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Objective: Although controversial, there may be a positive correlation between the body mass index (BMI) of individuals with obesity in euthyroidism and serum levels of thyroid stimulating hormone (TSH). This study aimed to evaluate the correlation between BMI and serum levels of TSH in individuals with morbid obesity undergoing bariatric surgery. Patients and methods: The medical records of patients treated between the years 2012 and 2016 were used. A total of 96 patients with obesity, pre-surgery BMI ≥ 40 kg/m2, being followed up in the endocrinology unit, with mean age of 50 years, were evaluated pre and post operatively. In addition to the plasma TSH dosage by IRMA and plasma free T4 by RIE, age, BMI and biochemical parameters (glycaemia, total cholesterol and triglycerides) were analyzed. Results: Patients with BMI > 40 kg/m2 prior to surgery showed higher serum TSH than subjects with a BMI Conclusion: Bariatric surgery was efficient in controlling obesity, since 100% of the patients had their degree of obesity decreased with concomitant metabolic improvement. We found that BMI and TSH are positively related, as post-surgical patients had both BMI and TSH decreased.

The coronavirus disease 2019 (COVID-19), caused by the novel coronavirus, SARS-CoV-2, affects tissues from different body systems but mostly the respiratory system, and the damage evoked in the lungs may occasionally result in severe respiratory complications and eventually lead to death. Studies of human respiratory infections have been limited by the scarcity of functional models that mimic in vivo physiology and pathophysiology. In the last decades, organoid models have emerged as potential research tools due to the possibility of reproducing in vivo tissue in culture. Despite being studied for over one year, there is still no effective treatment against COVID-19, and investigations using pulmonary tissue and possible therapeutics are still very limited. Thus, human lung organoids can provide robust support to simulate SARS-CoV-2 infection and replication and aid in a better understanding of their effects in human tissue. The present review describes methodological aspects of different protocols to develop airway and alveoli organoids, which have a promising perspective to further investigate COVID-19.

Background: Thyroid hormones play a significant role in bone development and maintenance, with triiodothyronine (T3) particularly being an important modulator of osteoblast differentiation, proliferation, and maintenance. However, details of the biological processes (BPs) and molecular pathways affected by T3 in osteoblasts remain unclear.Methods: To address this issue, primary cultures of human adipose-derived mesenchymal stem cells were subjected to our previously established osteoinduction protocol, and the resultant osteoblast-like cells were treated with 1 nm or 10 nm T3 for 72 h. RNA sequencing (RNA-Seq) was performed using the Illumina platform, and differentially expressed genes (DEGs) were identified from the raw data using Kallisto and DESeq2. Enrichment analysis of DEGs was performed against the Gene Ontology Consortium database for BP terms using the R package clusterProfiler and protein network analysis by STRING.Results: Approximately 16,300 genes were analyzed by RNA-Seq, with 343 DEGs regulated in the 1 nm T3 group and 467 upregulated in the 10 nm T3 group. Several independent BP terms related to bone metabolism were significantly enriched, with a number of genes shared among them (FGFR2, WNT5A, WNT3, ROR2, VEGFA, FBLN1, S1PR1, PRKCZ, TGFB3, and OSR1 for 1nM T3; and FZD1, SMAD6, NOG, NEO1, and ENG for 10 nm T3). An osteoblast-related search in the literature regarding this set of genes suggests that both T3 doses are unfavorable for osteoblast development, mainly hindering BMP and canonical and non-canonical WNT signaling.Conclusions: Therefore, this study provides new directions toward the elucidation of the mechanisms of T3 action on osteoblast metabolism, with potential future implications for the treatment of endocrine-related bone pathologies.

Made available in DSpace on 2020-12-12T02:32:13Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-03-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Adiponectin and leptin, important for metabolic regulation, are synthesized and secreted by adipose tissue and are influenced by triiodothyronine (T3) that activates the MAPK/ERK and integrin αVβ3 pathways, modulating gene expression. Adipocytes were treated with T3 (10 nM), for 1 h, in the absence or presence of PD98059 (PD) and tetraiodothyroacetic acid (Tetrac), which are pathways inhibitors. The cells were incubated with Adipo Red/Oil Red O reagents, and intracellular lipid accumulation [glycerol and triacylglycerol (TAG)], MTT, 8-hydroxideoxyguanosine (8-OH-dG), and mRNA and protein expression were assessed. T3 increased leptin mRNA and protein expression, and, in contrast, there was a decrease in the Tetrac + T3 group. Adiponectin mRNA expression was not altered by T3, though it had increased its protein expression, which was terminated by inhibitors PD + T3 and Tetrac + T3. However, T3 did not alter PPARγ protein expression, lipid accumulation, TAG, glycerol, and DNA damage, but PD + T3 and Tetrac + T3 reduced these parameters. T3 activated the MAPK/ERK pathway on adipocytes to modulate the adiponectin protein expression and integrin αvβ3 to alter the leptin gene expression. São Paulo State University (UNESP) Botucatu Medical School São Paulo State University (UNESP) Botucatu Medical School FAPESP: 2010/16911-4

Made available in DSpace on 2020-12-12T02:35:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-04-15 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Triiodothyronine (T3) and irisin (I) can modulate metabolic status, increase heat production, and promote differentiation of white adipose tissue (WAT) into brown adipose tissue (BAT). Herein, human subcutaneous white adipocytes were treated with 10 nM T3 or 20 nM I for 24 h to evaluate intracellular lipid accumulation, triglyceride, and glycerol levels, oxidative stress, DNA damage, and protein levels of uncoupling protein 1 (UCP1), adiponectin, leptin, peroxisome proliferator-activated receptor gamma (PPARγ), and fibronectin type III domain-containing protein 5 (FNDC5). T3 and irisin improved UCP1 production, lipid profile, oxidative stress, and DNA damage. T3 elevated adiponectin and leptin levels with a concomitant decrease in PPARy and FNDC5 levels. However, irisin did not alter adipokine, PPARy, and FNDC5 levels. The results indicate that T3 may be used to increase leptin and adiponectin levels to improve insulin sensitivity, and irisin may be used to prevent obesity or maintain weight due to its impact on the lipid profile without altering adipokine levels. Department of Internal Clinic Botucatu Medicine School São Paulo State University (UNESP) Department of Morphology Federal University of Espírito Santo Department of Internal Clinic Botucatu Medicine School São Paulo State University (UNESP) FAPESP: 2016/03242-3 CAPES: 409438/2016

Made available in DSpace on 2019-10-06T15:44:33Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-05-01. Added 1 bitstream(s) on 2019-10-09T18:34:54Z : No. of bitstreams: 1 S2359-39972019000200142.pdf: 239355 bytes, checksum: bb698bc40f032f4356658ecb11d9c169 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Objective: To verify the physiological action of triiodothyronine T3 on the expression of transforming growth factor α (TGFA) mRNA in MCF7 cells by inhibition of RNA Polymerase II and the MAPK/ERK pathway. Materials and methods: The cell line was treated with T3 at a physiological dose (10-9M) for 10 minutes, 1 and 4 hour (h) in the presence or absence of the inhibitors, α-amanitin (RNA polymerase II inhibitor) and PD98059 (MAPK/ERK pathway inhibitor). TGFA mRNA expression was analyzed by RT-PCR. For data analysis, we used ANOVA, complemented with the Tukey test and Student t-test, with a minimum significance of 5%. Results: T3 increases the expression of TGFA mRNA in MCF7 cells in 4 h of treatment. Inhibition of RNA polymerase II modulates the effect of T3 treatment on the expression of TGFA in MCF7 cells. Activation of the MAPK/ERK pathway is not required for T3 to affect the expression of TGFA mRNA. Conclusion: Treatment with a physiological concentration of T3 after RNA polymerase II inhibition altered the expression of TGFA. Inhibition of the MAPK/ERK pathway after T3 treatment does not interfere with the TGFA gene expression in a breast adenocarcinoma cell line. Departamento de Medicina Interna Faculdade de Medicina de Botucatu Universidade Estadual Paulista (UNESP) Universidade Estadual Paulista (UNESP) Departamento de Fisiologia e Biofísica Instituto de Ciencias Biomédicas Universidade de São Paulo (USP) Departamento de Medicina Interna Faculdade de Medicina de Botucatu Universidade Estadual Paulista (UNESP) Universidade Estadual Paulista (UNESP) FAPESP: 2013/05629-4 FAPESP: 2014/15209-5 FAPESP: 2015/09686-8

Made available in DSpace on 2019-10-06T16:34:57Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-08-15 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Triiodothyronine (T3) and estrogen (E2) play important roles in the bone remodeling process and signaling of receptor activator of the nuclear factor-kappa β (RANKL) and osteoprotegerin (OPG) expressed by osteoblasts. However, little is known of the molecular action of these hormones in conditions of hyperthyroidism and associated E2 in human cells. AIMS: This study evaluated the effects of the physiological concentration of E2 (10 nM), alone or in association with physiological (1 nM) and supraphysiological (10 nM) concentrations of T3, on RANKL and OPG gene expression in human osteoblasts. MAIN METHODS: Alkaline phosphatase and osteocalcin assays were performed to verify the presence of mature osteoblasts. After mimicking the experimental hyperthyroidism in osteoblasts untreated or treated with E2, RANKL and OPG gene expression was analyzed by real-time PCR and protein expression by western Blot and ELISA. Alizarin Red staining analyzed the amount of bone matrix after hormonal treatments. KEY FINDINGS: E2 enhanced the gene expression of OPG when associated with 1 nM and 10 nM T3. E2 was able to restore the bone matrix after an initial decrease using 1 nM and 10 nM T3. The protective effect of E2 on the RANKL and OPG signaling pathway was demonstrated. E2 restored the bone matrix induced by experimental hyperthyroidism. SIGNIFICANCE: The data highlight the importance of E2 to maintain OPG expression and osteoblast activity against possible loss of bone mass, especially in conditions where T3 is in excess. Department of Internal Medicine Botucatu Medical School Univ Estadual Paulista - UNESP Laboratory Biochemistry and Biophysics Butantan Institute, 1500, Avenue Vital Brazil Department of Internal Medicine Botucatu Medical School Univ Estadual Paulista - UNESP FAPESP: 2014/16406-9

Made available in DSpace on 2018-12-11T17:08:19Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-01-01. Added 1 bitstream(s) on 2019-10-09T18:30:42Z : No. of bitstreams: 1 S2359-39972016000600582.pdf: 208346 bytes, checksum: d4024e8a967f9479c5370d43b346a841 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Objective: The current study was aimed at analyzing sarcoplasmic reticulum Ca2+ ATPase (Serca2) and ryanodine receptor type 2 (Ryr2) gene expression in rats subjected to surgery that induced HF and were subsequently treated with T4 using physiological doses. Materials and methods: HF was induced in 18 male Wistar rats by clipping the ascending thoracic aorta to generate aortic stenosis (HFS group), while the control group (9-sham) underwent thoracotomy. After 21 weeks, the HFS group was subdivided into two subgroups. One group (9 Wistar rats) with HF received 1.0 µg of T4/100 g of body weight for five consecutive days (HFS/T4); the other group (9 Wistar rats) received isotonic saline solution (HFS/S). The animals were sacrificed after this treatment and examined for signs of HF. Samples from the left ventricles of these animals were analyzed by RT-qPCR for the expression of Serca2 and Ryr2 genes. Results: Rats with HF developed euthyroid sick syndrome (ESS) and treatment with T4 restored the T3 values to the Sham level and increased Serca2 and Ryr2 gene expression, thereby demonstrating a possible benefit of T4 treatment for heart function in ESS associated with HF. Conclusion: The T4 treatment can potentially normalize the levels of T3 as well elevated Serca2 and Ryr2 gene expression in the myocardium in heart failure rats with euthyroid sick syndrome. Departamento de Clínica Médica Unidade de Pesquisa Experimental (Unipex) Faculdade de Medicina de Botucatu Universidade Estadual Paulista (Unesp) Instituto de Ciências da Saúde Universidade Federal de Mato Grosso (UFMT) Departamento de Bioestatística Instituto de Biociências Universidade Estadual Paulista (Unesp) Departamento de Clínica Médica Unidade de Pesquisa Experimental (Unipex) Faculdade de Medicina de Botucatu Universidade Estadual Paulista (Unesp) Departamento de Bioestatística Instituto de Biociências Universidade Estadual Paulista (Unesp) FAPESP: 2007/55902-8

Made available in DSpace on 2018-12-11T17:02:58Z (GMT). No. of bitstreams: 0 Previous issue date: 2016-06-01 High expression levels of hypoxia inducing factor 1 alpha are related to mammary carcinogenesis. In previous studies, we demonstrated that expression of transforming growth factor alpha increases upon treatment with triiodothyronine, but this expression does not occur in cellular models that do not express the estrogen receptor, or when cells are co-treated with the anti-estrogen, tamoxifen. The aim of this study was to determine the effect of the hormone triiodothyronine on the expression of the genes HIF1A and TGFA in the breast cancer cell line MCF7. The cell line was subjected to treatment with triiodothyronine at the supraphysiological dose of 10- 8 M for 10 min, 30 min, 1 h, and 4 h in the presence or absence of actinomycin D, the gene expression inhibitor, cycloheximide, the protein synthesis inhibitor, and LY294002, the phosphoinositide 3 kinase inhibitor. HIF1A and TGFA mRNA expression was analyzed by reverse transcription polymerase chain reaction. For data analysis, we used analysis of variance complemented by Tukey test and an adopted minimum of 5% significance. We found that HIF1A and TGFA expression increased in the presence of triiodothyronine at all times studied. HIF1A expression decreased in triiodothyronine-treated cells when gene transcription was also inhibited; however, TGFA expression decreased after 10 and 30 min of treatment even when transcription was not inhibited. We found that activation of PI3K was necessary for triiodothyronine to modulate HIF1A and TGFA expression. Department of Internal Medicine Botucatu Medical School Univ Estadual Paulista - UNESP, Distrito de Rubi�o Jr s/n Department of Pathology Botucatu Medical School Univ Estadual Paulista - UNESP Department of Morphology Bioscience Institute Univ Estadual Paulista - UNESP Department of Internal Medicine Botucatu Medical School Univ Estadual Paulista - UNESP, Distrito de Rubi�o Jr s/n Department of Pathology Botucatu Medical School Univ Estadual Paulista - UNESP Department of Morphology Bioscience Institute Univ Estadual Paulista - UNESP

An impressive percentage of biomedical knowledge and advances were achieved through animal research and cell culture investigations. For drug testing and disease researches, both animal models and preclinical trials with cell cultures are extremely important, but they present some limitations, such as ethical concern and lack of representatively of human tissues and organs. Most cells are currently cultured using two-dimensional (2D) methods, but new and improved methods that implement three-dimensional (3D) cell culture techniques suggest convincing evidence that much more advanced experiments can be performed with more complex information. The environment and cell types in 3D culture can be manipulated to mimic tissue in vivo and provide more accurate data on cell-to-cell interactions; the cultivation techniques are based on a scaffold, which can be based on hydrogel or polymeric material, in addition there are techniques without using scaffold, such as suspended microplates, magnetic levitation and microplates for spheroids with ultra-low fixation coating. Even though 3D culturing is clearly incapable of replacing other current research types, they might continue to replace some unnecessary animal experimentation, as well as improve monolayer cultures. It is not even recommended or expected that 3D models substitute all other research types, but in regard to animal testing, they come in hand for the 3 Rs: Reduction, Refinement, Replacement. In this aspect, 3D culture emerges as valuable alternatives to the investigation of functional, biochemical and molecular aspects of human pathologies.

Obesity is a worldwide health problem which have reached pandemic proportions, now also including low and middle-income countries. Excessive or abnormal fat deposition in the abdomen especially in the visceral compartment is tightly associated with a high metabolic risk for arterial hypertension, type II diabetes, cardiovascular diseases, musculoskeletal disorders (especially articular degeneration) and some cancers. Contrariwise, accumulation of fat in the subcutaneous compartment has been associated with a neutral metabolic impact, favoring a lower risk of insulin resistance. Obesity results more often from an avoidable imbalance between food consumption and energy expenditure. There are several recommended strategies for dealing with obesity, including pharmacological therapies, but their success remains incomplete and may not compensate the associated adverse effects. Purinergic signaling operated by ATP and its metabolite, adenosine, has attracted increasing attention in obesity. The extracellular levels of purines often reflect the energy status of a given cell population. Adenine nucleotides and nucleosides fine tuning control adipogenesis and mature adipocytes function via the activation of P2 and P1 purinoceptors, respectively. These features make the purinergic signaling cascade a putative target for therapeutic intervention in obesity and related metabolic syndromes. There are, however, gaps in our knowledge regarding the role of purines in adipocyte precursors differentiation and mature adipocytes functions, as well as their impact among distinct adipose tissue deposits (e.g. white vs. brown, visceral vs. subcutaneous), which warrants further investigations before translation to clinical trials can be made.

Obesity patients are more susceptible to develop COVID-19 severe outcome due to the role of angiotensin-converting enzyme 2 (ACE2) in the viral infection. ACE2 is regulated in the human cells by different genes associated with increased (TLR3, HAT1, HDAC2, KDM5B, SIRT1, RAB1A, FURIN and ADAM10) or decreased (TRIB3) virus replication. RNA-seq data revealed 14857 genes expressed in human subcutaneous adipocytes, including genes mentioned above. Irisin treatment increased by 3-fold the levels of TRIB3 transcript and decreased the levels of other genes. The decrease in FURIN and ADAM10 expression enriched diverse biological processes, including extracellular structure organization. Our results indicate a positive effect of irisin on the expression of multiple genes related to viral infection in the adipose tissue; furthermore, translatable for other tissues and organs targeted by the novel coronavirus and present, thus, promising approaches for the treatment of COVID-19 infection as therapeutic strategy to decrease ACE2 regulatory genes., Highlights • Genes associated with increased virus, SARS-CoV-2, replication are diminished, whereas, the gene that decreases virus replication is elevated by irisin in human subcutaneous adipocytes. • TRIB3 is increased by irisin. • Irisin diminished FURIN and ADAM10 in human subcutaneous adipocytes.

A considerable increase in endocrine abnormalities has been reported over the last few decades worldwide. A growing exposure to endocrine-disrupting chemicals (EDCs) can be one of the causes of endocrine disorders in populations, and these disorders are not only restricted to the metabolic hormone system but can also cause abnormal functions. Thyroid hormone (TH) disruption is defined as an abnormal change in TH production, transport, function, or metabolism, which results in some degree of impairment in body homeostasis. Many EDCs, including organotin compounds (OTCs), are environmental contaminants that are commonly found in antifouling paints used on ships and in several other industrial procedures. OTCs are obesogenic and can disrupt TH metabolism; however, abnormalities in thyroid function resulting from OTC exposure are less well understood. OTCs, one of the most prevalent EDCs that are encountered on a daily basis, modulate the thyroid axis. In most toxicology studies, it has been reported that OTCs might contribute to hypothyroidism.

Adipose tissue (AT), an endocrine organ that modulates several physiological functions by synthesizing and releasing adipokines such as adiponectin, is a metabolic target of triiodothyronine (T3). T3 and adiponectin play important roles in controlling normal metabolic functions such as stimulation of fatty acid oxidation and increase in thermogenesis. The phosphatidylinositol 3-kinase (PI3K) pathway is important for the differentiation of preadipocytes into adipocytes and can be activated by T3 for the transcription of specific genes, such as adiponectin. We examined the role of PI3K in adiponectin modulation by T3 action in murine adipocytes (3T3-L1). The 3T3-L1 adipocytes were treated with 1000 nM T3 for 1 h in the presence or absence of 50 μM LY294002 (LY), a PI3K inhibitor. Then, we assessed the expression of adiponectin and the phosphorylated serine/threonine kinase Akt (pAkt), a PI3K signaling protein, in the adipocytes. Adiponectin and pAKT levels were higher in the T3-adipocyte cells, whereas in the LY group adiponectin was elevated and pAKT was decreased compared to the control (C). PI3K pathway inhibition for 1 h and posterior treatment with T3, in LY + T3, reduced the adiponectin level and increased pAKT levels compared to those in LY. T3 stimulated adiponectin levels by PI3K pathway activation and T3 can compensate alteration in the PI3K pathway, because with inhibition of the pathway it is able to maintain the basal levels of adiponectin and pAKT.

Made available in DSpace on 2018-12-11T17:21:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-06-01. Added 1 bitstream(s) on 2019-10-09T18:31:56Z : No. of bitstreams: 1 S2359-39972018000300366.pdf: 123932 bytes, checksum: 884f7b3ea5771ca0dc08ec15ac8f5036 (MD5) Made available in DSpace on 2018-12-11T17:21:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-06-01. Added 1 bitstream(s) on 2019-10-09T18:31:56Z : No. of bitstreams: 1 S2359-39972018000300366.pdf: 123932 bytes, checksum: 884f7b3ea5771ca0dc08ec15ac8f5036 (MD5) Made available in DSpace on 2018-12-11T17:21:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-06-01. Added 1 bitstream(s) on 2019-10-09T18:31:56Z : No. of bitstreams: 1 S2359-39972018000300366.pdf: 123932 bytes, checksum: 884f7b3ea5771ca0dc08ec15ac8f5036 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Objective: Graves’ ophthalmopathy (GO) is an autoimmune disease that leads to ocular proptosis caused by fat accumulation and inflammation, and the main treatment is corticosteroid therapy. Retinoid acid receptor-alpha (RARα) seems to be associated with inflammation and adipocyte differentiation. This study aimed to assess the effect of glucocorticoid treatment on orbital fibroblasts of GO patient treated or not with different glucocorticoid doses. Materials and methods: Orbital fibroblasts collected during orbital decompression of a female patient with moderately severe/severe GO were cultivated and treated with 10 nM and 100 nM dexamethasone (Dex). RARα gene expression in the treated and untreated cells was then compared. Results: Fibroblast RARα expression was not affected by 100 nM Dex. On the other hand, RARα expression was 24% lower in cells treated with 10 nM Dex (p < 0.05). Conclusions: Orbital fibroblasts from a GO patient expressed the RARα gene, which was unaffected by higher, but decreased with lower doses of glucocorticoid. Divisão de Endocrinologia Departamento de Medicina Interna Faculdade de Medicina de Botucatu Universidade Estadual de São Paulo (Unesp) Instituto Federal de Educação Ciência e Tecnologia do Estado de São Paulo (IFSP) Departamento de Oftalmologia Otorrinolaringologia e Cirurgia de Cabeça e Pescoço Faculdade de Medicina de Botucatu Universidade Estadual de São Paulo (Unesp) Divisão de Endocrinologia Departamento de Medicina Interna Faculdade de Medicina de Botucatu Universidade Estadual de São Paulo (Unesp) Departamento de Oftalmologia Otorrinolaringologia e Cirurgia de Cabeça e Pescoço Faculdade de Medicina de Botucatu Universidade Estadual de São Paulo (Unesp) FAPESP: #:03/03651-0 FAPESP: 2011/18464-8

Made available in DSpace on 2015-11-03T15:27:32Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-11-01. Added 1 bitstream(s) on 2015-11-04T10:15:38Z : No. of bitstreams: 1 S0004-27302014000800833.pdf: 270985 bytes, checksum: 17593a871f9249ed70d7043e87455a9f (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Objective: The present study aimed to examine the effects of thyroid hormone (TH), more precisely triiodothyronine (T3), on the modulation of TH receptor alpha (TR proportional to) mRNA expression and the involvement of the phosphatidyl inositol 3 kinase (PI3K) signaling pathway in adipocytes, 3T3-L1, cell culture. Materials and methods: It was examined the involvement of PI3K pathway in mediating T3 effects by treating 3T3-L1 adipocytes with physiological (P = 10nM) or supraphysiological (SI = 100 nM) T3 doses during one hour (short time), in the absence or the presence of PI3K inhibitor (LY294002). The absence of any treatment was considered the control group (C). RT-qPCR was used for mRNA expression analyzes. For data analyzes ANOVA complemented with Tukey's test was used at 5% significance level. Results: T3 increased TR proportional to mRNA expression in P (1.91 +/- 0.13, p < 0.001), SI (2.14 +/- 0.44, p < 0.001) compared to C group (1 +/- 0.08). This increase was completely abrogated by LY294002 in P (0.53 +/- 0.03, p < 0.001) and SI (0.31 +/- 0.03, p < 0.001). To examine whether TRa is directly induced by T3, we used the translation inhibitor cycloheximide (CHX). The presence of CHX completely abrogated levels TRa mRNA in P (1.15 +/- 0.05, p > 0.001) and SI (0.99 +/- 0.15, p > 0.001), induced by T3. Conclusion: These results demonstrate that the activation of the PI3K signaling pathway has a role in T3-mediated indirect TRa gene expression in 3T3-L1 adipocytes. Institute of Health Sciences, Universidade Federal do Mato Grosso (UFMT), Sinop, MT, Brazil Department of Internal Medicine, Botucatu School of Medicine, University of São Paulo State (Unesp), Botucatu, SP, Brazil FAPESP: 2010/16911-4

Made available in DSpace on 2014-12-03T13:10:29Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-01-01Bitstream added on 2014-12-03T13:23:11Z : No. of bitstreams: 1 WOS000330657200001.pdf: 1661891 bytes, checksum: 9392d9393b4792008534c4842bd77bd7 (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) It has been well established that estrogen plays an important role in the progression and treatment of breast cancer. However, the role of triiodothyronine (T-3) remains controversial. We have previously shown its capacity to stimulate the development of positive estrogen receptor breast carcinoma, induce the expression of genes (PR, TGF-alpha) normally stimulated by estradiol (E-2), and suppress genes (TGF-beta) normally inhibited by E-2. Since T-3 regulates growth hormones, metabolism, and differentiation, it is important to verify its action on other genes normally induced by E-2. Therefore, we used DNA microarrays to compare gene expression patterns in MCF-7 breast adenocarcinoma cells treated with E-2 and T-3. Several genes were modulated by both E-2 and T-3 in MCF-7 cells (Student's t-test, P < 0.05). Specifically, we found eight genes that were differentially expressed after treatment with both E-2 and T-3, including amphiregulin, fibulin 1, claudin 6, pericentriolar material 1, premature ovarian failure 1B, factor for adipocyte differentiation-104, sterile alpha motif domain containing 9, and likely ortholog of rat vacuole membrane protein 1 (fold change > 2.0, pFDR < 0.05). We confirmed our microarray results by real-time PCR. Our findings reveal that certain genes in MCF-7 cells can be regulated by both E-2 and T-3. Univ Sao Paulo State UNESP, Botucatu Sch Med, Dept Internal Med, BR-18618000 Botucatu, SP, Brazil Univ Sao Paulo, Sch Med, Dept Radiol, BR-01246903 Sao Paulo, Brazil AC Camargo Hosp, Res Ctr, BR-01509900 Sao Paulo, Brazil Univ Sao Paulo State, Dept Clin Med, BR-18618000 Botucatu, SP, Brazil Univ Sao Paulo State UNESP, Botucatu Sch Med, Dept Internal Med, BR-18618000 Botucatu, SP, Brazil FAPESP: 02/09798-0

OBJETIVO: Verificar o perfil dos hormônios tireóideos (HTs) em pacientes pós-menopausa portadoras de carcinoma de mama (CaM). SUJEITOS E MÉTODOS: Participaram 12 pacientes com CaM em estádio I ou II sem intervenções que pudessem interferir na progressão tumoral e um grupo controle com 18 pacientes em pós-menopausa sem CaM. Foram dosados os níveis séricos de anticorpo antitiroperoxidase (TPOAB), hormônio estimulante da tireoide (TSH), tiroxina livre (T4L), estradiol (E2), hormônio folículo estimulante (FSH) e hormônio luteinizante (LH) antes e após a cirurgia, e realizada a imunoistoquímica dos receptores de estrógeno (ER) e progesterona (PR). RESULTADOS: Quatro pacientes com CaM apresentaram alterações do perfil hormonal tireoidiano: dois hipertireoidismo, um hipotireoidismo e positividade TPO-AB, todas com ER e PR positivos. Os níveis de TSH dessas pacientes não foram diferentes dos níveis encontrados no grupo controle (1,89 ± 1,56 vs. 2,86 ± 3,12 mUI/mL), porém os níveis de T4L nas pacientes com CaM foram estatisticamente maiores que o controle (1,83 ± 0,57 vs. 1,10 ± 0,20 ng/dL). CONCLUSÃO: Esses resultados reforçam a necessidade de avaliação do status tireoidiano em pacientes com CaM, uma vez que, na ausência de E2, mudanças clínicas nos HTs podem atuar em vias controladas pelo E2. OBJECTIVE: The aim of this study was to determine thyroid hormone (TH) profile in postmenopausal patients with breast cancer (BC). SUBJECTS AND METHODS: 12 CaM patients stages I or II, without interventions that could interfere with tumor progression were selected, as well as and a control group with 18 postmenopausal women without CaM. We measured serum anti-thyroperoxidase antibody (TPOAB), thyroid-stimulating hormone (TSH), free thyroxine (T4L), estradiol (E2), follicle-stimulating hormone (FSH), and luteinizing hormone (LH), before and after surgery, besides immunohistochemistry for estrogen (ER) and progesterone (PR) receptors. RESULTS: Four patients with CaM showed changes in thyroid hormone profile: two had hyperthyroidism, one hypothyroidism, and one was positive for TPO-AB. All of them positive for ER and PR. TSH levels in breast cancer patients were not different from levels found in the control group (1.89 ± 1.56 vs. 2.86 ± 3.12 mIU/mL), but the levels of T4L in patients with CaM were statistically higher than those of the control group (1.83 ± 0.57 vs. 1.10 ± 0.20 ng/dL). CONCLUSION: These results reinforce the need for assessment of thyroid status in CaM patients, since in the absence of E2, changes in clinical HTs can act in E2-controlled processes.

Thyroid hormones regulate energy balance and act on adipokines. However, while it is unclear what the effects are of calorie restriction and high doses of triiodothyronine (T 3 ) on adipokines in obesity, thyroid hormones are illicitly administered in isolation or in association with a hypocaloric diet as an obesity treatment. The present study determined the effect of T 3 on serum concentrations and gene expression of the adipokines leptin, resistin, and adiponectin in calorie-restricted obese rats. Male Wistar rats received a hypercaloric diet for 20 weeks followed by calorie restriction for 8 weeks. The animals were then randomly divided into 3 groups: calorie restriction (OR), OR with 5 μg of T 3 /100 g BW (RS1), and OR with 25 μg of T 3 /100 g BW (RS2) for 2 weeks. Blood and adipose tissue samples were collected for biochemical, hormonal, and gene expression analyses. Serum concentrations of leptin (OR: 3.7±0.6, RS1: 3.8±1, RS2 0.2±0.07 ng/dl) and resistin (OR: 2.5±0.6, RS1: 2.5±0.5, RS2 1.6±0.3 ng/dl) were diminished at the higher dose, while serum adiponectin (OR: 31±7, RS1: 24±5, RS2 26±7 ng/dl) levels were lower in the low dose group. Administration of T 3 reduced leptin gene expression (OR: 0.91±0.1, RS1: 0.95±0.1, RS2 0.22±0.1) only at the higher dose, resistin expression (OR: 1.06±0.2, RS1: 1.04±0.1, RS2 0.88±0.2) was not influenced by T 3 treatment, and adiponectin expression (OR: 1.55±0.5, RS1: 0.95±0.15, RS2 0.97±0.13) was diminished independent of the T 3 dose. These results indicate that T 3 , directly or indirectly, inhibits the expression of leptin and adiponectin in calorie restricted obese animals.

Obesity has become a major public health problem, most commonly treated via dietary restriction to promote weight loss. Although leptin and thyroid hormones are involved in the regulation of energy balance, the role of these hormones after the stabilization of weight loss remains unclear. This study was designed to analyze the effect of thyroid hormone on sustained weight loss and leptin gene expression in obese animals after a loss of 5% to 10% of body weight. Thirty-day–old male Wistar rats were separated into 4 groups: control, obese, calorie restriction (CR), and calorie restriction with triiodothyronine administration (CRT). The obese group had increased weight and adiposity, leptin and insulin levels, and leptin gene expression. Dietary restriction in the CR group resulted in decreased body weight and adiposity, diminished leptin, and increased thyroid hormone receptor β expression. The CRT group, submitted to dietary restriction with concomitant administration of a physiologic triiodothyronine dose, had thyroid hormone receptor β expression at levels comparable with those observed in the control group and simultaneously increased leptin expression as compared with that in the CR group, suggesting that thyroid hormone modulates leptin expression under conditions of calorie restriction. Increased leptin expression in the CRT group did not result in increased circulating leptin or a statistically significant reduction in body weight during the treatment period. These data provide impetus for further study, as a longer treatment period may result in increased circulating leptin and, thus, further reduction in body weight during calorie restriction in an obesity model.

Made available in DSpace on 2015-02-02T12:39:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-10-01Bitstream added on 2015-02-02T13:07:53Z : No. of bitstreams: 1 S0004-27492014000500339.pdf: 579808 bytes, checksum: e8fd687573ee4954ed41efeed3136c7f (MD5) Made available in DSpace on 2015-02-02T12:39:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-10-01Bitstream added on 2015-02-02T13:07:53Z : No. of bitstreams: 1 S0004-27492014000500339.pdf: 579808 bytes, checksum: e8fd687573ee4954ed41efeed3136c7f (MD5) Made available in DSpace on 2015-02-02T12:39:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-10-01Bitstream added on 2015-02-02T13:07:53Z : No. of bitstreams: 1 S0004-27492014000500339.pdf: 579808 bytes, checksum: e8fd687573ee4954ed41efeed3136c7f (MD5) Made available in DSpace on 2015-02-02T12:39:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-10-01Bitstream added on 2015-02-02T13:07:53Z : No. of bitstreams: 1 S0004-27492014000500339.pdf: 579808 bytes, checksum: e8fd687573ee4954ed41efeed3136c7f (MD5) Made available in DSpace on 2015-02-02T12:39:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-10-01Bitstream added on 2015-02-02T13:07:53Z : No. of bitstreams: 1 S0004-27492014000500339.pdf: 579808 bytes, checksum: e8fd687573ee4954ed41efeed3136c7f (MD5) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Universidade Estadual Paulista Botucatu Medical School Internal Medicine Department Universidade Estadual Paulista Botucatu Medical School Ophthalmology, Otorhinolaryngology and Head and Neck Department Universidade Estadual Paulista Botucatu Medical School Internal Medicine Department Universidade Estadual Paulista Botucatu Medical School Ophthalmology, Otorhinolaryngology and Head and Neck Department

Made available in DSpace on 2015-08-26T19:19:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-06-01. Added 1 bitstream(s) on 2015-08-31T13:05:04Z : No. of bitstreams: 1 S2359-39972015000300273.pdf: 130120 bytes, checksum: 6dc2ae5dfe61f973ea9aecc7248da395 (MD5) Made available in DSpace on 2015-08-26T19:19:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-06-01. Added 1 bitstream(s) on 2015-08-31T13:05:04Z : No. of bitstreams: 1 S2359-39972015000300273.pdf: 130120 bytes, checksum: 6dc2ae5dfe61f973ea9aecc7248da395 (MD5) Made available in DSpace on 2015-08-26T19:19:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-06-01. Added 1 bitstream(s) on 2015-08-31T13:05:04Z : No. of bitstreams: 1 S2359-39972015000300273.pdf: 130120 bytes, checksum: 6dc2ae5dfe61f973ea9aecc7248da395 (MD5) Made available in DSpace on 2015-08-26T19:19:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-06-01. Added 1 bitstream(s) on 2015-08-31T13:05:04Z : No. of bitstreams: 1 S2359-39972015000300273.pdf: 130120 bytes, checksum: 6dc2ae5dfe61f973ea9aecc7248da395 (MD5) Made available in DSpace on 2015-08-26T19:19:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-06-01. Added 1 bitstream(s) on 2015-08-31T13:05:04Z : No. of bitstreams: 1 S2359-39972015000300273.pdf: 130120 bytes, checksum: 6dc2ae5dfe61f973ea9aecc7248da395 (MD5) Graves’ ophthalmopathy (GO) is one of the most severe clinical manifestations of Graves’ disease (GD), and its treatment might involve high-dose glucocorticoid therapy. The higher incidence of GO among females, and the reported association between polymorphisms of estrogen receptor (ER) and GD susceptibility have led us to question the role of estrogen and its receptor in GO pathogenesis. We, thus, assessed estrogen receptor-alpha (ERA) gene expression in cultures of orbital fibroblasts from a patient with GO before (controls) and after treatment with 10 nM and 100 nM dexamethasone (DEX). Orbital fibroblasts showed ERA gene expression. In the cells treated with 10 nM and 100 nM DEX, ERA gene expression was, respectively, 85% higher and 74% lower, than in the control group. We concluded that ERA gene expression is found in the orbital fibroblasts of patient with GO, which may be affected by glucocorticoids in a dose-related manner. Arch Endocrinol Metab. 2015;59(3):273-6 Sao Paulo State University Botucatu Medical School Internal Medicine Department Unesp Sao Paulo State University Botucatu Medical School Internal Medicine Department Unesp

Objective To study the effect of different doses of triiodothyronine on gene expression of the adipokines leptin and adiponectin, at different times, and to evaluate the difference in expression between the two adipokines in each group. Methods 3T3-L1 adipocytes were incubated with triiodothyronine at physiological dose (10nM) and supraphysiological doses (100nM or 1,000nM), or without triiodothyronine (control, C) for 0.5, 6, or 24 hours. Leptin and adiponectin mRNA was detected using real-time polymerase chain reaction (RT-PCR). One-way analyses of variance, Tukey’s test or Student’s t test, were used to analyze data, and significance level was set at 5%. Results Leptin levels decreased in the 1,000nM-dose group after 0.5 hour. Adiponectin levels dropped in the 10nM-dose group, but increased at the 100nM dose. After 6 hours, both genes were suppressed in all hormone concentrations. After 24 hours, leptin levels increased at 10, 100 and 1,000nM groups as compared to the control group; and adiponectin levels increased only in the 100nM group as compared to the control group. Conclusion These results demonstrated fast actions of triiodothyronine on the leptin and adiponectin expression, starting at 0.5 hour, at a dose of 1,000nM for leptin and 100nM for adiponectin. Triiodothyronine stimulated or inhibited the expression of adipokines in adipocytes at different times and doses which may be useful to assist in the treatment of obesity, assuming that leptin is increased and adiponectin is decreased, in obesity cases. Objetivo Examinar o efeito de diferentes doses de triiodotironina sobre a expressão gênica das adipocinas leptina e adiponectina, em diferentes períodos de tempo, além de avaliar a diferença de expressão entre as duas adipocinas em cada grupo. Métodos Adipócitos 3T3-L1 foram incubados com triiodotironina nas doses fisiológica (10nM) e suprafisiológicas (100nM ou 1.000nM), ou na ausência de triiodotironina (controle, C) durante 0,5, 6 ou 24 horas. O mRNA das adipocinas foi analisado em tempo real, utilizando a reação em cadeia de polimerase. Para as análises dos dados, foi utilizada a análise de variância, complementada com o teste de Tukey, ou o teste t de Student com 5% de significância. Resultados Os níveis de leptina diminuíram no grupo com dose de 1.000nM em 0,5 hora. A adiponectina também diminuiu no grupo com dose de 10nM, porém se elevou com a dose de 100nM. Após 6 horas, ambos os genes foram suprimidos em todas concentrações de hormônio. Em 24 horas, os níveis de leptina foram elevados em 10, 100 e 1.000nM, em relação ao grupo controle. No que concerne à adiponectina, observou-se aumento apenas no grupo cuja dose foi de 100nM, em comparação ao controle. Conclusão Foram demonstradas ações rápidas da triiodotironina sobre a expressão da leptina e da adiponectina, iniciando em 0,5 hora na dose de 1.000nM, para a primeira, e na dose de 100nM, para a segunda. A triiodotironina estimulou ou inibiu a expressão de adipocinas em adipócitos em diferentes tempos e doses, o que pode auxiliar no tratamento da obesidade, levando em consideração que, nesta, a leptina está aumentada e adiponectina, diminuída.

The thyroid hormones (THs), triiodothyronine (T3) and thyroxine (T4), are essential for survival; they are involved in the processes of development, growth, and metabolism. In addition to hyperthyroidism or hypothyroidism, THs are involved in other diseases. The role of THs in the development and differentiation of mammary epithelium is well established; however, their specific role in the pathogenesis of breast cancer (BC) is controversial. Steroid hormones affect many human cancers and the abnormal responsiveness of the mammary epithelial cells to estradiol (E2) in particular is known to be an important cause for the development and progression of BC. The proliferative effect of T3 has been demonstrated in various types of cancer. In BC cell lines, T3 may foster the conditions for tumor proliferation and increase the effect of cell proliferation by E2; thus, T3 may play a role in the development and progression of BC. Studies show that T3 has effects similar to E2 in BC cell lines. Despite controversy regarding the relationship between thyroid disturbances and the incidence of BC, studies show that thyroid status may influence the development of tumor, proliferation and metastasis.

Made available in DSpace on 2015-12-07T15:30:30Z (GMT). No. of bitstreams: 0 Previous issue date: 2014. Added 1 bitstream(s) on 2015-12-07T15:52:57Z : No. of bitstreams: 1 PMC3950583.pdf: 2375661 bytes, checksum: fa0182a0b8ea0d24d60647be62a0a9d1 (MD5) We investigated thyroid hormone levels in menopausal BrC patients and verified the action of triiodothyronine on genes regulated by estrogen and by triiodothyronine itself in BrC tissues. We selected 15 postmenopausal BrC patients and a control group of 18 postmenopausal women without BrC. We measured serum TPO-AB, TSH, FT4, and estradiol, before and after surgery, and used immunohistochemistry to examine estrogen and progesterone receptors. BrC primary tissue cultures received the following treatments: ethanol, triiodothyronine, triiodothyronine plus 4-hydroxytamoxifen, 4-hydroxytamoxifen, estrogen, or estrogen plus 4-hydroxytamoxifen. Genes regulated by estrogen (TGFA, TGFB1, and PGR) and by triiodothyronine (TNFRSF9, BMP-6, and THRA) in vitro were evaluated. TSH levels in BrC patients did not differ from those of the control group (1.34 ± 0.60 versus 2.41 ± 1.10 μ U/mL), but FT4 levels of BrC patients were statistically higher than controls (1.78 ± 0.20 versus 0.95 ± 0.16 ng/dL). TGFA was upregulated and downregulated after estrogen and triiodothyronine treatment, respectively. Triiodothyronine increased PGR expression; however 4-hydroxytamoxifen did not block triiodothyronine action on PGR expression. 4-Hydroxytamoxifen, alone or associated with triiodothyronine, modulated gene expression of TNFRSF9, BMP-6, and THRA, similar to triiodothyronine treatment. Thus, our work highlights the importance of thyroid hormone status evaluation and its ability to interfere with estrogen target gene expression in BrC samples in menopausal women. Department of Biological Science, São Paulo Federal Institute (IFSP), 18136-540 São Roque, SP, Brazil ; Department of Internal Medicine, Division of Endocrinology and Metabolism, UNESP, 18618-000 Botucatu, SP, Brazil. Department of Internal Medicine, Division of Endocrinology and Metabolism, UNESP, 18618-000 Botucatu, SP, Brazil. Department of Biological Science, São Paulo Federal Institute (IFSP), 18136-540 São Roque, SP, Brazil ; Department of Internal Medicine, Division of Endocrinology and Metabolism, UNESP, 18618-000 Botucatu, SP, Brazil. Department of Internal Medicine, Division of Endocrinology and Metabolism, UNESP, 18618-000 Botucatu, SP, Brazil.

Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:30:42Z No. of bitstreams: 0Bitstream added on 2014-05-27T14:38:01Z : No. of bitstreams: 1 2-s2.0-84884264499.pdf: 942276 bytes, checksum: 17045a1e8ee4d958b7e3d559fc56068d (MD5) Made available in DSpace on 2014-05-27T11:30:42Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-09-18 The present study aimed to examine the effects of thyroid hormone (TH), more precisely triiodothyronine (T3), on the modulation of leptin mRNA expression and the involvement of the phosphatidyl inositol 3 kinase (PI3K) signaling pathway in adipocytes, 3T3-L1, cell culture. We examined the involvement of this pathway in mediating TH effects by treating 3T3-L1 adipocytes with physiological (P=10nM) or supraphysiological (SI=100 nM) T3 dose during one hour (short time), in the absence or the presence of PI3K inhibitor (LY294002). The absence of any treatment was considered the control group (C). RT-qPCR was used for mRNA expression analyzes. For data analyzes ANOVA complemented with Tukey's test was used at 5% significance. T3 increased leptin mRNA expression in P (2.26 ± 0.36, p< 0.001), SI (1.99 ±0.22, p< 0.01) compared to C group (1± 0.18). This increase was completely abrogated by LY294002 in P (1.31±0.05, p< 0.001) and SI (1.33±0.31, p< 0.05). Western blotting confirmed these results at protein level, indicating the PI3K pathway dependency. To examine whether leptin is directly induced by T3, we used the translation inhibitor cycloheximide (CHX). In P, the presence of CHX maintained the levels mRNA leptin, but was completely abrogated in SI (1.14±0.09, p> 0.001). These results demonstrate that the activation of the PI3K signaling pathway has a role in TH-mediated direct and indirect leptin gene expression in 3T3-L1 adipocytes. © 2013 Oliveira et al. Department of Internal Medicine Botucatu School of Medicine University of São Paulo State (UNESP), Botucatu, São Paulo Department of Physiology Federal University of São Paulo (UNIFESP), São Paulo Department of Internal Medicine Botucatu School of Medicine University of São Paulo State (UNESP), Botucatu, São Paulo

This study aimed evaluate whether estrogen modulates amphiregulin (AREG) gene expression in breast tumor cells. For this, MCF-7 cells were treated for 1 or 4 hours with estrogen (E2 10−7 M) in the ...

Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:28:33Z No. of bitstreams: 0Bitstream added on 2014-05-27T14:32:18Z : No. of bitstreams: 1 2-s2.0-84874511518.pdf: 447094 bytes, checksum: b79744f190efab28202a9a9e0a1d266c (MD5) Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:28:33Z No. of bitstreams: 0Bitstream added on 2014-05-27T14:32:18Z : No. of bitstreams: 1 2-s2.0-84874511518.pdf: 447094 bytes, checksum: b79744f190efab28202a9a9e0a1d266c (MD5) Made available in DSpace on 2014-05-27T11:28:33Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-02-28 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:28:33Z No. of bitstreams: 0Bitstream added on 2014-05-27T14:32:18Z : No. of bitstreams: 1 2-s2.0-84874511518.pdf: 447094 bytes, checksum: b79744f190efab28202a9a9e0a1d266c (MD5) Made available in DSpace on 2014-05-27T11:28:33Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-02-28 Made available in DSpace on 2014-05-27T11:28:33Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-02-28 Made available in DSpace on 2014-05-27T11:28:33Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-02-28 This study was designed to determine the genotoxicity of a supraphysiological dose of triiodothyronine (T3) in both obese and calorie-restricted obese animals. Fifty male Wistar rats were randomly assigned to one of the two following groups: control (C; n = 10) and obese (OB; n = 40). The C group received standard food, whereas the OB group was fed a hypercaloric diet for 20 weeks. After this period, half of the OB animals (n = 20) were subjected to a 25%-calorie restriction of standard diet for 8 weeks forming thus a new group (OR), whereas the remaining OB animals were kept on the initial hypercaloric diet. During the following two weeks, 10 OR animals continued on the calorie restriction diet, whereas the remaining 10 rats of this group formed a new group (ORS) given a supraphysiological dose of T3 (25 μg/100 g body weight) along with the calorie restriction diet. Similarly, the remaining OB animals were divided into two groups, one that continued on the hypercaloric diet (OB, n = 10), and one that received the supraphysiological dose of T3 (25 μg/100 g body weight) along with the hypercaloric diet (OS, n = 10) for two weeks. The OB group showed weight gain, increased adiposity, insulin resistance, increased leptin levels and genotoxicity; T3 administration in OS animals led to an increase in genotoxicity and oxidative stress when compared with the OB group. The OR group showed weight loss and normalized levels of adiposity, insulin resistance, serum leptin and genotoxicity, thus having features similar to those of the C group. On the other hand, the ORS group, compared to OR animals, showed higher genotoxicity. Our results indicate that regardless of diet, a supraphysiological dose of T3 causes genotoxicity and potentiates oxidative stress. © 2013 de Sibio et al. Department of Internal Medicine Botucatu Medical School - University of Sao Paulo State (UNESP), Botucatu, SP Department of Pathology Botucatu Medical School - University of Sao Paulo State (UNESP), Botucatu, SP Department of Biostatistics Biosciences Institute - University of Sao Paulo State (UNESP), Botucatu, SP Department of Internal Medicine Botucatu Medical School - University of Sao Paulo State (UNESP), Botucatu, SP Department of Pathology Botucatu Medical School - University of Sao Paulo State (UNESP), Botucatu, SP Department of Biostatistics Biosciences Institute - University of Sao Paulo State (UNESP), Botucatu, SP

This study was designed to determine the genotoxicity of a supraphysiological dose of triiodothyronine (T3) in both obese and calorie-restricted obese animals. Fifty male Wistar rats were randomly assigned to one of the two following groups: control (C; n = 10) and obese (OB; n = 40). The C group received standard food, whereas the OB group was fed a hypercaloric diet for 20 weeks. After this period, half of the OB animals (n = 20) were subjected to a 25%-calorie restriction of standard diet for 8 weeks forming thus a new group (OR), whereas the remaining OB animals were kept on the initial hypercaloric diet. During the following two weeks, 10 OR animals continued on the calorie restriction diet, whereas the remaining 10 rats of this group formed a new group (ORS) given a supraphysiological dose of T3 (25 µg/100 g body weight) along with the calorie restriction diet. Similarly, the remaining OB animals were divided into two groups, one that continued on the hypercaloric diet (OB, n = 10), and one that received the supraphysiological dose of T3 (25 µg/100 g body weight) along with the hypercaloric diet (OS, n = 10) for two weeks. The OB group showed weight gain, increased adiposity, insulin resistance, increased leptin levels and genotoxicity; T3 administration in OS animals led to an increase in genotoxicity and oxidative stress when compared with the OB group. The OR group showed weight loss and normalized levels of adiposity, insulin resistance, serum leptin and genotoxicity, thus having features similar to those of the C group. On the other hand, the ORS group, compared to OR animals, showed higher genotoxicity. Our results indicate that regardless of diet, a supraphysiological dose of T3 causes genotoxicity and potentiates oxidative stress.

Made available in DSpace on 2016-04-01T18:44:23Z (GMT). No. of bitstreams: 0 Previous issue date: 2013. Added 1 bitstream(s) on 2016-04-01T18:49:52Z : No. of bitstreams: 1 ISSN0004-2730-2013-57-05-368-374.pdf: 218443 bytes, checksum: 5474d376f1638e12ee42afe0365fd7b3 (MD5) Objetivo: Examinar o efeito de diferentes doses de triiodotironina (T3) sobre a expressão gênica dos receptores TRα e TRβ em diferentes tempos. Materiais e métodos: Adipócitos, 3T3-L1, foram incubados com T3 nas doses fisiológica (F, 10nM) e suprafisiológicas (SI, 100nM ou SII, 1000nM) ou veículo (controle, C) durante 0,5, 1, 6 ou 24h. mRNA dos TRs foram detectados utilizando PCR em tempo real. Resultados: Níveis de TRβ aumentaram em F em 0,5h. Após 1h, níveis de TRα aumentaram em F e SI comparado ao C, enquanto TRβ diminuiu no SII comparado com C, F, e SI. Após 6h, ambos os genes foram suprimidos em todas concentrações. Em 24h, níveis de TRα e TRβ retornaram aos do C. Conclusões: Ação do T3 em F iniciou-se em 1h para TRα e 0,5h para TRβ. Esses resultados são importantes para determinar tempo inicial e dose de T3 em que os receptores de HT são ativados em adipócitos. Objective: To examine the effect of different doses of triiodothyronine (T3) on mRNA levels of thyroid hormone receptors, TRα and TRβ, at different times. Materials and methods: 3T3-L1 adipocytes were incubated with T3 (physiological dose: F; supraphysiological doses: SI or SII), or without T3 (control, C) for 0.5, 1, 6, or 24h. TRα and TRβ mRNA was detected using real-time polymerase chain reaction. Results: F increased TRβ mRNA levels at 0.5h. After 1h, TRα levels increased with F and SI and TRβ levels decreased with SII compared with C, F, and SI. After 6h, both genes were suppressed at all concentrations. In 24h, TRα and TRβ levels were similar to those of C group. Conclusions: T3 action with F began at 1h for TRα and at 0.5h for TRβ. These results suggest the importance of knowing the times and doses that activate T3 receptors in adipocytes. Department of Physiology, São Paulo Federal University (Unifesp), São Paulo, SP, Brazil Department of Internal Clinic, Botucatu Medicine School, São Paulo State University (Unesp), Botucatu, SP, Brazil Department of Biostatistics, Biosciences Institute, Unesp, Botucatu, SP, Brazil

The primary function of adipose tissue is storing energy in triacylglycerol (TG) form, neutralizing the excess of circulating lipids and saving non-adipose tissues of a fat overload. Under normal conditions, in the postprandial state, there is lipogenic endocrine system stimulation, allowing that positive energy balance can be stored as TG in adipose tissue, a process called lipogenesis. In contrast, the mobilization of fat in adipocytes occurs through the hydrolysis of TG by hormone sensitive lipase (HSL), a phenomenon called lipolysis. At the center of this interface lipolysis and lipogenesis is the insulin hormone, which exerts a potent inhibitory role on the HSL, allowing lower rates of lipolysis and hence, highest fat mass [1]. However, adipose tissue is not only a passive stock organ of triacylglycerol, being currently recognized as an endocrine organ with multiple functions [1, 2]. Produces several biologically active substances called adipokines, among them tumor necrosis factor- (TNF-), monocyte chemoattractant protein 1 (MCP-1), interleukin-6 (IL-6), leptin, resistin and adiponectin. These substances actively participate in, among others, body energy regulation, mainly, by endocrine, paracrine and autocrine signals, which allow the adipocyte play a metabolic role in other tissues [3-5].

There are several breast cancer experimental models including cell lines, which are commonly used due to ease of handling and storage. However, the continued propagation of cell lines and distribution among laboratories results in genetic drift and distancing from the in-vivo model. Primary organ culture of breast cancer slices may produce biological responses with high standard deviation for different samples, reflecting the heterogeneity of different tumors. Thus, the organ culture model system offers a new perspective to the results obtained in the cell lines and offers an alternative for studies that seek to individualize treatment for each patient, an increasingly prominent concern in current cancer therapy.

Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:26:51Z No. of bitstreams: 0Bitstream added on 2014-05-27T14:31:39Z : No. of bitstreams: 1 2-s2.0-84862333929.pdf: 1588872 bytes, checksum: d84eef3b9c43a00d6f4a6b79900182ab (MD5) Made available in DSpace on 2014-05-27T11:26:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-06-21 Aims. To analyze the influence of hyperthyroidism on the gene expression and serum concentration of leptin, resistin, and adiponectin in obese animals. Main Methods. Male Wistar rats were randomly divided into two groups: control (C)fed with commercial chow ad libitumand obese (OB)fed with a hypercaloric diet. After group characterization, the OB rats continued receiving a hypercaloric diet and were randomized into two groups: obese animals (OB) and obese with 25g triiodothyronine (T3)/100BW (OT). The T3 dose was administered every day for the last 2 weeks of the study. After 30 weeks the animals were euthanized. Samples of blood and adipose tissue were collected for biochemical and hormonal analyses as well as gene expression of leptin, resistin, and adiponectin. Results. T3 treatment was effective, increasing fT3 levels and decreasing fT4 and TSH serum concentration. Administration of T3 promotes weight loss, decreases all fat deposits, and diminishes serum levels of leptin, resistin, and adiponectin by reducing their gene expression. Conclusions. Our results suggest that T3 modulate serum and gene expression levels of leptin, resistin, and adiponectin in experimental model of obesity, providing new insights regarding the relationship between T3 and adipokines in obesity. Copyright © 2012 Renata de Azevedo Melo Luvizotto et al. Department of Internal Medicine Botucatu School of Medicine University of São Paulo State, 18618-000 Botucatu, SP Department of Sports Center of Physical Education and Sports Federal University of Espirito Santo (UFES), 29075-910 Vitória, ES

Objectives: To examine the effects of triiodothyronine (T3), 17β-estradiol (E2), and tamoxifen (TAM) on transforming growth factor (TGF)-α gene expression in primary breast cancer cell cultures and interactions between the different treatments. Methods and results: Patients included in the study (no.=12) had been newly diagnosed with breast cancer. Fresh human breast carcinoma tissue was cut into 0.3-mm slices. These slices were placed in six 35-mm dishes on 2-ml organ culture medium. Dishes received the following treatments: dish 1: ethanol; dish 2: T3; dish 3: T3+TAM; dish 4: TAM; dish 5: E2; dish 6: E2+TAM. TGF-α mRNA content was normalized to glyceraldehyde-3-phosphate dehydrogenase mRNA levels. All tissues included in this study were positive for estrogen receptor (ER) and thyroid hormone receptor expression. Treatment with T3 for 48 h significantly increased TGF-α mRNA levels compared to controls (15-fold), and concomitant treatment with TAM reduced expression to 3.4-fold compared to controls. When only TAM was added to the culture medium, TGF-α mRNA expression increased 5.3-fold, significantly higher than with all other treatment modalities. Conclusion: We demonstrate that TGF-α mRNA expression is more efficiently upregulated by T3 than E2. Concomitant treatment with TAM had a mitigating effect on the T3 effect, while E2 induced TGF-α upregulation. Our findings show some similarities between primary culture and breast cancer cell lines, but also some important differences: a) induction of TGF-α, a mitogenic protein, by TAM; b) a differential effect of TAM that may depend on relative expression of ER α and β; and c) supraphysiological doses of T3 may induce mitogenic signals in breast cancer tissue under conditions of low circulating E2.