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Research Center in Natural Resources, Environment and Society

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Aloysia citrodora extract as a chemopreventive agent against HPV16-induced lesions: findings from K14-HPV16 mice
Publication . Medeiros-Fonseca, Beatriz; Faustino-Rocha, Ana; Silva, Jessica; Silva, Mónica; Pires, Maria João; Neuparth, Maria João; Vala, Helena; Vasconcelos-Nóbrega, Cármen; Dias, Maria Inês; Barros, Lillian; Gonçalves, Lio; Gaivão, Isabel; Bastos, Margarida M.S.M.; Félix, Luís; Venâncio, Carlos; Medeiros, Rui; Costa, Rui Miguel Gil; Oliveira, Paula A.
Aloysia citrodora has a long history of traditional use in treating various ailments. This study evaluated the in vivo chemopreventive efficacy and systemic toxicity of an extract of A. citrodora in a transgenic mouse model of HPV16 (human papillomavirus type 16)-induced cancer. Methods: The experiment involved six groups (n = 5): group 1 (G1, wild-type (WT), water), group 2 (G2, HPV, water), group 3 (G3, WT, 0.013 g/mL), group 4 (G4, HPV, 0.006 g/mL), group 5 (G5, HPV, 0.008 g/mL), and group 6 (G6, HPV, 0.013 g/mL). Throughout the assay, humane endpoints, body weight, food, and water consumption were recorded weekly. The internal organs and skin of the mice were collected for analysis after they were sacrificed. Toxicological parameters that were studied included hematological and biochemical blood markers, splenic and hepatic histology, and hepatic oxidative stress. Results: A. citrodora extract seems to reduce the incidence of dysplastic and in situ carcinoma skin lesions induced by HPV16 in this model, suggesting that dietary supplementation with concentrations of 0.008 g/mL and 0.013 g/mL may have beneficial chemopreventive effects. Conclusions: The extract did not induce any concentration-dependent toxicological effects on any of the parameters included in the study, indicating a favorable toxicological profile under these experimental conditions.
Evaluation of FT-Raman and FTIR-ATR spectroscopy for the quality evaluation of Lavandula spp. Honey
Publication . Anjos, Ofélia; Guiné, Raquel P.F.; Santos, António J.A.; Paula, Vanessa B.; Pereira, Helena; Estevinho, Leticia M.
Monofloral Lavandula spp. honey is very appreciated by consumers due to its characteristic and pleasant aroma and flavor. Given the economic importance of this type of honey, it is important to develop a rapid and non-expensive methodology that allows certifying its quality. In this context, this study aimed to compare the applicability and accuracy of FTIR-ATR and FT-Raman techniques for the quality evaluation of Lavandula spp. honey. Calibration models, with PLS regression models, were obtained for both methodologies concerning the following parameters: total acidity, reducing sugars, hidroximetilfurfural (HMF), electrical conductivity, ash, proline content, diastase activity, apparent sucrose, total flavonoids, and total phenolic contents. The calibration models had high regression coefficients, r2 (FTIR-ATR: 0.965–0.996; FT-Raman: 0.983–0.999), high ratios of performance to deviation, RPD (FTIR-ATR: 5.4–15.7; FT-Raman: 7.6–53.7), and low root mean square errors (RMSEs; FTIR-ATR: 0.005–3.0; FT-Raman: 0.004–1.02). These results corroborate the potentiality of FTIR-ATR and FT-Raman for quality evaluation and evaluation of the chemical properties of Lavandula spp. honey even though FT-Raman technique provided more accurate models.
Pulmonary and Gastrointestinal Parasitic Infections in Small Ruminant Autochthonous Breeds from Centre Region of Portugal—A Cross Sectional Study
Publication . Pereira, Maria; Vila-Viçosa, Maria João M.; Coelho, Catarina; Santos, Carla; Esteves, Fernando; Cruz, Rita; Gomes, Liliana; Henriques, Diogo; Vala, Helena; Nóbrega, Carmen; Mega, Ana; Melo, Carolina de; Malva, Madalena; Braguez, Joana; Mateus, Teresa
The production of small ruminant autochthonous breeds in the Centre region of Portugal is practiced in a semi-extensive husbandry system, exposing animals to parasitic infections. The main objective of this study was to estimate the prevalence of lungworm infection and identify risk factors. Fecal samples of 203 goats and 208 sheep from 30 herds were collected per rectum and subjected to the modified Baermann test. The overall prevalence of infection was 57.7%, significantly higher in goats (95.6%) than in sheep (20.7%) (p < 0.001). According to the binary logistic regression model, sheep dewormed with albendazole, mebendazole plus closantel, or ivermectin plus clorsulon presented a risk of Protostrongylidae infection 29.702, 7.426, or 8.720 times higher, respectively, than those dewormed with eprinomectin. Additionally, the presence of gastrointestinal parasites was investigated in 307 fecal samples using Mini-FLOTAC (R). The overall prevalence of infection was 86.3%, also significantly higher in goats (93.2%) than in sheep (79.9%) (p < 0.001). Strongyle-type eggs were the most frequently identified, both in sheep (69.8%) and goats (87.8%), followed by Eimeria oocysts (40.3% in sheep and 68.9% in goats). Considering the high prevalence and the burden of lungworm parasitic infection, it is urgent to determine its economic impact and the repercussions in animal health in the Centre region of Portugal to establish appropriate therapeutic guidelines.
Effect of moisture on the characteristics of hazelnut kernel during storage
Publication . Correia, Paula; Filipe, Ana Filipa; Ferrão, Ana Cristina; Ramalhosa, Elsa; Guiné, Raquel P.F.
The organoleptic and nutritional quality of hazelnuts during storage can be greatly influenced by environment conditions. The aim of this study was to evaluate the effect of moisture on the physicochemical characteristics of hazelnut kernels when stored under controlled conditions of temperature (25 °C) and different relative humidities (RH) (57.6, 68.9, and 78.6%). To obtain these RH in the hazelnut storage atmospheres, the salts NaBr, KI, and NH4Cl were used, respectively. The following properties were evaluated: moisture, water activity (aw), color, texture, and fat oxidative stability, both in the initial sample and after 1, 2, 4 and 6 months of storage. The results showed that there was a progressive increase over time in moisture and water activity, with growth being most pronounced from 4 months onwards. Moreover, hazelnuts stored at a RH of 78.6%, at 6 months of storage were completely spoiled, making their evaluation of impossible. The hardness and friability of the hazelnuts decreased over the storage time, with the differences being more pronounced for the samples subjected to higher RH. In general, the color of the skin and core of the fruits showed little variation over the storage time for all parameters evaluated (L*, a*, b*) and for all RH conditions tested. The hazelnuts stored at a RH of 78,6% could only be preserved until the 4th month, but presented low oxidative stability, being comparable to the hazelnuts with 6 months at RH of 57,6%, meaning that with a reduction of about 10% of RH it is possible to increase the oxidative stability about 2 months. Thus, it can be concluded that high relative humidities lead to a lower conservation of the quality characteristics of hazelnuts. Moreover, during the 6 months of storage the color of the fruits was little affected for the different RH tested, however all the other evaluated characteristics presented significant alterations, namely the texture and the oxidative stability of the fats.
Exploring the therapeutic potential of Quercus ilex acorn extract in papillomavirus-induced lesions
Publication . Medeiros-Fonseca, Beatriz; Faustino-Rocha, Ana; Pires, Maria João; Neuparth, Maria João; Vala, Helena; Vasconcelos-Nóbrega, Cármen; Gouvinhas, Irene; Barros, Ana Novo; Dias, Maria Inês; Barros, Lillian; Bastos, Margarida M.S.M.; Gonçalves, Lio; Félix, Luís; Venâncio, Carlos; Medeiros, Rui; Costa, Rui Miguel Gil; Oliveira, Paula A.
Papillomaviruses (PVs) infections have been documented in numerous animal species across different regions worldwide. They often exert significant impacts on animal health and livestock production. Scientists have studied natural products for over half a century due to their diverse chemical composition, acknowledging their value in fighting cancer. Acorns (Quercus ilex) are believed to have several unexplored pharmacological properties. This study aimed to evaluate the in vivo safety and cancer chemopreventive activity of an infusion extract of Q. ilex in a transgenic mouse model of human PV (HPV)-16, which developed squamous cell carcinomas through a multistep process driven by HPV16 oncogenes. Q. ilex extract was prepared by heating in water at 90°C and then characterized by mass spectrometry. Phenolic compounds from this extract were administered in drinking water to female mice in three different concentrations (0.03, 0.06, and 0.09 g/mL) over a period of 28 consecutive days. Six groups (n = 6) were formed for this study: group 1 (G1, wildtype [WT], water), group 2 (G2, HPV, water), group 3 (G3, WT, 0.09 g/mL), group 4 (G4, HPV, 0.03 g/mL), group 5 (G5, HPV, 0.06 g/ mL), and group 6 (G6, HPV, 0.09 g/mL). Throughout the experiment, humane endpoints, body weight, food intake, and water consumption were recorded weekly. Following the experimental period, all mice were sacrificed, and blood, internal organs, and skin samples were collected. Blood was used to measure glucose and microhematocrit and later biochemical parameters, such as creatinine, urea, albumin, alanine aminotransferase, and total proteins. Histological analysis was performed on skin and organ samples. The administration of Q. ilex extract resulted in a statistically significant increase in relative organ weight among HPV transgenic animals, indicating adaptive biological response to the tested concentrations. Moreover, a reduction in characteristic skin lesions was observed in animals treated with the 0.06 and 0.09 g/mL extract. These results provide a favorable chemopreventive profile for Q. ilex extract at concentrations of 0.06 and 0.09 g/mL. This study highlights the potential of Q. ilex extract as a safe and effective therapeutic strategy against HPV16- associated lesions in transgenic mouse models. The limitation of our study was the durability of transgenic animals. As a more sensitive species, we must always be careful with the durability of the test. We intend to study concentrations of 0.06 and 0.09 g/mL for longer to further investigate their possible effects.

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Funding agency

Fundação para a Ciência e a Tecnologia

Funding programme

6817 - DCRRNI ID

Funding Award Number

UIDB/00681/2020

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