H-Ferritin Is Essential for Macrophages' Capacity to Store or Detoxify Exogenously Added Iron

Macrophages are central cells both in the immune response and in iron homeostasis. Iron is both essential and potentially toxic. Therefore, iron acquisition, transport, storage, and release are tightly regulated, by several important proteins. Cytosolic ferritin is an iron storage protein composed of 24 subunits of either the L- or the H-type chains. H-ferritin differs from L-ferritin in the capacity to oxidize Fe2+ to Fe3+. In this work, we investigated the role played by H-ferritin in the macrophages' ability to respond to immune stimuli and to deal with exogenously added iron. We used mice with a conditional deletion of the H-ferritin gene in the myeloid lineage to obtain bone marrow-derived macrophages. These macrophages had normal viability and gene expression under basal culture conditions. However, when treated with interferon-gamma and lipopolysaccharide they had a lower activation of Nitric Oxide Synthase 2. Furthermore, H-ferritin-deficient macrophages had a higher sensitivity to iron-induced toxicity. This sensitivity was associated with a lower intracellular iron accumulation but a higher production of reactive oxygen species. These data indicate that H-ferritin modulates macrophage response to immune stimuli and that it plays an essential role in protection against iron-induced oxidative stress and cell death.Tis work was fnanced by FEDER - Fundo Europeu de Desenvolvimento Regional funds through the COMPETE2020 - Operacional Programme for Competitiveness and Internationalization (POCI), Portugal 2020, and by Portuguese funds through FCT - Fundação para a Ciência e a Tecnologia/Ministério da Ciência, Tecnologia e Ensino Superior in the framework of the project PTDC/IMI-MIC/1683/2014 (POCI-01-0145-FEDER-016590). PFO and MGA acknowledge FCT for the Investigador FCT 2015. We thank the valuable collaboration of the following i3S Scientifc Platforms: Cell Culture and Genotyping Core Facility (CCGen), [Histology and Electron Microscopy Service (HEMS), and BioSciences Screening], member of the PPBI (PPBI-POCI-01-0145-FEDER-022122)], Animal Facility, and Flow Cytometry Unit (TraCy). We acknowledge Lukas Kuhn (Swiss Institute for Experimental Cancer Research, Lausanne, Switzerland) for kindly providing the frst breeding pairs of Fth1−/− mice. Te authors also acknowledge Marisa Castro, from Departamento de Biologia Molecular from ICBAS, Clara Bento, from i3S, and Edgar Pinto from LAQV – REQUIMTE for technical assistance at diferent stages of the project

Circunfer?ncia do pesco?o e risco cardiovascular em 10 anos : diferen?as por sexo. an?lise seccional da linha de base do Estudo Longitudinal de Sa?de do Adulto (ELSA-Brasil).

Programa de P?s-Gradua??o em Sa?de e Nutri??o. Escola de Nutri??o, Universidade Federal de Ouro Preto.INTRODU??O: A circunfer?ncia do pesco?o (CP) ? uma estimativa da gordura do pesco?o e da gordura subcut?nea da parte superior do corpo, cujo aumento parece conferir risco cardiovascular adicional ?quele conferido pela adiposidade geral e abdominal. O Framingham Global Risk Score (FGRS) fornece uma estimativa do risco de desenvolver DCV em 10 anos, utilizado para identificar indiv?duos sob maior risco de DCV, inclusive na pr?tica cl?nica. OBJETIVO: O objetivo da presente disserta??o foi verificar a associa??o entre a circunfer?ncia do pesco?o e o risco de evento cardiovascular em 10 anos em homens e mulheres participantes da linha de base do Estudo Longitudinal de Sa?de do Adulto (ELSA-Brasil). M?TODOS: Trata-se de estudo de corte transversal com participantes da linha de base do Estudo Longitudinal de Sa?de do Adulto (ELSABrasil) (2008-2010). O ELSA-Brasil ? uma coorte multic?ntrica com 15.105 servidores p?blicos, ativos e aposentados de institui??es de ensino e pesquisa de seis capitais de estados brasileiros. Para a presente an?lise, foram exclu?dos os indiv?duos com relato de DCV (n=738) e sem informa??es para DCV (n=26), CP (n=11), FGRS (n=28) e covari?veis (n=382), permanecendo 13.920 participantes. As caracter?sticas da popula??o do estudo e dos componentes do FGRS foram descritas por meio de frequ?ncias absolutas e relativas, m?dias e desvio padr?o (?DP) ou medianas (1? e 3? quartis) e utilizados testes compara??o dessas medidas. A associa??o entre a CP (utilizada como vari?vel cont?nua e agrupda em quartis) e o risco DCV em 10 anos, mensurado pelo FGRS, foi estimada por meio de Modelos Lineares Generalizados (MLG), com distribui??o gama e fun??o logar?tmica, cujo exponencial do coeficiente de regress?o fornece a Raz?o da M?dias Aritm?ticas com intervalo de 95% de confian?a (RMA, IC95%). Foram realizados ajustes por idade, escolaridade, ra?a/cor autorreferida, consumo de ?lcool, atividade f?sica no lazer, ?ndice de massa corporal (IMC) e circunfer?ncia da cintura (CC). RESULTADOS: A m?dia de idade dos participantes foi de 51,7 anos (DP?7,6), sendo 55% mulheres. A m?dia da CP aumentou com o incremento do risco de DCV em 10 anos agrupado em categorias de risco (risco baixo 20%) em ambos os sexos. Ap?s todos os ajustes, incluindo as demais medidas de adiposidade corporal, a CP permaneceu independentemente associada a um aumento de 3% na m?dia aritm?tica do risco DCV em 10 anos (RMA= 1,03; IC 95% 1,01-1,03) nos homens e de 5% (RMA= 1,05; IC 95% 1,04-1,06) nas mulheres. Nos modelos de regress?o utilizando a CP agrupada em quartis, ap?s todos os ajustes, observamos que comparados ao primeiro quartil todos os demais apresentaram aumento gradual na m?dia aritm?tica no risco de DCV em 10 anos, que chegou a um incremento de 18% entre os que estavam no ?ltimo quartil (IC95%: 1,13-1,24) entre os homens e a 35% (IC95%: 1,28-1,43) entre as mulheres. Foram realizadas an?lises de sensibilidade com a exclus?o de participantes em uso de hipolipemiantes, uso de corticoides, e de mulheres em uso de anticoncepcional ou em reposi??o hormonal e essas exclus?es n?o levaram a altera??o nos resultados observados. CONCLUS?O: Nossos resultados sugerem uma associa??o entre a circunfer?ncia do pesco?o e o risco DCV em 10 anos, mensurado pelo Framingham Global Risk Score em ambos os sexos, mas indicando maior for?a de associa??o entre as mulheres. Contudo, novos estudos em outras popula??es e an?lises longitudinais s?o necess?rios.INTRODUCTION: The effect of adipose tissue on the development of CVD varies between different fat deposits. Neck Circumference (NC) provides an estimate of neck fat and upper body subcutaneous fat that has been singled out as the sole body fat deposit that confers additional cardiovascular risk above and beyond general and central body fat. OBJECTIVE: The objective of this dissertation was to verify the association between neck circumference and cardiovascular risk in 10 years in men and women participants from the baseline of the Longitudinal Study of Adult Health (ELSA-Brazil). METHODS: A cross-sectional study was conducted with participants from the Study of the Longitudinal Study of Adult Health (ELSA-Brazil) baseline (2008-2010). ELSABrazil is a multicentre cohort composed of 15,105 civil serventes from institution of education and research institutions in six Brazilian state capitals. For the present analysis, individuals with a history of CVD (acute myocardial infarction, heart failure, stroke and coronary artery bypass grafting) (n = 738), missing information for CVD (n = 26), for NC = 11), for the Framingham Global Risk Score (n = 28) and for covariables (n = 382) were excluded. At the end the sample analytic was composed by 13,920 participants.The characteristics of the study population and the FGRS components were described by means of absolute and relative frequencies (categorical variables) and means and standard deviation (? SD) or medians (1st and 3rd quartiles) (continuous variables). Pearson's Chisquare test was used for comparison of frequencies, Student's t-test for comparison of means and Kruskal-Wallis test for comparison of medians. The association between NC (continous variable and agrouped in quartiles) and the10-years CDV risk, measured by the FGRS, was estimated using Generalized Linear Models (MLG), with gamma distribution and logarithmic function, whose exponential regression coefficient gives the Arithmetic Mean Ratio with 95% confidence interval (RMA, 95% CI). Potential confounding factors were age, schooling, self-reported race / color, alcohol consumption, leisure time physical activity, body mass index (BMI) and waist circumference (WC). RESULTS: The mean age of participants was 51.7 years (SD ? 7.6), being 55% female. The mean NC increased according to risk categories of the 10-year CVD risk (low risk 20%) in both sexes. In the crude analysis, it was observed that the increase of one centimeter in NC was associated with an increase of 5% (RMA = 1.05, 95% CI: 1.04-1.05) in the mean 10-year CVD risk in men and 11% (RMA = 1.11, 95% CI: 1.10-1.12) in women. After all the adjustments, including the other measures of body adiposity, NC remained independently associated with a 3% increase in the arithmetic mean of CVD risk in 10 years (RMA = 1.03, 95% CI, 1.01- 1.03) in men and 5% (RMA = 1.05, 95% CI 1.04-1.06) in women. In the regression models using NC grouped in quartiles, after all the adjustments, we observed that, compared to the first quartile, all the others presented a gradual increase in the arithmetic mean in the risk of the 10-years CVD risk, which reached an increase of 18% in men who were in the last quartile (95% CI: 1.13-1.24) and 35% (95% CI: 1.28-1.43) in women. Sensitivity analyzes were performed with the exclusion of participants taking lipidlowering drugs, corticosteroids, and women taking contraceptives or hormone replacement, and these exclusions did not lead to alteration in the observed results. CONCLUSION: Our results suggest an association between NC and 10-years CVD risk as measured by the Framingham Global Risk Score in both sexes, but indicating a greater strength of association among women

ESTIMATIVA DA IDADE CRONOLÓGICA HUMANA POR MEIO DA AVALIAÇÃO DO ESTÁGIO DE MINERALIZAÇÃO DOS 3º MOLARES

Apesar  de  existirem  métodos  eficazes  para  estimar  a  idade cronológica de indivíduos vivos ou mortos, nem sempre esses métodos podem ser executados. Apesar de alguns estudos apontarem uma possível aplicação da  avaliação  do  desenvolvimento  dos  terceiros  molares  na  identificação cronológica,  sua  a  utilização  ainda  necessita  de  maiores  comprovações

Serum amyloid A proteins reduce bone mass during mycobacterial infections

IntroductionOsteopenia has been associated to several inflammatory conditions, including mycobacterial infections. How mycobacteria cause bone loss remains elusive, but direct bone infection may not be required. MethodsGenetically engineered mice and morphometric, transcriptomic, and functional analyses were used. Additionally, inflammatory mediators and bone turnover markers were measured in the serum of healthy controls, individuals with latent tuberculosis and patients with active tuberculosis. Results and discussionWe found that infection with Mycobacterium avium impacts bone turnover by decreasing bone formation and increasing bone resorption, in an IFN gamma- and TNF alpha-dependent manner. IFN gamma produced during infection enhanced macrophage TNF alpha secretion, which in turn increased the production of serum amyloid A (SAA) 3. Saa3 expression was upregulated in the bone of both M. avium- and M. tuberculosis-infected mice and SAA1 and 2 proteins (that share a high homology with murine SAA3 protein) were increased in the serum of patients with active tuberculosis. Furthermore, the increased SAA levels seen in active tuberculosis patients correlated with altered serum bone turnover markers. Additionally, human SAA proteins impaired bone matrix deposition and increased osteoclastogenesis in vitro. Overall, we report a novel crosstalk between the cytokine-SAA network operating in macrophages and bone homeostasis. These findings contribute to a better understanding of the mechanisms of bone loss during infection and open the way to pharmacological intervention. Additionally, our data and disclose SAA proteins as potential biomarkers of bone loss during infection by mycobacteria.This article is a result of the project HEALTH-UNORTE: Setting-up biobanks and regenerative medicine strategies to boost research in cardiovascular, musculoskeletal, neurological, oncological, immunological and infectious diseases (NORTE-01-0145-FEDER-000039), supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the PORTUGAL 2020 Partnership Agreement, through the European Regional Development Fund (ERDF). This work was supported by KOG-202108-00929 from the European Haematology Society, awarded to AG. Work in the MS lab was financed by FEDER - Fundo Europeu de Desenvolvimento Regional funds through the COMPETE 2020 - Operacional Programme for Competitiveness and Internationalisation (POCI), Portugal 2020, and by Portuguese funds through FCT - Fundacao para a Ciencia e a Tecnologia/Ministerio da Ciencia, Tecnologia e Ensino Superior in the framework of the project POCI-01-0145-FEDER-028955 (PTDC/SAU-INF/28955/2017). AG and MS are supported by an Individual Scientific Employment contract (CEECIND/00048/2017; CEECIND/00241/2017 respectively). DS acknowledges the Portuguese Foundation for Science and Technology (FCT) for the Post-Doc fellowship (SFRH/BPD/115341/2016). RP, DS and AF have PhD grants (SFRH/BD/145217/2019; SFRH/BD/143536/2019; 2020.05949.BD, respectively) financed by FCT

Brazilian gamma detection device for sentinel lymph node biopsy

OBJECTIVE: To test the effectiveness of an intra-operative gamma detection Brazilian device (IPEN) on sentinel lymph node biopsy (SLNB) procedures. METHODS: Forty melanoma or breast cancer patients with indication for undergoing SLNB were studied. Lymphoscintigraphy was done 2 to 24 hours prior to surgery. Lymphatic mapping with vital dye and gamma detection were performed intraoperatively. For gamma detection Neoprobe ® 1500 was used followed by IPEN (equipment under test) in the first 20 patients and for the remaining half IPEN was used first to verify its ability to locate the sentinel node (SN). Measurements were taken from the radiopharmaceutical product injection site, from SN (in vivo and ex vivo) and from background. It was recorded if the SN was stained or not and if it was found easily by surgeon. RESULTS: There were 33 (82.5%) breast cancer and 7 (17.5%) melanoma patients. Ages varied from 21 to 68 year-old (median age of 46). Sex distribution was 35 (87.5%) women and 5 (12.5%) men. Sentinel node was found in all but one patient. There was no statistical difference between the reasons ex vivo/ background obtained with the measures of both equipments (p=0, 2583-ns). The SN was easily found by the surgeon with both devices. CONCLUSION: The SLNB was successfully performed using either equipment. It was possible to do SLNB with the Brazilian device developed by IPEN without prejudice for the patient.OBJETIVO: Testar a eficácia de equipamento de detecção gama intra-operatória (DGI) desenvolvido pelo IPEN (Brasil), em procedimentos de biópsia de linfonodo sentinela (BLS) no melanoma e no câncer de mama. MÉTODOS: Foram estudados 40 pacientes portadores de melanoma ou câncer de mama com indicação para realização de BLS.Todos pacientes foram submetidos à linfocintilografia e a BLS ocorreu entre 2 a 24 horas após a mesma. Concomitantemente à DGI, realizou-se o mapeamento linfático com corante vital. Foram feitas leituras com o equipamento convencional Neoprobe® 1500 e com o equipamento em teste (IPEN) dos valores de captação do sítio de injeção do radiofármaco, do LS in vivo e ex vivo e da captação de fundo. Foi registrado se o LS estava corado e se o cirurgião teve facilidade para encontrá-lo. Nos primeiros 20 pacientes utilizou-se o equipamento convencional e depois o de teste; nos outros 20, utilizou-se primeiro o equipamento em teste, com objetivo de verificar se o mesmo identificava primariamente o LS. RESULTADOS: Dos quarenta pacientes, 33 eram portadores de tumor de mama e sete de melanoma cutâneo; variação da idade: 21 a 68 anos (mediana= 46 anos); 35 mulheres e 5 homens. Em apenas um paciente o LS não foi encontrado, nem pela DGI nem pelo corante vital. Não houve diferença estatística entre as razões ex vivo/fundo obtidas com os dois equipamentos (p=0, 2583-ns). CONCLUSÃO: É possível realizar o procedimento de BLS com o equipamento brasileiro desenvolvido pelo IPEN, com facilidade e sem prejuízo para o paciente.Hospital Israelita Albert EinsteinIPENUniversidade Federal de São Paulo (UNIFESP)Faculdade de Medicina Anhembi-MorumbiUNIFESPSciEL

2-Methoxyestradiol-3,17-O,O-bis-sulfamate (STX140) inhibits proliferation and invasion via senescence pathway induction in human BRAFi-resistant melanoma cells

The endogenous estradiol derivative 2-Methoxyestradiol (2-ME) has shown good and wide anticancer activity but suffers from poor oral bioavailability and extensive metabolic conjugation. However, its sulfamoylated derivative, 2-methoxyestradiol-3,17-O,O-bis-sulfamate (STX140), has superior potential as a therapeutic agent, acts by disrupting microtubule polymerization, leading to cell cycle arrest and apoptosis in cancer cells and possesses much better pharmaceutical properties. This study investigated the antiproliferative and anti-invasive activities of STX140 in both SKMEL-28 naïve melanoma (SKMEL28-P) cells and resistant melanoma cells (SKMEL-28R). STX140 inhibited cell proliferation in the nanomolar range while having a less pronounced effect on human melanocytes. Additionally, STX140 induced cell cycle arrest in the G2/M phase and sub-G1, reduced migration, and clonogenic potential in monolayer models, and inhibited invasion in a 3D human skin model with melanoma cells. Furthermore, STX140 induced senescence features in melanoma and activated the senescence machinery by upregulating the expression of senescence genes and proteins related to senescence signaling. These findings suggest that STX140 may hold potential as a therapeutic agent for melanoma treatment

Effect of Weight Loss after Bariatric Surgery on Thyroid-Stimulating Hormone Levels in Patients with Morbid Obesity and Normal Thyroid Function

Background: Several studies have reported that morbid obesity is associated with increased thyroid-stimulating hormone (TSH) levels. However, it is not clear what is the impact of bariatric surgery on postoperative thyroid function. The aim of this study was to evaluate the effect of weight loss after bariatric surgery on TSH levels in euthyroid patients with morbid obesity. Methods: We performed a retrospective observational study of 949 euthyroid patients (86.1% female; age 42.0 ± 10.3 years, BMI 44.3 ± 5.7 kg/m2) with morbid obesity submitted to bariatric surgery (laparoscopic adjustable gastric band, Roux-en-Y gastric bypass, or sleeve gastrectomy). Patients were subdivided in two groups: normal TSH group (TSH <2.5 mU/L) and high-normal TSH group (TSH ≥2.5 mU/L). The impact of anthropometric parameters, comorbidities, TSH, free thyroxine (FT4), free triiodothyronine (FT3), type of surgery, and excessive body weight loss (EBWL) on TSH variation 12 months after surgery was evaluated. Results: The high-normal TSH group (24.3% of patients) included more women, presented a higher BMI, higher systolic blood pressure, and higher FT3 levels. There was a significant decrease of TSH 12 months after surgery that was more marked in the high-normal TSH group (normal TSH group: 1.57 ± 0.49 to 1.53 ± 0.69 mIU/L, p = 0.063; high-normal TSH group: 3.23 ± 0.59 to 2.38 ± 0.86 mIU/L, p < 0.001). In a multivariate analysis, after adjusting for relevant covariates, EBWL, baseline BMI, and baseline FT3 were significantly associated with TSH decrease 12 months after bariatric surgery. Conclusion: Bariatric surgery promotes a decrease of TSH that is significantly greater in patients with high-normal TSH and is independently associated with EBWL after surgery