Browsing by Author "Lima, Ana"
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- Actinic keratosis treated by topical Aloe barbadensis Mill. (Aloe Vera) leaf gelPublication . Direito, Rosa; Ferreira, João Boavida; Ferreira, Ricardo Boavida; Lima, AnaAloe 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]
- An up-scalable and cost-effective methodology for isolating a polypeptide matrix metalloproteinase-9 inhibitor from Lupinus albus seedsPublication . Mota, Joana; Figueira, Maria E.; Ferreira, Ricardo Boavida; Lima, AnaOne of the most challenging problems with food-borne bioactive compounds is that there are commonly no cost-effective, generally recognized as safe (GRAS) methods for obtaining gram quantities of their purified forms. Here we aimed at developing a method to isolate deflamin, an oligomeric protein from lupin seeds with anti-inflammatory and anticancer activity through matrix metalloprotease (MMP)-9 inhibition. Our goal was to develop a GRAS method that could be easily up-scalable whilst maintaining deflamin’s activity. A sequential precipitation methodology was developed, using an aqueous extraction, followed by heat denaturation, acid precipitation and solubilization in ethanol. A final precipitation with 90% ethanol yielded a purified protein which was sequenced through mass spectrometry and tested for its MMP inhibitory activity using the Dyequenched (DQ) gelatin assay and the standard wound healing assay in HT29 cells. The developed method yielded a purified oligomer, which represented 0.1% (w/w) of total dry seed weight and was positively confirmed to be deflamin. It further showed to effectively reduce MMP-9 gelatinolytic activity as well as colon cancer cell migration, hence corroborating the effectiveness of our method. Overall, this is the first reported method for isolating an MMP-9 inhibitor from legume seeds, which is up-scalable to an industrial level, in a cost-effective manner
- Azorean Cryptomeria japonica immature female cones essential oil: Effect of hydrodistillation fractionation on the chemical composition and in vitro antifungal activity against Thielaviopsis paradoxaPublication . Arruda, Filipe; Lima, Ana; Janeiro, Alexandre; Rodrigues, Tânia; Baptista, José; Rosa, José S.; Machado, Alexandra; Figueiredo, Ana Cristina; Lima, ElisabeteCryptomeria japonica's wood production in Azores generates large amounts of underutilized biomass residues, such as immature female cones (Az–CJIFC), that can be used to produce essential oils (EOs). Hydrodistillation (HD) can be used both to obtain and to fractionate EOs. In this study, EOs from Az–CJIFC, grinded (GR) and non-grinded (NGr) fresh samples, were obtained via HD over 4 h, yielding 1.0 % and 0.5 %, w/w, respectively. Thus, GR Az–CJIFC was chosen to obtain six EO fractions (Frs. 1–6), collected at sequential HD timeframes (HDTs: 0–2, 2–10, 10–30, 30–60, 60–120, and 120–240 min). The obtained EO samples (crude EOs and fractions) were evaluated for their chemical composition (GC–FID/GC–MS analyses) and antifungal activity (micro-atmosphere method) against phytopathogenic fungi (Penicillium italicum, P. digitatum and Thielaviopsis paradoxa). Results indicated that all samples were active only towards T. paradoxa, however, with a differential efficacy, due to their specific composition. Fraction 4 vapor treatment displayed the strongest activity, but lower than that of (–)-terpinen-4-ol, a key oxygen-containing monoterpene (OCM) of Az–CJIFC EO, peaking its concentration in Fr3 (14.5 %) and Fr4 (13.8 %). This latter fraction was the richest in the OCM α-terpineol (a minor Az–CJIFC EOs' component). On the other hand, Frs.1–3 and EOs were dominated by monoterpene hydrocarbons (65.0–96.5 %), mainly α-pinene (19.0–28.4 %) and sabinene (19.9–50.5 %), while Frs.5 and 6 were the richest in oxygen-containing sesquiterpenes (47.1–70.8 %; chiefly elemol plus α-, β- and γ-eudesmol) and diterpene hydrocarbons (5.2–6.4 %; mostly phyllocladene). In conclusion, new high value-added products can now be targeted in Az–CJIFC EO by adjusting the HDT, with potential importance in pineapple fruit black rot disease management caused by T. paradoxa on Ananas comosus in the Azores, and also contributing for the local C. japonica’s EO industry development and sustainable circular bio-economy.
- Combination of trans-resveratrol and E-viniferin induces a hepatoprotective effect in rats with severe acute liver failure via reduction of oxidative stress and MMP-9 expressionPublication . Fernandes, João C.; Schemitt, Elizangela; Silva, Juliana; Marroni, Norma P.; Lima, Ana; Ferreira, Ricardo BoavidaStilbenes are a major grapevine class of phenolic compounds, known for their biological activities, including anti-inflammatory and antioxidant, but never studied in combination. We aimed to evaluate the effect of trans-resveratrol + "-viniferin as an antioxidant mixture and its role in inflammatory development an in vivo model of severe acute liver failure induced with TAA. Trans-resveratrol + trans-"-viniferin (5 mg/kg each) was administered to Wistar rats. Resveratrol + "- viniferin significantly decreased TBARS and SOD activity and restored CAT and GST activities in the treated group. This stilbene combination reduced the expression of TNF , iNOS, and COX-2, and inhibited MMP-9. The combination of resveratrol + "-viniferin had a hepatoprotective effect, reducing DNA damage, exhibiting a protective role on the antioxidant pathway by altering SOD, CAT, and GST activities; by downregulating TNF , COX-2, and iNOS; and upregulating IL-10. Our results suggested that adding viniferin to resveratrol may be more effective in hepatoprotection than resveratrol alone, opening a new perspective on using this stilbene combination in functional diets
- Deflamin Attenuated Lung Tissue Damage in an Ozone-Induced COPD Murine Model by Regulating MMP-9 Catalytic ActivityPublication . Baltazar-García, Elia Ana; Vargas-Guerrero, Belinda; Lima, Ana; Ferreira, Ricardo Boavida; Mendoza-Magaña, María Luisa; Ramírez-Herrera, Mario Alberto; Baltazar-Díaz, Tonatiuh Abimael; Domínguez-Rosales, José Alfredo; Salazar-Montes, Adriana María; Gurrola-Díaz, Carmen MagdalenaChronic obstructive pulmonary disease (COPD) is comprised of histopathological alter- ations such as pulmonary emphysema and peribronchial fibrosis. Matrix metalloproteinase 9 (MMP-9) is one of the key enzymes involved in both types of tissue remodeling during the development of lung damage. In recent studies, it was demonstrated that deflamin, a protein component extracted from Lupinus albus, markedly inhibits the catalytic activity of MMP-9 in experimental models of colon adenocarcinoma and ulcerative colitis. Therefore, in the present study, we investigated for the first time the biological effect of deflamin in a murine COPD model induced by chronic exposure to ozone. Ozone exposure was carried out in C57BL/6 mice twice a week for six weeks for 3 h each time, and the treated group was orally administered deflamin (20 mg/kg body weight) after each ozone exposure. The histological results showed that deflamin attenuated pulmonary emphysema and peribronchial fibrosis, as evidenced by H&E and Masson’s trichrome staining. Furthermore, deflamin administration significantly decreased MMP-9 activity, as assessed by fluorogenic substrate assay and gelatin zymography. Interestingly, bioinformatic analysis reveals a plausible interaction between deflamin and MMP-9. Collectively, our findings demonstrate the therapeutic potential of deflamin in a COPD murine model, and suggest that the attenuation of the development of lung tissue damage occurs by deflamin-regulated MMP-9 catalytic activity.
- Development of Healthy and Clean-Label Crackers Incorporating Apple and Carrot Pomace FloursPublication . Salari, Saeed; Castigliego, Thomas; Ferreira, Joana; Lima, Ana; Sousa, IsabelThe valorization of fruit and vegetable side-streams from the juice industry is an important contribution to the optimization of food resources and is an environmentally friendly practice in line with the concepts of circular economy and sustainability. The aim of this work is to incorporate them back into the food value chain by adding them as ingredients in staple foods like crackers. This is also important in terms of food fortification, as they are rich in nutrients and bioactive compounds. Crackers are popular snacks with a huge global market value, enjoyed by consumers of all ages. The current study aims to integrate flour from dried apple and carrot pomaces, resulting from juice processing, as natural ingredients with potential health benefits. The incorporation levels ranged from 20 to 40% dry weight in crackers, and their impact on physicochemical and mechanical properties was evaluated, as well as bioactivity (potential impact on health) and sensory acceptance. The addition of pomaces resulted in significant changes in texture and color, as well as enhancing the antioxidant activity of the crackers. Crackers containing pomace flours, except for the cracker with 40% carrot pomace, showed a high overall sensory acceptability and good intentions to buy.
- Differential inhibition of gelatinase activity in human colon adenocarcinoma cells by Aloe vera and Aloe arborescens extractsPublication . Lima, Ana; Batista-Santos, Paula; Veríssimo, Eduarda; Rebelo, Patrícia; Ferreira, Ricardo BoavidaBackground: Aloe’s reported bioactivities (anticancer, anti-inflammatory and wound healing) suggest they might inhibit a subgroup of matrix metalloproteinases (MMPs) called gelatinases (MMP-2 and MMP-9). The goal of the present study was to compare the MMP inhibitory potential of two Aloe species, A. vera and A. arborescens. Methods: Different types of extraction were tested and specific bioactive compounds were quantified. Cancer cell invasion inhibitory activities were measured in vitro using the wound healing assay in human colon cancer cells (HT29). Effects on gelatinase activities were further assessed by dye-quenched gelatin and gelatin zymography. Results: Different types of extraction yielded significantly different levels of bioactivities and of bioactive compounds, which might be due to a greater amount of extractable bioactive compounds such as anthraquinones. Both A. arborescens and A. vera have potential as inhibitory agents in cancer cell proliferation via MMP-9 and MMP-2 enzymatic activity inhibition, being able to reduce colon cancer cell proliferation and migration but A. arborescens showed to be a more effective inhibitor of cancer cell migration than A. vera. Conclusion: This work opens novel perspectives on the mode of action of Aloe species in cancer cell migration and may provide clues as to why there are so many conflicting results on Aloe’s activities
- Effects of particle size on physicochemical and nutritional properties and antioxidant activity of apple and carrot pomacesPublication . Salari, Saeed; Ferreira, Joana; Lima, Ana; Sousa, IsabelThe food processing industry is growing rapidly and producing large amounts of by- products, such as pomaces, which are considered as no-value waste and cause significant environ- mental pollution. The main by-products of fruit juice processing companies are apple and carrot pomaces, which can be used to create new functional food products. In the present study, the effects of particle size (PS) on the proximate composition, nutritional properties, and antioxidant activity of apple pomace flour (APF) and carrot pomace flour (CPF) were determined. Four different PS fractions, PS > 1 mm, 1 > PS > 0.71 mm, 0.71 > PS > 0.18 mm, and 0.18 > PS > 0.075 mm were used for the present study. Their vitamin, carotenoid, organic acid, and reducing sugar contents were determined using HPLC. The proximate compositions of each PS fraction of the AP and CP flours were determined using recommended international standard methods. DPPH, FRAP, and Folin- Ciocalteu methods were used to measure their antioxidant activity and total phenolic compounds, respectively. The moisture content (around 12.1 mg/100 g) was similar in all PS fractions and in both flours. The APF had lower protein (4.3–4.6 g/100 g dw) and ash (1.7–2.0 g/100 g dw) contents compared to the CPF, with protein contents ranging from 6.4–6.8 g/100 g dw and ash contents ranging from 5.8–6.1 g/100 g dw. Smaller particles, regardless of flour type, exhibited higher sugar and phenolic contents and antioxidant activity, while vitamins were more abundant in particles larger than 1 mm. In the APF, larger particles had a higher fiber content than smaller particles, while their fat content was the lowest. PS also had an impact on the results of the carotenoid contents. This study underscores the direct impact of PS on the distribution of sugars, crude fiber, fat, carotenoids, vitamins, total phenolic compounds, and antioxidant activity in pomaces.
- Exploring the Bioactive Properties and Therapeutic Benefits of Pear PomacePublication . Ferreira, Joana; Karolina Tkacz, Karolina; Turkiewicz, Igor Piotr; Santos, Isabel; Silva, Mariana Camoesas e; Lima, Ana; Sousa, IsabelThe fruit juice industry generates a significant amount of waste, with a strong impact on the environment and the economy. Therefore, researchers have been focusing on the characterization of resources considered as food waste. This work provides information about the lipophilic and polar metabolites of pear pomace flours (PPFs) as a tool that can shed more light on the bioactive potential of this residue. Using UPLC-PDA, UPLC-FLR, and GC-MS, the study identified and quantified PPF’s polar and non-polar metabolites. Essential, conditional, and non-essential amino acids were found, with asparagine being the most abundant. Isoprenoids, including lutein, zeaxanthin, and carotene isomers, ranged from 10.8 to 22.9 mg/100 g dw. Total flavonoids and phenolic compounds were 520.5–636.4 mg/100 g dw and 536.9–660.1 mg/100 g dw, respectively. Tocotrienols and to- copherols were identified, with concentrations of 173.1–347.0 mg/100 g dw and 468.7–913.4 mg/ 100 g dw . Fatty acids were the major non-polar compounds. All fractions significantly reduced matrix metalloproteinase-9 (MMP-9) activity. Although PPF had lower antioxidant potential (3–6 mmol Trolox/100 g dw), it inhibited AChE and BuChE by 23–30% compared to physostigmine salicylate. These findings suggest that pear pomace waste can be repurposed into functional products with valuable bioactive properties by re-introducing it in the food chain.
- Extended cheese whey fermentation produces a novel casein-derived antibacterial polypeptide that also inhibits gelatinases MMP-2 and MMP-9Publication . Santos, Maria Isabel; Lima, Ana; Mota, Joana; Rebelo, Patrícia; Ferreira, Ricardo Boavida; Pedroso, Laurentina; Ferreira, Maria Adélia; Sousa, IsabelOur previous works produced a whey fermentation methodology that yielded antibacterial activity and potential inhibition of matrix metalloproteases (MMP)-2 and -9. Here, we evaluated if these activities were due to fermentation-produced peptides. Prolonged fermentation was carried out in the presence of our specific lactic acid bacteria (LAB) consortium. LAB fermentation yielded a total of 11 polypeptides, which were predominantly produced after 6 days of fermentation. One which was derived from beat casein presented a particularly high antibacterial activity against food pathogenic bacteria and was more effective than standard food disinfectants. This polypeptide was further studied and was also found to be active against several strains of pathogenic bacteria, including methicillin-resistant Staphylococcus aureus (MRSA), in a dose-dependent manner. It also inhibited MMP-2 and MMP-9 whilst reducing HT29 cancer cell migration in vitro. Overall, this novel whey-derived polypeptide presents dual antibacterial and anti-inflammatory activity, revealing a strong potential to be used in functional foods or as a nutraceutical. Its identification and further characterization can open novel perspectives in the field of preventive/curative diets related to gut microbiota, gut inflammation, and cancer prevention, particularly if used in in vivo studies
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