Characterisation of epicardial adipose tissue and myocardial fat infiltration in humans

L\u2019obesit\ue0 ed il diabete mellito di tipo 2 sono caratterizzati da un basso grado di infiammazione sistemica, aumento di dimensione del tessuto adiposo (TA), rilascio incontrollato di acidi grassi liberi nella circolazione; sono entrambi fortemente correlati con patologie metaboliche e cardiovascolari, come la dislipidemia, la patologia coronarica, l\u2019ipertensione e l\u2019infarto miocardico. Il TA bianco \ue8 stato accettato come essere molto pi\uf9 di un semplice deposito statico di energia. Le sue capacit\ue0 di regolare meccanismi omeostatici e metabolici dell\u2019intero organismo sono stati evidenziati negli ultimi 20 anni; molti studi hanno proposto il TA come un organo endocrino, secernente ormoni, adipochine ed altre sostanze biologicamente attive localmente o a livello sistemico. I depositi di TA non sono uniformi; le loro caratteristiche cambiano in diverse aree corporee, partecipando in modo diverso alla possibile patogenesi delle diverse patologie. Il TA epicardico (EAT) \ue8 un particolare deposito di TA, che \ue8 stato recentemente al centro di molti studi. Si trova localizzato principalmente sulla parete destra libera del cuore e circonda le coronarie, entrando a diretto contatto con il miocardio. Si \ue8 visto che EAT secerne molte differenti adipochine e si suppone quindi abbia un ruolo nella generazione e progressione della patologia coronarica. Lo scopo di questa tesi \ue8 quello di meglio caratterizzare EAT e lo strato di miocardio ad esso sottostante. Nell\u2019articolo 1 abbiamo analizzato le dimensioni cellulari adipocitarie e la secrezione di adiponectina confrontando EAT, il TA viscerale (VAT) e sottocutaneo (SAT). EAT \ue8 risultato avere cellule pi\uf9 piccole con ridotta secrezione di adiponectina. La dimensione adipocitaria sia in EAT che SAT \ue8 positivamente correlata con l\u2019insulino resistenza, e mostra associazione negativa con l\u2019espressione locale di adiponectina. I soggetti affetti da patologia cardiovascolare presentano inoltre adipociti significativamente pi\uf9 grandi in EAT. Nell\u2019articolo 2 abbiamo focalizzato la nostra attenzione sugli effetti della patologia diabetica su EAT, mostrando maggiore espressione genica di MCP-1, CD-68 e ridotta di adiponectina, cos\uec come dimensioni adipocitarie maggiori in soggetti diabetici rispetto ai non diabetici. In alcuni dati ancora non pubblicati abbiamo inoltre valutato le caratteristiche immunoistochimiche di infiltrazione lipidica e macrofagica del miocardio della stessa popolazione di pazienti. Alla tesi \ue8 stato inoltre aggiunto come articolo 3 il risultato di uno studio condotto nel corso di un\u2019esperienza all\u2019estero della durata di un anno presso il Lipid Laboratory KI di Stoccolma Svezia. Lasciando il soggetto di EAT, il lavoro si concentra su TA e gli aspetti genetici dell\u2019obesit\ue0. Niemann-Pick C1 (NPC-1) \ue8 stato recentemente implicato come gene di suscettibilit\ue0 per l\u2019obesit\ue0 attraverso uno studio genome wide. Abbiamo approfondito quindi il rapporto tra NPC-1 e l\u2019obesit\ue0 nell\u2019uomo. Le analisi mRNA hanno mostrato aumentata espressione nel paziente obeso sia in SAT che in VAT e riduzione in seguito a calo di peso. L\u2019espressione genica di NPC-1 era maggiore nelle cellule adipocitarie isolate, rispetto al tessuto in toto, sia in SAT che in VAT, ma non era modificata nel corso della differenziazione adipocitaria. L\u2019espressione proteica di NPC-1 rispecchiava l\u2019espressione genica.Obesity and type 2 diabetes are characterized by mild systemic inflammation, enlargement of fat depots, and uncontrolled release of free fatty acids into the circulation; they are both strongly associated with metabolic and cardiovascular disorders, such as dyslipidemia, coronary heart disease, high blood pressure and myocardial infarct. White adipose tissue has been widely accepted to be much more than a static fuel storage organ. Its capability of regulating homeostatic and metabolic mechanisms has been underlined during the last 20 years; several studies have proposed adipose tissue as an endocrine organ, secreting hormones, adipokines and other biologically active agents acting locally or in a systemic manner. Adipose tissue depots are not uniform; their characteristics change in different areas of the body, displaying distinct structural and functional properties and having different putative roles in pathologies. Epicardial adipose tissue (EAT) is a peculiar adipose tissue depot, which has recently been the center of many studies. It\u2019s located predominantly on the right free wall of the heart, surrounding coronary arteries and being directly in contact with the myocardial layer. EAT has been shown to secrete many different adipokines and is supposed to have a role in the generation and progression of coronary artery diseases. The aim of this thesis was to better characterize EAT and the underlying myocardial layer. In paper 1 we investigated adipocyte cell size and adiponectin secretion comparing EAT, visceral AT (VAT) and subcutaneous AT (SAT). EAT resulted to have smaller adipocytes and lower adiponectin secretion levels. Adipocyte size, both in EAT and in SAT, is positively related with insulin resistance, shows negative association with local adiponectin gene expression, and bigger in subjects with coronary artery disease. Adiponectin gene expression is significantly lower in EAT than in SAT. In paper 2 we focused our attention on the effect of diabetic state on EAT, showing higher MCP-1, CD-68, lower adiponectin level, and bigger adipocytes in subjects with than those without diabetes. We also analyzed thorough immunohistochemistry and present as unpublished data, the characteristics of fat and macrophagic infiltration of the myocardium of the same cohort of patients. Additionally, the result of a one year internship conducted at Lipid Laboratory in KI Sweden, has been included in the thesis, as paper 3. Leaving the subject of EAT, the paper is focused on adipose tissue and it\u2019s molecular and genetic aspects in obesity. Since the gene Niemann-Pick C1 (NPC-1) has recently being implicated in susceptibility to obesity, through a genome wide association study, we dig into the relationship between NPC-1 and obesity in humans. The analysis of NPC-1 mRNA ad protein in obesity, showed that NPC1 mRNA was significantly increased in obese individuals in SAT and VAT and down-regulated by weight loss. NPC-1 mRNA was enriched in isolated fat cells of WAT, in SAT versus VAT, but not modified during adipocyte differentiation. NPC-1 protein mirrored expression of mRNA in lean and obese individual

Effect of moderate weight loss on hepatic, pancreatic and visceral lipids in obese subjects

OBJECTIVE: To compare the effects of weight loss on visceral and subcutaneous abdominal fat, liver and pancreas lipid contentand to test the effects of these changes on metabolic improvement observed after weight loss.DESIGN: Weight-loss program designed to achieve a loss of 7--10% of the initial weight.SUBJECTS: 24 obese subjects (13 males and 11 females) with age ranging from 26 to 69 years and body mass index (BMI)30.2 -- 50.5 kgm2. Measurements: weight, BMI, waist circumference, body composition as assessed by dual-energy X-rayabsorptiometry, metabolic variables, leptin, adiponectin, visceral and subcutaneous abdominal fat, liver and pancreas lipidcontent as assessed by magnetic resonance were evaluated before and after weight loss achieved by hypocaloric diet.RESULTS: After a mean body weight decrease of 8.9%, BMI, waist circumference, fat mass, all metabolic variables, homeostasismodel assessment of insulin resistance (HOMA), alanine amino transferase, gamma glutamyl transpeptidase, high-sensitivityC-reactive protein (hs-CRP) and leptin, but not adiponectin and high-density lipoprotein-cholesterol, significantly decreased (allPo0.01). Visceral and subcutaneos abdominal fat, liver and pancreas lipid content significantly decreased (all Po0.01). Percentchanges in liver lipid content were greater (84.1\ub13%) than those in lipid pancreas content (42.3\ub129%) and visceral abdominalfat (31.9\ub115.6%). After weight loss, percentage of subjects with liver steatosis decreased from 75 to 12.5%. Insulin resistanceimprovement was predicted by changes in liver lipid content independently of changes in visceral fat, pancreas lipid content,systemic inflammation, leptin and gender.CONCLUSION: Moderate weight loss determines significant decline in visceral abdominal fat, lipid content in liver andpancreas. Reduction of liver lipid content was greater than that of pancreas lipid content and visceral fat loss. Liver lipidcontent is the strongest predictor of insulin resistance improvement after weight loss

Sobre a violência sexual no esporte olímpico envolvendo mulheres: um estudo a partir de reportagens em diferentes sites esportivos

The purpose of this research was to map the cases of sexual violence in olympic sports, publicized in different sports sites (FolhaEsporte, ESPN, GazetaEsportiva e El País, GloboEsporte, Lance!,SporTV e Terra), between the years of 2013 and 2018. To accomplish that, a quest for the expressions “sexual violence” and “sexual abuse” was made within the sites, such as the words “Olympic” and “olympism” were also verified. The results show 97 reports in total, mostly in the year of 2018 (56.72% of the total). The number of new reports grows expressively in 2016, that had 11 cases, whereas 2015 had only 3. The numbers kept growing in the following years (2017 showed 26 cases and in 2018, 55 cases were found).  Another thing we should draw attention to, is the number of reports in which the victim sex wasn't identified (44 cases or 45.36% of the reports). Of the 71.13% mapped reports involved Olympic gymnastics, 16.49% were about swimming and 4.12% were related to basketball. A possible explanation for the increasing of report news, could be the Rio’s Olympics of 2016 which gave visibility to several sporting issues, such as denouncing cases of sexual violence in different modalities. This article expects to contribute with the visibility of the subject within the scope of sports, making space for new articles to explore this theme.El objetivo de esta investigación fue mapear los casos de violencia sexual en el deporte olímpico, que fueron divulgados en diferentes sitios deportivos (Folha Deporte, ESPN, GazetaDeportiva , El País, Globo Deporte, Lance !, SporTV y Tierra), entre los años de 2013 y 2018. Para ello se realizó una búsqueda por reportajes sobre el tema en los sitios citados, que contenían en su cuerpo los términos "violencia sexual" o "abuso sexual" teniendo también una de las siguientes palabras: "olímpico", "olímpica", "olímpica" "olimpiadas" o "olimpismo". Los resultados mostraron un total de 97 reportajes, predominantemente en el año 2018 (56,72% del total). Hay un crecimiento expresivo en el número de reportajes a partir del año 2016 (saliendo de 3 casos en 2015 a 11 casos en 2016), tal número sub aun en los años subsiguientes (26 casos en 2017 y 55 casos en 2018). Nos llamo la atención el número de reportajes en que no hay la exposición del sexo de la víctima (44 casos, el 45,36% del total de reportajes). De los reportajes mapeados 71,13% corresponden a casos envolviendo la Gimnasia Artística, 16,49% a Natación y 4,12% el Baloncesto. El aumento de los reportajes tal vez pueda ser explicado en función de las Olimpiadas Río 2016, que dio visibilidad a diversas situaciones del deporte, tales como, las denuncias de los casos de violencia sexual en distintas modalidades. Se espera que este trabajo pueda agregar a la visibilidad del tema en el ámbito del deporte, abriendo caminos para que nuevas pesquisas puedan explorar la temática.O objetivo desta pesquisa foi mapear os casos de violência sexual nos esportes olímpicos divulgados em diferentes sites esportivos (Folha Esporte, ESPN, Gazeta Esportiva e El País, Globo Esporte, Lance!, SporTV e Terra), entre os anos de 2013 e 2018. Para tal, as pesquisadoras buscaram reportagens que tratavam sobre tal temática nos sites citados. Essas notícias precisavam conter em seus corpos os termos “violência sexual” ou “abuso sexual”, bem como uma das seguintes palavras: “olímpico”, “olímpica”, “olimpíadas” ou “olimpismo”. Os resultados mostraram o total de 97 reportagens, predominantemente no ano de 2018 (56,72% do total). Percebeu-se que há um crescimento expressivo do número de reportagens a partir do ano de 2016 (saindo de 3 casos em 2015 para 11 casos em 2016), tal número aumenta ainda nos anos subsequentes (26 casos em 2017 e 55 casos em 2018). Chamou-nos a atenção o número de reportagens em que não há a exposição do sexo da vítima (44 casos, 45,36% do total de reportagens). Das reportagens mapeadas, 71,13% correspondem a casos envolvendo a Ginástica Olímpica, 16,49% a Natação e 4,12% o Basquete. O aumento das reportagens se deu, provavelmente, em função das Olimpíadas no Rio de Janeiro, em 2016, que deu visibilidade para diversas questões do esporte, tal como as denúncias dos casos de violência sexual em diferentes modalidades. Espera-se que este trabalho possa contribuir para a visibilidade do tema no âmbito do esporte, abrindo caminhos para que novas pesquisas possam explorar a temática

Ultrasound B-lines for detection of late lung fibrosis in breast cancer patients after radiation therapy

Background and purpose. Radiation therapy (RT) for breast cancer after conservative surgery can be life-saving but remains associated with significant late side effects, including lung fibrosis, detected by chest CT. Aim of this study was to assess whether lung ultrasound (LUS) may detect late lung fibrosis through the biomarker of B-lines. Materials and methods. We evaluated 30 women (median age 67 years, range 46-80 years) about 3-8 years after RT (follow up 38-101 months, median 58 months) for left (n = 12) or right (n = 18) breast cancer (stage 1, n = 24; stage 2, n = 6), treated with total dose 40.5 – 50.00 Gy with/without boost dose). In all, both treated and contralateral hemithorax were evaluated. LUS was performed and B-lines evaluated with a 28-region antero-lateral scan, from second to fifth intercostal spaces, along the mid-axillary, anterior axillary, mid-clavicular, and arasternal lines. In each space, the B-lines were counted from 0 = black lung to 10 = white lung. The sum of B-lines in all spaces generated the B-line score of each hemithorax. Results. Median B-line score was higher in the irradiated site than in the contralateral control hemithorax (9, 1st-3rd quartiles: 2-23 vs 3, 1st-3rd quartiles: 1-4; P median value of 2.7 Gy) were associated with more B-lines when compared to lower doses (< 2.7 Gy): 9 vs 5, p < 0.001. Conclusion. RT in female breast cancer survivors is associated with increase in B-lines in the targeted hemithorax, likely due to lung fibrosis, and related to the lung mean dose. LUS can provide a simple “echo-marker” of lung fibrosis.

Validation of Reference Genes for the Relative Quantification of Gene Expression in Human Epicardial Adipose Tissue

BACKGROUND: Relative quantification is a commonly used method for assessing gene expression, however its accuracy and reliability is dependent upon the choice of an optimal endogenous control gene, and such choice cannot be made a priori. There is limited information available on suitable reference genes to be used for studies involving human epicardial adipose tissue. The objective of the current study was to evaluate and identify optimal reference genes for use in the relative quantification of gene expression in human epicardial fat depots of lean, overweight and obese subjects. METHODOLOGY/PRINCIPAL FINDINGS: Some of the commonly used reference genes including 18S, ACTB, RPL27, HPRT, CYCA, GAPDH, RPLPO, POLR2A and B2M were quantified using real-time PCR analysis. The expression stability of these genes was evaluated using Genorm, Normfinder and Bestkeeper algorithms. In addition, the effect of sample size on the validation process was studied by randomly categorizing subjects in two cohorts of n = 2 and n = 33. CONCLUSIONS/SIGNIFICANCE: CYCA, GAPDH and RPL27 were identified as the most stable genes common to all three algorithms and both sample sizes. Their use as reference gene pairs might contribute to the enhanced robustness of relative quantification in the studies involving the human epicardial adipose tissue

Active Whey Protein Edible Films and Coatings Incorporating Lactobacillus buchneri for Penicillium nordicum Control in Cheese

Fungal contamination of food is responsible for health issues and food waste. In this work, the incorporation of a lactic acid bacteria (LAB) with antifungal properties (Lactobacillus buchneri UTAD104) into whey protein-based films and coatings was tested for the control of an ochratoxigenic fungi (Penicillium nordicum) in a cheese matrix. The incorporation of L. buchneri cells resulted in thicker films with less luminosity than control films and colour alteration. Nevertheless, cells inclusion did not alter moisture content, water vapour permeability, mechanical properties, hydrophobicity and chemical structure of the films. Whey protein films were able to maintain the viability of L. buchneri UTAD104 cells in 105 CFU/mL after 30 days of storage at 25 \textdegreeC. When applied in cheese, films and coatings containing L. buchneri cells prevented fungal contamination for at least 30 days, while control cheeses with films and coatings either without LAB or with Lactobacillus casei UM3 (a strain without antifungal ability) showed fungal contamination during that period. Ochratoxin A was not found in cheeses treated with films and coatings containing L. buchneri UTAD104. Results showed that the inclusion of a LAB with antifungal properties in edible films and coatings can help to reduce or eliminate P. nordicum contamination in cheeses.This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. Ana Guimarães received support through grant SFRH/BD/103245/2014 from the Portuguese FCT.info:eu-repo/semantics/publishedVersio