Altered expression of G proteins in thyroid gland adenomas obtained from hyperthyroid cats

OBJECTIVE: To determine whether expression of G proteins (G(i) and G(s)) is altered in thyroid gland adenomas obtained from hyperthyroid cats. SAMPLE POPULATION: Adenomatous thyroid glands obtained from 8 hyperthyroid cats and thyroid glands obtained from 4 age-matched euthyroid cats. PROCEDURE: Expression of G(i) and G(s) was quantified in enriched membrane preparations of thyroid gland tissue, using immunoblotting with G(i) and G(s) antibodies and toxin-catalyzed ADP-ribosylation. RESULTS: Expression of G(i) was significantly reduced in thyroid gland adenomas from hyperthyroid cats, compared with normal thyroid gland tissue from euthyroid cats. Expression of G(s) was similar between the 2 groups. CONCLUSIONS AND CLINICAL RELEVANCE: A decrease in expression of G in adenomatous thyroid glands of cats may reduce the negative inhibition of the cAMP cascade in thyroid cells, leading to autonomous growth and hypersecretion of thyroxine. Understanding the molecular mechanisms for hyperthyroidism in cats may lead to better treatment or, ultimately, prevention of the disease.     

Relationship between semi-quantitative thyroid palpation and total thyroxine concentration in cats with and without hyperthyroidism

In 155 cats, both with and without clinical signs of hyperthyroidism, total thyroxine (TT4) concentrations were compared to a sensitive, semi-quantitative thyroid palpation technique. On the basis of TT4 concentrations, 23 of the 155 cats were classified as hyperthyroid. The size of individual thyroid glands was scored between '0' (non-palpable) and a maximum of '6'. One or more enlarged thyroid glands (score >0) were palpated in 22 of the 23 hyperthyroid cats and in 78 of the 132 euthyroid cats. However, none of the 132 euthyroid cats had a thyroid lobe score of greater than '3' whereas 18 of the 23 hyperthyroid cats had a thyroid lobe score of '4' or greater, and in two of the five that had scores below '4' there was evidence of intrathoracic functional thyroid tissue on scintigraphy.     

INVESTIGATION OF TWO METHODS FOR ASSESSING THYROID-LOBE ASYMMETRY DURING PERTECHNETATE SCINTIGRAPHY IN SUSPECTED HYPERTHYROID CATS

Our aim was to investigate thyroid:thyroid (T:T) ratio and visual inspection for assessing thyroid-lobe asymmetry in suspected hyperthyroid cats. Although thyroid-salivary asymmetry is a preferred test, inherent thyroid symmetry may assist image interpretation. Association was determined using a scatter plot and Spearman's rank correlation. Agreement was assessed using the kappa (K) statistic. Accuracy was assessed by sensitivity and specificity. Hyperthyroidism was diagnosed in 33/48 (69%) cats based on elevated serum total thyroxine level. Using two Wilcoxan rank-sum tests, a significant difference (P < 0.0001) was detected between cats with and without hyperthyroidism for both methods of assessing thyroid symmetry. For the 18 cats with T:T ratios < or = 1.5, there was poor correlation between the two methods (r(s) = 0.39). Using a cut-point of 1.5 for the T:T ratio, the test accurately predicted hyperthyroidism in 28/33 cats (sensitivity, 85%; 95% confidence interval (CI), 71-99%) and correctly predicted that hyperthyroidism was absent in 14/15 cats (specificity, 93%; CI, 77-100%). For visual inspection, agreement for diagnosing hyperthyroidism was excellent between methods (kappa = 0.82), within the same examiner (weighted kappa = 0.85) and between examiners (weighted kappa = 0.89). Considering cats with only definitely asymmetric thyroid lobes as positive, visual inspection accurately predicted hyperthyroidism in 28/33 cats (sensitivity, 85%; CI, 71-99%) and correctly predicted that hyperthyroidism was absent in 11/15 cats (specificity, 73%; CI, 48-99%). Thyroid-lobe asymmetry occurs more frequently in hyperthyroid than in euthyroid cats but caution should be exercised because some euthyroid cats have asymmetric thyroid glands.     

Expression of inhibitory G proteins in adenomatous thyroid glands obtained from hyperthyroid cats

OBJECTIVE: To identify within guanosine triphosphate-binding proteins (G proteins) the subset of inhibitory G proteins (G) that have decreased expression in adenomatous thyroid glands obtained from hyperthyroid cats. SAMPLE POPULATION: Adenomatous thyroid glands obtained from 5 hyperthyroid cats and normal thyroid glands obtained from 3 age-matched euthyroid cats. PROCEDURE: Expression of G(i1), G(i2), and G(i3) in enriched membrane preparations from thyroid glands was quantified by use of immunoblotting with G(i) subtype-specific antibodies. RESULTS: Expression of G(i2) was significantly decreased in tissues of hyperthyroid glands, compared with expression in normal thyroid tissue. Expression of G(i1) and G(i3) was not significantly different between normal thyroid tissues and tissues from hyperthyroid glands. CONCLUSIONS AND CLINICAL RELEVANCE: A decrease in G(i2) expression decreases inhibition of adenylyl cyclase and allows a relative increase in stimulatory G protein expression. This results in increased amounts of cAMP and subsequent unregulated mitogenesis and hormone production in hyperthyroid cells. Decreased G(i2) expression may explain excessive growth and function of the thyroid gland in cats with hyperthyroidism.     

Nodular thyroid hyperplasia in a cynomolgus monkey

Nodular thyroid hyperplasia was found in a 4-year-old male cynomolgus monkey (Macaca fascicularis). The monkey was clinically normal; however, necropsy revealed multiple variably sized nodules in both lobes of the thyroid gland. In contrast to the fairly uniform diameter of the lumen of follicles in the surrounding gland, the diameter of the follicular lumen within the hyperplastic nodules was highly variable and ranged from nonexistent to cystlike. Occasionally, in the larger follicles there were papillary infoldings of epithelium. The hyperplastic nodules were partially encapsulated by a fibrous capsule and showed little compression of the surrounding tissue. The follicular cells and colloid comprising the hyperplastic nodule were immunohistochemically positively stained with the antibody for thyroglobulin. Ultrastructurally, the cells forming follicles had numerous microvilli along the luminal surface, and lysosomal bodies and dilated rough endoplasmic reticulum in the cytoplasm. All these morphologic findings are consistent with nodular thyroid hyperplasia, which is rare in nonhuman primates.     

p53 expression and environmental tobacco smoke exposure in feline oral squamous cell carcinoma

The purpose of this study was to determine the prevalence of p53 overexpression in feline oral squamous cell carcinomas (SCC) and to determine, if any, the association between p53 overexpression and lifestyle factors and environmental exposures, including exposure to environmental tobacco smoke (ETS). Questionnaires concerning exposure to ETS and other environmental factors were sent to owners of cats presenting to the Harrington Oncology Program with a diagnosis of oral SCC between 1991 and 2000. Additionally, 23 formalin-fixed biopsy samples from these cats, with information regarding ETS, were evaluated immunohistochemically for p53 expression using the CM-1 clone and the avidin-biotin-horseradish peroxidase method. Of the 23 samples evaluated, 15 (65%) showed positive nuclear staining for the CM-1 clone. Tumor biopsy samples from cats exposed to any ETS were 4.5 times more likely to overexpress p53 than were tumors from unexposed cats (P = 0.19). Among cats with any ETS exposure, those with 5 years or longer of exposure were 7.0 times more likely to overexpress p53 (P = 0.38). Longhaired cats (P = 0.18) and female cats (P = 0.35) were also more likely to show p53 expression in their tumors. These results provide additional support for a relationship between oral SCC development and exposure to household ETS and may implicate p53 as a potential site for carcinogen-related mutation in this tumor.     

FELINE HYPERTHYROIDISM: EFFICACY OF TREATMENT USING VOLUMETRIC ANALYSIS FOR RADIOIODINE DOSE CALCULATION

Hyperthyroidism was diagnosed in 80 cats with thyroid scintigraphy using technetium pertechnetate. These cats were subsequently treated with radioiodine using a modified fixed dose method based on the volume of hyperfunctioning thyroid tissue calculated from the pertechnetate scans. The medical records and thyroid scintigrams were evaluated retrospectively. Follow-up was obtained on the cats to evaluate treatment success. Several parameters were evaluated in an attempt to identify a difference between treatment success and failure. Cats that failed to become euthyroid after one dose of radioiodine had a significantly higher pretreatment serum thyroxine level, had a significantly larger volume of hyperfunctioning thyroid tissue on scintigrams, and cats receiving oral versus intravenous radioiodine were over represented. Based on our results we conclude: 1) the administration of a dose of radioiodine based solely on the volume of hyperfunctioning thyroid tissue as estimated from the pertechnetate scan may be inadequate for those patients with extremely elevated serum thyroxine levels or large thyroid glands, and 2) oral administration of radioiodine is not recommended for the treatment of feline hyperthyroidism.     

Association of Iatrogenic Hypothyroidism with Azotemia and Reduced Survival Time in Cats Treated for Hyperthyroidism

Background: Iatrogenic hypothyroidism can occur after treatment of hyperthyroidism, and is correlated with a reduced glomerular filtration rate in humans and dogs. Hypothesis: Cats with iatrogenic hypothyroidism after treatment for hyperthyroidism will have a greater incidence of azotemia than euthyroid cats. Animals: Eighty client owned cats with hyperthyroidism. Methods: Two retrospective studies. (1) Longitudinal study of 12 hyperthyroid cats treated with radioiodine (documented as euthyroid after treatment), to assess changes in plasma thyroid stimulating hormone (TSH) concentration over a 6‐month follow‐up period, (2) Cross‐sectional study of 75 hyperthyroid cats (documented as euthyroid) 6 months after commencement of treatment for hyperthyroidism to identify the relationship between thyroid status and the development of azotemia. Kaplan‐Meier survival analysis was performed to identify relationships between thyroid and renal status and survival. Results: Plasma TSH concentrations were not suppressed in 7 of 8 cats with hypothyroidism 3 months after radioiodine treatment. The proportion of cats with azotemia was significantly (P= .028) greater in the hypothyroid (16 of 28) than the euthyroid group (14 of 47). Twenty‐eight of 41 cats (68%) with plasma TT4 concentration below the laboratory reference range had an increased plasma TSH concentration. Hypothyroid cats that developed azotemia within the follow‐up period had significantly (P= .018) shorter survival times (median survival time 456 days, range 231–1589 days) than those that remained nonazotemic (median survival time 905 days, range 316–1869 days). Conclusions and Clinical Importance: Iatrogenic hypothyroidism appears to contribute to the development of azotemia after treatment of hyperthyroidism, and reduced survival time in azotemic cats.     

Thyroidectomy in the cat

Thyroidectomy in cats is most commonly indicated to treat hyperthyroidism because of adenomatous hyperplasia of the thyroid glands. Preoperative stabilization of the hyperthyroid cat with antithyroid drugs is preferred to minimize anesthetic and surgical complications. Multiple surgical techniques for thyroidectomy have been reported, and results of surgery and complications differ between techniques. The extracapsular technique offers the most complete removal of abnormal thyroid tissue, while still being associated with a low incidence of postoperative hypoparathyroidism. Important postoperative considerations are to carefully monitor serum calcium concentrations and treat hypocalcemia if necessary, and to monitor the cat for evidence of relapse of hyperthyroidism.     

Percutaneous ultrasound-guided radiofrequency heat ablation for treatment of hyperthyroidism in cats

OBJECTIVE: To determine efficacy and safety of percutaneous radiofrequency heat ablation for treatment of hyperthyroidism in cats. DESIGN: Prospective study. ANIMALS: 9 cats. PROCEDURE: Hyperthyroidism was diagnosed via clinical signs and high serum total (TT4) and free thyroxine (fT4) concentrations. One or 2 hyperfunctional cervical thyroid nodules were detected by use of scintigraphy and ultrasonography. If cats had 1 abnormal thyroid lobe, heat ablation was performed on that lobe; if cats had 2 abnormal lobes, heat ablation was applied to the larger lobe. Overall, heat ablation was performed 14 times in the 9 cats. Clinical signs and serum TT4, fT4, and calcium concentrations were monitored daily for 2 days after the procedure, weekly for the first month, and then monthly. Laryngeal function was evaluated and cervical ultrasonography and thyroid scintigraphy were also performed. Monitoring continued for as long as 9 months after heat ablation if a cat became euthyroid or until an owner chose an alternative treatment because of recurrence of hyperthyroidism. RESULTS: Serum TT4 and fT4 concentrations transiently decreased after all 14 heat ablation procedures (< or = reference range after 10 of 14 treatments) within 2 days after the procedure. Cats were euthyroid for 0 to 18 months (mean, 4 months). Hyperthyroidism recurred in all cats. Adverse effects included transient Horner's syndrome (2 cats) and laryngeal paralysis without clinical signs (1 cat). CONCLUSIONS AND CLINICAL RELEVANCE: Percutaneous heat ablation as a treatment for hyperthyroidism in cats is effective transiently but not permanently.     

Diagnostic tests for hyperthyroidism in cats

The diagnosis of hyperthyroidism, one of the most common disorders affecting elderly cats, is usually straightforward and considered routine by most practitioners. Nowadays, however, most cats suffering from hyperthyroidism tend to be diagnosed earlier and at a milder stage of the disease than those cats diagnosed 10 to 25 years ago. There are, in fact, a growing number of cats with clinical signs of hyperthyroidism and palpably large thyroid glands whose baseline serum total thyroid hormone concentrations are within the normal or borderline range, making diagnosis problematic. This paper reviews the available tests used to confirm a diagnosis of hyperthyroidism in cats and discusses their overall usefulness.     

ULTRASONOGRAPHIC EXAMINATION OF THE THYROID GLAND OF HYPERTHYROID CATS: COMPARISON TO 99mTcO−4 SCINTIGRAPHY

High-resolution ultrasonography was evaluated as an alternative to ^99mTcO^-₄ scintigraphy for examining size and appearance of thyroid glands in hyperthyroid cats. Thyroid ultrasound examinations were performed on 6 normal cats and 14 cats with hyperthyroidism. Thyroid lobe volume was estimated from ultrasound images using the equation for a prolate ellipsoid, π/6 (length * height * width). Total thyroid volume was estimated by adding the volume estimations of the left and right lobes. Thyroid lobes of hyperthyroid cats were considered abnormal if estimated volume exceeded the 99% confidence interval for normal thyroid volume determined from the control group. Scintigraphic examinations performed on hyperthyroid cats were evaluated for unilateral versus bilateral disease and for the presence of ectopic activity. Mean thyroid lobe volume and total thyroid volume for normal cats was 85 and 169 mm³, respectively. Mean thyroid lobe volume and total thyroid volume for hyperthyroid cats was 578 and 889 mm³. There was a significant difference in mean estimated total thyroid volume of normal and hyperthyroid cats. Thyroid lobes with greater than normal TcO^-₄ uptake on scintigraphy were larger and had variable homogeneity, echogenicity, and margination on ultrasound examination. There also was an 85.7% agreement of scintigraphy and ultrasonography in differentiating normal from abnormal thyroid lobes. A fair correlation between estimated total thyroid volume of hyperthyroid cats and most recent pretherapy serum thyroxine values were also found. This preliminary study indicates that thyroid ultrasound examination may provide information that is useful for diagnosis and treatment of feline hyperthyroidism. Although ultrasound provides accurate evaluation of the thyroid glands, it cannot replace ^99mTcO^-₄ scintigraphy for screening of metastatic lesions and ectopic glands.     

Serum troponin I levels in hyperthyroid cats before and after treatment with radioactive iodine

A raised concentration of serum cardiac troponin I (cTnI) is a sensitive marker of cardiac myocyte injury in the cat and assays developed for its measurement in human patients have been validated in the cat. Raised levels have been associated with a number of cardiac insults including hypertrophic cardiomyopathy and trauma. Hyperthyroidism is a common disease of older cats and excess thyroid hormone is known to produce significant cardiovascular effects in this species. This study evaluated the effect of treatment for hyperthyroidism with radioactive iodine on cTnI concentration, assessed the association between thyroxin levels and glomerular filtration rate (GFR) and cTnI concentration in cats treated for hyperthyroidism and described changes in echocardiographic parameters following treatment. Prior to the treatment serum cTnI was measured and echocardiography performed, thyroxin, cTnI, and echocardiography were then repeated at various time points following radioisotope therapy. The results show that higher thyroxin levels were significantly (P=0.002) associated with a higher likelihood of the cat presenting with detectable levels of cTnI. No significant association was found between GFR and presence of detectable levels of cTnI. Furthermore the results indicate that the effects of hyperthyroidism on echocardiographic parameters appear considerably less in this study than in previous studies and that the main outcome of treatment on these parameters is a significant reduction in fractional shortening (P=0.006). These results suggest that chronic exposure to excess thyroid hormone may induce myocyte damage of sufficient severity to raise serum cTnI concentration in a proportion of cats that resolves following establishment of a euthyroid state.     

Morphology of thyroid follicular cells of methimazole-treated rats

Thyroids of rats treated with methimazole (MMI) were analysed histologically, immunohistochemically and electron microscopically. Thyroids of MMI-treated rats displayed greater hypertrophy and were heavier than those of control rats. Thyroids of MMI-treated rats were composed of variant or small follicles and contained more columnar cells than those of control rats. Strong staining of thyroid granules by azocarmine G was evident in the apical regions of follicular cells from MMI-treated rats. Anti-thyroglobulin positivity was detected in the apical regions of follicular epithelial cells, especially those of MMI-treated rats. After application of a post-embedding method to detect thyroglobulin, colloidal gold particles were often observed in apical vesicles of MMI-treated rats. Our study suggests that apical vesicles may serve as markers of hyperthyroidism. When the functional state of the thyroid gland is known from histological analysis, use of the Azan stain could provide useful additional information.     

Lack of circulating thyroid stimulating immunoglobulins in cats with hyperthyroidism

Although feline hyperthyroidism has become a commonly diagnosed disorder of older cats, the underlying etiology remains unknown. Pathological findings of adenomatous hyperplasia involving both thyroid lobes in most hyperthyroid cats suggests the possibility that feline hyperthyroidism may be similar to human Graves' disease, which results from high circulating levels of thyroid stimulating immunoglobulins (TSIs). To exclude high circulating levels of TSIs as the cause of feline hyperthyroidism, we measured intracellular concentrations of cyclic adenosine monophosphate (cAMP) in functioning rat thyroid cells (FRTL-5) incubated with IgG extracted from hyperthyroid cat serum. Since TSIs stimulate thyroid hormone secretion through activation of cAMP, their presence can be evidenced in vitro by generation of high cAMP concentrations in cultured thyroid cells. No significant difference was found in intracellular cAMP concentrations in FRTL-5 cells incubated with IgG from normal versus hyperthyroid cats. In contrast, IgG from a human patient with Graves' disease caused substantially more cAMP generation than either normal human IgG or IgG from the cats of this study. These results indicate that feline hyperthyroidism does not result from high circulating concentrations of TSI and, in that respect, is not analogous to Graves' disease.     

Radio-iodine treatment of hyperthyroid cats

Thirty-two elderly domestic shorthaired cats (mean age 12.9 years) were treated with radioiodine (131I). The dose of 131I administered ranged from 39 mBq to 134 mBq. Twenty-eight cats became euthyroid after treatment, one became hypothyroid and three remained hyperthyroxaemic. Two of the hyperthyroxaemic cats were successfully re-treated with 131I. Five cats died from concurrent diseases within one year of treatment. The administration of a dose of 131I selected by assessing the severity of the clinical signs, the size of the thyroid gland(s) and the serum level of thyroxine was an effective treatment for hyperthyroidism.     

Thyroid enlargement and its relationship to clinicopathological parameters and T(4) status in suspected hyperthyroid cats

To relate thyroid size to routine blood parameters and T(4) status the ventral neck of 161 cats with clinical signs consistent with hyperthyroidism was examined by two independent observers using a semi-quantitative palpation system. Thyroid gland size of each side was scored from 0 (non-palpable) to a maximum of 6 (>25 mm). In 127 of the 161 cats, at least one thyroid gland was palpable. The palpation score was significantly correlated with the T(4) concentration. The 17 hyperthyroid cats had significantly higher palpation scores than the 110 euthyroid cats. Euthyroid animals with a palpation score >or=3 were significantly older, had higher body weights, lower alkaline phosphatase, alanine aminotransferase, phosphate, and urine specific gravity, but higher lipase and creatinine concentrations than hyperthyroid cats. Our study demonstrates that although no reliable conclusion on the functional status of the thyroid can be drawn based on its size the likelihood of hyperthyroidism increases with increasing size of the gland.     

Liver function in cats with hyperthyroidism before and after 131I therapy

BACKGROUND: The clinical significance of high serum concentration or activity of markers of liver damage in cats with hyperthyroidism is unknown. OBJECTIVE: To evaluate serum markers of liver function and damage, and ultrasonographic changes in cats with hyperthyroidism and with high liver enzymes, and to determine if abnormalities resolve after treatment with 131I. ANIMALS: Nineteen cats with hyperthyroidism (15 with high serum activities of liver enzymes) and 4 age-matched healthy control cats. METHODS: Serum bile acids, albumin, ammonia, cholesterol, and blood urea nitrogen concentrations, and activities of liver-derived enzymes, and blood glucose concentrations were measured before and after 131I therapy. These values were compared with those of cats that were euthyroid. In addition, gross liver parenchymal changes detected by abdominal ultrasonographic examination, before and after 131I therapy were evaluated. RESULTS: High serum liver enzyme activities were not associated with abnormalities in hepatic parenchyma and liver functional variables, regardless of the degree of increase. Serum liver enzyme activities return to normal after control of hyperthyroidism with 131I therapy. Cats with hyperthyroidism have a significantly higher serum fasting ammonia concentration than cats who were euthyroid (P = .019). Cats with hyperthyroidism also have significantly lower serum cholesterol (P = .005) and glucose (P = .002) concentrations before compared with after 131I therapy. Nine of 19 cats with hyperthyroidism had trace ketonuria. CONCLUSIONS AND CLINICAL IMPORTANCE: These results demonstrate that extensive examination for hepatobiliary disease in most cats with hyperthyroidism is unnecessary.