Plasma Thyroid Hormone Research Articles

Thyroid hormone manipulation influences development of endothermy and hatching in white leghorn chickens (Gallus gallus)

Chickens experience rapid change in their physiology and metabolism during hatching. We propose that thyroid hormones play a major role in regulating the developmental changes associated with attaining endothermy. To better understand the role thyroid hormones play in hatch timing and development of thermogenic capacity and metabolic rate we manipulated plasma thyroid hormone levels in chicken embryos beginning at 80% development (day 17 of a 21-day incubation) with either a single dose of triiodo-L-thyronine (T3) or the thyroperoxidase inhibitor methimazole (MMI). Manipulation of thyroid hormones altered the timing of hatching, accelerating hatching under hyperthyroid conditions, and prolonging hatching with hypothyroid conditions. Effect sizes comparisons of morphological variables between treatment groups revealed larger heart and body masses in hyperthyroid 1-day post hatch animals. Thyroid hormone manipulation influenced the thermal neutral zone for O2 consumption and body temperature during gradual cooling from 35 to 15 °C of externally pipped embryos and 1-day post hatch chicks. Hyperthyroid EP animals had a wider thermal neutral zone during cooling when compared to control animals. At the temperatures tested, the hypothyroid animals did not exhibit a thermal neutral zone. Similar differences between treatments in the breadth of the thermal neutral zone carried through to 1-day post hatch chickens. These findings suggest that thyroid manipulations influence the timing and development of the animal’s thermogenic response to cooling.

Identifying elevated plasma free triiodothyronine levels: age-adapted reference intervals for pediatrics.

Elevated free T3 (FT3) is an important feature for the early diagnosis of several diseases among which Grave's disease or Allan-Hernon-Dudley syndrome. However, there is a lack of age-adapted reference intervals for plasma thyroid hormones in children. We conducted a study to define reference values of peripheral FT3 in children using a commonly used automated immunoassay. All thyroid function test (TFT) results from our lab collected during 9 months were extracted anonymously, and reference intervals establishment followed recommendations validated by International Federation of Clinical Chemistry (IFCC). We defined five reference intervals covering the whole pediatric period. Overall, 26.1% of peripheral FT3 measured in children with normal TSH are out of the adult reference range, and 22.2% are upper it leading to misinterpretation. In a 9-month old patient with severe neurodevelopmental disorders, a pathological elevated FT3 has been securely interpreted using the newly established interval. The study highlights the poor relevance of adult intervals in pediatric cares, as it confirms that plasmatic FT3 is higher during the whole pediatric period. This work reports useful age-adapted reference intervals for free T3 in pediatrics using a widely used electrochemiluminescent Immunoassay (ECLIA) kit.

Divergent Thyroid Hormone Levels in Plasma and Left Ventricle of the Heart in Compensated and Decompensated Cardiac Hypertrophy Induced by Chronic Adrenergic Stimulation in Mice.

Chronic hemodynamic overload of the heart induces ventricular hypertrophy that may be either compensatory or progress to decompensation and heart failure. The gradual impairment of ventricular function is, at least in part, the result of a reduction of cardiac thyroid-hormone (TH) action. Here, we examined the proposed roles of increased cardiac expression of the TH-inactivating enzyme deiodinase type 3 (D3) and reduced plasma TH levels in diminishing cardiac TH levels. Using minipumps, mice were infused for one and two weeks with isoproterenol (ISO) alone or in combination with phenylephrine (PE). Remodeling of the heart induced by these adrenergic agonists was assessed by echocardiography. Left ventricular (LV) tissue and plasma TH levels (T4 and T3) were determined using liquid chromatography-tandem mass spectrometry. LV D3 activity was determined by conversion of radiolabeled substrate and quantification following HPLC. The results show that ISO induced compensated LV hypertrophy with maintained cardiac output. Plasma levels of T4 and T3 remained normal, but LV hormone levels were reduced by approximately 30% after two weeks, while LV D3 activity was not significantly increased. ISO + PE induced decompensated LV hypertrophy with diminished cardiac output. Plasma levels of T4 and T3 were substantially reduced after one and two weeks, together with a more than 50% reduction of hormone levels in the LV. D3 activity was increased after one week and returned to control levels after two weeks. These data show for the first time that relative to controls, decompensated LV hypertrophy with diminished cardiac output is associated with a greater reduction of cardiac TH levels than compensated hypertrophy with maintained cardiac output. LV D3 activity is unlikely to account for these reductions after two weeks in either condition. Whereas the mechanism of the mild reduction in compensated hypertrophy is unclear, changes in systemic TH homeostasis appear to determine the marked drop in LV TH levels and associated impairment of ventricular function in decompensated hypertrophy.

Plasma T3 and T4 Hormone Levels as Reference Values in Rehabilitated Common Buzzard (Buteo buteo), Long-Legged Buzzard (Buteo rufinus) and Golden Eagle (Aquila chrysaetos)

Applying appropriate criteria for raptors at rehabilitation stages including release and post release determines how healthy the animal can survive in its natural habitat for any length of time. In determining release criteria, clinical examinations of animals as well as clinical data should be evaluated. In this study, plasma T3 and T4 levels of 16 Common Buzzard (Buteo buteo), 7 Long-legged Buzzard (Buteo rufinus) and 7 Golden Eagles (Aquila chrysaetos) were determined at Kafkas University/ Kafkas Wildlife Conservation, Rescue, Rehabilitation Practice and Research Center between 2016-2018. All raptors were kept at the rehabilitation center for 1-3 months in bird care rooms and flight tunnels specially arranged for each species. Rehabilitated birds blood were collected before released to wild. Plasma T3 and T4 hormone levels were analysed and determined in raptors that considered suitable for release. Although small differences were detected between T3 and T4 values as a result of the analysis, no statistical difference was found. These small differences in values may be due to the metabolic responses and environmental conditions of each raptor. As a result, these plasma thyroid hormone levels in rehabilitated raptors can be accepted as reference values.

TOXIC EFFECTS OF AROCLOR 1254 ON RAT HEART AND THYROID AND MODIFYING ROLE OF SELENIUM STATUS

Objective: Polychlorinated biphenyls (PCBs) were very widely used in industrial products in past. These chemicals were banned in the 1970s due to their toxic effects. PCBs can still affect human health, as they are persistent in the environment. Aroclor 1254 (A1254) is a commercial PCB congener which was used in electrical transformers, fluorescent lighting fixtures and old appliances such as televisions or refrigerators. In this study, we aimed to evaluate the toxic effects of A1254 on heart and thyroid in male Sprague-Dawley rats. In addition, the modifying role of selenium status was also evaluated.
 Material and Method: 8-week-old male Sprague-Dawley (SD) rats were used in the experiment. The animals were separated randomly into 6 groups (n=6) as control; selenium supplemented (SeS); selenium deficient (SeD); A1254 exposed (A); selenium supplemented A1254 exposed (ASeS) and selenium deficient A1254 exposed (ASeD). A1254 was applied by gavage during the last 15 days of feeding period. Heart and thyroid weights and relative weights, plasma thyroid hormone levels, as well as thyroid and heart tissue oxidative/antioxidative parameters were evaluated.
 Result and Discussion: Results showed that A1254 exposure and selenium deficiency caused oxidative stress on both heart and thyroid. Plasma fT3 and fT4 levels markedly changed in ASeD group. In conclusion, it can be stated that A1254 exposure can cause lead to oxidative/antioxidative imbalance in both thyroid and heart and can disrupt functioning of thyroid hormones. Selenium seems to have a modifying role in A1254 toxicity in both organs, the importance of which should be evaluated with further mechanistic experiments.

Manipulating plasma thyroid hormone levels alters development of endothermy and ventilation in nestling red-winged blackbirds.

Thyroid hormones are key regulators of development and metabolism in vertebrates. During the nestling period, young of altricial species transition from an ectothermic phenotype to an endothermic phenotype. Red-winged blackbirds are an altricial species that exhibit an increase in plasma 3,3', 5-triiodo-L-thyronine (T3) levels during the first 5days post-hatch (dph), begin to develop endothermic metabolic responses by 7 dph, and fledge within 10 days of hatching. We propose that thyroid hormones play an important role in regulating development of endothermy during the nestling period in altricial birds. To better understand the effects of thyroid hormones on endothermic metabolic development in an altricial species, we treated nestling red-winged blackbirds on 2, 3, and 5 dph with either methimazole (MMI) to induce hypothyroidism or supplemental T3 to induce hyperthyroidism. We then measured on 5, 7, and 9 dph morphology and whole animal O2 consumption ( ) and ventilation in the thermal neutral zone and during gradual cooling. Treatment of nestlings with MMI resulted in lower plasma T3 levels on 5 dph that recovered by 7 dph, while supplementing with T3 did not affect plasma T3 levels on 5, 7 and 9 dph. Treatment with MMI resulted in smaller nestlings with smaller hearts and structural characters such as wing chord and femur length, but larger lungs and kidneys. Treatment with T3 produced smaller nestlings with smaller body masses and shorter femur and tarsus lengths. The development of and ventilation endothermic responses to gradual cooling in MMI treated nestlings were delayed when compared with control nestlings. In 9 dph nestlings, hypothyroidism resulted in alterations in the responses of ventilation frequency and tidal volume to cooling when compared with the control nestlings. Supplemental T3 had no effect on the development of and ventilation in the thermal neutral zone or in response to cooling. Our data suggest plasma thyroid hormone levels play an active role in the systemic development of endothermic capacity and the development of ventilatory control. In the nestling avian, multiple systems develop in concert to produce an endothermic phenotype, but reduced thyroid hormone delays maturation of endothermic capacity.

Artificial Dim Light at Night during Pregnancy Can Affect Hormonal and Metabolic Rhythms in Rat Offspring.

Artificial light at night (ALAN) is considered an environmental risk factor that can interfere with the circadian control of the endocrine system and metabolism. We studied the impact of ALAN during pregnancy on the hormonal and biochemical parameters in rat pups at postnatal (P) days P3, P10, and P20. Control dams (CTRL) were kept in a standard light-dark regime, and ALAN dams were exposed to dim ALAN (<2 lx) during the whole pregnancy. A plasma melatonin rhythm was found in all CTRL groups, whereas in ALAN pups, melatonin was not rhythmic at P3, and its amplitude was lowered at P10; no differences were found between groups at P20. Plasma corticosterone was rhythmic at P20 in both groups, with decreased mesor in ALAN pups. Plasma thyroid hormones exhibited an inconsistent developmental pattern, and vasopressin levels were suppressed at the beginning of the dark phase at P20 in ALAN compared to CTRL. Glucose and cholesterol showed significant daily rhythms in CTRL but not in ALAN offspring at P3. Exposure to ALAN during pregnancy disturbed the development of daily rhythms in measured hormones and metabolites, suggesting that ALAN during pregnancy can act as an endocrine disruptor that can interfere with the normal development of the progeny.

Transplantable human thyroid organoids generated from embryonic stem cells to rescue hypothyroidism

The thyroid gland captures iodide in order to synthesize hormones that act on almost all tissues and are essential for normal growth and metabolism. Low plasma levels of thyroid hormones lead to hypothyroidism, which is one of the most common disorder in humans and is not always satisfactorily treated by lifelong hormone replacement. Therefore, in addition to the lack of in vitro tractable models to study human thyroid development, differentiation and maturation, functional human thyroid organoids could pave the way to explore new therapeutic approaches. Here we report the generation of transplantable thyroid organoids derived from human embryonic stem cells capable of restoring plasma thyroid hormone in athyreotic mice as a proof of concept for future therapeutic development.

Tau protein is differentially phosphorylated in young- and old-aged rats with experimentally induced hyperthyroidism.

Aging involves progressive physiological changes, including thyroid dysfunction; thus, changes in plasma thyroid hormone (TH) level may affect neuronal function such as synaptic plasticity and Tau phosphorylation. However, how Tau protein is modulated in hyperthyroidism with aging is not clear. To clarify this issue, long-term potentiation (LTP) and accompanying phosphorylation of Tau protein in different residues were investigated in the hippocampus of young and old rats with experimentally induced hyperthyroidism. The study was performed in vivo under urethane anesthesia on 2- and 12-month-old Wistar albino male rats. Field potentials, composed of a field of excitatory postsynaptic potential (fEPSP) and a population spike (PS), occurring in the hippocampal dentate gyrus region, were recorded by applying high-frequency stimulation (HFS) to the perforant pathway (100 Hz, four times at 5-min intervals) to induce LTP. Total-Tau and phosphorylated-Tau were measured in HFS-induced hippocampus by using western blotting. The TH suppressed hippocampal somatic LTP (PS) was suppressed with aging, and treatment improved this suppression in aged rats without any changes in synaptic LTP (fEPSP). The phosphorylation of Tau at Ser202/Thr205 and Thr231 residues was increased in aged control rats. Treatment of aged rats with l-thyroxine reduced the phosphorylation of Tau at these residues to the young control condition. Impaired LTP that occurs with aging may be among the underlying causes of dementia in relatively older ages, and l-thyroxine treatment restores this impaired LTP. In addition, the phosphorylation level of Tau epitopes known to play a role in the pathogenesis of Alzheimer's disease may support a critical role in the modulation of synaptic plasticity in hyperthyroidism.

Thyroid Hormones Are Not Associated with Plasma Osteocalcin Levels in Adult Population with Normal Thyroid Function.

Thyroid hormones (THs) play an indispensable role in skeletal development and bone remodeling. Some studies have reported associations of THs with serum osteocalcin (OC) levels, but the results are quite inconsistent and the molecular mechanism of their simultaneous or interdependent activity on bone is almost unknown. Therefore, the aim of this study was to determine the possible associations of plasma THs with plasma OC levels and the possible mediating effect of OC on the relationship between THs and bone mineral density (BMD). For this purpose, out of the initial 1981 participants, we selected healthy euthyroid participants controlled for available confounding factors that can affect thyroid function and bone metabolism (N = 694). Given our results, we could not confirm any associations of THs with plasma OC levels nor the mediating effect of OC on the relationship between THs and BMD in euthyroid population. In the group of women controlled for menopause status (N = 396), we found a significant negative association of body mass index (BMI) with OC levels (β = −0.14, p = 0.03). We also found a negative association of free triiodothyronine (fT3) (β = −0.01, p = 0.02) and age (β = −0.003, p < 0.001) with BMD, and a positive association of BMI (β = 0.004, p < 0.001) and male gender (β = 0.1, p < 0.001) with BMD. In addition, we found significantly higher plasma OC levels and lower values of BMD in postmenopausal euthyroid women compared with premenopausal euthyroid women. In our opinion, the results of previous studies suggesting an association between circulating THs and serum OC levels may be influenced by an inconsistent selection of participants and the influence of confounding factors.

A rare case report of renal ewing sarcoma/primitive neuroectodermal tumor with ACTH production

BackgroundEwing sarcoma/primitive neuroectodermal tumor (PNET) of the renal is extremely rare. The common cause of ectopic ACTH syndrome is pulmonary neuroendocrine tumors, such as small cell carcinomas and carcinoid tumors. Here, we present an unusual case of ectopic ACTH syndrome and hypothyroidism caused by Ewing sarcoma/PNET of the right kidney.Case presentationA 19-year-old girl presented with a history of right lumbar pain and discomfort for 2 months, aggravated for 2 days. Abdominal contrast-enhanced computed tomography and computed tomography angiography showed an upper pole occupancy of the right kidney occupancy with subepithelial hemorrhage. Preoperative hormone levels including plasma total cortisol (PTC), adrenocorticotrophic hormone (ACTH) and thyroid hormone measurements were abnormal, indicating that the patient had Cushing syndrome and hypothyroidism. The patient underwent right radical nephrectomy. Histopathological analysis revealed a renal small round blue cell tumor (consistent with a primitive neuroectodermal tumor), with positive immunohistochemistry for CD99 and Ki67 (about 10%) and molecular pathology for EWSR1 gene fusions. PTC, ACTH and thyroid hormone returned to normal after surgery.ConclusionsWe report a rare ectopic ACTH syndrome and hypothyroidism due to renal Ewing sarcoma/PNET. The clinical manifestation of renal Ewing sarcoma/PNET is non-specific and the diagnosis relies on pathological morphology, immunohistochemistry and fusion gene detection. At present, surgery combined with radiotherapy and chemotherapy is used in the treatment, but the prognosis is still not optimistic.

Assessment of Joint Impact of Iodine, Selenium, and Zinc Status on Women's Third-Trimester Plasma Thyroid Hormone Concentrations

ABSTRACTBackgroundIodine is essential for synthesizing thyroid hormones, but other micronutrients are also required for optimal thyroid function. However, there is a lack of data on combined micronutrient status in relation to thyroid hormones in pregnancy.ObjectivesWe aimed to assess the joint associations of iodine, selenium, and zinc status with plasma concentrations of thyroid hormones and thyroid-stimulating hormone (TSH) in pregnancy.MethodsWe included 531 pregnant women (aged 22–40 y) participating in a Swedish birth cohort who provided blood and spot urine samples in gestational weeks 27–33 (mean: 29). Associations of urinary iodine concentration (UIC), plasma selenium concentration, and plasma zinc concentration (measured by inductively coupled plasma mass spectrometry) with plasma hormone concentrations [total and free thyroxine (tT4, fT4), total and free triiodothyronine (tT3, fT3), and TSH] were explored with Bayesian kernel machine regression (BKMR; n = 516; outliers excluded) and multivariable-adjusted linear regression (n = 531; splined for nonlinear associations).ResultsMedian (IQR) micronutrient concentrations were 112 μg/L (80–156 μg/L) for UIC, 67 μg/L (58–76 μg/L) for plasma selenium, and 973 μg/L (842–1127 μg/L) for plasma zinc; the former 2 median values were below recommended concentrations (150 μg/L and 70 μg/L, respectively). Mean ± SD TSH concentration was 1.7 ± 0.87 mIU/L, with 98% < 4 mIU/L. BKMR showed a positive trend of joint micronutrient concentrations in relation to TSH. Plasma zinc was most influential for all hormones but tT3, for which plasma selenium was most influential. In adjusted linear regression models, zinc was positively associated with tT4, tT3, and TSH, and <1200 μg/L also with fT4 and fT3. Selenium was inversely associated with fT3, and <85 μg/L with tT3.ConclusionsPregnant women's plasma TSH concentrations in the early third trimester increased with increasing joint status of iodine, selenium, and zinc. Zinc and selenium were more influential than iodine for the hormone concentrations. Multiple micronutrients need consideration in future studies of thyroid hormone status.

Thyroid and Extrathyroid Effects of Methimazole in Modeling Hypothyroidism with Different Doses of Antithyroid Drug: Morfofunctional Study

Aims: Methimazole (MMI) is widely used in experimental thyroidology. However, the effect on thyroid function can vary widely depending on the route of administration and doses of the antithyroid drug. In some cases, it is difficult to determine whether the results obtained are a manifestation of thyroid dysfunction or a direct toxic effect of xenobiotic. The aim of the study was to establish the optimal dose of methimazole for the induction of hypothyroidism with minimal toxic effects on tissues.&#x0D; Study Design: The effect of various doses of methimazole administered intragastrically to rats on thyroid status, histological structure of thyroid, liver and kidney was studied.&#x0D; Place and Duration of Study: Bashkir institute of Technology and Management (branch) of Moscow State University of Technology and Management n.a. K. G. Razumovsky, city of Meleuz, Bashkir State Medical University, Ufa, Russia, 2018-2019.&#x0D; Methodology: Experiments were conducted on 60 nonlinear white male rats intragastrically administered different doses of methimazole. To assess the thyroid status, the concentration of circulating thyroid hormones in blood plasma and body temperature rhythms were determined. Along with the indicators of thyroid status, morphological changes in the thyroid gland, liver and kidneys were compared in rats that were administered different doses of MMI.&#x0D; Results: Hormonal profile of rats received MMI in the dose 2,5 mg/100 g of body weight. for 3 weeks revealed decrease in mean free T4 (10,8±2,14 against 16,2±0,57 in control, P&lt;0,001), total T3 (3,12±0,57 against 2,36±0,19, P&lt;0,001 with the increase of TSH (0, 12±0,01 against 1,96±0,18, P&lt;0,001). The hypothyroid state in this group of animals is also confirmed by decrease in the arithmetic mean values of body temperature.&#x0D; Conclusion: The results obtained indicate that a dose of 2.5 mg/100 g of b. w. for 3 w. leads to the development of symptoms characteristic of endemic thyroid dysfunction, and is accompanied by minimal toxic effects on tissues.

An animal model for Pierpont syndrome: a mouse bearing the Tbl1xr1Y446C/Y446C mutation.

Pierpont syndrome is a rare disorder characterized mainly by global developmental delay, unusual facial features, altered fat distribution in the limbs and hearing loss. A specific mutation (p.Tyr446Cys) in TBL1XR1, encoding a WD40 repeat-containing protein, which is a component of the SMRT/NCoR (silencing mediator retinoid and thyroid hormone receptors/nuclear receptor corepressors), has been reported as the genetic cause of Pierpont syndrome. Here, we used CRISPR-cas9 technology to generate a mutant mouse with the Y446C mutation in Tbl1xr1, which is also present in Pierpont syndrome. Several aspects of the phenotype were studied in the mutant mice: growth, body composition, hearing, motor behavior, thyroid hormone state and lipid and glucose metabolism. The mutant mice (Tbl1xr1Y446C/Y446C) displayed delayed growth, altered body composition with increased relative lean mass and impaired hearing. Expression of several genes involved in fatty acid metabolism differed in white adipose tissue, but not in liver or muscle of mutant mice compared to wild-type mice (Tbl1xr1+/+). No difference in thyroid hormone plasma concentrations was observed. Tbl1xr1Y446C/Y446C mice can be used as a model for distinct features of Pierpont syndrome, which will enable future studies on the pathogenic mechanisms underlying the various phenotypic characteristics.

Effect of elevated carbon dioxide on chicken eggs during the early and late incubation periods

Carbon dioxide (CO2) is always maintained at ambient levels by ventilation in commercial egg incubators. However, elevated CO2 levels during the early and late periods have been reported to improve the quality of chicks and shorten the hatch window. This study investigated the effect of precise CO2 supplementation during the early and late periods of incubation on embryo growth and incubation performance by developing and using a CO2 supplementation system to increase the CO2 level in an experimental egg incubator. The CO2 level was maintained at 1% in the early period (from the beginning to the 10th day of incubation, E0–E10) and in the late period (from internal pipping (IP) to the 21st day of incubation (E21), IP–E21) in an incubator for the treatment group, whereas the CO2 level was maintained at the ambient level in the other incubators for the control group. A comparative assessment of embryonic development, hatching characteristics, and hormone and nutrient levels was conducted for each trial. The experiment comprised three trials, with 300 Jing Hong No. 1 breeding eggs in each incubator. The elevated CO2 treatment significantly shortened the chick hatching time (H0) by 4 h (P < 0.05) and the hatch window by 3 h (P < 0.05) without affecting hatchability, chick weight at 1 d of age, brooding period, or quality score. At external pipping (EP), the heart weight, intestinal weight, relative intestinal weight, and relative heart weight in the treatment group were significantly higher than those in the control group (P < 0.05). In addition, the embryonic intestine, relative intestine, and relative heart weights of the newly hatched chicks in the treatment group were significantly higher than those in the control group (P < 0.05) at H0. The treatment significantly increased the concentration of corticosterone in the embryonic plasma during the period from IP to EP (P < 0.05), promoted the secretion of triiodothyronine and tetraiodothyronine (P < 0.05), and increased the glycogen content of the embryonic liver on E21 (P < 0.05). This result indicates that elevated CO2 (1%) during the early and late periods of incubation accelerated embryonic organ development and shortened the chick hatching time and hatch window without affecting hatchability or hatchling quality, which can be explained by the synergistic functions of the secretion of plasma corticosterone and thyroid hormones and the accumulation of liver glycogen between the early and late periods of incubation.

Thyroid Hormone Concentrations in Testudo spp. by Season and Sex

Thyroid hormones and the factors influencing them are rarely studied in tortoises. This study therefore aimed to calculate reference intervals (RI) for thyroid hormones and iodine concentrations by species, sexes, and seasons in four species of adult Mediterranean tortoises (Testudo spp.) and to evaluate possible correlations between plasma thyroid hormones, serum iodine, plasma protein, and increased liver and kidney values. Thyroid hormones (total tetraiodothyronine [tT4], free tetraiodothyronine [fT4], total triiodothyronine [tT3], and free triiodothyronine [fT3]) were measured in plasma from adult, healthy Hermann's (Testudo hermanni, n = 255), spur-thighed (Testudo graeca, n = 89), marginated (Testudo marginata, n = 72), and Russian tortoises (Testudo horsfieldii, n = 30). Species, sex, and seasonal variations were determined by Bayesian information criterion (BIC), and correlations between thyroid hormones, protein, iodine, and increased liver/kidney values were evaluated by Spearman's rank correlation test. Total T4 did not reveal any species, sex, or seasonal differences (RI: 0.102–0.455 µg/dl), while seasonal differences were found for fT4 (RI: spring 0.624–9.012; summer 0.379–5.476; fall 0.376–5.426 pmol/L). The tT3 concentrations differed significantly depending on species, season, and the interaction of species × season. Seasonal differences were also found for fT3 and iodine. Several significant (P ≤ 0.05) correlations were detected between the analytes, including a positive correlation between tT4 and fT4. These results provide a tool for the evaluation of thyroid hormone concentrations in Mediterranean tortoises and confirm the influence of season on thyroid function in these animals.

Effects of Standardized Exercise Tests on Plasma Thyroid Hormones’ Kinetics in Standardbred Racehorses

This study examined how a standard exercise test (SET) affected (1) thyroid hormones (THs) of horses and (2) the relationship between the VLa4 of horses and TH responses to the exercise in trained Standardbred racehorses (VLa4 is the velocity run at defined conditions at which a blood lactate concentration of 4 mmol/L is determined). Twelve trained Standardbred racehorses (six stallions and six mares) performed SETs until the horses’ blood lactate concentration was at or above 4 mmol/L. The horses were divided into three age groups (2, 3, and 4 years old); each group consists of 4 horses respectively (two male and two female), to evaluate the effects of age and sex on hormonal responses to SET. During each SET, blood samples were taken at rest and after each interval and at the end of SET. Blood was analysed for total and free triiodothyronine (T3, fT3) as well as total and free thyroxine (T4, fT4). The statistical model included three fixed factors (SET, sex and age) and their main interactions. ANOVA analysis revealed that T3 and fT3 were significantly influenced by SETs. Plasma T3 and fT3 concentrations were higher in 4-year-old horses compared to the other age groups. All plasma THs concentrations were higher in mares than in stallions. Correlations revealed that a higher VLa4 was negatively related to all THs responses in 2-year-old Standardbred mares only. The SET used to determine VLa4 increased selected THs (T3, fT3); these increases were inversely related to VLa4 and affected by age and sex of the horses. The correlation of VLa4 with thyroid exercise’ response might provide some additional information for performance evaluation of Standardbred racehorses, especially for evaluating training adaptation, according to sex, and age. Further studies are necessary to provide support on the value of measuring THs in Standardbred racehorses of different sex and age.

Thyroid hormone levels in Tortoises (Testudo spp.) depending on Season and Sex

Thyroid hormones and the factors influencing them are rarely studied in tortoises. This study therefore aimed to calculate reference intervals (RI) for specific species, sexes, and seasons for thyroid hormones and iodine levels in blood of four adult Mediterranean tortoise species and to evaluate possible correlations between thyroid hormones, serum iodine, plasma protein and increased liver and kidney values. Thyroid hormones (total tetraiodothyronine [tT4], free tetraiodothyronine [fT4], total triiodothyronine [tT3], and free triiodothyronine [fT3]) were measured in plasma from adult, healthy Hermann´s (Testudo hermanni, n = 255), spur-thighed (Testudo graeca, n = 89), marginated (Testudo marginata, n = 72), and Russian tortoises (Testudo horsfieldii, n = 30). Species, sex and season specific variations were determined by Bayesian information criterion (BIC) and correlations between plasma thyroid hormones, protein, iodine and increased liver/kidney values were evaluated by Spearman’s rank correlation test. Total T4 did not reveal any species, sex, or seasonal differences (RI 0.102 to 0.455 µg/dL), while seasonal differences were found for fT4 (RI spring 0.624 to 9.012; summer 0.379 to 5.476; fall 0.376 to 5.426 pmol/L). The tT3 levels differed significantly depending on species, season, and the interaction of species x season. Seasonal differences were also found for fT3 and iodine. Several significant (p &amp;lt; 0.05) correlations were detected between the tested analytes, especially positive correlations between tT4 and fT4. These results provide a tool for the evaluation of thyroid hormone levels in Mediterranean tortoises and indicate the influence of season on the thyroid in these animals.

Developing potency factors for thyroid hormone disruption by PFASs using TTR-TRβ CALUX® bioassay and assessment of PFASs mixtures in technical products.

Over the last decade, per- and polyfluoroalkyl substances (PFASs) have become one of the most heavily investigated persistent organohalogen compound class of environmental concern. However, knowledge about their toxicology is still scarce, although PFASs as individual compounds and their industrial mixtures were shown to exert effects on the thyroid hormone system. In vitro toxicity potency factors were established for thyroid hormone transport disruption potential using the novel TTR-TRβ CALUX® bioassay for major PFASs. We assessed technical PFASs mixtures, including aqueous film-forming foam (AFFF) surfactants and chromium mist suppressants (CMS) applications with and without total oxidizable precursor (TOP) by TTR-TRβ CALUX® assay for their thyroid hormone transport disrupting potential. All PFASs listed in the German guideline for drinking water (German Environment Agency, 2017) affected the T4 binding to TTR, an important plasma thyroid hormone transport protein. For all tested PFASs, potency factors based on PC80 values relative to PFOA could be obtained and ranged between PFBA (0.0018) and PFOS (2.0). Applying in vitro potency factors obtained from the present in vitro TTR-TRβ CALUX® assay study and recently reported in vivo potency factors (Zeilmaker et al., 2018; Bil et al., 2021) on the above-mentioned German guideline for PFAS in drinking water, showed that the cumulative effect-based trigger values (in vivo and in vitro) are comparable (3.0 vs. 2.9 to 4.6μg PFOA-EQ/l). Additionally, AFFF surfactants and CMS with and without TOP assay were tested. Highest activities were found in the older AFFF surfactants (2013/2014) due to higher PFOS/PFOA levels, which were already substituted with 6:2 FTS in 2019, resulting in much lower PFOA-EQ levels. As expected also the PFOA-EQ levels increased in the samples with TOP treatment compared to the original AFFF surfactants and CMS as confirmed here by biological and chemical PFOA-equivalents (PFOA-EQ) analysis. Additionally, CMS (which have been used in the electroplating chromium industry since the 1950s) as well as PFOS-free, but not PFAS-free fume suppressants (such as Fumetrol® 21) have been tested in the TTR-TRβ CALUX® assay and showed much lower activity levels then the AFFFs, confirmed by the similar potency determination based on chemical PFASs analysis followed by transformation to PFOA-EQ for comparison. The potency factor of 6:2 FTS, which is the main substitute for PFOS in CMS, indicates that it is approximately 100-times less potent as a thyroid hormone disruptor as compared to PFOS. Potency factors based on PC80 values from TTR-TRβ CALUX® relative to PFOA have been developed for major PFASs. In AFFF surfactants and CMS a trend of higher activities with higher amounts of PFOS and PFOA have been found. PFOA and PFOS showed high responses in the TTR-TRβ CALUX® assay and had the largest contributions to the PFOA-EQs in the AFFF surfactants and CMS applications. Using potency factors as determined in the TTR-TRβ CALUX® to convert PFASs assessed by chemical analysis to PFOA-EQ led to comparable results as compared to the results from PFASs measured directly by the TTR-TRβ CALUX® assay. This study supports the claim that semiquantitative effect- and group-based in vitro CALUX bioanalysis tools can be applied effectively to assess industrial products containing complex mixtures with PFAS compounds for which no instrumental analysis are established, and for many compounds where in vitro toxicity data are not yet available.

Seasonal Changes in Plasma Hormones, Sex-Related Genes Transcription in Brain, Liver and Ovary during Gonadal Development in Female Rainbow Trout (Oncorhynchus mykiss)

The purpose of this study was to investigate the periodic seasonal changes in endocrine activity and gonadal development of female rainbow trout (Oncorhynchus mykiss) in a high-altitude cold-water environment. The fish were sampled monthly from January to November and the levels of plasma hormones (estradiol (E2), cortisol and thyroid hormones (THS)) and vitellogenin (VTG) were measured by ELISA. Moreover, the transcriptions of sex-related genes in the ovary, brain, and liver were detected by qRT-PCR. The results showed a seasonal fluctuation of plasma hormones and VTG together with the development of the ovary, which reached a peak from August to October. Similarly, the transcription of hypothalamic gonadotropin-releasing hormone-2 (cgnrh-2), hypothalamic gonadotropin-releasing hormone receptors (gnrhr) and follicle-stimulating hormone (fsh) in the brain varied from January to September, but the highest level was detected in September to November. In addition, the transcription of sex-related genes located in the ovary and liver increased significantly during August to October, accompanied by a continuous increase in the gonadosomatic index (GSI) and a decrease in the hepatosomatic index (HSI). Therefore, plasma hormones and sex-related genes regulate the development and maturation of O. mykiss oocytes with the change in seasons and peaked in November. The results of this study provide a reference for improving the efficiency of the artificial reproduction of O. mykiss.