Impaired local deiodination of thyroxine to triiodothyronine in dogs with symmetrical truncal alopecia

Thyroid dysfunction causes certain dermatological alterations in dogs. Insufficient delivery of thyroid hormone to the skin may originate not only from inadequate thyroid function but also from impaired local activation of thyroxine in the target organ. Thyroid parameters and deiodination were investigated in healthy dogs (group C) and in dogs with cutaneous lesions associated with hypothyroidism (group H) or with a low-T₃ syndrome (group LT). The ability of the skin to convert T₄ to T₃ was impaired in both groups H and LT but not in the controls. It is concluded that impaired local deiodination may contribute to skin problems in dogs.Abbreviations bwt body weight - DTT dithiothreitol - PBS phosphate-buffered saline (pH 7.5, 0.05 mol/L) - PTU propylthiouracyl - RIA radioimmunoassay - TRH thyrotropin-releasing hormone - TSH thyroid-stimulating hormone     

Evaluation of an enzymatic immunoassay for free thyroxine determination in canine serum

Free thyroxine (FT4) and cholesterol were measured in 400 dogs with either suspected hypothyroidism or dermatological signs such that hypothyroidism needed to be ruled out. Hypothyroidism was diagnosed in 68 dogs from the history, physical examination and stated lower reference limit (<7 pmol/L) for FT4 in euthryoid dogs. Dogs with FT4 concentrations in the range 6–9 pmol/L were finally categorized as hypo- or euthyroid either on the basis of retesting after 2 months or on their clinical response to thyroid replacement therapy over at least 2 months.The enzyme immunoassay evaluated in this paper is considered to be of clinical value and offers many advantages compared with radioimmunoassays.Abbreviations FT4 free thyroxine fraction - MEIA microparticle enzyme immunoassay - RIA radioimmunoassays - T4 thyroxine - TBG thyroxine-binding globulin     

Concentrations of thyroid hormones and iodothyronine binding proteins in serum of the koala (Phascolarctos cinereus)

Objective To examine circulating total and free thyroid hormone (T₃ and T₄) concentrations, determine serum iodothyronine binding characteristics and estimate thyroid stimulating hormone (TSH) activity in sera of coastal and inland koalas.
Design A prospective study.
Procedure Koala serum T₃ and T₄ were measured by radioimmunoassay. T₄ binding parameters were determined by radioligand binding and electrophoresis. Koala TSH values were determined by bioassay.
Results Mean total T₄ concentrations were 3.2 ± 2.1 nM although values were significantly higher in inland-dwelling females in comparison to coastal-dwelling males. Free T₄ was 3.3 ± 2.1 pM. Total and free T₃ were 0.4 ± 0.2 nM and 1.4 ± 0.9 pM respectively, although these values were at the lower end of the assay detection limit and should be viewed with reservation. Electrophoresis of [¹²⁵I]-T₄-labelled serum revealed only two proteins of electrophoretic mobility similar to human transthyretin (TTR) and albumin. Scatchard analysis of T₄ binding to serum gave a curvilinear plot, which could be resolved into two binding sites with affinities identical to that of TTR and albumin but both of low concentration. The bioactivity of the TSH present in the sera was measured using a cell line (JP09) transfected with the human TSH receptor. The mean level of stimulation found in the sera corresponded to a bovine TSH activity of less than 10 mU/L.
Conclusion These results suggest that the serum concentrations of free and total thyroid hormones in koalas are low compared to other marsupials and very low compared to eutherian mammals. The mechanism of maintenance of euthyroidism in this species remains to be determined.     

铜缺乏对奶甲状腺功能的影响

为阐明铜缺乏奶牛甲状腺激素的变化特征及补铜对奶牛甲状腺功能的影响，本实验选择缺铜地区全血铜低于NRC下限（0.8mg/kg)但无明显摇摆症状的犊牛25头作为亚临床铜缺乏组；并对其中15头进行补铜试验（饲料中添加硫酸酮100mg/kg干重），以15头临床健康奶牛作为对照组，分别于40天及80天采集血样并检测T3、T4及TSH；同时测试了有明显摇摆症状的6头发病奶牛及健康奶牛的肝脏中T4－5’脱碘酶的活性。结果表明：铜缺乏奶牛血清T3显著低于亚临床及健康对照组；发病组及亚临床组奶牛血清T4含量显著高于对照组；对照组奶牛血清TSH含量显著高于发病组及亚临床组；发病奶牛肝脏中的T4－5’脱碘酶活性也显著低于健康奶牛；试验组奶牛在补铜后40天血清T3含量显著上升；在补铜后80天，亚临床组奶牛血清T4含量显著下降，TSH含量显著高于补铜前。结论：铜缺乏能够影响奶牛甲状腺激素的调节及代谢过程。     

Ultrasound-guided core needle biopsy in dogs with thyroid carcinoma

Currently, a histological diagnosis of highly vascularized canine (c) thyroid carcinoma (TC) is primarily obtained following excisional biopsy (EB) through thyroidectomy. Non-EBs are contraindicated in unresectable invasive cTCs due to their highly vascularized nature, which subsequently, lack histological diagnosis. We hypothesised ultrasound-guided core needle biopsy (UGCNB) to be a safe biopsy technique to obtain an accurate histological diagnosis in unresectable TCs. Nine client-owned dogs with suspected naturally occurring TC, presented for surgical excision, were included. First, a UGCNB was taken from the cervical tumour, followed by EB. Haemorrhage following UGCNB was evaluated preoperatively and once the tumour was surgically exposed by visual inspection and ultrasonography. Histological analysis, including cell organisation, tumour capsular and vascular invasion, and immunohistochemistry were performed and compared between both biopsy specimens (i.e., UGCNB and EB) of the same dog. Pre- and peroperative visual inspection revealed minor, localised haemorrhage, subsequent to the UGCNB, in 7/9 dogs. Histology of the EBs confirmed TC in 8/9 dogs and was inconclusive in 1/9 dogs. Histology of the UGCNBs revealed neoplastic thyroid tissue in 7/9 UGCNBs and was inconclusive in 1/9 UGCNBs. The remaining UGCNB contained no mass related tissue and was, therefore, excluded. Histological parameters (i.e., cell organisation, tumour capsular and vascular invasion) were not concordant between 6/8 included UGCNBs and their respective EB. Immunolabelling for thyroglobulin and calcitonin was concordant between all eight included UGCNBs and their respective EB. The remaining evaluated immunohistochemical markers (i.e., cyclooxygenase-2 [COX-2], P-glycoprotein and vascular endothelial growth factor [VEGF]) were concordant between the included UGCNBs and the EBs in 6/8 dogs. To conclude, UGCNBs can be safely obtained in suspected cTCs and enable a reliable diagnosis of the thyroid origin, thyroid cell origin and potential therapeutic markers such as COX-2, P-glycoprotein and VEGF. Subsequently, UGCNB enables clinicians to establish an individually tailored treatment plan in dogs with unresectable TC.     

Plasma and urine pharmacokinetics of intravenously administered flunixin in greyhound dogs

Medication control in greyhound racing requires information from administration studies that measure drug levels in the urine as well as plasma, with time points that extend into the terminal phase of excretion. To characterize the plasma and the urinary pharmacokinetics of flunixin and enable regulatory advice for greyhound racing in respect of both medication and residue control limits, flunixin meglumine was administered intravenously on one occasion to six different greyhounds at the label dose of 1 mg/kg and the levels of flunixin were measured in plasma for up to 96 hr and in urine for up to 120 hr. Using the standard methodology for medication control, the irrelevant plasma concentration was determined as 1 ng/ml and the irrelevant urine concentration was determined as 30 ng/ml. This information can be used by regulators to determine a screening limit, detection time and a residue limit. The greyhounds with the highest average urine pH had far greater flunixin exposure compared with the greyhounds that had the lowest. This is entirely consistent with the extent of ionization predicted by the Henderson–Hasselbalch equation. This variability in the urine pharmacokinetics reduces with time, and at 72 hr postadministration, in the terminal phase, the variability in urine and plasma flunixin concentrations are similar and should not affect medication control.     

Investigation of thyroid function in dogs treated with the tyrosine kinase inhibitor toceranib

Tyrosine kinase inhibitors are widely utilized in veterinary oncology for the treatment of mast cell and solid tumours. In man, these drugs are associated with thyroid dysfunction: however, to date only one study has investigated this in dogs. The aim of this study was to prospectively assess thyroid function in a group of dogs with cancer receiving toceranib. Thirty‐four dogs were prospectively enrolled at two referral hospitals into two groups; those receiving toceranib with prednisolone and those receiving toceranib alone. Total thyroxine (TT4) and thyroid stimulating hormone (TSH) was monitored at regular time points during treatment. Follow‐up data was available for 19 dogs. Overall, 12 incidences of elevated TSH occurred but none of these dogs had concurrent low TT4 concentrations. There was a significant difference in median TSH at week six compared with baseline. Hypothyroidism was not diagnosed in any patient during the study period. Patient drop‐out was higher than anticipated which prevented the assessment of longer term toceranib administration on thyroid function. Toceranib therapy was not associated with hypothyroidism in this study but did result in elevations in TSH which confirms what has been previously reported. Toceranib should be considered to cause thyroid dysfunction in dogs and monitoring is advised.     

Evaluation of Serum Thyroid‐Stimulating Hormone Concentration as a Diagnostic Test for Hyperthyroidism in Cats

Background

In humans, measurement of serum thyroid‐stimulating hormone (TSH) concentration is commonly used as a first‐line discriminatory test of thyroid function. Recent reports indicate that canine TSH (cTSH) assays can be used to measure feline TSH and results can help diagnose or exclude hyperthyroidism.

Objectives

To investigate the usefulness of cTSH measurements as a diagnostic test for cats with hyperthyroidism.

Animals

Nine hundred and seventeen cats with untreated hyperthyroidism, 32 euthyroid cats suspected of having hyperthyroidism, and 131 clinically normal cats.

Methods

Prospective study. Cats referred to the Animal Endocrine Clinic for suspected hyperthyroidism were evaluated with serum T₄, T₃, free T₄ (fT ₄), and TSH concentrations. Thyroid scintigraphy was used as the gold standard to confirm or exclude hyperthyroidism.

Results

Median serum TSH concentration in the hyperthyroid cats (<0.03 ng/mL) was significantly (P < .001) lower than concentrations in clinically normal cats (0.05 ng/mL) or euthyroid cats with suspected thyroid disease (0.06 ng/mL). Only 18 (2.0%) hyperthyroid cats had measurable TSH concentrations (≥0.03 ng/mL), whereas 114 (69.9%) of the 163 euthyroid cats had detectable concentrations. Combining serum TSH with T₄ or fT ₄ concentrations lowered the test sensitivity of TSH from 98.0 to 97.0%, but markedly increased overall test specificity (from 69.9 to 98.8%).

Conclusions and Clinical Importance

Serum TSH concentrations are suppressed in 98% of hyperthyroid cats, but concentrations are measurable in a few cats with mild‐to‐moderate hyperthyroidism. Measurement of serum TSH represents a highly sensitive but poorly specific test for diagnosis of hyperthyroidism and is best measured in combination with T₄ and fT ₄.     

Effect of age on serum chemistry profile, electrophoresis and thyroid hormones in beagle dogs two weeks to one year of age

Chemistry tests were conducted on serum from young Beagle dogs in order to deter mine the effect of age on these parameters. Blood was collected at regular intervals from 25 normal Beagle puppies (13 males and 12 females) at ages ranging from 2 weeks to 12 months. Serum chemistry profiles, protein electrophoresis and radioimmunoassays for thyroxine and triiodothyronine were included. Rapidly changing age related differences were observed in several parameters. Urea nitrogen, cholesterol, triglycerides, lactate dehydrogenase, thyroxine, glucose, gamma glutamyl transpeptidase, and total bilirubin values were elevated early in life, and decreased during the first 6 to 8 weeks, while alanine aminotransferase activity was low initially and increased during this period. Lactate dehydrogenase, thyroxine, gamma glutamyl transpeptidase, total bilirubin and alanine aminotransferase attained stability by 3 months, but the remaining parameters showed slight changes subsequently, gradually approaching adult values. More gradual age related changes were observed in other parameters. These included alkaline phosphatase, inorganic phosphorus and calcium values, which were higher in younger dogs, and creatinine, aspartate aminotransferase and total protein values, which were lower in younger dogs. Creatinine and aspartate aminotransferase values were stable by approximately 6 months; alkaline phosphatase, inorganic phosphorus, calcium and total protein values continued to change gradually up to 1 year.     

Post‐radiotherapy hypothyroidism in dogs treated for thyroid carcinomas

Hypothyroidism is a common adverse event after head and neck radiotherapy in human medicine, but uncommonly reported in canine patients. Records of 21 dogs with histologically or cytologically confirmed thyroid carcinoma receiving definitive or hypofractionated radiotherapy were reviewed. Nine cases received 48 Gy in 12 fractions, 10 received 36 Gy in 4 fractions and 2 received 32 Gy in 4 fractions. Seventeen cases had radiotherapy in a post‐operative setting. Ten cases developed hypothyroidism (47.6%) after radiotherapy. The development of hypothyroidism was not associated with the radiotherapy protocol used. Median time to diagnosis of hypothyroidism was 6 months (range, 1–13 months). Hypothyroidism is a common side effect following radiotherapy for thyroid carcinomas. Monitoring of thyroid function following radiotherapy is recommended. No specific risk factors have been identified.     

Protein and mRNA expression of follicle‐stimulating hormone receptor and luteinizing hormone receptor during the oestrus in the yak (Bos grunniens)

Follicle‐stimulating hormone (FSH) and luteinizing hormone (LH) have a central role in follicle growth, maturation and oestrus, but no clear pathway in the seasonal oestrus of yak (Bos grunniens) has been found. To better understand the role of FSH and LH in seasonal oestrus in the yak, six yaks were slaughtered while in oestrus, and the pineal gland, hypothalamus, pituitary gland, and gonads were collected. Using real‐time PCR and immunohistochemical assays, we determined the mRNA and protein expression of the FSH and LH receptors (FSHR and LHR) in these organs. The analysis showed that the FSHR mRNA expression level was higher in the pituitary gland tissue compared with LHR (p < .01) during oestrus. By contrast, there was low expression of FSHR and LHR mRNA in the pineal gland and hypothalamus. FSHR mRNA expression was higher than that of LHR (p < .05) in the ovary, whereas LHR mRNA expression was higher than that of FSHR (p < .01) in the uterus. FSHR and LHR proteins were located in the pinealocyte, synaptic ribbon and synaptic spherules of the pineal gland and that FSH and LH interact via nerve fibres. In the hypothalamus, FSHR and LHR proteins were located in the magnocellular neurons and parvocellular neurons. FSHR and LHR proteins were localized in acidophilic cells and basophilic cells in the pituitary gland, and in surface epithelium, stromal cell and gland epithelium in the uterus. In the ovary, FSHR and LHR protein were present in the ovarian follicle. Thus, we concluded that FSHR and LHR are located in the pineal gland, hypothalamus, pituitary and gonad during oestrus in the yak. However, FSHR was mainly expressed in the pituitary gland and ovaries, whereas LHR was mainly expressed in the pituitary gland and uterus.     

Heifers express G‐protein coupled receptor 153 in anterior pituitary gonadotrophs in stage‐dependent manner

We recently found that orphan G‐protein‐coupled receptor (GPR)153 is expressed in the anterior pituitary (AP) of heifers, leading us to speculate that GPR153 colocalizes with gonadotropin‐releasing hormone receptor (GnRHR) in the plasma membrane of gonadotrophs and is expressed at specific times of the reproductive cycle. To test this hypothesis, we examined the coexpression of GnRHR, GPR153, and either luteinizing hormone or follicle‐stimulating hormone in AP tissue and cultured AP cells by immunofluorescence microscopy. GPR153 was detected in the gonadotrophs, and was colocalized with GnRHR in the plasma membrane. GPR153 was also detected in the cytoplasm of cultured gonadotrophs. Real‐time PCR and western blot analyses found that expression was lower (P < 0.05) in AP tissues during early luteal phase as compared to pre‐ovulation or late luteal phases. The 5′‐flanking region of the GPR153 gene contained a consensus response element sequence for estrogen, but not for progesterone. These data suggest that some, but not all GPR153 colocalizes with GnRHR in the plasma membrane of gonadotrophs, and its expression changes stage‐dependently in the bovine AP.     

Thyroid hormones alter estrous cyclicity and antioxidative status in the ovaries of rats

To expand our understanding of the roles of thyroid hormones on female reproduction, we induced hypo‐ and hyper‐T rat models to investigate the roles of thyroid hormones on estrous cyclicity, as well as the antioxidative status in the ovaries of rats. In the current study, our data show that hypothyroidism (hypo‐T) and hyperthyroidism (hyper‐T) led to significantly reduced body weights and ovarain weights and delayed vaginal opening day. For hyper‐T, thyroxine (T4), tri‐iodothyronine (T3), progesterone (P4) and follicle‐stimulating hormone (FSH) were significantly increased, while estradiol (E2) and luteinizing hormone (LH) were significantly decreased. For hypo‐T rats, serum levels of total T4 and T3, E2, P4, FSH and LH were significantly increased, while concentrations of E2 and LH were significantly decreased. For ovary morphology, the numbers of secondary and antral follicles were significantly decreased with more atretic antral follicles and less corpora lutea in both hyper‐ and hypo‐T groups. Both hyper‐T and hypo‐T treatment significantly decreased the expressions of thyroid hormone receptor α1 in the ovary. Hypo‐T significantly reduced nitric oxide (NO), total NO synthase (tNOS), inducible NOS and constitutive NOS activities, but hyper‐T increased them. For antioxidative parameters, hypo‐T and hyper‐T treatment significantly increased malondialdehyde (MDA) contents. The activities of both glutathione peroxidase (GSH‐Px) and catalase (CAT) significantly decreased in the hypo‐T group but increased in the hyper‐T group. Total superoxide dismutase (T‐SOD) activity was significantly increased in the hyper‐T group. In summary, thyroid hormones alter estrous cyclicity and antioxidative status in the ovary of the rat may act through the NOS signaling pathway.     

Vitamin D status before and after hypophysectomy in dogs with pituitary-dependent hypercortisolism

Both spontaneous hypercortisolism and chronic glucocorticoid treatment are associated with osteoporosis and low circulating concentrations of 1,25-dihydroxy-vitamin D in humans. Pituitary-dependent hypercortisolism (PDH) is a common disorder in dogs, but little is known about the vitamin D status of affected dogs. Pituitary-dependent hypercortisolism in dogs can be treated effectively by hypophysectomy and subsequent hormone supplementation. Because hormone supplementation does not include GH, dogs that have undergone hypophysectomy have low circulating concentrations of GH and IGF-1, which may result in low plasma 1,25-dihydroxy-vitamin D concentrations and consequently increased parathyroid hormone secretion. The aim of this study was to determine whether dogs with PDH need vitamin D supplementation before and/or after hypophysectomy. To this end, we measured plasma concentrations of GH, IGF-1, parathyroid hormone, calcium, phosphate, and vitamin D metabolites in 12 dogs with PDH before and 8 wk after hypophysectomy and in 12 control dogs. Although plasma GH concentrations were lower in dogs with PDH than in control dogs both before and after hypophysectomy, the vitamin D status was similar. In conclusion, there is no need for vitamin D supplementation in dogs with PDH, either before or after hypophysectomy.     

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.