Avaliação do papel do extrato de diospyros kaki L. : (diospiro) na prevenção da inflamação e cancro

Tese de doutoramento, Farmácia (Bromatologia), Universidade de Lisboa, Faculdade de Farmácia, 2019Vários estudos têm mostrado que os processos oxidativos e inflamatórios estão fortemente implicados na etiologia das doenças crónicas. A ingestão de frutos e produtos hortícolas, fontes de vitaminas e compostos fenólicos (CF) com ações antioxidantes e outras, podem ter um papel primordial na proteção da saúde e prevenção da doença. O dióspiro (Diospyros kaki L.) é um fruto rico em CF especialmente proantocianidinas. O objetivo deste trabalho é caracterizar quimicamente o extrato de dióspiro, em especial a sua composição fenólica, averiguar as suas atividades antioxidante, antiinflamatória e antiproliferativa e ainda preparar uma formulação farmacêutica oral deste extrato. A caracterização química do extrato por HPLC-DAD-MS/MS mostrou a sua riqueza em glucose e frutose, L-triptofano e ácidos málico e cítrico. Foram ainda identificados ácidos fenólicos benzóicos e derivados do ácido gálhico e do ácido vanílico bem como procianidinas e flavanóis, derivados glicosilados de flavonas e derivado de flavonol. A atividade antioxidante do extrato avaliada por ensaios de ORAC e HORAC é menor que a de alguns frutos vermelhos mas superiores à de maçãs e morangos. Os ensaios de CAA e burst oxidativo dos neutrófilos mostraram atividade antioxidante superior à das laranjas, peras ou maçãs. A dose oral de extrato de 15mg EAG/Kg mostrou atividade anti-inflamatória em modelo animal de inflamação aguda (edema da pata induzido pela carragenina) e nos modelos animais de inflamação crónica de artrite reumatóide e de colite ulcerosa. O extrato fenólico de dióspiro reduziu a invasão e proliferação de células de adenocarcinoma do cólon HT-29 e inibiu ligeiramente as atividades de MMPs com efeito dependente da dose. Os fitossomas com extrato, de dimensão inferior a 300nm, conseguem encapsular 97,4% do total de CF e são monodispersos e com potencial zeta negativo e apresentam maior atividade antioxidante durante 6 meses sob condições de armazenamento de T e RH estabelecidas do que o extrato livre. Assim, o extrato fenólico de dióspiro mostrou propriedades antioxidantes, antiinflamatórias e efeito anti-proliferativo nas células de cancro de cólon. A incorporação em fitossomas é uma alternativa para produção de suplemento alimentar.Several studies have shown that oxidative and inflammatory processes are strongly implicated in the etiology of chronic diseases. Ingestion of fruits and vegetables, sources of vitamins and phenolic compounds (PC) with antioxidant and other actions, may play a primary role in protecting health and preventing disease. The persimmon (Diospyros kaki L.) is a fruit rich in PC, especially proanthocyanidins. The aim of this work is to chemically characterize the persimmon extract, in particular its phenolic composition, ascertain its antioxidant, anti-inflammatory and antiproliferative activities and to prepare an oral pharmaceutical formulation of this extract. The chemical characterization of the extract by HPLC-DAD MS/MS showed its richness in glucose and fructose, L-tryptophan and malic and citric acids. Also identified were benzoic phenolic acids and derivatives of gallic acid and vanillic acid as well as procyanidins and flavanols, glycosylated derivatives of flavones and flavonol derivative. The antioxidant activity of the extract evaluated by ORAC and HORAC assays is lower than that of some red fruits but higher than that of apples and strawberries. The CAA and oxidative burst assays of neutrophils showed antioxidant activity superior to that of oranges, pears or apples. The oral dose of 15mg GAE/kg extract showed anti-inflammatory activity in an animal model of acute inflammation (carrageenan-induced paw edema) and in animal models of chronic inflammation of rheumatoid arthritis and ulcerative colitis. The phenolic persimmon extract reduced the invasion and proliferation of HT29 colon adenocarcinoma cells and slightly inhibited MMPs activities with dosedependent effect. Phytosomes with a size smaller than 300nm can encapsulate 97.4% of total PC and are monodisperse and with negative zeta potential and present higher antioxidant activity for 6 months under established T and RH storage conditions than the free extract. Thus, the persimmon phenolic extract showed antioxidant, anti inflammatory and anti-proliferative effect in colon cancer cells. The incorporation in phytosomes is an alternative for food supplement production.Universidade de Lisboa, REITORIA/BD/FF01/201

Actinic keratosis treated by topical Aloe barbadensis Mill. (Aloe Vera) leaf gel

Aloe vera is a cactus-resembling, succulent, watery plant, extensively used by the cosmetic and food industries[1]. Actinic keratosis, a dysplastic skin lesion commonly found in low Fitzpatrick skin type individuals[2] associated with chronic UV exposure[3,4], is defined by a scaly, keratotic or pigmented papule on an erythematous base[5]. The precursor to skin field cancerization is actinic keratosis, which is an area of photodamaged skin containing subclinical genetic changes, a direct consequence of intraepithelial UV-induced damage[6,7]. Actinic keratosis, as a precancerous lesion, can develop into an invasive squamous cell carcinoma (ISCC)[6]. There is no way to predict which lesions will progress to ISCC, with guidelines recommending treatment of all actinic keratosis lesions[8- 11]. Field-directed therapies are used to treat multiple actinic keratosis lesions and contiguous field cancerization subclinical lesions[12]info:eu-repo/semantics/publishedVersio

Attenuation of colonic injury and inflammation by administration of a phenolic extract of summer savory (Satureja hortensis l.) in experimental inflammatory bowel disease in mice

UIDB/04077/2020Summer Savory (Satureja hortensis L.) is a plant traditionally used as a food spice in the Mediterranean region. Surprisingly, not much is known about the health beneficial effects of its phenolic-rich extracts. The majority of publications have always focused on the properties of their essential oil. One of the main phenolic compounds of Summer Savory is rosmarinic acid, which has demonstrated anti-inflammatory outcomes in several animal models of inflammatory-mediated diseases. Inflammatory Bowel Disease (IBD) is a chronic inflammatory disease, in addition to Ulcerative Colitis and Crohn’s Disease, frequently related with increased morbidity and even mortality due to the complications associated, including colorectal cancer. Our work has shown, to our knowledge, for the first time, that administration of a phenolic extract of Summer Savory in a mouse model of Ulcerative Colitis led to the reduction of several markers for intestinal injury, including reduction of inducible nitric oxide synthase (iNOS) and Cyclooxygenase-2 (COX-2 or prostaglandin-endoperoxide synthase) expression, two well-known mediators of tissue inflammation and progression to cancer and led also to a reduction of the mortality. Given the chemical constitution found in the extract and the preclinical evidence of a beneficial effect of polyphenols in inflammatory processes, an opportunity arises for pharmacological modulation of pathways relevant for IBD and progression to cancer with phenolic-rich extracts.publishersversionpublishe

Preparation and preliminary demonstration of in vivo tolerability

This research was funded by Universidade de Lisboa PhD grant: REITORIA/BD/FF01/2015. supported in part through Grant UID/DTP/04138/2019 from Fundação para a Ciência e a Tecnologia (FCT), Portugal.Persimmon (Diospyros kaki L.), a fruit rich in phenolic compounds (PCs), has been considered effective in mitigating oxidative damage induced by an excess of reactive oxygen species. Due to large molecular weight and intrinsic instability in some physiological fluids, PCs’ passage through biological membranes is very limited. Carriers like phytosomes are promising systems to optimize oral absorption of encapsulated extracts. This work prepared and fully characterized phytosomes containing bioactive phenolic extracts from persimmon in terms of size, surface charge, encapsulation efficiency and stability over six months. These phytosomes were orally dosed to Wistar rats during a 15-day period. Afterwards, haematological and biochemical analyses were performed. Monodisperse phytosomes were successfully prepared, with size less than 300nm (PI < 0.3) and high encapsulation efficiency (97.4%) of PCs. In contrast to free extract, extract-loaded phytosomes had higher antioxidant activity after 6 months storage. Oral administration of extract-loaded phytosomes and free extract did not lead to lipidic profile changes and were within referenced normal ranges, as well as glycaemia levels and urine parameters. The results highlighted the potential of persimmon PCs as food supplements or pharmacological tools, suggesting a promising and safe phytosomal formulation containing bioactive agents of persimmon that could lead to health benefits.publishersversionpublishe

Reduction of inflammation and colon injury by a Pennyroyal phenolic extract in experimental inflammatory bowel disease in mice

Purpose: Little is known about the pharmacological effects of the phenolic compounds of Pennyroyal (Mentha pulegium). This Mediterranean aromatic plant, used as a gastronomic spice and as food preservative by the food industry has been studied mainly due to its essential oil antibacterial properties, composed primarily by monoterpenes. With this work, we aimed to evaluate the effects of a phenolic extract of pennyroyal in the impairment of inflammatory processes in Inflammatory Bowel Diseases (IBD) and in the potential inhibition of progression to colorectal cancer (CRC). Methods: To that purpose, we evaluated the effect of pennyroyal extract administration in a model of TNBSinduced colitis in mice and further determined its effect on human colon carcinoma cell proliferation and invasion. Results: The phenolic extract of pennyroyal exhibited antioxidant properties in in vitro assays and administration of the extract in a rat model of carrageenan-induced paw oedema led to significant anti-inflammatory effects. Further results evidenced a beneficial effect of the phenolic extract in the attenuation of experimental colitis and a potential antiproliferative effect on cultured colon cancer cells, effects not previously described, to our knowledge. A reduction in several markers of colon inflammation was observed following administration of the extract to colitis-induced mice, including functional and histological indicators. A successful inhibition of cancer cell invasion and proliferation was also observed in in vitro studies with HT-29 cells. Furthermore, the extract also led to a reduced expression of iNOS/COX-2 in the colon of colitis-induced mice, both being crucial mediators of intestinal inflammation. Conclusions: Taking into consideration the central role of inflammation in the pathophysiology of CRC and the recognised connection between inflammatory events and cancer, these results enlighten the relevance of the phenolic constituents of pennyroyal as important pharmacological sources in the investigation of new treatment options for patients with inflammatory bowel diseasesinfo:eu-repo/semantics/publishedVersio

Reduction of Inflammation and Colon Injury by a Spearmint Phenolic Extract in Experimental Bowel Disease in Mice

Background: Inflammatory Bowel Diseases (IBD) encompasses both Crohn’s Disease and Ulcerative Colitis, known to be connected to an enlarged risk for developing colorectal cancer (CRC). Spearmint (Mentha spicata L.) is a Mediterranean plant used as an aromatic agent, and studies have mainly focused on the essential oil suggesting an anti-inflammatory activity. This work aimed to perform a preliminary screening of the in vivo anti-inflammatory effects of a spearmint phenolic extract in an acute inflammation model, in a chronic inflammation model of colitis, and also study the effects in vitro on a colon cancer model. Methods: Spearmint extract was administered to rats of a paw oedema model (induced by carrageenan) and to mice from a TNBS-induced colitis model in parallel with studies using HT-29 CRC cells. Results: Administration of the extract led to reduced paw inflammation, reduction of colon injury and inflammation, with attenuation of histological markers, and reduction of iNOS expression. It repressed the in vitro movement of HT-29 cells in a wound healing assay. Conclusions: These findings suggest that spearmint extract exhibits acute and chronic anti-inflammatory activity and is able to inhibit migration of cancer cells, suggesting a potential role in the supplementary therapy of IBD patients.publishe

Caracterização química e avaliação da actividade biológica da framboesa (Rubus Idaeus L.) : Contribuição para o desenvolvimento de uma alegação de saúde

info:eu-repo/semantics/publishedVersio

Prototype ATLAS IBL Modules using the FE-I4A Front-End Readout Chip

The ATLAS Collaboration will upgrade its semiconductor pixel tracking detector with a new Insertable B-layer (IBL) between the existing pixel detector and the vacuum pipe of the Large Hadron Collider. The extreme operating conditions at this location have necessitated the development of new radiation hard pixel sensor technologies and a new front-end readout chip, called the FE-I4. Planar pixel sensors and 3D pixel sensors have been investigated to equip this new pixel layer, and prototype modules using the FE-I4A have been fabricated and characterized using 120 GeV pions at the CERN SPS and 4 GeV positrons at DESY, before and after module irradiation. Beam test results are presented, including charge collection efficiency, tracking efficiency and charge sharing.Comment: 45 pages, 30 figures, submitted to JINS