <3 months did not alter the results of the second FNAB, nor change the rate of malignancy on final pathology compared to repeat FNA ≥3 months. Overall rate of malignancy was 15%.

Atypical FNAB cytology constitutes a management dilemma. Current practice relies mainly on established prognostic factors such as gender, age, and cytological features. There seems to be no difference in outcome whether repeat biopsy is performed within or later then 3 months.

1 Institute for Experimental Endocrinology, Charité-Universitätsmedizin Berlin, Berlin, Germany 2 Basic Research Laboratory, National Cancer Institute, NIH, Bethesda, MD Oral 64 Targeted Disruption of Selenoprotein Biosynthesis in Thyrocytes

Thyroid Hormone Metabolism and Regulation Thursday Oral Basic 1:45 PM

Thyroid hormone biosynthesis requires oxidative iodination of tyrosine residues. Thus, thyrocytes need antioxidant protection against hydrogen peroxide. Oxidative DNA damage is implicated in the formation of thyroid nodules and cancer. Low dietary supply with the trace element selenium (Se) is associated with increased thyroidal damage and exacerbates inflammatory thyroid conditions. Conversely, Se supplementation ameliorates Graves' and Hashimoto's disease. The effects of Se are likely mediated by the expression of selenoproteins, proteins containing the rare amino acid, selenocysteine (Sec).

We have genetically abrogated selenoprotein biosynthesis in thyrocytes by conditional deletion of the gene encoding tRNA(Sec), Trsp. Recombination during thyroid differentiation was mediated by Pax8-Cre. Tamoxifen-inducible Trsp inactivation in the adult was achieved by Tg-CreER. Thyroids were dissected for histological examination, including morphometry of epithelial extension and follicle size. Expression of selenoproteins Gpx1 and Dio1 was assessed by immunohistochemistry and enzyme assay, respectively. The thyroid hormone axis was characterized according to serum hormone levels and T3-regulated transcripts in target organs.

Expression of Gpx1 and Dio1 activity were significantly reduced in the thyroids in both models. Despite the loss of thyroidal Dio1 expression, thyroid hormone serum levels or pituitary TSH-beta were not changed. Likewise, hepatic Dio1 expression was not altered in the Trsp-mutant mice. Thyroid morphology was not changed up to 6 months after recombination, but epithelial extension was diminished in Trsp-deficient thyroids and the mean follicle size increased significantly. In order to increase oxidative stress in the thyroids, animals were treated with perchlorate together with a low iodine diet. However, no indication of increased cell death or tissue disintegration were found.

We conclude from our experiments that, unexpectedly, selenoproteins including Dio1, are not essential for thyroid function. Possibly, non Se-dependent redox systems maintain protection of thyrocytes in the mutants. Reported beneficial Se effects may rest with the immune system and not with the thyrocytes.

1 Medicine, Baystate Medical Center, Springfield, MA 2 Pediatrics, Boston Children Hospital, Boston, MA 3 Medicine, Brigham and Women's Hospital, Boston, MA Oral 65 Mouse Skeletal Muscle Microsomes have Type-2 Iodothyronine Deiodinase (D2) MRNA and Activity. Possible Role in Hypermetabolism of Trα Deficient Mice

Thyroid Hormone Metabolism and Regulation Thursday Oral Basic 2:00 PM

We reported that mice with deletion of TRα gene (Thra-0/0) have a cold-dependent increase in energy expenditure. Wild type (WT) and Thra-0/0 mice have D2 mRNA in muscle. In both, levels were ≈10-fold higher in aerobic (Type-I) than fast muscle, but D2 mRNA was 2–3xhigher in Thra0/0, while activity in mouse muslcle has been questioned.

We used microsomes prepared by differential centrifugation. Initial experiments with 2 nM ¹²⁵I-rT3+1 mM PTU showed activity in proportion to RNA levels, with highest levels in soleus and red fibers of gastrocnemious, and lower levels in Type-II muscle fibers. Because spurious deiodination of ¹²⁵I-rT3 has been a concern, we validated the assay measuring ¹²⁵I-T3 production from outer-ring labeled ¹²⁵I-T4 by HPLC. Results showed production of ¹²⁵I-T3 in equimolar amounts with ¹²⁵I-. Activity was resistant to 1 mM PTU.

Using ¹²⁵I-rT3 as substrate, ¹²⁵I- production was inhibited in a concentration-dependent manner by T4 in vitro, and activity was reduced by in vivo T4 injections. Activity was increased about 2 fold in Thra-0/0 mice (P < 0.001); it was stimulated about 3-fold by cold and about 8-fold by 2 days of oral cholic acid. The effect of cold was reduced by prazosin or propranolol. Using outer-ring ¹²⁵I-T4, fractional ¹²⁵I- production was reduced by increasing concentrations of unlabeled T4 and by T4 administered in vivo. Since D3 activity has been reported in muscle, we added 100nM T3 to reduce T3 degradation. Apparent Km was 2.38 nM, in agreement with measurements in BAT and brain homogenates or microsomes, while Vmax was 240 fmol/h/mg microsomal protein (Lineweaver-Burk plot). Two-day treatment of WT and Thra-0/0 with 0.8 μg T4/100 g every 12h nearly normalized UCP3 mRNA in Thra-0/0 soleus but not in WT.

D2 is expressed in skeletal muscle of mice, substantially more in Type-I than in Type-II fiber muscle. With a sensitive method, D2 activity is demonstrable in muscle microsome with either ¹²⁵I-rT3 or ¹²⁵I-T4. Activity is proportional to the mRNA, and its manipulation and responses of T3-dependent genes in muscle to T4 suggest that D2 contributes to the hypermetabolism of Thra-0/0 mice and is a local source of T3 for muscle thermogenesis.

1 Medicine, Otolaryngology, Pathology and Laboratory Medicine, Mount Sinai Hospital, Toronto, Canada 2 Centre de Recherche Hotel-Dieu de Quebec, Universite Laval, Quebec City, Canada Oral 67 PIAS3 (Protein Inhibitor of Activated STAT3) acts as a TH-Dependent Co-Activator Which Is Dependent upon the Recruitment of an Intact SRC1 and HTRβ1 to Form a Trimeric Transcriptional Complex

Thyroid Hormone Action Thursday Oral Basic 2:30 PM

In a previous report, we have shown that PIAS3 (protein inhibitor of activated STAT3) with putative sumoylation properties is a thyroid hormone (TH)- dependent coactivator of hTRβ1 (79^th Annual Meeting of the American Thyroid Association, Abstract #88). In the present report, we have performed further studies to elucidate the molecular mechanisms accounting for its coactivator functional effects.

Protein-protein interaction assays were performed in vitro and in vivo using GST pull-down and mammalian two-hybrid assays respectively to determine the interaction of PIAS3 with hTRβ1 in the presence or absence of TH. In vivo studies were performed in HeLa cells for reporter gene activation effects and dual immunofluorescence labeling analyses, while coimmunoprecipitation analysis utilized 293T cells. The role of several post-translational modifications of phosphorylation by protein kinase A (PKA), histone deacetylation inhibition by trichostatin A (TSA), and histone acetyltransferase inhibition by Garcinol were also studied in HeLa cells. The specific roles of SRC1 and PIAS3 were determined by siRNA knockdown and replacements in HeLa cells.

We show that the hormone binding domain of hTRβ1 is essential for a TH-dependent interaction with PIAS3 and that intact RING finger and serine-rich domains are required for PIAS3 binding and coactivator function. PIAS3 complexes in vivo with hTRβ1 and is severely impaired by a siRNA knockdown of PIAS3, but can be reversed by replacement of wild-type PIAS3 coexpression. In wild-type HeLa cells, coexpression of PKA or addition of TSA increases PIAS3 mediated transcriptional activity, whereas added Garcinol dramatically inhibits increased PIAS3 induced coactivator function by SRC-1 but not TIP60. Similarly, a SRC1 knockdown prevents PIAS3 coactivator function. In vitro GST pull-down assays demonstrate that a SRC1/PIAS3/hTRβ1 trimeric complex is induced by the presence of TH.

1) PIAS3 functions as a potent TH-dependent coactivator which facilitates the formation of a trimeric complex with SRC1 and hTRβ1. 2) This TH-dependent trimeric complex can also be modulated by post-translational changes in phosphorylation, histone acetyltransferase and deacetylase pathways.

1 Division of Endocrinology and Metabolism, University of Pittsburgh, Pittsburgh, PA 2 Department of Pathology, University of Pittsburgh, Pittsburgh, PA 3 Section of Endocrine Surgery, University of Pittsburgh, Pittsburgh, PA 4 Department of Otolaryngology, University of Pittsburgh, Pittsburgh, PA 5 Department of Radiology, University of Pittsburgh, Pittsburgh, PA 6 Division of Endocrinology, Stanford University, Stanford, CA 7 Division of General Oncologic Surgery, City of Hope, Duarte, CA Oral 68 RAS Mutations are a Potent Predictor of Thyroid Carcinoma in Fine Needle Aspiration (FNA) Specimens

Thyroid Cancer Thursday Oral Clinical 1:45 PM

RAS is the most frequently mutated oncogene in human cancer and strongly contributes to neoplastic transformation. The prevalence, clinical significance and predictive utility of RAS mutations in the diagnosis of thyroid cancer have not been established. We report results of prospective molecular analysis performed between April, 2007 and May 2008 on 485 thyroid nodules tested for the most common RAS mutational hotspot codons: NRAS 61, HRAS 61 and KRAS 12/13.

Molecular evaluation was routinely performed on thyroid aspirates from cytologically malignant, indeterminate and non-diagnostic nodules and on selected benign specimens using real-time PCR and fluorescence melting curve analysis.

RAS mutations were found in 39 of 485 nodules (8%) from 37 patients. Of these, 29 were positive for NRAS 61, 8 for HRAS 61 and 2 for KRAS 12/13. Cytopathologic diagnoses included: 2 benign, 20 follicular lesion of undetermined significance (FLUS), 5 atypical or suspicious, 11 non-diagnostic, and 1 malignant nodule. Lobectomy or total thyroidectomy was subsequently performed in 30 of the 37 patients with RAS-positive nodules. Final histopathology was benign in 3 patients (10%) demonstrating 2 follicular adenomas and 1 hyperplastic nodule. Malignancy was confirmed in 27 patients (90%) consisting of 1 follicular carcinoma, 1 oncocytic (Hürthle cell) carcinoma, and 25 papillary thyroid carcinomas (PTC) (21 follicular variant (FV), 4 conventional type). Histologic tumor size ranged from <0.1 to 4 cm. None of the 25 PTCs demonstrated angiolymphatic invasion or extracapsular spread.

In this study we show that detection of a RAS mutation in a thyroid aspirate predicts malignancy in 90% of cases. This includes specimens with non-diagnostic and indeterminate cytology such as FLUS that comprised over 50% of our cohort. Malignant tumors were most commonly FVPTCs and typically occurred at an early stage without extensive invasion or extrathyroidal extension. However, 1 follicular and 1 oncocytic carcinoma were also present. The presence of RAS-positivity is therefore a highly reliable predictor of malignancy and should prompt consideration of surgical management even when the cytologic diagnosis is not definitively malignant.

1 Internal Medicine, University of Pisa, Pisa, Italy 2 Pathology, University of Verona, Verona, Italy 3 Surgery, University of Pisa, Pisa, Italy 4 Morphological and Biomedical Sciences, University of Verona, Verona, Italy Oral 70 Role of CLAUDIN-1 Expression, BRAFV600E Mutation and P2X7 Receptor Polymorphism Combined Analysis in the Appraisal of Papillary Thyroid Cancer Patients

Thyroid Cancer Thursday Oral Clinical 2:15 PM

BRAFV600E gene mutation represents the most common genetic event in papillary thyroid cancer (PTC). Claudin 1 (CLDN1) gene expression is frequently altered in several human cancers, including thyroid carcinoma. The modulation of the purinergic receptor P2X7 may be involved in human carcinogenesis. Recently, we observed a relationship between the 1513A/C loss-of-function polymorphism of the P2X7 receptor gene and either the PTC follicular variant (PFV) or an aggressive cancer behavior. The aim of the current study was to analyze CLDN1 expression and the frequency of both BRAFV600E mutation and the 1513A/C polymorphism of P2X7 receptor in different thyroid carcinoma samples and, to evaluate their potential role as suitable markers of the disease.

57 PTC [34 classical variant (PCV), 12 PFV, 7 tall cell variant (PTCV) and 4 PTC with poorly differentiated areas (PD)], with diverse TNM staging (class I/II: 50.9%; class III: 32.1%; class IV: 17.0%) were immunostained using a polyclonal CLDN1 antibody. 1513A/C polymorphism was evaluated by PCR amplification followed by RFLP analysis and BRAFV600E mutation by PCR-single-strand conformational polymorphism and sequencing. Non-neoplastic thyroid tissue was also investigated.

CLDN1 expression was significantly reduced in PTCV and PD samples (p = 0.01 vs PCV and PFV). 1513A/C polymorphism resulted significantly increased in PFV, PTCV and PD samples (p = 0.003 vs PCV). A significant relationship was found between 1513A/C polymorphism and either CLDN1 expression (p = 0.04) or BRAFV600E mutation (p = 0.0006). BRAFV600E mutation and 1513A/C polymorphism were associated with an advanced TNM stage (p = 0.04 for both), just BRAFV600E relating to lymph node metastasis (p = 0.05). Moreover, considering the joint presence of reduced CLDN1 expression and BRAFV600E mutation, a significant correlation was found with the histological aggressiveness (p = 0.03).

Overall, our preliminary data suggest that the combined use of these molecular markers may represent a potential tool in the early selection of PTC patients with aggressive clinical outcome and, in planning their therapeutic approach.

1 R&D, Diagnostic Hybrids, Inc. , Athens, OH 2 R&D, Interthyr Corporation, Athens, OH Poster 73 Development of a Novel Reporter Cell Line and a Standardized Protocol for the Detection of Thyroid Stimulating Immunoglobulin (TSI)

Autoimmunity Friday Poster Basic

Current methods for the detection of Thyroid Hormone Receptor Autoantibodies (TRAbs), such as the KRONUS TSH Receptor Autoantibody Coated-Tube RIA Kit, do not distinguish between TSI and other TRAbs that bind but do not stimulate. TSI are of particular importance because of their role in Grave's hyperthyroidism, where they mimic the function of thyroid stimulating hormone. The development of a cell line and standardized protocol that could be used to specifically detect and differentiate TSI from other TRAbs would be a powerful tool to assist physicians in the diagnosis of Grave's disease.

This novel cell line was developed by stably transfecting Chinese hamster ovary cells with a human thyrotropin/rat luteinizing hormone (LH) chimeric receptor and a firefly luciferase reporter gene. Selection of the clone was based on luciferase expression in response to TSI and results from cell passage stability data. A standardized assay protocol was developed based on results from analytical performance studies which included precision and reproducibility studies.

Stability studies showed that the cell line retained a constant level of luciferase activity in response to TSI over twenty-six passages regardless of the presence or absence of selection media. Results from our cross-reactivity and interference studies showed that that follicle stimulating hormone, LH, and other hormones did not interfere or cross-react at clinically relevant levels. A reproducibility study was conducted at 3 sites between 4 four technicians in order to ascertain the coefficient of variation (CV) for a panel of specimens containing various levels of TSI. The average CV was shown to be 20.8%. In a precision study involving 1 technician who performed the assay over 20 days, the average CV of a panel of specimens was 12%. The experimental CHO cell line was compared to the KRONUS TSH Receptor Autoantibody Coated-Tube RIA Kit using a series of clinical specimens. This set of clinical specimens showed an overall agreement between the two methods of 94.6%.

Our results show that this cell based reporter system is stimulated by TSI, is stable, and performs significantly similar to an FDA cleared predicate device.

1 Dept. of Medicine I, Gutenberg University, Mainz, Germany 2 Dept. of Research and Development, Diagnostic Hybrids Inc. , Athens, OH Poster 74 A Novel Cell-Based Assay of Thyroid Stimulating Immunoglobulins Correlates with Severity/Activity of Graves Hyperthyroidism and Orbitopathy

Autoimmunity Friday Poster Clinical

Discrimination between immunoglobulins that bind TSH-R from those with functional autoantibodies that play a relevant role in Graves disease (GD) is warranted. Thus, a validation trial was performed to compare the anti-TSH-R binding assays (TRAb) vs. a novel thyroid stimulating immunoglobulin (TSI) bioassay.

Sera of 200 GD patients (median age 46 yrs/range 13–80 yrs, 162 female) and of 40 euthyroid controls (21 yrs/13–68 yrs, 20 fem) were twice tested for TRAb (RIA and ELIA) and for TSIs in the Mc4CHOluc, a reporter cell line designed to measure the immunoglobulins that bind the extracellular domain of TSH-R and transmit the signals for cAMP-dependent activation of luciferase gene expression. The amino acid residues 262–335 of human TSH-R are replaced by the rat luteinizing receptor to confer greater sensitivity to TSIs than wild type TSH-R. The cAMP/CREB/CRE complex induces luciferase that is quantified after lysis of the cells in the presence of luminescent substrate.

Overall agreement of 91% was reached between the anti-TSH-R binding assays and the TSI Mc4CHOluc bioassay with a positive and a negative predictive agreement of 92% and 87%, respectively. All control sera were TSI negative. The results of the TRAb-confirmed discordant reactivity were 11/200 (5.5%) TSI pos/TRAb-neg as well as 6/200 (3%) TSI neg/TRAb-pos. Patients testing TSI pos/TRAb-neg were hyperthyroid and all had extra-thyroidal manifestations of GD: e.g. orbitopathy (GO, dermopathy, acropachy, and myxomatous cardiac valves. TSIs positively correlated with the clinical severity and activity scores of GO (p < 0.001). Also, TSIs were significantly more prevalent in patients with optic neuropathy and in smokers. Patients testing TSI neg/TRAb-pos were euthyroid and did show a mild course of GD.

TSIs closely correlate with clinical severity and activity of GD and GO. The utility of TSIs for both the monitoring of drug therapies as well as for the prediction of relapse in GD is now prospectively evaluated.

1 Dept. Bioregulation, LRC, NIID, Tokyo, Japan 2 Dept. Physiology, Keio Univ. , Tokyo, Japan 3 Dept. Medicine, USHUS, Bethesda, MD 4 Dept. Genetics, Tottori University, Tottori, Japan 5 Dept. Pathology, Yamanashi University, Yamanashi, Japan Poster 76 Molecular Cloning and Characterization of a Novel Thyroglobulin (TG) Variant in Mouse Kidney

Cell Biology Friday Poster Basic

Thyroglobulin (Tg) is a major product of the thyroid gland and is an essential substrate for thyroid hormone biosynthesis. It was shown that in addition to its classical function, Tg regulates thyroid follicular activity by modulating thyroid gene transcription. Tg also regulates cell growth in thyroid cells, lung epithelial cells, and kidney mesangial cells in culture. We have previously identified a fragment of Tg mRNA in cultured mesangial cells.

We have cloned and characterized a novel Tg variant expressed in mouse kidney.

Transcription of kTg starts in the middle of intron 41 of the Tg gene and results in the generation of a novel 13 amino acid segment. Real-time PCR and Western blot analysis indicated that kTg is expressed in the kidney and heart, but not in the thyroid. The presence of a predicted signal peptide in the kTg sequence was supported by the demonstration of a secreted protein from HEK293 cells expressing this gene. Sera from patients with Hashimoto's thyroiditis possessed IgG that recognized kTg.

The present study suggests that organs other than thyroid express a variant form of Tg that could serve both physiological and pathological roles, including that of an autoantigen in tissues where the variant is expressed.

1 Medicine and Medical Science, University College Dublin, Dublin, Ireland 2 Department of Obstetrics and Gaenacology, National Maternity Hospital, Dublin, Ireland Poster 78 Hormonal Control of Iodide Transport in Placenta Primary Cultures

Iodine Uptake and Metabolism Friday Poster Basic

Iodide transporter in the in human placental is known to be at least in part dependent on the anion transporters sodium iodide symporter (NIS) and Pendrin (PDS). A role for the placenta in iodine accumulation and storage and the former's possible control in a dose dependent manner by varying concentrations by of the pregnancy related hormones hCG (up to 60% increase), Oxytocin (up to 45% increase) and Prolactin (up to 32% increase) has been demonstrated by our group. The effect of these hormones in combination was even greater with the most effective combination of 17β Estradiol-Progesterone-Prolactin-Oxytocin increasing uptake by 82%. The aim of this study was to determine the mechanism by which these increases in uptake occur.

RNA was isolated from placental trophoblasts which had been pre-incubation with various hormones both alone and in combination using the quiagen RNA extraction kit. Taqman real time RT-PCR with NIS and PDS probes was carried out to determine the levels of expression of both transporters. NIS and PDS were normalised against the 18s RNA endogenous control.

In all treatments where iodide uptake was increased there were corresponding significant increases in expression level of NIS mRNA as compared with control non-treated trophoblast mRNA (hCG 1.42 ; Oxytocin 1.28 & Prolactin 1.26). There were also increases in NIS expression levels in the combination hormone treated cells(17β Estradiol -Progesterone-Prolactin-Oxytocin 1.56). There were no significant increases in the expression levels of Pendrin in any of the hormone treated cells indicating that the increased iodide uptake observed was solely due to increased NIS expression.

This study suggests a role for pregnancy associated hormones in promoting placental iodide uptake and indicates that modulation of NIS expression may provide protection against iodine deficiency in the fetus.

1 Endocrinology, Walter Reed Army Medical Center, Washington, DC 2 Endocrinology, National Naval Medical Center, Bethesda, MD 3 Dermatology, DeWitt Army Community Hospital, Fort Belvoir, VA Poster 79 Thyroid Protection Gone AWRY: A Rare Case of Iododerma

Iodine Uptake and Metabolism Friday Poster Clinical

Iododerma is a rare skin eruption that was more commonly reported in the first half of the 20th century when iodide was used as an expectorant, in asthma remedies, and as an antithyroid agent. It was later infrequently reported after the systemic use of iodide-containing radiographic contrast medium or treatment with oral potassium iodide. It has been rarely reported to occur following topical application of iodine. We herein report a 34 year old male with an adrenal mass, who developed an acneiform reaction over the chest and back with subjective swelling of the throat, shortly after treatment with supersaturated potassium iodide (SSKI).

SSKI containing 1g/mL (Upsher-Smith Labs, MN) was administered for protection of the thyroid gland during I123-metaiodobenzylguanidine scintigraphy to evaluate for pheochromocytoma. Though instructed to take only 2 drops twice daily, the patient administered 0.6mL twice per the manufacturer's instructions, with symptom onset the next day. Cessation of SSKI was accompanied by regression of all lesions. The patient's history showed no previous reaction to multiple iodide-containing compounds. As such, Lugol's solution was started, as it was speculated the syndrome was due to an additive in the SSKI, vice iodide itself. Lugol's solution was well tolerated without syndrome recurrence through the procedure and for 3 days post imaging. On the 3rd night, the patient enjoyed a crab and shrimp dinner. The next morning, the acneiform rash recurred, and the patient discontinued the medication. 3 weeks later, he was rechallenged with SSKI to evaluate for recurrence, and within 6 hours developed the same syndrome.

The diagnosis of iododerma was made. A skin punch biopsy was performed to confirm the diagnosis.

This case not only illustrates an adverse effect rarely encountered by endocrinologists, but reiterates the need for clear dosing instructions. Patients should receive explicit SSKI dosing instructions as the manufacturer label states an alternate dose.

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A rarely seen iododermic pustular eruption on the trunk one day following SSKI ingestion. This is thought to represent a hypersensitivity reaction to iodine. Resolution is usually spontaneous following cessation of iodide exposure.

1 Pharmacy Practice and Pharmaceutical Sciences, University of Minnesota Duluth, Duluth, MN 2 Biochemistry and Molecular Biology, University of Minnesota Duluth, Minneapolis, MN 3 Pediatrics, University of Minnesota, Minneapolis, MN Poster 80 Iron and Copper Deficiencies Lead to Reduced Thyroid Hormone Levels During Rodent Neonatal Development

Thyroid and Development Friday Poster Basic

Micronutrients play a critical role during late mammalian brain development. Copper (Cu), iron (Fe), and iodine deficiencies lead to similar defects in late brain developmental processes including reduced myelination of axons, and aberrant hippocampal structure and function, suggesting that these micronutrient deficiencies share a common mechanism that contributes to these derangements. Studies in both rodents and humans indicate that Cu and Fe deficiencies affect the hypothalamic-pituitary-thyroid (HPT) axis, leading to altered thyroid hormone (TH) status. Importantly, the relationship between Fe or Cu deficiencies and thyroidal status has not been assessed during the TH-sensitive period of perinatal mammalian life. We hypothesized that Cu and Fe deficiencies lead to reduction in circulating and brain TH levels during development and that this reduction contributes to the defects in brain development associated with these deficiencies.

To test this hypothesis we rendered pregnant Sprague-Dawley rat dams Cu-, Fe-, or TH-deficient from early gestation through weaning. Metal deficiencies were induced by feeding the dams Cu- or Fe-deficient diets and were confirmed by serum, liver, and brain metal analysis. TH deficiency was induced by addition of 10 ppm PTU in the drinking water. Thyroxine (T4) and triiodothyronine (T3) levels were subsequently measured in the serum and brains (T3 only) of rat pups at postnatal (PN) day 12.

Copper deficiency reduced serum total T3 by 48% (p < 0.05), serum total T4 by 21% (p = 0.06), and whole brain T3 by 10% at PN12. Iron deficiency reduced serum total T3 by 43% (p < 0.05), serum total T4 by 67% (p < 0.05), and whole brain T3 by 25% (p < 0.05) at PN12.

These results indicate that some of the brain defects associated with Fe and Cu deficiency may be mediated through reduced circulating and brain TH levels. We are currently assessing the expression of TH-responsive genes in PN12 Fe and Cu deficient brains to determine the physiological effect of the measured T3 reductions.

1 Otolaryngology, Head/Neck & Facial Plastic Surgery, HELIOS Kliniken Leipziger Land, Borna, Germany 2 General, Visceral & Vascular Surgery, HELIOS Kliniken Leipziger Land, Borna, Germany Poster 81 Endoscopic Minimal-Invasive Thyroidectomy (EMIT): Anatomical Considerations, Experimental Results and First Clinical Applications

Thyroid and Development Friday Poster Clinical

Refinements in the surgical technique and intraoperative neuromonitoring have lowered the complication rates of thyroidectomy in the past decades dramatically. Nevertheless the risk of transient recurrent laryngeal nerve palsy (RLNP) is 3.9%, those for permanent RLNP 1.9% respectively; local immediate and intermittent haemorrhage occurs in 1.5%, wound infections in 0.4% as well as postoperative dysphagia still count for 1.3% of the postoperative complications. Therefore in the last 20 years attempts were undertaken to reduce surgical access trauma by minimal-invasive techniques (MIVAT, extracervical accesses: axillary, ABBA). Due to extensive dissection especially in the extracervical routes these procedures do not compile with the term minimal-invasive.

Endoscopic minimal-invasive thyroidectomy (eMIT) should access the surgical field nearby in anatomical defined surgical spaces with minimal operative trauma (Img. 1). The eMIT-technique is defined by a sublingual (optic trocar) and bi-vestibular (working trocars) access to the subplatysmal space with standard instruments (3.7 mm diameter) for minimal-invasive surgery. Under endoscopic magnification the thyroid is reached after opening of the pretracheal muscles. The isthmus is dissected and the thyroid mobilized from medial to lateral. Vessels are identified, separated and dissected by the harmonic scalpell. The recurrent laryngeal nerve is tested by neuromonitoring for integrity. Resected thyroid is removed through the sublingual incision in the midline. Surgical plane is controlled finally.

Anatomical studies (8 embalmed humans) and an experimental study on 5 pigs with short-time survival showed the safety and feasibility of the method. Clinical application in the first 5 humans (first operation on 18th of March 2009) were successful and will be presented.

Benefits of the eMIT-technique are: direct access to anatomically performed spaces (subplatysmal plane), preparation under endoscopic magnification, expected lower RLNP-rate comparable to microscopic techniques (0.6%), lower local infection rates, avoidance of postoperative dysphagia caused by local scar formation and at least removal of the thyroid pathology without visible scar formation.

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1 Pediatrics, Uniformed Services University of the Health Sciences, Bethesda, MD 2 Surgery, Hospital for Endocrine Surgery, Kiev, Ukraine Poster 83 Trem Signaling is Activated in Metastatic Thyroid Cancer Cells

Thyroid Cancer Friday Poster Basic

Triggering receptor expressed on myeloid cells (TREM) molecule is a mediator of inflammatory response and is considered as a biomarker for sepsis. TREM signaling plays an important role in the regulation of tumor microenvironment, development of metastases and is considered as a prognostic marker for lung metastases from solid malignancies. In this study we examined the possible involvement of TREM signaling in thyroid cancer metastases.

We examined thyroid cancer cell lines that derived from primary thyroid cancer (FTC133) and metastases (FTC236 and FTC238) by real time PCR, Western blot and immunostaining. Gene silencing experiments and pharmacological inhibition of TREM signaling were performed. Human thyroid cancer samples were examined by immunostaining.

TREM expression and activation of TREM downstream target SYK (spleen tyrosine kinase) was detected in all examined cells with highest level in metastases-derived thyroid cancer cell lines (FTC236 and FTC238). Consistently, expression of genes coding for pro-inflammatory cytokines, chemokines and reactive oxygen species (ROS) were increased in FTC236 and FTC236 cells compared to FTC133 cells. Activation of TREM signaling was associated with increased resistance to serum deprivation inducible apoptosis in the metastatic cell lines. Treatment with a SYK pharmacological inhibitor (Emodin) decreased Akt activation and was associated with inhibition of cancer cell growth. In human thyroid tissue samples, TREM expression was increased in cancer compared to normal thyroid tissue and correlated with the presence of metastases as well as multifocal patterns of growth. SYK expression correlated with the level of Akt activation and staining of serial tissue samples showed co-localization of SYK and activated Akt.

Activation of TREM signaling occurs in metastatic cancer cells and correlates with activation of anti-apoptotic signaling. Targeting TREM/SYK pathway using currently available pharmacological compounds could represent a new therapeutic strategy for the treatment of metastatic thyroid cancer.

1 Medicine, Endocrine Division, Mount Sinai Hospital and University of Toronto Medical School, Toronto, Canada 2 Otolaryngology, Head & Neck Surgery Program, Mount Sinai Hospital and University of Toronto Medical School, Toronto, Canada Poster 84 Differential Expression of Epithelial Cell Adhesion Molecule Is a Potential Surface Biomarker for Aggressive Thyroid Carcinomas

Thyroid Cancer Friday Poster Basic

Identification of surface biomarkers for predicting aggressive cancers could assist in the improved management of thyroid carcinoma and the designing of relevant diagnostic and therapeutic strategies. In this report, we have performed studies using human thyroid cancer derived cell lines to assess the utility of epithelial cell adhesion molecule (EpCAM) as a potential surface biomarker in the detection of aggressive human thyroid carcinoma.

Using immunocytochemistry, Western blot, FACS and qRT-PCR technology, we have studied the differential expression of EpCAM in several cultured human thyroid cancer derived cell lines consisting of aggressive follicular (WRO, MRO), medullary (TT), anaplastic colon cancer (ARO) compared to a non-aggressive papillary thyroid carcinoma cell line (TPC-1). Additionally, we have compared the results of these studies to the effects in these cultured cell lines of in vitro administration of an immunotoxin (generously supplied by Vivientia Biotechnologies Inc) which links the Fab fragment of EpCAM monoclonal antibody to Pseudomonus exotoxin A (Vicinium^R) versus the effects of a full-length EpCAM monoclonal antibody MOC-31 (purchased from AbD seroTec).

Using these investigational techniques, we demonstrate that EpCAM is over-expressed in WRO, MRO and TT thyroid carcinoma and ARO cell lines but not in TPC-1. Treatment with Vicinium^R in increasing concentrations added to cell cultures dramatically impaired the growth of not only WRO, MRO and TT, but also ARO cell lines (IC50 = 5pM) whereas much higher concentrations of Vicinium^R to 125pM failed to significantly inhibit TPC-1. Addition of full-length MOC-31 EpCAM monoclonal antibody had no significant growth-inhibiting effects on any of the cultured cell lines at concentrations as high as 250pM.

1) These studies in cultured human carcinoma cell lines demonstrate that EpCAM is a potential surface biomarker which can predict aggressive thyroid cancer. 2) Further studies are warranted to determine the role of EpCAM as a potential diagnostic and therapeutic target in the management of aggressive thyroid carcinomas unresponsive to traditional surgery and radioactive iodine therapy.

1 Research and Development, Diagnostic Hybrids Inc. , Athens, OH 2 Research and Development, Columbus Nanoworks, Columbus, OH 3 College of Osteopathic Medicine, Ohio University, Athens, OH Poster 85 Evaluation of TSHR Expression in Peripheral Blood of Normal Subjects

Thyroid Cancer Friday Poster Basic

In 2007, Chia et al. developed a qRT-PCR assay which detects TSHR mRNA in peripheral blood. This assay has since been shown to be highly sensitive and specific for the detection of both preoperative and recurrent DTC and could provide an alternative method for detection and diagnosis. While previous studies have demonstrated that TSHR mRNA is expressed in white blood cells, we wanted to identify the particular white blood cell fractions that express TSHR as a potential means for reducing that background. In addition, we needed to determine the baseline level of TSHR expression in samples obtained from subjects with thyroids, and no known thyroid disease, in addition to subjects without thyroids.

A total of 31 subjects with thyroids and 33 subjects whose thyroids had been removed as a result of thyroid disease were analyzed for TSHR mRNA. Total RNA was prepared from whole blood and 250 ng was analyzed in our qRT-PCR assay using target specific primers. Flow cytometry was used to determine whether lymphocytes, granulocytes or monocytes express TSHR when compared to the TSHR positive control cell line.

Our preliminary results show that using a real time qRT-PCR protocol the mean for TSHR mRNA expression in normal subjects with thyroids (29.9+/−0.3 Cts; 95% CI) was approximately the same as that from subjects without thyroids (30.1+/−0.3Cts). Results from the flow cytometry studies show that the TSHR mAbs recognize this marker in the thyroid cancer control cells. However, the lymphocyte, granulocyte and monocyte fractions were negative for TSHR.

These data demonstrate that a baseline can be established for TSHR mRNA expression in peripheral blood and that this baseline is approximately the same in the two subject populations we studied. Our studies also indicate that while TSHR mRNA is expressed in peripheral blood, the receptor either exists in a form that is not recognized by our mAbs or at a significantly lower concentration than the positive control. In future studies, fluorescent activated cell sorting (FACS) will be used to isolate lymphocyte, granulocyte and monocytes cells followed by preparation of total RNA and testing in our qRT-PCR assay.

1 Section of Endocrinology, Diabetes, and Nutrition, Boston University Medical Center, Boston, MA 2 Division of Graduate Medical Sciences, Boston University Medical Center, Boston, MA, ³Internal Medicine, Harvard Vanguard Medical Associates, Boston, MA 4 Endocrinology, Harvard Vanguard Medical Associates, Boston, MA Poster 87 Factors Determining the Persistence or Recurrence of Well-Differentiated Thyroid Cancer in the Boston Area

Thyroid Cancer Friday Poster Clinical

Predictors of well-differentiated thyroid cancer (WDTC) persistence or recurrence have been inconsistent. No single scoring system is superior in predicting WDTC outcomes.

We conducted a retrospective chart review. Patients with ICD-9 code 193 (thyroid carcinoma) seen between 1979–2007 in a large Boston-area multispecialty group practice (Harvard Vanguard Medical Associates) were identified. Of these 1,025 patients, 670 WDTC subjects met study eligibility criteria. Cox proportional hazards models were used to assess predictors of WDTC persistence/recurrence. Independent predictors were gender, age, ethnicity, tumor size, surgical histology, extent of surgery, use of radioiodine, and clinician (endocrinologist or other) managing follow-up. Local extension of disease and lymph node involvement were not assessed. The primary outcome was WDTC persistence/recurrence, defined by either elevated thyroglobulin (Tg) levels in Tg-antibody negative patients or positive imaging. Study entry was the date of surgery. Follow-up was accrued until the first occurrence of the primary outcome or study termination.

Table 1 describes the cohort. There were 5051 total person-years of follow-up; 146 subjects (21.8%) had WDTC persistence/recurrence at a median of 4.2 years (range 0.02–42.5 years) after thyroid surgery. In multivariate regression models, age, sex, ethnicity, number of tumors, surgical histology, radioiodine use, and type of managing clinician were not predictive of the primary outcome. Tumor size ≥1 cm had a protective effect on WDTC persistence/recurrence (hazard ratio 0.49; 95% CI, 0.31 to 0.78; p = 0.002), which persisted after adjustment for extent of surgery and post-operative radioiodine use (hazard ratio 0.56; 95% CI, 0.3 to 0.98; p = 0.04). More complete surgery was also protective (hazard ratio 0.56; 95% CI, 0.34 to 0.91; p = 0.02).

Despite more complete surgery in subjects with tumors ≥1 cm, tumor size <1 cm was a predictor of WDTC thyroid cancer persistence/recurrence. Contrary to other studies, age and sex were not significant predictors of a worse prognosis. Further research is needed to better understand which factors and staging systems predict differentiated thyroid cancer persistence/recurrence.

1 Medical University Vienna, Vienna, Austria 2 Medicine, Nephrology and Dialysis, Vienna, Austria Poster 89 Reversibility of Increased Basal Calcitonin Concentrations in 133 Patients with Chronic Kidney Disease Stage 5 After Kidney Transplantation

Thyroid Cancer Friday Poster Clinical

Secondary hypercalcitoninemia, the occurrence of elevated calcitonin concentrations in patients with chronic kidney disease in absence of medullary thyroid carcinoma, appears with a prevalence of up to 46%. Up to 54% men with chronic kidney disease stage 5 on dialysis presents with elevated calcitonin concentrations, while in women only up to 28% show hypercalcitoninemia. It has been seen that kidney transplantation leads to a decrease of elevated calcitonin concentrations, but an investigation of a large cohort of chronic kidney disease patients has not been conducted before.

We investigated a group of 133 patients with chronic kidney disease stage 5 awaiting kidney transplantation. We studied whether elevated calcitonin concentrations would decrease one, two, or three years after transplantation. Responding to earlier results in gender-specific analysis, we divided the cohort in two subgroups, separating men from women.

48 (54%) of the men and 12 (28%) of the women with chronic kidney disease stage 5 presented secondary hypercalcitoninemia. Three years after successful kidney transplantation around 82% of men returned to normal calcitonin concentrations. In women secondary hypercalcitoninemia completely resolved within that time. 18% of men continued to have hypercalcitoninemia, although they presented rather low calcitonin levels compared to pre-transplantation values.

The present data show an almost complete reversibility of secondary hypercalcitoninemia in patients with CKD stage 5 after kidney transplantation. All patients with elevated calcitonin concentrations above 30 pg/ml presented declining calcitonin concentrations after kidney transplantation.

1 Endocrine Surgery, Cleveland Clinic, Cleveland, OH 2 Clinical Pathology, Cleveland Clinic, Cleveland, OH Poster 90 Understanding False Negative Values of TSH Receptor MRNA, a New Molecular Marker for Thyroid Cancer

Thyroid Cancer Friday Poster Clinical

The detection of circulating TSH receptor (TSHR) mRNA is indicative of thyroid cancer (TC). This bloodtest has shown early promise for differentiated TC diagnosis and long-term surveillance, but false negative results do occur. The purpose of this study was to describe a series of TC patients with negative TSHR-mRNA values.

From 251 samples in 176 patients, 30 false negative results in 28 patients were identified. Their clinical records were reviewed to determine TC-related diagnostic tests, operative findings and pathology.

Of 30 false-negative samples, 23 were collected preoperatively and 7 were collected during surveillance <6 months (n = 2) or ≥6 months (n = 5) after surgery. In the preoperative group, 11 patients (48%) had papillary microcarcinomas <1 cm, 7 patients (30%) had aggressive histologies (tall cell, angioinvasive follicular or poorly-differentiated cancers), and only 5 (22%) had classical papillary TC. All patients had clear indications for surgical treatment either by abnormal biopsy results or symptomatic goiter, hence negative mRNA did not impact treatment decisions. These 23 patients contributed a 50% NPV value to a cohort of 94 patients where preoperative TSHR mRNA had sensitivity 67%, specificity 96%, and PPV 98%. In the surveillance group, 5 patients had 7 negative TSHR mRNA samples in a cohort of 60 patients with 96% specificity, 95% NPV, and 50% sensitivity and PPV. Aggressive histology was present in 80% (4 of 5 patients). Recurrence was evident by biopsy-confirmed locoregional recurrence, stimulated thyroglobulin elevation, and either US, CT scan or FDG-PET imaging.

False negative TSHR mRNA values are associated with small papillary microcarcinomas preoperatively, but during surveillance typically occur in patients with aggressive cancers. This pattern may reflect unique determinants of circulating tumor cell biology and bears further study. False negative results did not deter appropriate clinical management, thus TSHR mRNA should continue to be viewed as a useful component of multimodality thyroid cancer care.

1 Tiroides, Fundación Dr. José Luis Novelli, Rosario, Argentina 2 Tiroides, Centro de Tiroides Dr. José luis Novelli, Rosario, Argentina 3 Ecografía, Instituto Cardiovascular de Rosario, Rosario, Argentina Poster 92 Neck Nodes Presurgical Evaluation with Doppler Color Echography and Angio Power in Papillary Thyroid Cancer

Thyroid Cancer Friday Poster Clinical

Papillary thyroid carcinoma (PTC) accounts for 80% of this gland's cancers. It shows very good prognosis, with a 10-year survival in 95% of patients. However, a specific feature is that it develops early metastasis in cervical ganglia.

55 patients operated on between March 2007 and May 2009 with PTC diagnosis. Two groups of patients were studied. Group A: 39 patients without palpable cervical adenopathies and Group B: 16 patients with palpable adenopathies. All patients' necks were examined with DCAP before initial treatment. The echographic features suspicious of malignity were the following: a) cephalo-caudal diameter similar to anteroposterior diameter and >10 mm in size, b) anomalous vascularization, c) heterogeneous cystic parenchyma, d) microcalcifications. Aspiration puncture with fine needle (PAAF) was performed under echography in patients with ecographically suspicious nodes.

DCAP reported suspicious nodes in 7/39 patients in Group A (18%) and in 6/16 patients in Group B (37,5%). PAAF found positive cytology for PTC in 4/7 in Group A (57%) and in 6/6 in Group B (100%), all confirmed by deferred biopsy.

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The use of DCAP enabled neck treatment in the first surgery in 10,3% more patients (4/39) without palpable nodes. Not all palpable nodes are metastatic, in Group B only 6/16 had suspicious nodes for DCAP, confirmed cytologically. This prevented the overtreatment of patients.

1 International Centre of Thyroid Disease, The International Fund “Help for Patients with Radiation-Induced Thyroid Cancer ‘Arnica’”, Minsk, Belarus 2 Clinic and Polyclinic of Nuclear Medicine, University of Wuerzburg, Wuerzburg, Germany Poster 94 Use of RHTSH for Follow-Up of Radiation-Induced Thyroid Cancer in Young Belarusian Women in the Period of Pregnancy Planning

Thyroid Cancer Friday Poster Clinical

Since 1992, 247 young Belarusian patients with advanced radiation-induced thyroid cancer (TC) after the Chernobyl accident have received radioiodine therapy in Germany. Today, 84% of them are in complete disease remission, and 79 patients (32%) already have their own children. Other patients will wish to have children, and the confirmation of the disease-free status before planning pregnancy is therefore very important.The goal of this study is to validate for the first time the use of Recombinant Human Thyrotropin (rhTSH) in Belarus during follow up of young women with TC in the period of pregnancy planning.

The study group consisted of 30 females under observation in the National Dispensary of Medical Rehabilitation (Belarus). Inclusion criteria for the cohort-study were: well-differentiated TC; complete surgery; more than 1 yr since the last course of radioiodine therapy, undetectable (<0.2 ng/mL) basal serum Tg and no clinical or US evidence of residual disease. Patients were given rhTSH, 0.9 mg (Thyrogen; Genzyme Corp., Cambridge, MA) as im. injections on 2 consecutive days on levothyroxine therapy. On day 1, 3 and 5, blood samples were drawn for determination of baseline and stimulated serum TSH (DPC, Germany), Tg (Brahms, Germany), and Tg-antibody levels (Pharmacia, Germany). An urine pregnancy test was performed a few days before rhTSH administration.

The mean age of patients at the time of the study has been 23.9 ± 1.8 yr, at the time of the Chernobyl accident 1.6 ± 1.6 yr and 12.7 ± 3.6 yr at the time of thyroid surgery. The peak TSH level after two injections of rhTSH was 111.5 ± 37,1 mU/L sharply decreasing to 10,2 ± 4,3 mU/L on day 5. All 30 patients had baseline Tg below 0.2 ng/ml. Serum Tg increased slightly after rhTSH stimulation to 0.4–0.9 ng/ml in three patients and was stable below 0,2 ng/ml in 27 patients.

Our data have shown that rhTSH can be successfully used in young female patients with TC for proof of complete remission in the phase of pregnancy planning. Under this regimen patients need no thyroid hormone withdrawal and so hypothyroidism with its possible unfavorable impact on the course of the disease and later cognitive development of the offspring can be prevented.

1 Thyroid Unit, Massachusetts General Hospital-Harvard Medical School, Boston, MA 2 Dept. Of Surgery, Beth Israel Deaconess Medical Center, Boston, MA 3 Dept. Of Surgery, Massachusetts General Hospital-Harvard Medical School, Boston, MA 4 Biostatistics Center, Massachusetts General Hospital, Boston, MA Poster 95 Remnant Lobe Ablation with I-131 is an Effective Alternative to Completion Thyroidectomy in Follicular Thyroid Cancer: Long Term Follow-Up of 39 Cases

Thyroid Cancer Friday Poster Clinical

Follicular thyroid carcinoma (FTC) is often diagnosed only after hemithyroidectomy for suspicious cytology of a thyroid nodule. FTC patients usually need completion thyroidectomy to prepare for radioiodine scan and treatment.

In this retrospective study, we have analyzed the long term outcome for 39 FTC patients treated with I-131 (30–32 mCi) to ablate the remnant thyroid lobe (Group LA), as an alternative to completion thyroidectomy. This group was compared to 73 patients (CT) who underwent completion thyroidectomy and to 29 patients who underwent total thyroidectomy (TT) initially. Age, gender, tumor size and grading were not significantly different among groups. All groups underwent I-131 WBS while hypothyroid, after either lobe ablation or total thyroidectomy. Select cases then underwent standard I-131 ablation, based on clinical findings.

After lobe ablation or total thyrodectomy, neck uptake was significantly lower in LA (0.6%) than in CT (2.0%-P < 0.005) and in TT (1.3%-P = 0.054). TSH and Tg levels were not significantly different between groups. TSH was >25 mcU/mL in all patients. Seventy-two percent of LA patients received I-131 ablation, compared with 88% in CT and 93% in TT. Median follow-up was 87 months in LA, 47 in CT, 53 in TT. There was one FTC related death each in LA and TT and no deaths in CT. At the end of follow-up there were 2 living patients with detectable disease in CT, 2 in TT and none in LA (P = NS). As patients without detectable disease were followed over decades, not all had follow-up stimulated Tg's. Of those measured, stimulated Tg's were <2 in 92% of LA (n = 12) and 100% of CT (n = 25) and TT (n = 11). Stimulated or suppressed Tg's <2 were found in 97% of LA (n = 30), 94% of CT (n = 65) and 100% of TT (n = 22). However, in 40% LA patients low level Tg (<2 ng/mL) was detectable at the end of follow-up, compared to 9% in CT and 15% in TT (p < 0.05).

Remnant lobe ablation with I-131 is an effective alternative to completion thyroidectomy in patient with follicular thyroid carcinoma, with similar long-term outcome in terms of survival and disease-free status. Persistence of very low level Tg is more common after this procedure, but this does not indicate an adverse clinical outcome.

1 Internal Medicine, School of Medicine, Catholic University of Daegu, Daegu, South Korea 2 Pathology, School of Medicine, Catholic University of Daegu, Daegu, South Korea 3 Surgery, School of Medicine, Catholic University of Daegu, Daegu, South Korea Poster 96 The Result of Microarray Analysis in Korean Papillary Thyroid Cancers

Thyroid Cancer Friday Poster Clinical

Papillary thyroid cancer (PTC) is the most common malignancy in the thyroid. Several molecular mechanisms are involved in the development and progression of PTC. BRAF V600E point mutation has been identified in the majority of PTC. BRAF mutation is more prevalent in Korea than other country. We investigated distinct genetic profiles in Korean PTC using cDNA microarrays analysis.

Transcriptional profiles of five PTCs and corresponding normal tissues were generated using cDNA microarrays. The tumors were genotyped for BRAF V600E point mutation. The results of gene expression of cDNA microarrays were confirmed by real-time PCR in 36 PTCs.

Four patients had BRAF V600E point mutation. Expression was increased for 93 genes, was decreased for 16 genes in cDNA microarrays of five PTCs. Realtime-PCR showed high expression of SLC34A2, TM7SF4, COMP, KLK7, KCNJ2 genes, showed low expression of FOXA2, SLC4A4, LYVE-1, TFCP2L1 genes in 36 PTCs. Among these genes, COMP, LYVE-1 KLK7 were previously unidentified genes in microarray of PTC. TM7SF4 gene was previously known to be overexpressed in microarray of PTC with BRAF V600E point mutation.

These findings demonstrate our result is similar to previous result, but show some different results compared with previous result. These findings may reveal clues to the molecular pathways involved in Korean PTC.

1 Department of Otorhinolayngology-Head and Neck Surgery, Korea University College of Medicine, Seoul, South Korea 2 Department of Pathology, Korea University College of Medicine, Seoul, South Korea Poster 99 The Value of Frozen Section for Detecting Extracapsular Spread in Papillary Thyroid Carcinomas

Thyroid Cancer Friday Poster Clinical

In papillary thyroid carcinomas (PTCs), extracapsular spread (ECS) is seems to be one of the indications of total thyroidectomy. Preoperative radiologic findings may be helpful for detecting ECS,. Many patients, however, undergo completion thyroidectomy according to the results of postoperative permanent section (PS), especially ECS. We investigated the value of frozen section (FS) for detecting ECS.

Medical records of 81 patients with PTC who underwent thyroid surgery and between June 2005 and June 2007 were reviewed. 19 patients were males and 62 were females. Median age was 52 years (range, 17–87). We compared the ECS results of FS to PS.

In 47 patients with ECS negative results on FS, only 18 patients (38.3%) were ECS negative on permanent section. In 34 patients with ECS positive results on FS, all patients had capsular invasion of cancer in final PS. (sensitivity: 54%; specificity: 100%, positive predictive value: 100%, negative predictive value: 38.3%).

Positive result of FS for ECS should be a reliable indicator to determine the extent of thyroid surgery. FS for detecting ECS is thought to be a helpful method to decrease the rate of completion thyroidemtomy.

1 Surgery, UMDNJ-Robert Wood Johnson Medical School, New Brunswick, NJ 2 Pathology, UMDNJ-Robert Wood Johnson Medical School, New Brunswick, NJ Poster 100 Prostate Adenocarcinoma Metastatic to the Thyroid in the Setting of Concomitant Papillary Thyroid Cancer: A Case Report

Thyroid Cancer Friday Poster Clinical

Metastatic disease to the thyroid is a rare clinical occurrence. The most common primary sites include the lung, breast, and kidney. Prostate cancer metastasis to the thyroid has only rarely been reported in the literature as it most frequently metastasizes hematogenously to the bone, lung, liver, pleura, and adrenals.

We present a case of an 87 year old man that presented with a palpable thyroid mass and significant weight loss. Fine needle aspiration of the left thyroid mass was suggestive of papillary thyroid carcinoma. A total thyroidectomy was performed showing a 2 cm papillary carcinoma with a second separate small focus of what appeared to be follicular variant papillary carcinoma.

Because of suspicion of prostate cancer, the slides were stained for thyroglobulin and prostate-specific antigen. The smaller focus was a prostate adenocarcinoma metastasis to the thyroid gland.

To our knowledge, this is the first report of a patient with a papillary thyroid carcinoma and a prostate cancer metastasis.

1 School of Clinical and Experimental Medicine, University of Birmingham, Birmingham, United Kingdom 2 School of Health and Population Sciences, University of Birmingham, Birmingham, United Kingdom Poster 104 Symptoms and Signs of Hyperthyroidism are Dependent on Patients' Age, Smoking History and Disease Severity: A Cross-Sectional Study of 3049 Patients

Thyroid Diseases Friday Poster Clinical

Hyperthyroidism is common and the clinical presentation of patients varies widely. Absence of symptoms and signs may result in delayed diagnosis and treatment in 0.5–1.5% of cases.

We investigated 3049 patients with hyperthyroidism simultaneously analyzing the influence of patients' age & gender, disease etiology & severity, symptom duration, and smoking on clinical symptoms and signs. All patients were evaluated using a standard proforma noting classical manifestations of hyperthyroidism. Patients were divided into quartiles according to age [16–32y (n = 766); 33–44y (n = 772); 45–60y (n = 779); ≥61y (n = 732)].

There was a higher prevalence of males (18.4%; 19.2%; 21.3%; 26.6%, p < 0.001) and toxic nodular disease (2.1%; 5.1%; 13.9%; 28.1%, p < 0.001) in older patients. Younger patients had more severe hyperthyroidism (mean fT4: 56.6; 51.7; 45.5; 40.6 pmol/l, p < 0.001). Overall, weight loss (60.7%), heat intolerance (54.9%), tremor (53.9%), palpitation (50.8%) and anxiety (41%) were the most common symptoms. The majority of patients ≥61y were asymptomatic (35.6%; 32.4%; 29.8%; 54.4%, p < 0.001) and the lowest proportion of subjects reporting ≥5 symptoms was found in the elderly (28.3%; 34.9%; 33.9%; 14.8%, p < 0.001). Increasing age was independently associated with reduced odds ratios for the presence of most classical symptoms except weight loss (AOR:1.02, p < 0.001) and shortness of breath (AOR:1.02, p < 0.001). Older age (AOR:1.08, p < 0.001), higher fT4 (AOR:1.01, p = 0.03), male gender (AOR:1.73, p = 0.03) and toxic nodular disease (AOR:3.13, p = 0.01) were independently associated with risk of atrial fibrillation. Higher fT4 and smoking were associated with tremor (AOR:1.01, p < 0.001 & AOR:1.27, p = 0.02) and goiter (AOR:1.01, p < 0.001 & AOR:1.29, p = 0.03) whereas older subjects were less likely to present with goiter (AOR:0.96, p < 0.001). Symptoms and signs of ophthalmoplegia were associated with increasing age (AOR:1.02, p = 0.006 & AOR:1.02, p = 0.001) and smoking (AOR:1.63, p = 0.03 & AOR:1.57, p = 0.002).

Symptoms and signs of hyperthyroidism are more prevalent in younger patients, smokers and those with more severe biochemical hyperthyroidism. We propose a lower threshold for performing thyroid function tests especially in the elderly.

1 Endocrinology, Mayo Clinic, Rochester, MN 2 Cardiovascular Diseases, Mayo Clinic, Rochester, MN Poster 105 Cohort Study of Amiodarone-Induced Thyrotoxicosis in Adults with Congenital Heart Disease

Thyroid Diseases Friday Poster Clinical

Amiodarone-induced thyrotoxicosis (AIT) is a well recognized complication of treatment with this anti-arrhythmic agent. It occurs abruptly, often after many months of treatment. Unfortunately, management options for AIT are limited. In contrast to amiodarone-induced hypothyroidism, no predisposing factors have thus far been identified, yet a higher prevalence was reported in the group of patients with congenital heart disease (CHD). We analyzed a cohort of adult CHD patients on amiodarone in order to determine the prevalence of AIT development and identify risk factors.

A historical cohort study was conducted. We included all Mayo Clinic patients seen in our CHD Clinic over the last 25 years that were treated with Amiodarone for ≥3 months. Baseline data were collected and patients were followed until diagnosis of AIT, discontinuation of amiodarone or last thyroid assessment on amiodarone. The diagnosis of AIT was made by identifying biochemical thyrotoxicosis during ongoing amiodarone therapy.

We evaluated 221 patients. Outcome assessment was possible in 172 patients. AIT developed in 22 patients, which represents 12.79% of the cohort (95% CI: 8.6 to 18.6%). AIT developed after a median of 1033 days on amiodarone (IQR 819–1463 days). The patients that did not develop AIT were followed on amiodarone for a median of 1098 days (IQR 415–2323 days). Women did not have a higher risk then men for AIT development, RR = 0.72 (95% CI: 0.34–1.51). Autoimmunity (any non-thyroidal autoimmune conditions or positive anti-thyroid antibodies) was not statistically associated with the development of AIT (RR = 1.64 with 95% CI: 0.46–5.82). In contrast a significantly lower BMI was present in the group that developed AIT (mean 21.91) by comparison with the rest of the cohort (mean BMI 25.31) for p = 0.0002.

AIT appears more likely to develop in the CHD population exposed to amiodarone than in patients with other varieties of cardiac disease. While autoimmunity does not appear to increase the risk for AIT in this population a lower BMI at initiation of treatment appears to be a strong risk factor. Therefore BMI may be a consideration in determining amiodarone dosing schedules.

1 Internal Medicine, University of Leipzig, Leipzig, Germany 2 Section of Endocrinology and Metabolism, Marmara Medical School, Istanbul, Turkey 3 Internal Medicine, Klinikum Innenstadt, Medizinische Klinik Muenchen, Munich, Germany 4 Institut für Medizinische Informatik, Statistik und Epidemiologie, University of Leipzig, Leipzig, Germany 5 General Surgery Department, Marmara Medical School, Istanbul, Turkey 6 Department of Pathology, Marmara Medical School, Istanbul, Turkey Poster 106 Comparison of Colour Flow Doppler Sonography (CFDS) and Immunostaining in the Detection of Intranodular Microvessels

Thyroid Diseases Friday Poster Clinical

The assessment of tumor vascularisation by colour flow doppler sonography (CFDS) has been suggested for the distinction between benign and malignant thyroid nodules. As microvessels are increased in cancer our objective was to investigate if the perfusion type determined by CFDS reflects the percentage of histologically determined microvessels in adenomas (A) & adenomatous nodules (AN) and papillary carcinoma (PC).

Tissue sections from 10 adenomas, 8 adenomatous nodules and 13 PC and surrounding tissue of 10 PCs and 2 benign nodules were immunostained for CD34. A computerized image analysis was used to determine the microvessel density in four hot spots and ten systematically selected fields. Preoperatively colour flow doppler sonography was performed for all thyroid nodules and classified according to Frates et al.

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A ROC analysis on the basis of the histologically determined number of microvessels demonstrated 70% microvessels as an optimal cutpoint for the diagnosis of PC with the highest sensitivity of 92% and highest specificity of 89%. The analysis of the CFDS-classification IV by Frates for the distinction between PC and A & AN showed a sensitivity of 62% with a specificity of 100%.

We found a consistent increase of the CD34 stained percentage of microvessels in PC (83% and 96%) as compared to adenomas & adenomatous nodules (38% and 49%) determined by the hot spot analysis and systematic field analysis. A ROC analysis on the basis of the histologically determined number of microvessels demonstrated 70% microvessels as an optimal cutpoint for the diagnosis of PC with the highest sensitivity of 92% and highest specificity of 89%. The analysis of the CFDS-classification IV by Frates for the distinction between PC and A & AN showed a sensitivity of 62% with a specificity of 100%.

The lower sensitivity of the CFDS-classification as compared with the immunohistologic determination of the microvessel density indicates that the CFDS-classification detects the pathognomonic intranodular microvessels only incompletely. The higher CFDS specificity is most likely due to the detection of other aspects of malignancy in addition to intranodular vascularisation.

1 Medicine, Obstetrics & Gynecology, Touro University College of Medicine, Hackensack, NJ 2 Division of Rheumatology, Johns Hopkins University School of Medicine, Baltimore, MD 3 Thyroid Research Unit, Mount Sinai School of Medicine, New York, NY Poster 107 High Prevalence of Thyroid Disease in Pregnant Women with Systemic Lupus Erythematosus

Thyroid Diseases Friday Poster Clinical

Postpartum thyroiditis (PPT), an autoimmune disease, occurs in 5–10% of all women. The incidence of PPT in women with Type 1 diabetes mellitus, another autoimmune disease is 25%. Individuals with systemic lupus erythematosus (SLE), yet another autoimmune disorder, have an increased incidence of autoimmune thyroid disease. The goal of the present study is to determine the incidence of thyroid disease during pregnancy and postpartum in women with SLE.

Sixty-three women who participated in the Hopkins Lupus Cohort and who had sera frozen during pregnancy and/or the first year postpartum were included in the study. As the study was retrospective, the timing of the samples during pregnancy and postpartum was not standardized. All samples were assayed for TSH, thyroid peroxidase and thyroglobulin antibodies. Extensive demographic and laboratory data from the Hopkins Lupus Cohort was available (i.e. sedimentation rate, steroid use, etc).

Twenty-four percent of the 63 women were positive for thyroid antibodies. Eight of the 63 women (13%) were on levothyroxine prior to becoming pregnant. Six of the remaining 55 women (11%) were diagnosed with hypothyroid based on their first pregnancy sample. Six of the remaining 49 women had a serum sample available only during pregnancy leaving 43 women who were evaluated for PPT. On average, each woman had 2 postpartum samples available for analysis. The incidence of PPT was 14% (6/43). Only one of the women who developed PPT was thyroid antibody positive either during pregnancy or the postpartum. Analysis of demographic and laboratory data revealed no correlation with the presence of thyroid disease.

The percentage of women with SLE who either had levothyroxine treated thyroid disease prior to pregnancy, who were diagnosed with hypothyroidism during pregnancy, or who developed PPT was 38%. It can be concluded, that women with SLE have a marked increased incidence of autoimmune thyroid disease. It is recommended that all women with SLE should have a TSH and thyroid antibodies performed prior to pregnancy.

1 Biyokimya, Acibadem University, Istanbul, Turkey 2 CNPRC, BMB, University of California, Davis, CA 3 California Animal Health & Food Safety Laboratory (CAHFS), University of California, Davis, CA 4 Division of Laboratory Sciences, Centers for Disease Control and Prevention, CDC, Atlanta, GA 5 Center of Health and Environment, University of California, Davis, CA 6 Western Institute for Food Safety and Security, University of California, Davis, CA Poster 108 Thyroid Status of Female Rhesus Monkeys in a Breeding Colony, Relationship to Reproductive Outcome and Preliminary Information on Impact of Perchlorate Administration on Thyroid Function

Thyroid Diseases Friday Poster

Thyroid disorders are known to disrupt fertility and reproduction in women and environmental contaminants that are seen as “endocrine disruptors” are thought to affect reproduction through their action on thyroid hormone systems. However, little is known about links between thyroid, reproductive function, and endocrine disrupting chemicals in nonhuman primates.Thyroid status was measured in adult female rhesus monkey breeders at the beginning of the breeding season.

Three related studies were conducted; screening of a group of indoor breeders for thyroid status; comparison of thyroid status to colony records on health and reproduction in a subsample of breeders, and administration of perchlorate and measurement of thyroid status in three lactating breeders. In additional work, three healthy lactating rhesus monkeys were given three different doses of environmental contaminant and thyroid iodine uptake inhibitor, ammonium perchlorate (0.006, 0.34, 12.8 mg/kg/day, respectively) in food for two weeks. Thyroid status variables (TSH, T4, T3, thyroid radioactive iodine uptake) were then measured.

The 95% confidence intervals for TSH, T4 and T3 (N = 66–80) were similar to those previously reported in smaller samples of macaque monkeys. Based on human criteria, 10 of 80 monkeys (12%) were hypothyroid (TSH > 2.0 μIU/mL. Because hypothyroxinemia can be a risk factor in pregnancy, T4 status was compared to past breeding history, breeding outcome for that season, and general health records in a subset of 42 breeders. T4 concentrations were significantly associated with the number of missed menstrual cycles during the previous breeding season. In the monkey receiving the highest perchlorate dose, iodine uptake was suppressed relative to baseline.

The study shows the availability of tools to study thyroid status in rhesus monkeys, the variability of thyroid status in the breeder colony, and the potential ability of environmental factors to influence thyroid status.

1 Department of Endocrinology and Metabolic Diseases, Rui-Jin Hospital, Shanghai Jiao-Tong University School of Medicine, Shanghai Institute of Endocrinology and Metabolic Diseases, Shanghai Clinical Center For Endocrine and Metabolic Diseases, Shanghai, China 2 Department of Surgery, Rui-Jin Hospital, Shanghai Jiao-Tong University School of Medicine, Shanghai, China 3 Department of Nuclear Medicine, Rui-Jin Hospital, Shanghai Jiao-Tong University School of Medicine, Shanghai, China 4 Department of Pathology, Rui-Jin Hospital, Shanghai Jiao-Tong University School of Medicine, Shanghai, China Poster 110 Successful Treatment with Short-Acting Octreotide of Two Unusual Cases of Intractable Hyperthyroidism Who Were Resistant to Antithyroid Drugs, Thyroidectomy and Radio-Iodine Therapy

Thyroid Diseases Friday Poster Clinical

We reported two unusual cases of hyperthyroidism who were unrespond to almost all kinds of antithyroid treatments: ATDs, corticosteroid hormone, radioiodine, thyroidectomy, lithium, etc. and were both finally effectively controlled by short-acting octreotide.

Case 1 and case 2 were both middle-age women. They presented with thyrotoxicosis with low serum concentration of TSH and very low RAIU values. The underlying causes, such as thyroiditis, iodine-induced or factitious hyperthyroidism, metastatic thyroid cancer and struma ovarii have been explored and ruled out. In spite of good compliance, their thyrotoxicosis could not be controlled with PTU (1000 mg/d in case 1) or MMI (60mg/d in case 2). B-blocker, lithium, methylprednisolone (in both cases) and radio-iodine therapy (in case 2) were all tried and failed. Even after total thyroidectomy in both cases, the hyperthyroidism recurred 5 months later in case 1 and 2 weeks later in case 2. Then short acting octreotide 0.1 mg q8h subcutaneously were first tried in case 2 and then in case 1.

The thyroid function improved greatly 3 days later in both cases. The doses of octreotide were decreased to 2 times a week gradually with consistent efficacy. After one month of octreotide treatment, case 1 developed hypothyroidism and needed L-T4 therapy. But 2 months later, L-T4 had to be discontinued and octreotide was re-started at the dose of 0.1mg qod because of the recurrence of hyperthyroidism in case 1. The thyrotoxicosis was again controlled and she again developed hypothyroidism. Under the preparation of this abstract, case 1 is treated with 100 ug L-T4, case 2 is under the therapy of octreotide 0.1mg twice a week with normal thyroid function.

Although the real cause of hyperthyroidism in these two patients is not identified, the mechanism of short acting octreotide in treating thyrotoxicosis with relative long term effect is not elucidated, and the definite conclusion on the usefulness of somatostatin analogue in treating hyperthyroidism could not be drawn, it provides us an alternative method to rapidly control thyrotoxicosis in patients resistant to different modalities of anti-thyroid therapy.

1 Neuroendocrinology, Cincinnati Cognitive Collaborative, Cincinnati, OH 2 Genetics, Institute for Chronic Diseases, Silver Spring, MD Poster 111 Metal Toxicity Determined by Bioassay of Urine Porphyrins Correlates with Thyroid Associated Cognitive Impairment and Psychiatric Disease

Thyroid Diseases Friday Poster Clinical

Toxic metals contribute to pathophysiology of dialysis dementia (Al), autism (Hg), neurodegeneration/neuropathy (As), Parkinson's/Lyme (Mn) and Alzheimer's disease (Al, Hg, Zn). Thyroid disease impairs cognition, likely via immunopathology after euthyroidism is achieved, and is associated with depression, especially bipolar. Urine porphyrins are associated with neuropathophysiology, perhaps via direct porphyrin toxicity. Symptomatic porphyria is rare; however, even low levels of toxic metals, particularly Hg, Pb or As, individually or in combination, often cause/s abnormal quantitative urine porphyrins whose association with thyroid disease or its related neuropsychiatry is unknown.

Urine samples were collected in dark containers, usually after achieving euthyroidism in thyroid patients. Brain SPECT used Tc-99m-HMPAO or Tc-99m-ECD IV in a quiet, dark room, to monitor cerebral pathophysiology with indices of cortical metabolism (CMi) and stimulated perfusion (CPi). Results were felt statistically significant if p < 0.05 using appropriate multiple component regression or standard two parameter comparisons, including thyroid disease or its subsets vs. other neuroendocrine patients, such as diabetics and traumatic brain injured (TBI).

Thyroid patients had significant (p < 0.05) correlation of abnormal corproporphyrins (likely due to Hg toxicity) and CPi which was more robust than noted for diabetics. Depressed patients were nonsystematically but confidently identified and especially thyroid disease associated bipolar patients trended toward marked Hg toxicity. Patients with history of Hg and other metal or other neuropathology were more severely affected. Patients treated with dimercapto-propane sulfonate (DMPS) and agents to enhance brain perfusion, such as omega 3 fish oil or cilostazol, and immune modulators such as vitamin D, statins and antihypertensives, stabalized or improved SPECT indices and clinical symptoms.

Toxic metals, particularly Hg body burden, identified by fractionated, quantitative urine prophyrins, correlate with thyroid associated cognitive impairment and possibly neuropsychiatric disorders, particularly bipolar depression.

1 Medicine, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Canada 2 Physiology and Pharmacology, Schulich School of Medicine, University of Western Ontario, London, Canada 3 Nuclear Medicine, Schulich School of Medicine and Dentistry, University of Western Ontario, London, Canada Poster 117 The 521C Variant of OATP1B1 Is Not Associated with a Change in Required Levothyroxine Dose

Thyroid Hormone Metabolism and Regulation Friday Poster Clinical

The individual levothyroxine (T4) dose requirement varies highly between patients and is difficult to predict. Recent studies indicate that transporters are required for thyroid hormone as shown by the role of Monocarboxylate Transporter 8 for levothyroxine uptake in the brain. We hypothesized that T4 transporters in the liver facilitate T4 uptake, and that variation in hepatic uptake due to genetic polymorphisms of the Organic Anion Transporter 1B1 (OATP1B1, gene SLCO1B1) may contribute to the interindividual T4 dose variation. In this study we focused on the SLCO1B1*5 (521T>C) variant of OATP1B1, which has been suggested to have decreased transporter activity.

We recruited patients with previous thyroidectomy for thyroid cancer. They were 18 years or older, and had no signs of persistent or recurrent disease. They had a stable T4 dose for at least 3 months and a TSH at target below 1 mIU/L. The T4 daily dose was adjusted per kg body weight, and a blood sample obtained to determine TSH, Free T4, Free T3 and thyroglobulin. Genotyping for OATP1B1 was performed using a TaqMan Real Time SNP Assay.

We included 112 patients, 83 women and 29 men. Genotyping identified 73 carriers of SLCO1B1 TT (wild-type), 34 of SLCO1B1 TC, and 5 of SLCO1B1 CC. The T4 dose/kg body weight (mean ±SD) in these subgroups was 2.15 ± 0.46, 2.08 ± 0.44, and 2.13 ± 0.32 microg/kg, respectively (P = NS). There was no difference in age, weight, waist, or thyroid function tests among groups.

The 521C variant is not associated with a change in T4 dose requirement. This suggests that OATP1B1 alone is not sufficient to alter systemic exposure to ingested T4, or possibly that expressed levels of other thyroid hormone uptake and efflux transporters compensate for the reduced OATP1B1-mediated hepatic T4 uptake among subjects who harbor the variant allele.

1 Department of Nuclear Medicine, Clinical Center, Niš, Serbia 2 Clinic for General Surgery, Clinical Center, Niš, Serbia 3 Department of Nuclear Medicine, Clinical Center, Kragujevac, Serbia 4 Clinic for Endocrinology, Clinical Center, Niš, Serbia 5 Institute of Oncology and Radiology, Clinical Center of Serbia, Belgrade, Serbia Poster 118 The Influence of 99MTC-EDDA/HYNIC-TOC Scintigraphy in Staging and Managing Strategy of Patients with Sporadic Medullary Thyroid Carcinoma

Thyroid Imaging Friday Poster Clinical

Medullary thyroid carcinoma (MTC) is a tumor of neuroendocrine origin with multiple somatostatin receptors (sst 2–5) expression, which represent the molecular basis for the clinical use of somatostatin analogues in diagnosis and treatment and their in vivo localisation. The aim of this study was to assess the diagnostic efficiency of 99mTc-EDDA/HYNIC-TOC (Tektrotyd) scintigraphy in the detection and staging of patients with MTC and to analyze its influence on managing strategy.

Nine patients with sporadic MTC, aged 53–70 years, were submitted to radionuclide investigation. Sequential scintigraphy using 99mTc-Tektrotyd was performed 2 and 24 hours after the injection of 740 MBq of the tracer. All the patients were imaged after the surgery during the follow up (2–17 years), whereas three of them were imaged before the surgery as well. Other imaging techniques were also applied and analyzed in individual cases and compared with 99mTc-Tektrotyd scan.

Out of the 3 patients who were imaged before the surgery, one was found to have increased uptake in primary tumor, while the scans of the other two showed cervical and axilar lymph node involvement beside the primary tumor. Scintigraphy using Tektrotyd detected a significantly greater number of lesions in other 5 out of 6 patients during the follow-up after thyreoidectomy, so that the patients were upstaged. In one of these patients, who had Tektrotyd avid metastases in bones, lung and lymph nodes were successfully managed using somatostatin analogue (Sandostatin LAR), while the other two with widespread metastases were selected for 90Y-DOTATOC therapy. In one patient Tektrotyd scan demonstrated mediastinal lymph nodes, but failed to show 4 bone lesions.

It can be concluded that 99mTc-Tektrotyd scan is a sensitive technique for diagnosis and follow-up of patients with MTC which can be used both for preoperative staging and for the detection of local recurrence and distant metastases. Moreover, the patients with widespread somatostatin avid metastatic lesions could be managed using either cold or labeled somatostatin analogues.

1 School of Clinical and Experimental Medicine, University of Birmingham, Birmingham, United Kingdom 2 ENT, University Hospitals Birmingham NHS Foundation Trust, Birmingham, United Kingdom Poster 119 Evidence of a Role for the PTTG Binding Factor (PBF) in Goitrogenesis

Thyroid Nodules and Goiter Friday Poster Basic

PTTG Binding Factor (PBF) is a poorly characterised transforming gene that is overexpressed in thyroid tumors. However, little is known about the precise role of PBF in vivo. In preliminary studies we assessed PBF expression in multinodular goiters (MNG) harvested at surgical excision. PBF mRNA expression was 2.6-fold higher in MNG tissue (n = 25, p < 0.01) than in normal thyroid tissue obtained during laryngectomy. Western blot analysis confirmed an approximate 2-fold increase in the level of PBF protein in MNG compared with normal thyroid.

We next generated a murine transgenic model of targeted overexpression of PBF in the thyroid to study its function in vivo. Transgenic mice were generated by pronuclear injection of a HA-tagged PBF transgene driven by the bovine thyroglobulin promoter, and founders identified by PCR.

Western blot analysis confirmed elevated levels of PBF-HA in thyroids from homozygote mice compared with wild-type (WT) FVB/N mice, with no significant expression in other tissues. Immunohistochemical staining using an anti-HA rabbit polyclonal antibody also detected extensive PBF-HA expression in thyroid follicular epithelial cells.The mean weight of thyroids from 6–12 week old homozygote mice (4.5 ± 0.8 mg, n = 18) was significantly greater than thyroids from WT mice (1.8 ± 0.2 mg, n = 15, p < 0.0001) of similar age. By 6 months of age the mean weight of thyroids from homozygote mice had increased further to 6.3 ± 1.1 mg (n = 11). Serum analysis by radioimmunoassays indicated no significant difference in thyroid function [serum T4 (3.1 ± 0.9 versus 3.2 ± 0.5 μg/dl, p = 0.207), T3 (118.4 ± 23.6 versus 140.5 ± 17.2 ng/dl, p = 0.128), and TSH (81.3 ± 20.3 versus 74.7 ± 14.7 ng/ml, p = 0.669)] between homozygote and WT mice, respectively.

These results demonstrate that targeted overexpression of PBF causes significant enlargement of the thyroid gland in vivo independent of thyroid hormone status. Given that PBF expression is induced both in human tumors and MNG, these results implicate PBF as having an important role in promoting goitrogenesis as well as thyroid tumorigenesis.

1 General Surgery, Cerrahpasa Medical Faculty, Istanbul, Turkey 2 General Surgery, Florence Nightingale Private Hospital, Istanbul, Turkey 3 Pathology, Nisantasi Pathology Laboratory, Istanbul, Turkey 4 Endocrinology, Istanbul Medical Faculty, Istanbul, Turkey Poster 120 The Curious Cases of Thyroid Follicular Lesions

Thyroid Nodules and Goiter Friday Poster Clinical

Fine needle aspiration biopsy (FNA)in thyroid nodules may not always give the correct result for the characteristics of a nodule. We used the diagnostic classification recommended by Papanicolau Society of Cytopathology for increasing the coherence of cytology and histology and for finding the resource of mistakes.

The results of frozen section(FS) and histologic analysis (HA) of 245 patients which had undergone FNA between August and December 2008 were analyzed. FNA results are classified 6 diagnostic categories.

Among the 96 patients who had benign results (BR) after FNA and FS, 69 patients (28%) had BR and 17 patients (7%) had follicular adenoma (FA) in the histological analysis (HA). 100 patients had follicular lesions (FL) or follicular neoplasm (FN). These patients were evaluated in 3 subgroups according to their histological types. I. group: 11 patients were diagnosed as FN and hurthle cell neoplasm (HCN) with FNA and FN(5), HCN(2), papillary thyroid cancer (PTC)(4) in FS and 5(2%) of them had FA in HA and 2 of patients (1%) had HCN and four of the patients (2%) had PTC.II. group:64 patients were diagnosed as FL by FNA and FN(59), HCN (3), hurhtle cell metaplasia in FS. 59 patients (24%) had FA, 3 patients (1%) had hurthle cell adenoma, 2 patients (1%) had BR in the HA.III.group:25 patients were diagnosed as FL by FNA and FN (23), benign (2) in FS. 3patients (1%) had folliculary thyroid cancer(FTC), 2 patients (1%) had PTC, 10 patients (4%) had PTC+FA, 8 patients (3%) had BR and 2 patients (1%) had BR+PTC in HA. 7 patients who had a suspect malignant FNAB result and FS result as PTC were accepted as PTC with histological analysis (2%). All of the 52 patients (21%) with malignant findings at FNAB and FS were diagnosed as PTC with HA.

The success of cytologic results of fine needle aspiration biopsy can be improved by knowledge, experience, technical equipment and by setting a standard terminology for pathology. The Bethesta Classification which is consisted of 6 diagnostic categories is a useful evaluation method for cytologic analysis. Among 6 of the groups the group with the most cytological diversity was the groups with follicular lesion and neoplasm.

1 Laval University, Quebec, Canada 2 Endocrinology, McGill University, Montreal, Canada 3 Otolaryngology, McGill University, Montreal, Canada 4 Pathology, McGill University, Montreal, Canada Poster 121 USFNAB Performed by Non-Radiologists in the Clinic on Previously Biopsied Non-Diagnostic Specimens: Does it Improve Patient Care in the Management of Thyroid Nodules?

Thyroid Nodules and Goiter Friday Poster Clinical

To assess the efficacy of ultrasound guided thyroid fine needle aspiration biopsies (USFNAB) performed in the clinic by non-radiologists in patients whose initial FNA were nondiagnostic with palpation guided FNA (PGFNAB). Second, to determine whether patient care can be enhanced by consolidating the diagnostic workup of nondiagnostic thyroid nodules by circumventing the inherent inefficiencies associated with radiological referral.

This is a retrospective analysis of 161 patients followed for thyroid nodules. All USFNABs were performed by a non-radiologist on consecutive patients with a previous non-diagnostic PGFNAB at a University based teaching hospital in Montreal, Canada. Biopsies were performed using a 20-gauge fine needle with a Mylab25 Biosound Esoate ultrasound (US) machine. Samples were then classified according to adequacy of sample and pathological findings.

One hundred and thirty-three patients underwent 161 USFNABs. The specimen adequacy rate was 91.3% (147/161). Among the adequate specimens, 7 were malignant (4.4%), 7 were suspicious for malignancy (4.4%), 86 were benign (53.3%), and 47 were indeterminate (29.2%). An analysis of thyroid USFNAB adequacy rates based on nodule consistency (cystic/complex vs. solid) proved statistically insignificant (P = 0.59). Likewise, there was no significant difference (P = 0.34) in the inadequacy rate between small (<1.5 cm) and large nodules (≥1.5 cm).

Our experience demonstrates that USFNABs performed in clinic by a non-radiologist is a promising tool for improving specimen adequacy for nodules initially classified as nondiagnostic. It also avoids the need for radiological consultation in the majority of cases thus improving efficacy in the work up of nodules. No statistically significant relationship was found between adequacy rate and nodule size or consistency.

1 Dept. of Experimental Medicine, University La Sapienza of Rome, II Faculty of Medicine, Rome, Italy 2 Centro Ricerche, Ospedale San Pietro Fatebenefratelli, Rome, Italy 3 Lab. of Molecular Oncogenesis, Regina Elena Cancer Inst. , Rome, Italy 4 Dept. of Pathology, Sant'Andrea Universitary Hospital, Rome, Italy Oral 124 Loss of HIPK-2 and the Paradoxical Co-Expression of WT-P53 and GAL-3 in Well-Differentiated Thyroid Tumors

Thyroid Cancer Friday Oral Basic 2:00 PM

Galectin-3 (Gal-3) is an anti-apoptotic molecule, essential for thyroid cells transformation in vitro, and useful as a preoperative diagnostic marker of thyroid malignancy. Recently, we demonstrated that Gal-3 gene expression is down-regulated by wt-p53, while it is stimulated by gain-of-function p53 mutants. However, in well-differentiated thyroid carcinomas there is a contradictory concomitant expression of Gal-3 and wt-p53. HIPK2 modulates p53 phosphorylation, acetylation, and localization and is necessary for the activation of p53 transcriptional properties, including Gal-3 repression. We, therefore, asked whether the loss of HIPK-2 expression could explain this paradoxical behaviour of p53 and Gal-3.

The expression of HIPK-2 was analyzed by immunohistochemistry in a total of 62 thyroid tissues, including 5 normal thyroid samples, 26 multinodular goiter (MNG), 26 papillary thyroid carcinomas (PTC) and 5 follicular thyroid carcinomas (FTC). To analyze the possible mechanism of HIPK-2 loss of expression we performed LOH analysis at the HIPK2 gene locus (7q32-34) on laser capture microdisseceted cells from 32 PTC, 7 FTC and 22 follicular adenomas (FA). Our results were also confirmed by in vitro experiments, using the PTC-derived K1 cell line, in which a stable RNA interference (RNAi) of the HIPK2 gene expression was generated. In these cells, the effect of the loss of HIPK2 gene expression on Gal-3 expression was evaluated by western blotting.

We found high levels of HIPK-2 expression in all normal tissues and MNG analyzed. HIPK2 expression was totally absent in almost all PTC (88.5%), and it was expressed at low levels in the remaining cases (11.5%). HIPK-2 expression was also absent in 40% of FTC. We found allelic loss at the HIPK-2 gene locus in 37.5% of PTC, in 13.5% of FTC and in 17.5% of FA. Interference with the HIPK-2 gene expression in K1 cells induced an up-regulation of Gal-3 mRNA and protein levels.

Our results indicate that HIPK2 expression is lost in PTCs and that this event is responsible for the lack of p53 inhibitory effect on Galectin-3 gene expression. HIPK-2 can be considered as a new tumor suppressor gene in well-differentiated thyroid cancers.

1 Medicine, Brigham & Women's Hospital, Boston, MA 2 Medicine, Newton Wellesley Hospital, Boston, MA Oral 127 Thyroid Hormone Early Adjustment in Pregnancy (the Therapy) Trial

Thyroid and Development Friday Oral Clinical 1:45 PM

Maternal thyroid hormone requirement increases approximately 30–40% during gestation. Women with known hypothyroidism must increase levothyroxine to prevent fetal harm, though how and when this should be accomplished is unclear.

We conducted a randomized, prospective, 4-year study enrolling 60 women with treated thyroid dysfunction seeking pregnancy. Once pregnant, women were randomized to proactively increase levothyroxine (LT4) by either 2 extra tablets/wk (29%; Group A), or 3 extra tablets/wk (43%; Group B) from their baseline dose. Thyroid function was tested every 2 wks through mid-pregnancy, and at 30 wks gestation. Levothyroxine was adjusted to maintain TSH between 0.5–5.0 uL/mL (0.1–5.0 uL/mL if cancer). We sought to determine which intervention would best prevent maternal hypothyroidism, while limiting hyperthyroidism.

48 women completed the protocol. Despite initial thyroid testing before 10wks gestation (mean 6.3wks), 13 women had elevated TSH values at study entry. Both interventions prevented further hypothyroidism later in pregnancy (1 of 25 patients - Group A; 1 of 23 patients - Group B). Because of TSH suppression, subsequent LT4 dose reduction was necessary in 8 of 25 (32%) women in group A, compared to 15 of 23 (65%) women in group B (p < 0.01). A higher pre-pregnancy LT4 dose predicted a greater risk for subsequent dose reduction (p = 0.03), while the gestational week of initial LT4 adjustment did not (p = 0.82). Subanalysis of Group A demonstrated that 24 of 25 subjects (95%) required LT4 testing no more frequent than once monthly; and 15 of 25 (60%) remained euthyroid through gestation with no further LT4 modification.

A 29% increase in levothyroxine (2 extra tablets weekly) initiated at confirmation of pregnancy prevents maternal hypothyroidism in 96% of subjects. 60% of subjects require no further LT4 adjustment, though 40% require subsequent dose modification, primarily predicted by higher pre-pregnancy LT4 requirements. Monthly evaluation of thyroid function is recommended through mid-pregnancy.

1 Neuroscience and Mental Health Program, Research Institute, The Hospital for Sick Children, Toronto, Canada 2 Psychology, University of Toronto, Toronto, Canada 3 Psychology, Toronto Western Hospital Research Institute, Toronto, Canada Oral 129 Using Functional Magnetic Resonance Imaging (FMRI) to Investigate Memory Functions in Children with Congenital Hypothyroidism

Thyroid and Development Friday Oral Clinical 2:15 PM

Extensive animal research has shown that thyroid hormone (TH) is essential for normal development of the hippocampus (H) and if lacking during late gestation or early postpartum, both H malformations and learning and memory impairments are seen. Children with congenital hypothyroidism (CH) detected by newborn screening undergo a brief period of TH insufficiency comparable to when experimental rodents lacked TH. We have previously shown these children exhibit (a) memory problems suggestive of H abnormalities, (b) H atrophy, particularly in posterior H region, (c) abnormalities in H chemical constitution, (d) abnormal integrity of H circuitry, and (e) correlations between memory weaknesses and structural H abnormalities. However H functioning itself has never been directly studied in CH. This study used fMRI, a noninvasive tool to examine brain function based on hemodynamic changes within specific regions during performance of different tasks, and two memory paradigms known to activate H in normal adults.

Studied were 15 children with CH and 15 controls aged 11–14 years. Each child underwent two separate scan sessions on the same day. Following extensive pretraining, two different task paradigms were performed in a 1.5 T scanner. One task used a verbal paired associates learning paradigm (VPAL) requiring learning and remembering novel word pairs and the other task, an object-place learning paradigm (OPL) requiring recall of objects in a grid. Data were processed and analyzed using SPM5.

Preliminary findings based on 9 CH and 12 controls indicate that although groups do not differ in performance during scanning, H activation on correct recognition was more diffuse and bilateral in CH. On VPAL, CH showed increased activation in left and right posterior H relative to controls (p = .018 and .032). On OPL location trials, CH showed increased activation in left and right middle H (p = .002 and p = .007) and on OPL object-pair trials, increased activation in left middle and right posterior H (p = .013 and p = .027).

Early TH insufficiency in children with CH alters how they process aspects of associative memory in H suggesting them may use compensatory, albeit less effective, H mechanisms to support such memory functions.

1 Endocrinology and Metabolism, Academic Medical Center, Amsterdam, Netherlands 2 INSERM U 577, Universite Victor Segalen Bordeaux 2, Bordeaux, France Oral 131 A Possible Role for the Thyroid Hormone Receptor α in Inflammation-Induced Changes in Pituitary Thyroid Hormone Metabolism

Thyroid Hormone Metabolism and Regulation Friday Oral Basic 4:30 PM

Acute illness in mice profoundly affects central thyroid hormone metabolism. We have recently shown a prominent role for the TRβ gene in the observed alterations. The thyroid hormone receptor α (TRα) is predominantly expressed in brain. It is unknown at present whether the TRα, like the TRβ, plays an important role in altered central thyroid hormone metabolism during acute illness. The aim of our study therefore is to evaluate the role of TRα in central thyroid hormone metabolism during acute illness by administration of a sub-lethal dose of bacterial endotoxin (LPS) to TRα0/0 and Wild Type (WT, 129/Sv) mice.

Mice were killed at 4, 8 and 24h after LPS; D2 and D3 mRNA expression was measured in the hypothalamic periventricular area (PE) and in the arcuate nucleus region (ARC). TSHβ, D1 and D2 mRNA expression was measured in the pituitary. Liver interleukin (IL)-1β mRNA was measured as a reflection of the inflammatory response.

The inflammatory response, indicated by increased liver IL-1β mRNA expression, was similar in WT and TRα0/0 mice. LPS increased D2 mRNA expression in the PE and ARC to a similar extent in TRα0/0 and WT mice. D3 mRNA decreased upon LPS administration in the PE which was also not different in both strains. D3 expression in the ARC region did not decrease in TRα0/0 mice and was significantly lower at 4h after LPS administration in WT compared to saline and LPS-treated TRα0/0 mice. Pituitary TSHβ and D1 mRNA expression decreased significantly after LPS in both genotypes, but the decrease of both mRNAs was more pronounced in TRα0/0 mice. Pituitary D2 mRNA increased shortly after LPS in TRα0/0 and WT but no difference was observed between genotypes.

The TRα gene is not involved in illness-induced alterations in hypothalamic and pituitary D2 mRNA expression. However, lacking the TRα gene intensifies the LPS-induced decrease of pituitary TSHβ and D1 mRNA despite a similar inflammatory response in TRα0/0 and WT mice. Our results implicate a possible role for TRα in selective LPS-induced alterations in pituitary thyroid hormone metabolism.

1 Endocrinology, Erasmus University Medical Center, Rotterdam, The Netherlands 2 Endocrinology and Metabolism, Odense University Hospital, Odense, Denmark 3 Institute of Public Health, University of Southern Denmark, Odense, Denmark 4 Statistics, University of Southern Denmark, Odense, Denmark 5 Epidemiology, Erasmus University Medical Center, Rotterdam, The Netherlands Oral 132 A Large Scale Association Analysis of 68 Thyroid Hormone Pathway Genes with TSH and FT4 Levels

Thyroid Hormone Metabolism and Regulation Friday Oral Basic 4:45 PM

Large variations in thyroid hormone (TH) concentrations exist between individuals. Approximately 65% of this variation is due to genetic factors. However, the causative genes and variants are largely unknown. We therefore performed a large scale association analysis of 68 TH pathway genes with TSH and FT4 levels.

68 genes with a role in TH synthesis, transport and metabolism were selected. Single nucleotide polymorphisms (SNPs) in these genes plus 10 kb flanking regions were selected from the Illumina HumanHap 550K SNP array (n = 1372 SNPs). Association analyses for these SNPs with TSH and FT4 were performed in a Dutch cohort of 1121 men and women (age: 69.0 ± 7.6 yrs). Suggestive SNPs (p < 0.01) were genotyped in an independent cohort of 700 Danish twins (age: 36.0 ± 10.6 yrs) for confirmation. All subjects were euthyroid with negative TPO antibodies. Analyses were adjusted for age and gender.

In the initial cohort, 32 SNPs in 11 genes were associated with TSH and 16 SNPs in 7 genes with FT4 (p < 0.01). Associations were replicated for SNPs in DIO1 (deiodinase), PDE8B (phosphodiesterase) and SLC16A10 (TH transporter). (see table).

We performed an association analysis of 68 TH pathway genes with serum TSH and FT4 levels in a large cohort. Genetic variation in 14 genes was associated with serum TSH and/or FT4 levels. Results could be replicated in an independent sample for DIO1, PDE8B and SLC16A10. Population characteristics (e.g. sample size, iodine status, age) might explain the absence of replication of the other associations. Therefore, additional cohorts will be studied and meta-analyzed for confirmation. These findings identify at least three causative genes and provide insight into the molecular basis of inter-individual variation in serum TH levels.

1 Laboratory of Comparative Endocrinology, K.U.Leuven, Leuven, Belgium 2 Division of Rheumatology, K.U.Leuven, Leuven, Belgium Oral 134 Thyroid Hormone Responsiveness in the Early Embryonic Brain

Thyroid and Development Friday Oral Basic 5:15 PM

Thyroid hormones (THs) play a central role in brain development. While experimental proof for their regulatory actions in late brain development continues to increase, experimental data for early brain development remain scarce. We used the chick embryo as a model to study the possible involvement of maternal THs in brain development during the first half of incubation (total duration 21 days).

We first measured the normal expression profile of TH-transporters OATP1c1 and MCT8, TH-deiodinases D1-D3, and TH-receptors TRα and TRβ in telen-, dien-, mesen- and rhombencephalon from 4- up to 12-day-old embryos (E4-E12). Subsequently we induced either hypothyroidism by treating laying hens with 0.05% methimazole or hyperthyroidism by injecting 0.5 μg T3+1 μg T4 into the egg yolk.

During early development OATP1c1 mRNA was mainly present in telen- and diencephalon and levels strongly decreased from E4 towards E12. mRNA levels of MCT8, D2, TRα, and TRβ varied between different brain regions but all expression profiles showed a gradual increase. D3 expression decreased from E4 to E12 while D1 expression showed no clear trend. The presence of active D2 and D3 protein was shown by in vitro deiodination tests while in vivo TH uptake in brain indicated the presence of active TH transporters. Microarray analysis of control and hypothyroid E4 and E8 telencephalon samples showed differential expression of a number of genes, including the homeobox genes Gbx2 and Six3, transcription factors involved in the organization of early brain. qRT-PCR confirmed increased expression of Gbx2 in telen-, dien- and rhombencephalon of hypothyroid E4 but not E8. Six3 expression was increased in hypothyroid E4 and E8 but only in diencephalon. Analysis of hyperthyroid telencephalon again showed increased expression of Gbx2 in E4 but not E8. This was accompanied by increased TRα, TRβ and D3 expression as well as decreased OATP1c1 expression in hyperthyroid E4.

Our results show that the early brain expresses the proteins needed for intracellular TH-uptake, -metabolism and -action in a region- and stage-specific way. They also show that both shortage and excess of THs change the expression of at least some genes involved in early brain organization.

1 Department of Investigational Cancer Therapeutics, [Phase I Clinical Trials Program], Division of Cancer Medicine, MD Anderson Cancer Center, Houston, TX 2 Department of Endocrine Neoplasia, Division of Internal Medicine, MD Anderson Cancer Center, Houston, TX 3 Head and Neck Medical Oncology Service, Memorial Sloan Kettering Cancer Center, New York, NY 4 Department of Medical Oncology, Fox Chase Cancer Center, Philadelphia, PA 5 Departments of Medcine and Oncology, Johns Hopkins University, Baltimore, MD 6 Department of Radiology, M.D. Anderson Cancer Center, Houston, TX 7 Clinical Development, Exelixis, South San Francisco, CA 8 Department of Medicine, Section of Hematology/Oncology, University of Chicago, Chicago, IL Oral 136 Preliminary Results of a Phase 1 Study of XL184 (BMS 907351)*, a MET, VEGFR2 and RET Receptor Tyrosine Kinase Inhibitor, Administered Orally to Patients (PTS) with Medullary Thyroid Cancer (MTC). *Co-developed by BMS and EXELIXIS

Thyroid Cancer Friday Oral Clinical 4:45 PM

XL184 is an oral inhibitor of MET, VEGFR2 and RET and exhibits anti-tumor and anti-angiogenic activity in xenograft tumor models.

Adults with advanced malignancies received XL184 in a 3 + 3 study design. Primary objectives include evaluation of safety and pharmacokinetic (PK) parameters and maximum tolerated dose (MTD) determination. RECIST response was assessed on day 28 and every 8 weeks. Pharmacodynamic and tumor markers, and RET mutational status are being analyzed. Adverse events (AEs) for 83 pts and PK results as of the 3/2009 data transfer are reported. Pharmacodynamic data and efficacy are reported for the 37-pt MTC subgroup.

85 pts were enrolled. XL184-related AEs were primarily Grade 1/2. Grade 3 related-AEs in ≥2 pts included palmar-plantar erythrodysesthesia (PPE, 11%); fatigue (10%); diarrhea (5%); increased AST, ALT, lipase and decreased weight (4% each); and hypertension and increased amylase (2% each). There were 2 related Grade 4 AEs; pulmonary embolism and increased lipase. The MTD of 175mg QD was based on DLTs of PPE, mucositis, and increased AST, ALT and lipase. The terminal half life is ∼100 hrs. Changes in PlGF, VEGF-A, sVEGFR2 and EPO were statistically significant and consistent with anti-angiogenic effects. Statistically significant increases in sMET occurred indicating MET pathway inhibition. Inhibition of XL184 targets and downstream signaling pathways was observed in skin samples. Of the MTC pts with measurable disease, 14 of 34 (41%) had ≥30% reduction in tumor measurements, with 9 of 34 (26%) achieving confirmed PR (cPR). The median time to response and preliminary duration of response for pts with a cPR were 28 days and 6.7 months respectively. The disease control rate (PR+SD >3 months) was 84%. Most MTC pts had reductions in plasma calcitonin and CEA. Response in MTC pts appears to be independent of RET mutational status.

XL184 is generally well tolerated; the MTD is defined. In MTC pts with measurable disease, 41% had ≥30% reduction in tumor measurements and of these, 26% achieved a cPR. The majority of MTC pts derived clinical benefit. A global phase 3 pivotal study of XL184 in MTC is ongoing.

1 Endocrinology, Mayo Clinic, Rochester, MN 2 Nuclear Medicine, Mayo Clinic, Rochester, MN 3 Biostatistics, Mayo Clinic, Rochester, MN Oral 137 Radioiodine Remnant Ablation (RRA) Did Not Improve Postoperative Outcome in 765 Adult Patients with Surgically Curable Macis <6 AJCC T1 Papillary Thyroid Carcinomas (PTC), Whether They Were Microcarcinomas (0.1–1.0 CM) or with Larger (1.1–2.0 CM) Diameters

Thyroid Cancer Friday Oral Clinical 5:00 PM

Neither the evidence base for the 2006 ATA Thyroid Cancer Guidelines nor a more recently published report (Thyroid 17: 1225–8, 2007) provided convincing data regarding a role for RRA in improving postoperative outcome in PTC patients with AJCC T1 tumors, especially those with diameters of 11 to 20 mm. If a prospective trial was to be designed to answer this question, it would likely involve “low-risk” (eg with MACIS scores <6) patients aged 21 yr or older, with surgically curable T1 disease (ie neither T4 nor M1) completely resected by initial bilateral lobar resection (BLR).

In this retrospective study, our aim was to define outcome in such a cohort of 765 adult patients with MACIS <6 AJCC T1 PTC treated during the 35 yr after the introduction of RRA to our institution, and before the current era of ultrasound-guided nodal biopsies, rhTSH-stimulated Tg testing and central compartment dissection. The 765 patients (545F; 220 M) underwent BLR during 1950–1985 for tumors that were completely resected and neither locally invasive or distant spread.

Median patient age was 45 yr (range 21–72), mean tumor size 12 mm (49% <11 mm); 24% were multicentric and 30% node-positive. Mean follow-up was 27 yr (longest 55); 23% for >35 yr. 170 (22%) received RRA. 35-yr occurrence rates for cause-specific mortality (CSM), local recurrence (LR), regional nodal metastases (RNM) and distant metastases (DM) were 1.1, 2.2, 5.0 and 1.6%, respectively. Comparable 35-yr rates for Group A, (n = 375 tumors <11 mm), were 0.6, 1.8, 4.5, and 0.5%. For Group B, (n = 390 tumors of 11–20 mm), rates were higher at 1.6, 4.3, 5.5, and 2.6%, respectively. RRA's impact was assessed by comparing survival to 4 endpoints in patients undergoing BLR alone versus those receiving BLR+RRA within 18 mos.The 35-yr CSM rates in Groups A and B were after BLR 0.6 and 1.6%, and after BLR+RRA insignificantly different (p > 0.75). Similarly, survival rates to LR, RNM and DM were no different in ablated patients than after BLR alone in both Groups A (p > 0.07) and B (p > 0.33).

These results confirmed the excellent prognosis of AJCC T1 tumors treated by BLR, and did not identify a significant reduction in mortality or recurrence rates in patients with T1 PTC tumors selected for RRA.

1 Medicine, Endocrine Division, Mount Sinai Hospital and University of Toronto Medical School, Toronto, Canada 2 Otolaryngology, Head and Neck Surgery, Mount Sinai Hospital and University of Toronto Medical School, Toronto, Canada Oral 138 Serial Stimulated Thyroglobulin Measurements in Low-Risk Well-Differentiated Thyroid Carcinoma Patients Not Receiving Radioactive Remnant Ablation

Thyroid Cancer Friday Oral Clinical 5:15 PM

We have previously reported on the application of a post-surgical, TSH-stimulated serum thyroglobulin (S-Tg), as a method for radioactive remnant ablation (RRA) selection in low-risk well-differentiated thyroid carcinoma (WDTC) (79th ATA Meeting, Abstract #79). Herein, we report on the serial S-Tg measurements of those low-risk WDTC patients who did not receive RRA.

Forty low-risk WDTC patients were followed after total thyroidectomy and an initial S-Tg ≤5 μg/L, without RRA. Low-risk was defined as tumors with papillary histology confined to the thyroid gland and without multi-level local lymph node involvement. All patients had negative anti-Tg antibodies by two independent assays. Patients underwent repeat S-Tg measurements, neck ultrasound and additional imaging when indicated. The mean follow-up interval was 3.4 years (range = 1.3 to 12.0 years).

All patients had an undetectable serum Tg on thyroid hormone suppression therapy for the duration of the follow-up period. Among the 40 patients, 23 patients (57.5%) had an undetectable S-Tg (<1 μg/L) after total thyroidectomy. Within this subgroup, all patients (100%) had subsequent S-Tg values ≤1 μg/L with no significant imaging abnormalities. The remaining 17 patients (42.5%) had a post-surgical S-Tg of 1–5 μg/L. Within this subgroup, 7 (41.2%) had repeat S-Tg values that declined, 3 (17.6%) had repeat S-Tg values that increased, but did not exceed 4 μg/L, and the remaining 7 (41.2%) had no change in their repeat S-Tg value. None of these 17 patients showed significant imaging abnormalities on follow-up routine ultrasound studies or selected CT scans when indicated.

To date, follow-up of low-risk WDTC patients who did not receive RRA by serial S-Tg measurements has revealed that (1) patients with a post-surgical S-Tg value of <1 μg/L have no evidence of residual/recurrent thyroid carcinoma, and (2) patients with a post-surgical S-Tg value of 1–5 μg/L have not had significant changes in their follow-up S-Tg and imaging studies to justify RRA intervention. (3) These observations add support to our previously proposed RRA selection guidelines in low-risk WDTC based on an initial post-surgical S-Tg value.

1 School of Clinical and Experimental Medicine, University of Birmingham, Edgbaston, Birmingham, United Kingdom 2 Institute of Human Genetics, International Centre for Life, University of Newcastle, Newcastle upon Tyne, United Kingdom 3 MRC Center for Causal Analyses in Translational Epidemiology, Department of Social Medicine, University of Bristol, Bristol, United Kingdom Poster 139 Extended Analysis of Nine Novel Nonsynonymous SNPS in a Large UK Graves' Disease Association Study

Autoimmunity Saturday Poster Basic

Recent genome-wide association studies in Graves' disease (GD) are beginning to reveal a number of novel genetic variants. The Wellcome Trust Case Control Consortium (WTCCC) in the UK recently investigated 900 GD cases and 1500 controls, using a genome-wide set of 14,500 nonsynonymous coding single nucleotide polymorphisms (nsSNPs). Whilst the strongest association signal was unsurprisingly identified in the HLA region (P value <10⁻²⁰), association was also confirmed at the previously reported thyroid stimulating hormone receptor and Fc receptor-like 3 genes, along with the identification of nine novel possible regions of association (P > 10⁻⁴).

The aim of this study was to try to confirm association of these nine novel regions in an extended cohort of 2478 GD cases and 3446 British Caucasian controls (including the original 900 cases and 1500 controls).

Weak association of nsSNPs in JSRP1, TEKT1 and HDLBP (P < 10⁻³) was determined in the extended cohort. To determine if these weak associations were in linkage disequilibrium with larger effects elsewhere in these genes, 24 Tag SNPs were employed to screen these three genes, with Tag SNPs in HDLBP and TEKT1 providing further weak evidence of association (P < 10⁻³). Association was not seen between GD and any of the other nsSNPs or Tag SNPs studied.

This study was unable to replicate six of the novel WTCCC associations in an enlarged GD dataset, suggesting these results were likely to be false positives within the original dataset. Weak association signals remain between GD and the JSRP1, TEKT1 and HDLBP nsSNPs in a large extended UK Caucasian cohort, suggesting that although a small effect can not be ruled out at these regions, as these regions did not reach genome wide significant (P < 10⁻⁷) replication in further adequately powered datasets is essential before determining if these nsSNPs truly represent novel susceptibility loci.

1 Endocrinology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China 2 Central Laboratory, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China Poster 140 Genotype Analysis of Killer Cell Immunoglobulin-Like Receptors in Graves' Disease Patients

Autoimmunity Saturday Poster Basic

Killer cell immunoglobulin-like receptor (KIR) can modulate the activity of NK and T lymphocyte. To investigate whether the KIR genotype possessing a susceptibility to Graves' disease (GD).

Using PCR-SSP to detect KIR genotype in 96 GD patients and 96 randomly selected healthy controls.

The genotype frequency of 2DS2-,2DL2-, 2DL3+, 2DL1+, 3DL1+, 3DS1-, 2DL5-, 2DS3-, 2DS5-, 2DS1-, 2DS4- was significantly higher in the patient group compared to that of the control group (6.25% vs 0%, P < 0.05). Genotype of 2DS2-, 2DL2-, 2DL3+, 2DL1+, 3DLl+, 3DS1-, 2DL5-, 2DS3-, 2DS5-, 2DS1-, 2DS4+ is the most prevalent in the controls (28.13% vs 10.42%, P < 0.01). Genotypes without activating KIR genes have higher frequency in patient group.

The difference of KIR genotypes between GD patients and healthy controls may explain the pathogenesis of GD.

1 Department of Biomedical Informatics, Division of Health Sciences, Osaka University Graduate School of Medicine, Suita, Japan 2 Translational Research Center, Kyoto University Hospital, Kyoto University School of Medicine, Kyoto, Japan Poster 141 Association of the −31C/T Functional Polymorphism in the Interleukin-1β Gene with the Intractability of Graves' Disease and the Proportion of TH17 Cells

Autoimmunity Saturday Poster Clinical

Interleukin (IL)-1β is a proinflammatory cytokine and has been implicated in the pathogenesis of several autoimmune diseases. IL-1β also has an important role for differentiation and proliferation of IL-17-producing helper T cells (Th17 cells) in human, which increases in patients with autoimmune thyroid diseases (AITD), especially in patients with intractable Graves' disease (GD). The region containing the −31T allele of functional −31C/T polymorphism (rs1143627) in the gene encoding IL-1β is the TATA box, and the change from −31T to −31C in this polymorphism causes potential disruption and decrease in expression of the IL-1β protein.

To evaluate the hypothesis that −31C/T functional polymorphism in IL1B gene is associated with the intractability and the severity of AITD, we genotyped this polymorphism in 64 GD patients who had been treated with methimazole or propylthiouracil for at least 5 years and remained positive for TRAb (intractable GD), 28 GD patients who had maintained a euthyroid state and had been negative for TRAb for more than 2 years without medication (GD in remission), 49 patients with Hashimoto's disease (HD) who developed hypothyroidism before 50 years of age, and were treated daily with thyroxine (severe HD), 28 untreated, euthyroid HD patients who were over 50 years of age (mild HD), and 59 healthy volunteers who were euthyroid and negative for thyroid autoantibodies. We also analyzed the proportion of peripheral Th17 cells in these patients by flow cytometry.

The −31T allele, which is related to high producibility of IL-1β, was significantly more frequent in patients with intractable GD than in those with GD in remission (p = 0.0017; odds ratio, 2.8; 95% confidence interval, 1.5−5.3), although there was no difference in this frequency between the 2 groups of HD patients. Furthermore, the proportion of Th17 cells, whose differentiation and proliferation are promoted by IL-1β, was higher in AITD patients with the T allele than in those with CC genotypes (p < 0.05).

Our data indicated that T allele of −31C/T polymorphism in the IL1B gene was involved in the intractability of GD, and this involvement could arise through the differentiation and proliferation of Th17 cells.

1 Department of Internal Medicine, Azienda USL Modena, Carpi (MO), Italy 2 Institute of Otolaryngology, University of Modena and Reggio Emilia, Modena, Italy 3 Chair of Endocrinology, Catholic University of Rome, Rome, Italy Poster 142 Association Between Autoimmune Chronic Thyroiditis and Benign Paroxysmal Positional Vertigo: Preliminary Results from a Perspective Study

Autoimmunity Saturday Poster Clinical

A higher prevalence of autoimmune chronic thyroiditis (ACT) has been demonstrated in pts with benign paroxysmal positional vertigo (BPPV) than in control subjects. The aim of the present study is to evaluate BPPV prevalence in pts with ACT and in controls.

100 consecutive ACT pts and 100 age- and sex-matched blood donors were recruited in the study. All pts underwent: written questionnaire on vertigo's signs/symptoms; measurement of serum TSH (n.v.: 0.3–4 mcIU/ml), free-T4 (FT4; n.v: 0.8–1.8 pg/ml) and Thyroglobulin (TG-Ab; n.v.: <100 UI/l) and Thyroperoxidase (TPO-Ab; n.v.: <40 IU/l) antibody concentrations; thyroid ultrasound (US). The diagnosis of BPPV was confirmed by Dix-Hallpike manoeuvre in pts with signs/symptoms peculiar to BPPV, as gathered from the questionnaire.

In ACT group, 81 (81%) pts were female and 19 (19%) male, aged 49 ± 6 yrs. Mean serum TSH and FT4 concentrations were 18.2 ± 8.9 mcIU/ml and 0.8 ± 0.3 pg/ml, respectively. Thirty-one (31%) ACT pts had overt hypothyroidism, 38 (38%) subclinical hypothyroidism, 31 (31%) were euthyroid. Mean serum TG-Ab and TPO-Ab levels were 344 ± 116 IU/l and 677 ± 210 IU/l, respectively. All ACT pts had a diffusely hypoechoic thyroid. In the control group (81 females and 19 males aged 48 ± 6 yrs), TSH and FT4 were 2.0 ± 0.5 mcIU/ml and 1.3 ± 0.3 pg/ml, respectively (P < 0.001 and <0.01 vs ACT group, respectively). Mean serum TG-Ab e TPO-Ab levels were 104 ± 5 IU/l and 43 ± 3 IU/l, respectively (P < 0.001 for both vs ACT group). Two pts had subclinical hypothyroidism and 1 Graves disease; high serum TG-Ab and TPO-Ab titres and a diffusely hypoechoic thyroid were detected in all these 3 pts. BPPV was diagnosed in 18 (18%) ACT pts - 13 females and 5 males, aged >40 yrs - and in 1 (1%) 47-yr old control subject, who was hypothyroid and showed high serum TG-Ab e TPO-Ab values.

ACT pts demonstrate a significantly higher BPPV prevalence than controls, strengthening the hypothesis that BPPV and ACT share a common immune-mediated origin.

1 Surgery, Tulane University, New Orleans, LA 2 Physiology, Tulane University, New Orleans, LA Poster 143 Understanding the Role of Cell Adhesion Molecule CD146 and its Downstream Signaling Gene in Mediating Thyroid Tumor Progression

Cell Biology Saturday Poster Basic

Molecular basis of thyroid carcinogenesis is recently expanded, however, CD146 role and its downstream targets have not been yet understood. CD146 is well known to promote certain cancers progression and proved to validate novel target genes. The aim of the current study is to understand the molecular mechanisms and the role of the CD146 and its downstream target tissue carboxypeptidase inhibitor, Latexin (LXN), in the process of invasion and metastasis of thyroid cancer.

We used human papillary, follicular and anaplastic thyroid cancer cell lines; in addition to melanoma SkMel-28, prostate cancer PC3 and vascular endothelial cell lines. Western blot, CD146 transfection, migration/invasion assay, siRNA knockdown were used to investigate the role of CD146 and LXN in thyroid cancer.

CD146 expression increased in SkMel-28, PC3 and endothelial cells. However, the expression of CD146 was significantly suppressed in all human thyroid cancer cell lines. In contrast, induction of CD146 expression in NPA and DRO cells, downstream target LXN expression also increased. CD146 expression additionally significantly suppressed the migration and invasion capacities of thyroid cancer cells. Knockdown of CD146 by siRNA in NPA cells returned the migration and invasion potential to the level of non-transfected cells.

CD146 and its downstream target LXN act as tumor suppressor gene in thyroid cancer and may play an important role in the progression of thyroid cancer. These highly significant findings may lead to validate novel target genes as a marker for thyroid cancer and may be used in therapeutic applications.

1 School of Clinical and Experimental Medicine, University of Birmingham, Birmingham, United Kingdom 2 Department of Pathology, University of Pittsburgh, Pittsburgh, PA Poster 144 Analysis of PTTG and PBF MRNA Expression in BRAF WT and V600E Thyroid Tumors

Iodine Uptake and Metabolism Saturday Poster Basic

The BRAF serine-threonine kinase is a component of the MAPK signaling pathway. The constitutively activating BRAF V600E mutation is found in approximately 45% of papillary thyroid cancers (PTC). This mutation has been associated with a reduction in iodide uptake, thought to be attributable to impaired expression and trafficking of the sodium iodide symporter (NIS). Pituitary tumor transforming gene (PTTG) and PTTG binding factor (PBF) are proto-oncogenes over-expressed in thyroid tumors. We have shown previously that, like BRAF V600E, PTTG and PBF reduce NIS expression and iodide uptake in thyroid cells.

We hypothesized that constitutively activated BRAF and high levels of PTTG and/or PBF may act either synergistically or in sequence to reduce iodide uptake in PTC. Gene expression analysis was performed in a series of PTC.

NIS mRNA expression was reduced by 36.6% in cancers with the BRAF V600E mutation (n = 22) compared with wild-type (WT) BRAF (n = 23), although this result was not statistically significant. No significant difference was detected in either PTTG or PBF mRNA expression. PTTG has been shown to upregulate FGF-2 and VEGF expression. A significant 1.8-fold increase in FGF-2 mRNA expression was detected in V600E cancers compared with BRAF WT (p = 0.04). VEGF mRNA expression was found to be significantly lower in the V600E cancers compared with BRAF WT (56% reduction, p = 0.001). Interestingly, FGF-2 expression was significantly correlated with PBF (r_s  = 0.742, p < 0.001) and PTTG (r_s  = 0.514, p = 0.012) expression in BRAF WT tumors but not in V600E cancers. Conversely, associations between VEGF expression and NIS, PTTG and PBF expression, and between PBF expression and NIS and PTTG expression, either became significant or increased in significance in the presence of the V600E mutation. This may indicate that the regulation of these gene interactions is altered by the constitutive activation of BRAF. Evaluation of clinical data identified a significant positive correlation between FGF-2 mRNA expression and patient age in V600E tumors (r_s  = 0.563, p = 0.007) not present in BRAF WT tumors (r_s  = 0.106, p = 0.627).

Further study is now required to determine whether PTTG and PBF function are altered in BRAF WT vs. V600E PTC.

1 Division of Otolaryngology Head and Neck Surgery, The George Washington University, Washington, DC 2 Department of Pathology, The George Washington University, Washington, DC Poster 145 Iatrogenic Hypothyroidism in Post-Surgical Head and Neck Cancer Patients

Iodine Uptake and Metabolism Saturday Poster Clinical

The incidence of hypothyrodism in head and neck surgical patients following total laryngectomy varies in the surgical literature. We present the interesting case of a head and neck cancer patient following who became profoundly hypothyroid while being treated for a wound dehiscence with povidone-iodine solution soaked gauze packing. To our knowledge, this is the first reported case of iatrogenic hypothyroidism in a surgical patient caused by supraphysiologic iodine levels.

The retrospective review of one patient's case, followed by a discussion of the literature.

A patient who underwent total layrngopharyngectomy developed a midline wound dehiscence with a superimposed infection on post-operative day 5. The wound was cultured, and treatment with antibiotics and povidone-iodine soaked gauze packing twice daily was begun. On post-operative day 12, the patient began complaining of profound depression and lethargy. A TSH level was obtained, and was noted to be 35 mIU/L. The pre-operative TSH level was within normal limits. The patient was immediately started on 75 mcg of parenteral levothyroxine. He subsequently went on to develop hypothyroidism induced renal-failure. After approximately 5 days of parenteral levothyroxine treatment, the patient's renal failure began to improve, and the patient began to show symptomatic improvement. Once stable, the patient was transitioned to oral levothyroxine, and discharged from the hospital.

The Wolff-Chaikoff effect is well documented for hyperthyroid patients who are treated with iodine for thyroid suppression. To our knowledge, this is the first reported case of surgically induced hypothyroidism after treatment of a wound dehiscence and infection with povidone-iodine packing. We advise caution when using povidone-iodine packing to treat wound related complications in surgical patients, and suggest that alternative materials be used for treatment when possible.

1 ENDOCRINOLOGY, PGIMER, Chandigarh, India 2 Endocrinology, Postgraduate Institute Of Medical Education and Research, Chandigarh, India Poster 147 Prevalence of Goiter in the Post Iodization Phase: Thyroid Autoimmunity or Micronutrient Deficiency?

Thyroid and Development Saturday Poster Clinical

The current study is aimed to survey the prevalence of goiter in the post iodization phase and the relationship of goiter with various micronutrient status and thyroid autoimmunity in school children of Chandigarh, an union territory of India.

Two phase study; the first phase is a cross sectional to find out goiter prevalence and the second phase is a case-control study of children with goiter and similar age and sex matched controls. In the first phase, 10,150 children in the age group of 6–16 years from seven schools of Chandigarh were randomized. 2148 children were screened for goiter by two independent observers as per the WHO grading system. In the second phase, 191 children with goiter and 165 children without goiter were compared. Urinary iodine, iodine content of salt, serum levels of T3, T4, TSH, hemoglobin, ferritin, selenium, and anti TPO antibody titer in children included in the case control study were measured.

The prevalence of goiter in the study population was 15.13%. It was more prevalent in girls than in boys (17.2% vs 13.8%, p = 0.02) and this sex difference was more marked in higher age group children (13–16 yrs, p = 0.01). The median urinary iodine excretion in both the groups were sufficient and comparable (137 vs 130 μg/L, p = 0.74). 31.2% children with goiter and 13.04% without goiter had subclinical hypothyroidism (p = 0.01) and 4.9% children in the goiter group and 1.9% of the control group were anti TPO antibody positive (≥34 IU/ml) (p = 0.22). The median serum selenium value were not different in the goiter and control group (181.93 ± 97.97 μg/L vs 193.5 μg/L, p = 0.45). More than half of the goitrous children had anemia (hemoglobin < 12 gm/dl) as compared to control group (52.75% vs 42.03%, p = 0.049) and goitrous children were severely depleted of tissue iron stores (serum ferritin < 12 μg/L) as compared to controls (18.8% vs 10.3%, p = 0.02). On multiple regression analysis, only serum ferritin was negatively correlated with goiter (correlation coefficient = − 0.160, p = 0.008).

Iron deficiency state per se might be playing a role in high prevalence of goiter in this iodine replete population, which needs to be further explored.

1 Department of Cancer Biology, Mayo Clinic Comprehensive Cancer Center, Mayo Clinic, Jacksonville, FL 2 Department of Laboratory Medicine/Pathology, Mayo Clinic, Rochester, MN 3 Department of Surgery, Mayo Clinic, Rochester, MN 4 Endocrine Malignancy Working Group, Mayo Clinic, Rochester, MN 5 Department of Medicine, Mayo Clinic, Rochester, MN 6 Department of Otolaryngology, Mayo Clinic, Rochester, MN 7 Department of Otolaryngology - Head & Neck Surgery, Eastern Virginia Medical School, Norfolk, VA 8 Department of Surgery, University of California San Francisco, Mt. Zion Medical Center, San Francisco, CA 9 Pharmaceutical Genomics Division, The Translational Genomics Research Institute, Scottsdale, AZ 10 Division of Endocrinology, Mayo Clinic, Jacksonville, FL Poster 149 Comprehensive Molecular Characterization of Four New Human Anaplastic Thyroid Carcinoma Cell Lines Reveals RHOB as a Molecular Target for Therapy

Thyroid Cancer Saturday Poster Basic

Anaplastic thyroid carcinoma (ATC) is a highly aggressive carcinoma with a mortality rate near 100% and is in need of new therapeutic options. One critical component of drug discovery is the availability of well-characterized cell lines derived from patient tumors for identification of molecular mechanisms related to tumor biology and drug responsiveness. A recent report indicates that up to 42% of thyroid cancer cell lines, including ATC, are redundant or not of correct tissue origin. We report four new ATC cell lines and the methodologies used to characterize the originating tumor tissue and derivative cell lines.

To validate the integrity of our newly derived cell lines, we confirmed histologically the original diagnosis of the ATC tumor tissues and demonstrated identical matching of the short tandem DNA repeats (STR) and mutational status to the tumor-derived cell lines. This is the first time that molecular descriptions of thyroid cell lines are matched to the originating tumor tissues. In addition, we fully characterized the cell lines for proliferative growth, mRNA expression of seven thyroid markers, four oncogenes and array CGH to identify novel deletions and amplifications.

Previously in other cell lines, we showed that RhoB is a key signaling node for the growth inhibition of ATC. Using our new four cell lines, we have now identified five classes of compounds (10 μM FTI-277, 10 μM GGTI-286, 1 μM lovastatin, 1 nM romidepsin, 1 μM UCN-01) that upregulate RhoB and induce inhibition of cell proliferation in a dose-responsive fashion.

In conclusion, we have developed four new molecular characterized ATC cell lines. Our results further implicate RhoB as a molecular target for therapy in genuine ATC cell lines.

1 Nuclear Medicine, Instituto Português de Oncologia, Lisbon, Portugal 2 Nuclear Medicine, Hospital Reina Sofia, Cordoba, Spain 3 Nuclear Medicine, University of Duisburg/Essen, Essen, Germany 4 Endocrine Surgery, 4th Medical Clinical Amalgamation, Dnepropetrovsk, Ukraine 5 Nuclear Medicine and Endocrine Oncology, Maria Sklodowska Curie Memorial Cancer Center and Institute of Oncology, Gliwice, Poland 6 Endocrine Surgery, Ukrainian Scientific and Practical Center of Endocrine Surgery, Kyiv, Ukraine 7 Nuclear Medicine, University of Würzburg, Würzburg, Germany 8 Endocrinology, Evgenidion Hospital, University of Athens, Athens, Greece 9 Thyroid Unit, Royal Marsden Hospital, London, United Kingdom 10 Nuclear Medicine, Shaukat Khanum Memorial Cancer Hospital & Research Center, Lahore, Pakistan 11 Nuclear Medicine, Hospital São João, Porto, Portugal 12 Nuclear Medicine, Institute of Oncology, Ljubljana, Slovenia 13 Endocrinology, Chaim Sheba Medical Center, Tel Hashomer, Israel 14 Thyroid and Mineral Bone Diseases, Clinical Center of Endocrinology, Medical University of Sofia, Sofia, Bulgaria 15 Nuclear Medicine, Hannover University School of Medicine, Hannover, Germany Poster 150 Radioiodine (RAI) Thyroid Remnant Ablation using Recombinant Human Thyrotropin (RHTSH) in Differentiated Thyroid Cancer (DTC) Patients: Preliminary Data from a Multinational, Multicenter Retrospective Study

Thyroid Cancer Saturday Poster Clinical

Publications on rhTSH-aided RAI thyroid remnant ablation (rhTSH-A) generally have involved single centers and not reported ablation success rates by T or N status or other potential predictive patient/disease characteristics.

This multinational, multicenter retrospective study seeks to further characterize safety/efficacy of rhTSH-A. The study will include multivariate statistical analysis of factors potentially predicting rhTSH-A success. Also, ablation success rates will be compared for rhTSH-A patients versus historical controls given thyroid hormone withholding-aided ablation (THW-A). Ablation success is defined by institutional criteria, based on diagnostic whole-body scan, serum thyroglobulin assay, or both. Patients must have received a large RAI activity primarily to ablate thyroid remnant and any microscopic residual DTC, and been followed ≥8 months post-ablation. Anonymized data are collected via a standardized Excel spreadsheet pursuant to local ethical requirements. Data collection will close circa 30 June 2009.

We report here selected descriptive statistics for our rhTSH-A group as of 11 May 2009. Data were available on 181 rhTSH-A patients in total from 15 European/Asian centers (64% F, median age 48 yrs, 66% papillary, 20% follicular, 9% mixed, 4% less differentiated histology). Proportions of patients by TNM stages (n = 180) were: T1–2, 62%, T3–4, 34%, TX or T not available, 3%; N0, 57%, N1, 39%, NX, 4%; M0, 83%, M1, 10%, MX, 7%. Median (range) ablative activities were 100 (60–300) mCi/3.7 (2.2–11.1) GBq. Table 1 gives rhTSH-A success rates overall and by tumor stage. Despite frequent locally advanced disease or nodal metastasis in our patients, rhTSH-A success rates appear to be high and within the range of those reported for rhTSH-A or THW-A (75%-100%) in comparative studies using rhTSH in the standard regimen (0.9 mg/day × 2, RAI given 24 hr post-second injection).