Effect Of T3 Research Articles

MRNA-miRNA networks identify metabolic pathways associated to the anti-tumorigenic effect of thyroid hormone on preneoplastic nodules and hepatocellular carcinoma.

BackgroundThyroid hormones (THs) inhibit hepatocellular carcinoma (HCC) through different mechanisms. However, whether microRNAs play a role in the antitumorigenic effect of THs remains unknown.MethodsBy next generation sequencing (NGS) we performed a comprehensive comparative miRNomic and transcriptomic analysis of rat hepatic preneoplastic lesions exposed or not to a short-term treatment with triiodothyronine (T3). The expression of the most deregulated miRs was also investigated in rat HCCs, and in human hepatoma cell lines, treated or not with T3.ResultsAmong miRs down-regulated in preneoplastic nodules following T3, co-expression networks revealed those targeting thyroid hormone receptor-β (Thrβ) and deiodinase1, and Oxidative Phosphorylation. On the other hand, miRs targeting members of the Nrf2 Oxidative Pathway, Glycolysis, Pentose Phosphate Pathway and Proline biosynthesis – all involved in the metabolic reprogramming displayed by preneoplastic lesions– were up-regulated. Notably, while the expression of most miRs deregulated in preneoplastic lesions was not altered in HCC or in hepatoma cells, miR-182, a miR known to target Dio1 and mitochondrial complexes, was down-deregulated by T3 treatment at all stages of hepatocarcinogenesis and in hepatocarcinoma cell lines. In support to the possible critical role of miR-182 in hepatocarcinogenesis, exogenous expression of this miR significantly impaired the inhibitory effect of T3 on the clonogenic growth capacity of human HCC cells.ConclusionsThis work identified several miRNAs, so far never associated to T3. In addition, the precise definition of the miRNA-mRNA networks elicited by T3 treatment gained in this study may provide a better understanding of the key regulatory events underlying the inhibitory effect of T3 on HCC development. In this context, T3-induced down-regulation of miR-182 appears as a promising tool.

Visualizing Hormonal Effects on Cardiomyocyte Hypertrophic Growth Dynamics Using Digital Holographic Microscopy: Does Size Matter?

Heart disease continues to be the leading cause of death in the United States. Humans are unable to regenerate their heart tissue following an injury. However, neonatal mice are able to regenerate their heart tissue when cardiomyocytes (CMs) proliferate. This regenerative ability is lost approximately one week after birth when proliferating mononucleated CMs become binucleated and can no longer complete the cell-cycle. Recent studies have shown the combined inhibition of thyroid hormone (T3) and norepinephrine (NE) increases CM proliferation, promotes heart regeneration, and reduces cell size in vivo. Using digital holographic microscopy, the aim of this study was to (1) validate the novel Halomonitor approach and (2) visualize and quantify the effects of T3 and NE on cardiomyocyte size. CMs were isolated from neonatal rats 1 to 2 days after birth which were then treated with NE and/or T3 in serum-free culture. Live cell imaging was visualized utilizing digital holographic microscopy and changes in CM dynamics and morphology were quantitatively tracked and analyzed using HoloMonitor software. The results from this study validate the HoloMonitor technology by demonstrating that NE induced hypertrophy in CMs after 12 hours. Our results also demonstrate the ability of HaloMonitor technology to differentiate between CMs and non-CMs in living, serum-free culture, while also studying and quantifying their dynamics in vitro. Lastly, our data shows that T3 has little effect on CM growth and that NE decreased CM motility.

Triiodothyronine-stimulated dendritic cell vaccination boosts antitumor immunity against murine colon cancer

Given the ability of dendritic cells (DCs) to modulate other immune players, their manipulation holds great potential for inducing efficient antitumor immunity. However, DC vaccine manufacturing deserve optimization since tumor cell cargo and DC functional state induced by maturation signals influence their in vivo immunogenic potential. We reported that triiodothyronine (T3) stimulates mice DCs’ maturation and their ability to promote pro-inflammatory and cytotoxic T-cell responses. This study aimed to evaluate the efficacy of T3-conditioned DC vaccination in a murine model of colon carcinoma, deciphering the molecular players involved; and to examine the effects of T3 on the maturation and activation of human DCs (huDCs). Bone marrow-derived DCs were exposed to T3 and MC38 cancer cells that underwent cell death (MC38-Apo/Nec) by UVB irradiation. Our results showed that MC38-Apo/Nec cells are efficiently uptaken by DCs and that T3 upregulates CD86 expression with increased production of the pro-inflammatory cytokines IL-12, IL-6 and TGF-β. In a colon cancer model, vaccination with T3-stimulated and tumor antigen-loaded DCs inhibited tumor growth in wild type mice, an effect that was eliminated in IL-17-deficient animals. Notably, secretion of high levels of IFN-γ and IL-17, induction of Th1, Th17 and tumor-specific cytotoxic T lymphocytes characterized the antitumor response upon vaccination. Moreover, our initial findings demonstrated a significant increase in CD86 expression and IL-12 production by huDCs induced by T3. Overall, these results reinforce the adjuvant properties of T3-conditioned DCs to potentiate T-cell-mediated antitumor immunity and support promising advances in the translation process to human oncotherapy.

Brief Research Report: Effect of Triiodothyronine on Hepatic Growth Hormone Receptor Expression in Primary Bovine Hepatocytes.

In previous studies, triiodothyronine (T3) was found to be lower in cows with ketosis and an effect of T3 on Growth Hormone Receptor (GHR) expression is described, e. g., in a human hepatoma cell line. Therefore, this study aimed to test whether T3 affects GHR messenger RNA (mRNA) expression in a well-established bovine hepatocyte model. Hepatocytes were kept in a sandwich culture and stimulated for 6 days with constant (10 μg/ml) or decreasing (from 10 to 5 μg/ml) T3 concentrations, and GHR, as well as IGF-1 mRNA expression, was measured using real-time polymerase chain reaction (RT-PCR). We could confirm in vitro that T3 has a stimulatory effect on GHR1A mRNA expression.

3,3′,5-triiodo-l-thyronine inhibits drug-induced liver injury through activation of PPARα as revealed by network pharmacology and biological experimental verification

Drug-induced liver injury (DILI) has increased in recent years, leading to acute liver failure. 3,3′,5-triiodo-l-thyronine (T3) has been reported to exert a potent hepatoprotective effect. However, the mechanism and efficacy of T3 on DILI remain undocumented. In this study, an MTT assay was used to detect the effect of T3 on hepatotoxicity of acetaminophen (APAP) in L02 cells. Then, we screened key targets and related biological pathways by network pharmacology. Finally, enzyme-linked immunosorbent assay (ELISA) and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) were used to verify the mechanism and key targets of T3 on DILI. The results of the MTT assay showed that T3 significantly decreased hepatocellular injury induced by APAP. Network pharmacology and bioinformatics analysis showed that 118 intersection targets of T3 and DILI were identified and the mechanism of T3 on DILI was related to cell proliferation and oxidative stress. ELISA results showed that T3 may be an effective treatment for DILI as biomarkers of hepatocellular injury such as AST, ALP were decreased compared to APAP only treated cells, and the mechanism of T3 may be mediated in part through improving redox balance. The topological parameter screening results suggested 12 key targets of T3 for DILI. Among them, PPARα is associated with DILI, and activation of PPARα can reduce oxidative stress and cell necrosis. Therefore, PPARα was identified as a target for verification. qRT-PCR analysis demonstrated that T3 could reverse the down-regulation of PPARα induced by APAP exposure. Taken together, we demonstrated for the first time that T3 could activate PPARα, promote cell proliferation and reduce oxidative stress, and play a vital role in the treatment of DILI, which provides a reference for T3 as a candidate treatment for DILI.

Remuscularization with triiodothyronine and β1-blocker therapy reverses post-ischemic left ventricular dysfunction and adverse remodeling

Renewal of the myocardium by preexisting cardiomyocytes is a powerful strategy for restoring the architecture and function of hearts injured by myocardial infarction. To advance this strategy, we show that combining two clinically approved drugs, but neither alone, muscularizes the heart through cardiomyocyte proliferation. Specifically, in adult murine cardiomyocytes, metoprolol, a cardioselective β1-adrenergic receptor blocker, when given with triiodothyronine (T3, a thyroid hormone) accentuates the ability of T3 to stimulate ERK1/2 phosphorylation and proliferative signaling by inhibiting expression of the nuclear phospho-ERK1/2-specific phosphatase, dual-specificity phosphatase-5. While short-duration metoprolol plus T3 therapy generates new heart muscle in healthy mice, in mice with myocardial infarction-induced left ventricular dysfunction and pathological remodeling, it remuscularizes the heart, restores contractile function and reverses chamber dilatation; outcomes that are enduring. If the beneficial effects of metoprolol plus T3 are replicated in humans, this therapeutic strategy has the potential to definitively address ischemic heart failure.

Triiodothyronine Improves Contractile Function of Hypoxic/Reoxygenated Human Right Atrial Tissue

BackgroundIn patients with acute myocardial infarction, the one‐year mortality remains at 7‐15% despite successful interventional reperfusion. Thus, innovative treatment strategies beyond reperfusion are needed. In isolated perfused rat hearts, triiodothyronine (T3), when given during early reperfusion in a supraphysiological concentration, improves left ventricular function and reduces infarct size (Basic Res Cardiol 2021;116:27).AimTo investigate whether or not such acute, non‐genomic effects of T3 also protect human myocardium.MethodsWe isolated right atrial trabeculae from patients undergoing routine coronary artery bypass grafting and investigated the effect of T3 on contractile function during hypoxia/reoxygenation. Trabeculae (≥ 3 mm length, ≤ 1 mm diameter) were isolated in cardioplegic buffer and transferred to Tyrode buffer. Electrically initiated (1 Hz) developed force of contraction (mN/mm2) was recorded for 10 min at baseline (95% O2 – 5% CO2, glucose). Hypoxia/reoxygenation (60 min/30 min) was induced by changing the buffer gas supply and composition and by increasing the stimulation rate (95% N2 – 5% CO2, choline chloride, 3 Hz) and returning back to baseline. T3 (500 µg/l) or NaOH (as solvent control), respectively, was administered A) during hypoxia and reoxygenation (n=11 vs. n=10) or B) 15 min before and during reoxygenation (n=12 vs. n=14).ResultsAt baseline, before T3 or NaOH treatment, the contractile function of trabeculae was comparable (mN/mm2: 15.4±3.5, 13.8±4.4 in protocol A, and 14.5±4.1, 13.7±4.0 in protocol B, respectively). T3 improved the contractile function during reoxygenation, irrespectively of whether given continuously (A) or just before and during reoxygenation (B; see Figure).ConclusionT3 protects patient myocardium: contractile function is improved during reoxygenation in right atrial trabeculae. This bioassay will now permit an identification of causally involved cardioprotective signaling molecules of such non‐genomic protection in human tissue by T3.

Pairing in the Hubbard model on the honeycomb lattice with hopping up to the third-nearest-neighbor

The electronic properties of graphene can be approximately described by the standard Hubbard model on the honeycomb lattice. It was argued that the d+id-wave pairing is the dominant pairing symmetry in such a system. However, in the standard Hubbard model, only the nearest-neighbor hopping t1 is considered, while the second-nearest-neighbor hopping t2 and third-nearest-neighbor hopping t3 are ignored. In this paper, by doing the determinant quantum Monte Carlo simulations of the Hubbard model with hopping up to the third-nearest-neighbor, we examine the effect of t2 and t3 on the pairing symmetries. We find that the second- and third-nearest-neighbor hopping amplitudes have strong effect on the d+id-wave pairing symmetry.

Direct Interaction of Mitochondrial Cytochrome c Oxidase with Thyroid Hormones: Evidence for Two Binding Sites.

Thyroid hormones regulate tissue metabolism to establish an energy balance in the cell, in particular, by affecting oxidative phosphorylation. Their long-term impact is mainly associated with changes in gene expression, while the short-term effects may differ in their mechanisms. Our work was devoted to studying the short-term effects of hormones T2, T3 and T4 on mitochondrial cytochrome c oxidase (CcO) mediated by direct contact with the enzyme. The data obtained indicate the existence of two separate sites of CcO interaction with thyroid hormones, differing in their location, affinity and specificity to hormone binding. First, we show that T3 and T4 but not T2 inhibit the oxidase activity of CcO in solution and on membrane preparations with Ki ≈ 100–200 μM. In solution, T3 and T4 compete in a 1:1 ratio with the detergent dodecyl-maltoside to bind to the enzyme. The peroxidase and catalase partial activities of CcO are not sensitive to hormones, but electron transfer from heme a to the oxidized binuclear center is affected. We believe that T3 and T4 could be ligands of the bile acid-binding site found in the 3D structure of CcO by Ferguson-Miller’s group, and hormone-induced inhibition is associated with dysfunction of the K-proton channel. A possible role of this interaction in the physiological regulation of the enzyme is discussed. Second, we find that T2, T3, and T4 inhibit superoxide generation by oxidized CcO in the presence of excess H2O2. Inhibition is characterized by Ki values of 0.3–5 μM and apparently affects the formation of O2●− at the protein surface. The second binding site for thyroid hormones presumably coincides with the point of tight T2 binding on the Va subunit described in the literature.

Identification and expression analysis of thyroid-stimulating hormone β subunit, and effects of T3 on gonadal differentiation-related gene expression in rice field eel, Monopterus albus.

Thyroid-stimulating hormone (TSH) is an important glycoprotein in hypothalamic-pituitary-thyroid axis, which plays a crucial role in the synthesis and release of thyroid hormones in vertebrates. Rice field eel, Monopterus albus, a protogynous hermaphroditic fish, which undergoes sex reversal from a functional female to a male, is an ideal model to investigate the regulation of sex differentiation. In this study, we obtained the cDNA sequence of thyroid-stimulating hormone β subunit (tshβ) from rice field eel, which contained a complete open reading frame and encoded a putative protein of 151 amino acids. Multiple alignment of protein sequences showed that tshβ was highly conserved in teleost. The tissue distribution indicated that tshβ showed high expression in the pituitary, moderate expression in the brain region, gonad, intestine and liver, and low expression in other peripheral tissues. During natural sex reversal, the expression of tshβ had no significant difference in the pituitary. Compared to that in the ovary, the expression of tshβ increased significantly in the gonad at late intersexual and male stages. After treatment by different doses of triiodothyronine (T3) (1μg/g, 10μg/g and 100μg/g body weight), serum T3 and free triiodothyronine (FT3) increased sharply, while the expression of tshβ were inhibited significantly in the pituitary. Although T3 had no significant effect on the levels of serum E2, it stimulated the release of serum 11-KT at high-dose group. We also detected the effects of T3 on the expression of gonadal differentiation-related genes in rice field eel. T3 treatment inhibited the expression of foxl2, cyp19a1a and dax1, while stimulated the expression of sox9a1. These results indicate that TSH may be involved in sex differentiation, and THs may play roles in the regulation of male development and sex reversal in rice field eel.

The Role of General and Study-Related Intraindividual Factors on Academic Learning Outcomes under COVID-19: A Cross-Sectional and Longitudinal Analysis

Little is known about the intraindividual dispositional factors related to cognitive, behavioral, and emotional academic learning outcomes under COVID-19. This study investigated (i) the associations of intraindividual factors, some related to studying (motivation to learn, self-regulated learning, and study resilience), others more general (soft skills, intolerance of uncertainty) with three situational academic learning outcomes (general distress, online self-regulated learning, study-related emotions), and (ii) the effect of time, intraindividual factors, online self-regulated learning, and study-related emotions on distress and achievement over the following three exam sessions. A total of 331 university students took part in the study during the first Italian nationwide lockdown (T1; March–May 2020). Of those, 121 also completed at least one follow-up (T2: August 2020; T3: September 2020; T4: February 2021). At T1, study-related dispositions and soft skills were positively associated with online self-regulated learning and study-related emotions, while study-related dispositions were also negatively associated with general distress. Intolerance of uncertainty was associated positively with general distress and negatively with study-related emotions. Longitudinal effects of T2 and T3 for intolerance of uncertainty and study-related emotions were observed for distress, while those for T4 were study-related dispositions for achievement. Nurturing intraindividual factors can help students cope with a prolonged stressful situation such as a pandemic.

Thyroid Hormone Receptor α Controls the Hind Limb Metamorphosis by Regulating Cell Proliferation and Wnt Signaling Pathways in Xenopus tropicalis.

Thyroid hormone (T3) receptors (TRs) mediate T3 effects on vertebrate development. We have studied Xenopus tropicalis metamorphosis as a model for postembryonic human development and demonstrated that TRα knockout induces precocious hind limb development. To reveal the molecular pathways regulated by TRα during limb development, we performed chromatin immunoprecipitation- and RNA-sequencing on the hind limb of premetamorphic wild type and TRα knockout tadpoles, and identified over 700 TR-bound genes upregulated by T3 treatment in wild type but not TRα knockout tadpoles. Interestingly, most of these genes were expressed at higher levels in the hind limb of premetamorphic TRα knockout tadpoles than stage-matched wild-type tadpoles, suggesting their derepression upon TRα knockout. Bioinformatic analyses revealed that these genes were highly enriched with cell cycle and Wingless/Integrated (Wnt) signaling-related genes. Furthermore, cell cycle and Wnt signaling pathways were also highly enriched among genes bound by TR in wild type but not TRα knockout hind limb. These findings suggest that direct binding of TRα to target genes related to cell cycle and Wnt pathways is important for limb development: first preventing precocious hind limb formation by repressing these pathways as unliganded TR before metamorphosis and later promoting hind limb development during metamorphosis by mediating T3 activation of these pathways.

Thyroid hormones regulate cardiac repolarization and QT-interval related gene expression in hiPSC cardiomyocytes

Prolongation of cardiac repolarization (QT interval) represents a dangerous and potentially life-threatening electrical event affecting the heart. Thyroid hormones (THs) are critical for cardiac development and heart function. However, little is known about THs influence on ventricular repolarization and controversial effects on QT prolongation are reported. Human iPSC-derived cardiomyocytes (hiPSC-CMs) and multielectrode array (MEA) systems were used to investigate the influence of 3,3′,5-triiodo-l-Thyronine (T3) and 3,3′,5,5′-tetraiodo-l-Thyronine (T4) on corrected Field Potential Duration (FPDc), the in vitro analog of QT interval, and on local extracellular Action Potential Duration (APD). Treatment with high THs doses induces a significant prolongation of both FPDc and APD, with the strongest increase reached after 24 h exposure. Preincubation with reverse T3 (rT3), a specific antagonist for nuclear TH receptor binding, significantly reduces T3 effects on FPDc, suggesting a TRs-mediated transcriptional mechanism. RNA-seq analysis showed significant deregulation in genes involved in cardiac repolarization pathways, including several QT-interval related genes. In conclusion, long-time administration of high THs doses induces FPDc prolongation in hiPSC-CMs probably through the modulation of genes linked to QT-interval regulation. These results open the way to investigate new potential diagnostic biomarkers and specific targeted therapies for cardiac repolarization dysfunctions.

Sperm associated antigen 7 is activated by T3 during Xenopus tropicalis metamorphosis via a thyroid hormone response element within the first intron.

Thyroid hormone (T3) affects many diverse physiological processes such as metabolism, organogenesis, and growth. The two highly related frog species, diploid Xenopus tropicalis and pseudo tetraploid Xenopus laevis, have been used as models for analyzing the effects of T3 during vertebrate development. T3 regulates T3-inducible gene transcription through T3 receptor (TR)-binding to T3-response elements (TREs). We have previously identified sperm associated antigen 7 (spag7) as a candidate T3 target gene that is potentially involved in adult stem cell development and/or proliferation during intestinal metamorphosis. To investigate whether T3 regulates spag7 directly at the transcriptional level via TR, we first conducted qRT-PCR to analyze its expression during natural and T3-induced metamorphosis and found that spag7 was up-regulated during natural metamorphosis in the intestine, tail, brain and hindlimb, peaking at the climax of metamorphosis in all those organs, and upon T3 treatment of premetamorphic tadpoles. Next, we demonstrated that an intronic TRE in spag7, first identified through bioinformatic analysis, could bind to TR in vitro and in vivo during metamorphosis. A dual luciferase assay utilizing a reconstituted frog oocyte transcription system showed that the TRE could mediate promoter activation by liganded TR. These results indicate that spag7 expression is directly regulated by T3 through the TRE in the first intron during metamorphosis, implicating a role for spag7 early during T3-regulated tissue remodeling and resorption.

The alleviative effect of thyroid hormone on cold stress-induced apotosis via HSP70 and mitochondrial apoptosis signal pathway in bovine Sertoli cells

Thyroid hormone was involved in gene expression and functional regulation in various signal pathways. Cold stress can increase triiodothyronine (T3) level in the blood. The aim of this study was to investigate the effect of T3 on HSP70 expression and apoptosis in Sertoli cells (SCs) under cold stress in vitro culture at 26 °C, and provide a theoretical and practical basis for improving the reproductive efficiency of bulls in cold areas. SCs were treated with different cold stress duration and different T3 concentrations for pre-screening. HSP70 inhibitor was added later, and the apoptotic rate was measured using flow cytometry. The expression of HSP70 and the main genes of mitochondrial apoptosis pathway were determined by means of real-time PCR and western-blot, respectively. The localization of HSP70 was assessed by immunofluorescence. The results showed that cold stress (26 °C, 6 h) played an inductive role in SCs apoptotic rate (P < 0.01) and the transfer of HSP70 into the nucleus. 100 nM T3 further promoted HSP70 expression and its transfer into the nucleus, which significantly inhibited the expression of vital genes (cyt-c, Caspase-9 and Caspase-3) in mitochondrial pathway (P < 0.05). Subsequently, higher survival and lower apoptotic rates of SCs (P < 0.01) were observed. When T3 and HSP70 inhibitor were added together, the expression of cyt-c, Caspase-9 and Caspase-3 were inhibited (P < 0.05), and then the declining apoptotic rate increased again (P < 0.01). In conclusion, T3 can regulate HSP70 expression and translocation to mediate mitochondrial apoptosis pathway to inhibit SCs apoptosis induced by cold stress.

The potential anti-amyloidogenic candidate, SPA1413, for Alzheimer's disease.

Recently, isoflavone derivatives have been shown to have neuroprotective effects against neurological disorders. For instance, genistein attenuated the neuroinflammation and amyloid-β accumulation in Alzheimer's disease animal models, suggesting the potential for use to prevent and treat Alzheimer's disease. Here, 50 compounds, including isoflavone derivatives, were constructed and screened for the inhibitory effects on amyloid-β42 fibrilization and oligomerization using the high-throughput screening formats of thioflavin T assay and multimer detection system, respectively. The potential neuroprotective effect of t3-(4-hydroxyphenyl)-2H-chromen-7-ol (SPA1413), also known as dehydroequol, idronoxil or phenoxodiol, was evaluated in cells and in 5xFAD (B6SJL) transgenic mouse, a model of Alzheimer's disease. SPA1413 had a potent inhibitory action on both amyloid-β fibrilization and oligomerization. In the cellular assay, SPA1413 prevented amyloid-β-induced cytotoxicity and reduced neuroinflammation. Remarkably, the oral administration of SPA1413 ameliorated cognitive impairment, decreased amyloid-β plaques and activated microglia in the brain of 5xFAD (B6SJL) transgenic mouse. Our results strongly support the repurposing of SPA1413, which has already received fast-track status from the US Food and Drug Administration (FDA) for cancer treatment, for the treatment of Alzheimer's disease due to its potent anti-amyloidogenic and anti-neuroinflammatory actions.

The Circadian Oscillator of the Cerebellum: Triiodothyronine Regulates Clock Gene Expression in Granule Cells in vitro and in the Cerebellum of Neonatal Rats in vivo.

The central circadian clock resides in the suprachiasmatic nucleus (SCN) of the hypothalamus, but an SCN-dependent molecular circadian oscillator is present in the cerebellar cortex. Recent findings suggest that circadian release of corticosterone is capable of driving the circadian oscillator of the rat cerebellum. To determine if additional neuroendocrine signals act to shape cerebellar clock gene expression, we here tested the role of the thyroid hormone triiodothyronine (T3) in regulation of the cerebellar circadian oscillator. In cultured cerebellar granule cells from mixed-gender neonatal rats, T3 treatment affected transcript levels of the clock genes Per2, Arntl, Nr1d1, and Dbp, suggesting that T3 acts directly on granule cells to control the circadian oscillator. We then used two different in vivo protocols to test the role of T3 in adult female rats: Firstly, a single injection of T3 did not influence clock gene expression in the cerebellum. Secondly, we established a surgical rat model combining SCN lesion with a programmable micropump infusing circadian physiological levels of T3; however, rhythmic infusion of T3 did not reestablish differential clock gene expression between day and night in SCN lesioned rats. To test if the effects of T3 observed in vitro were related to the developmental stage, acute injections of T3 were performed in mixed-gender neonatal rats in vivo; this procedure significantly affected cerebellar expression of the clock genes Per1, Per2, Nr1d1, and Dbp. Developmental comparisons showed rhythmic expression of all clock genes analyzed in the cerebellum of adult rats only, whereas T3 responsiveness was limited to neonatal animals. Thus, T3 shapes cerebellar clock gene profiles in early postnatal stages, but it does not represent a systemic circadian regulatory mechanism linking the SCN to the cerebellum throughout life.

Deiodinase variation and liothyronine treatment interaction to accelerate ECT response in major depression: Pilot data and implications for thyroid pharmacogenomic testing in mood disorders

BackgroundConcurrent liothyronine treatment has been shown to accelerate time to response in patients treated for major depression. However, the relationship between genetic variation in deiodinase and T3 treatment response has received little investigation. MethodsThis is a post-hoc analysis of a randomized, double-blind, placebo-controlled trial of concurrent liothyronine (T3-Cytomel®, n = 23; mean dose 46.3 ± 3.3 mcg) in patients with major depression where a clinical recommendation was made to pursue ECT. The primary outcome measure was time to 50% reduction in depressive symptoms. Multivariable linear models were used to examine effect of T3 and placebo and genetic variants as predictors of change in HAMD24 and number of ECT treatments. ResultsSurvival analysis did not show a significant effect for the T3 + ECT group achieving a faster time to response compared to placebo + ECT (p + ECT). Survival analysis for RUL lead placement alone showed a significant effect for the T3 + ECT group achieving a faster time to response compared to p + ECT (p = 0.01) with significantly fewer ECT treatments (p = 0.03). Carriers of the functional SNP rs11206244 gene DI01-C785T minor allele (TT and CT) vs. non-carriers (CC), showed fewer ECT treatments in the T3 + ECT group, compared to p + ECT (5.3 ± 1.0 vs. 8.3 ± 2.1, p = 0.045). ConclusionsThese pilot data identify clinically relevant genetic variation in DI01- C785T contributing to an accelerated response time, but not an overall greater symptom reduction. Exogenous T3 supplementation may be therapeutic by attenuating a baseline DI01 relative functional deficit which functionally reduces peripheral conversion of T4 to biologically active T3. Further work is encouraged to assess genetic variation of thyroid deiodinase enzymes in larger samples of depressed patients to better optimize accelerating strategies to ECT and more broadly, thyroid augmentation strategies in major depressive and bipolar disorders.

Effect of insect feces (Hermetia illucens) on rice growth and heavy metal migration from polluted soil to rice plant.

Insect feces are a new kind of biological organic fertilizer. Little is known about the influences of insect feces on rice growth and heavy metal migration from soil to rice plant. In this study, the effects of different amounts (CK (0%), T1 (2%), T2 (4%), T3 (6%), and T4 (8%)) of black soldier fly larvae (BSFL) feces on the rice growth and the migration/accumulation of heavy metals (Cd and Pb) were investigated by pot experiments within 2years. The application of insect feces remarkably increased the contents of soil pH, organic matter, ammonium nitrogen, available phosphorus, and potassium. Meanwhile, the insect feces application reduced the weak acid-soluble contents of soil Cd and Pb by 8.3-56.8%, but increased those in the oxidizable (by 22.4-165.7%) and residual (by 1.8-225.6%) states. Except for the T4 treatment in the first year, all fertilization treatments increased the rice yield (up to 43.7% and 195.5% higher than those of CK within 2years). Moreover, the insect feces application reduced the contents of Cd (8.3-66.7%) and Pb (6.4-61.8%) in different parts of rice. Under the same treatment, the metal contents in each part of rice in the second year were lower than those in the first year. The insect feces application decreased the absorption coefficients (24.4-57.5%) and secondary transport coefficients (3.6-44.1%) of Cd and Pb by rice plant. The findings implied that the insect feces might act as effective organic fertilizers for rice plants as well as reducing heavy metal accumulation in rice plants growing in polluted soil.

Smoothelin-Like Protein 1 Regulates Development and Metabolic Transformation of Skeletal Muscle in Hyperthyroidism.

Hyperthyroidism triggers a glycolytic shift in skeletal muscle (SKM) by altering the expression of metabolic proteins, which is often accompanied by peripheral insulin resistance. Our previous results show that smoothelin-like protein 1 (SMTNL1), a transcriptional co-regulator, promotes insulin sensitivity in SKM. Our aim was to elucidate the role of SMTNL1 in SKM under physiological and pathological 3,3′,5-Triiodo-L-thyronine (T3) concentrations. Human hyper- and euthyroid SKM biopsies were used for microarray analysis and proteome profiler arrays. Expression of genes related to energy production, nucleic acid- and lipid metabolism was changed significantly in hyperthyroid samples. The phosphorylation levels and activity of AMPKα2 and JNK were increased by 15% and 23%, respectively, in the hyperthyroid samples compared to control. Moreover, SMTNL1 expression showed a 6-fold decrease in the hyperthyroid samples and in T3-treated C2C12 cells. Physiological and supraphysiological concentrations of T3 were applied on differentiated C2C12 cells upon SMTNL1 overexpression to assess the activity and expression level of the elements of thyroid hormone signaling, insulin signaling and glucose metabolism. Our results demonstrate that SMTNL1 selectively regulated TRα expression. Overexpression of SMTNL1 induced insulin sensitivity through the inhibition of JNK activity by 40% and hampered the non-genomic effects of T3 by decreasing the activity of ERK1/2 through PKCδ. SMTNL1 overexpression reduced IRS1 Ser307 and Ser612 phosphorylation by 52% and 53%, respectively, in hyperthyroid model to restore the normal responsiveness of glucose transport to insulin. SMTNL1 regulated glucose phosphorylation and balances glycolysis and glycogen synthesis via the downregulation of hexokinase II by 1.3-fold. Additionally, mitochondrial respiration and glycolysis were measured by SeaHorse analysis to determine cellular metabolic function/phenotype of our model system in real-time. T3 overload strongly increased the rate of acidification and a shift to glycolysis, while SMTNL1 overexpression antagonizes the T3 effects. These lines of evidence suggest that SMTNL1 potentially prevents hyperthyroidism-induced changes in SKM, and it holds great promise as a novel therapeutic target in insulin resistance.