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Lavandula penduculata (Mill.) Cav., common English name French lavender, belongs to the Lamiaceae family and has been used as a medicinal plant in infusions for respiratory and digestive systems and as a therapeutic agent with antiseptic action for cleaning wounds [1,2]. The K14HPV16 mice is a skin squamous carcinoma model that can be used to test the antitumoral properties of several chemical and natural products [3]. The aim of this work was to evaluate the effect of the hydroethanolic French lavender extract (FLE) on body weight, relative organs weights, food and water consumption in an HPV-16-transgenic mice model. The extract was obtained from a maceration with ethanol/water (80:20, v/v), and the phenolic composition was determined through HPLC-DAD-ESI/MS. Twenty-eight male mice were randomly divided into four groups (n=7/group) according to their genotype: group I (HPV16- control); II (HPV16- FLE); III (HPV16+ control) and IV (HPV16+ FLE). The FLE was administrated orally in drinking water at 6.8 mg/10mL/animal to animals from groups II and IV and changed every 4 days. The animals were kept under controlled conditions such as temperature, light and humidity This work was supported by European Investment Funds by FEDER/ COMPETE/POCI - Operational Competitiveness and Internationalization Program and National Funds by FCT - Portuguese Foundation for Science and Technology, under the projects Project UIDB/04033/2020 (CITAB), and PhD fellowship SFRH/BD/136747/2018 and 2020.04789.BD. The authors are also grateful to FCT for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020). L. Barros thanks FCT, P.I for her contract through the institutional scientific employment program. This work was also funded by the European Regional Development Fund (ERDF) through the Regional Operational Program North 2020, within the scope of Project GreenHealth - Norte-01-0145-FEDER-000042. The authors would like to thank BPGV for the samples provided. info:eu-repo/semantics/publishedVersion

The K14HPV16 mice is a skin squamous carcinoma model that can be used to test antitumoral properties of several chemical and natural compounds1. Lavandula penduculata (Mill.) Cav., known as lavender, belongs to the Laminaceae family and has been used in traditional medicine as infusions to treat several conditions2. This work aimed to evaluate the effects of the hydroethanolic French lavender extract (FLE) in an HPV16-transgenic mice model lesions. The extract was obtained through a maceration with ethanol/water (80:20, v/v) and its phenolic composition was determined by HPLC-DADESI/ MS. The FLE was dissolved in drinking water at 6.8 mg/10mL/animal and the animals were supplemented during 29 consecutive days. Twentyeight male mice were randomly divided into four groups: (n=7/group): group I (HPV16- control); II (HPV16- FLE); III (HPV16+ control) and IV (HPV16+ FLE). After 29 days all animals were sacrificed by xylazineketamine overdose following cardiac puncture to obtain blood samples. Skin samples (chest and ear), kidney, liver and spleen were processed for histological analysis.A total of thirteen compounds were identified in the hydroethanolic extract, being salvianolic acid B and rosmarinic acid the main molecules present. Moreover, the compounds revealed to be stable in the drinking water for 5 days. Histological analyses of skin samples from wild-type mice exposed (group II) and not exposed (group I) to FLE showed normal skin histology. Group III showed skin chest epidermal hyperplasia in 100% of the mice while group IV showed less epidermal hyperplasia frequency (66.6%) (p>0.05). Concerning to liver, kidney, and spleen lesions there are no differences between groups (p>0.05). The lavender extract did not prevent the progression of HPV-16 induced cutaneous lesions in this model. These data deserve more investigation to clarify the effect of lavender extract on HPV-16 lesions. This work was supported by European Investment Funds by FEDER/ COMPETE/POCI - Operational Competitiveness and Internationalization Program and National Funds by FCT - Portuguese Foundation for Science and Technology, under the projects Project UIDB/04033/2020 (CITAB), and PhD fellowship SFRH/BD/136747/2018 and 2020.04789.BD. The authors are also grateful to FCT for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020). L. Barros thanks FCT, P.I for her contract through the institutional scientific employment program. This work was also funded by the European Regional Development Fund (ERDF) through the Regional Operational Program North 2020, within the scope of Project GreenHealth - Norte-01-0145-FEDER-000042. The authors would like to thank BPGV for the samples provided. info:eu-repo/semantics/publishedVersion

Infection by Human Papillomavirus (HPV) is the main cause of cervical cancer, highlighting the importance of studying compounds that may reduce viral activity and its lesions/symptoms. The aromatic herb spearmint (Mentha spicata) (MS) has proven anti-tumor properties [1]. Thus, this study aimed to evaluate the effects of an hydroethanolic extract obtained from spearmint in HPV16-transgenic (HPV+/-) mice. The extract was obtained through maceration with ethanol/water (80:20, v/v), and the phenolic composition was determined through HPLC-DAD-ESI/MS. Thirty-three female mice (16 HPV-/- and 17 HPV+/-) were randomly divided into six groups: Group (G) I – HPV-/- w/o Mentha (n=5); G II – HPV-/- w/ Mentha (0.5 mg/ml; n=6); G III – HPV-/- w/ Mentha (0.55 mg/ml; n=5); G IV – HPV+/- w/o Mentha (n=6); G V – HPV+/- w/ Mentha (0.5 mg/ml; n=6); G VI – HPV+/- w/ Mentha (0.55 mg/ml; n=5). The spearmint extract was administered in the animal’s drinking water for 28 days. During the study, water and food intake as well the animals’ weights were recorded weekly. Afterwards, the animals were sacrificed, and their organs were collected for oxidative stress and genetic damage analysis. A total of thirteen compounds were identified in the hydroethanolic extract, being salvianolic acid B, rosmarinic acid and luteolin-7-O-glucuronide, the main compounds found. Moreover, the compounds revealed to be stable in the drinking water during the 5 tested days. Results show that HPV+/- (GIV, V and VI) have lower weight but higher water and food intake, as expected. Analysis of HPV+/- mice GIV showed a significant increase of superoxide dismutase activity when compared with GV (p=0.0029) and VI (p=0.0011). This suggests that group IV was subjected to a higher oxidative stress, as expected. These results could also mean that MS is responsible for a decrease in the oxidative stress known to be induced by HPV, which in turn decreases SOD, further the role of MS as an antioxidant. Regarding genetic damage, no statistically significant changes between groups were found in the comet and micronucleus assays, implying that spearmint has no influence on genotoxicity at the concentrations employed. There were no significant differences concerning basal DNA damage between WT and transgenic individuals as seen in other studies [2]. Further studies are required to clarify the antioxidant and antigenotoxic effects of spearmint. Interact R&D project, NORTE-01-0145-FEDER-000017, in its ISAC research line, ERDF-NORTE 2020. The authors are also grateful to FCT, for financial support through national funds FCT/MCTES to CIMO (UIDB/00690/2020); L. Barros thanks the national funding by FCT, P.I., through the institutional scientific employment program-contract. info:eu-repo/semantics/publishedVersion

Em, 1959 Russel e Burch propuseram os 3Rs (Reduce, Refine e Replacement) em experimentação animal e desde então as diversas entidades envolvidas na temática têm feito esforços conjuntos para os aplicar (1). Nesse sentido foi gradualmente introduzida a avaliação e registo dos humane endpoints (HE) dos animais durante procedimentos experimentais, para monitorizar o seu sofrimento e tomar decisões sobre a necessidade, ou não, de finalizar o ensaio antes da data prevista (2). Contudo, apesar das exigências legais, observa-se grande escassez de publicações sobre a avaliação de HE nos procedimentos com animais de laboratório. Assim, é nosso objetivo com este trabalho apresentar os HE durante a aplicação diária de um creme com Cytinus hypocistis numa estirpe de murganhos transgénicos. Financiamento nacional pela FCT, Fundação para a Ciência e a Tecnologia, no âmbito da bolsa de doutoramento 2020.07675.BD de Beatriz Medeiros Fonseca. info:eu-repo/semantics/publishedVersion

The Human Papillomavirus (HPV) is the prime cause of cervix cancer in women info:eu-repo/semantics/publishedVersion

O Papiloma Vírus Humano (HPV) é responsável pelo desenvolvimento do carcinoma do colo do útero, estando ainda associado a outros carcinomas. Para compreender melhor os mecanismos associados às infeções causadas por este vírus e estudar novas terapias foi desenvolvido um modelo transgénico de murganho: K14HPV16. Assim, o objetivo deste trabalho foi avaliar o efeito da administração oral do extrato hidroetanólico (80:20, v/v) de tília (Tilia platyphyllos Scop.) em murganhos machos e fêmeas K14HPV16 e wild type. Utilizaram-se 43 animais divididos em grupos de machos e fêmeas. Cada um destes grupos foi ainda dividido em dois grupos de animais K14HPV16 com e sem exposição oral à tília (4,5 mg/animal, calculado com base em trabalhos prévios feitos in vitro) e dois grupos de animais wild type com e sem tratamento. O extrato hidroetanólico foi caracterizado por HPLC-DAD-ESI/MS quanto ao perfil em compostos fenólicos e a sua estabilidade foi estudada ao longo de 5 dias consecutivos. (-)-Epicatequina, quercetina-3-O-glucósido e ácido protocatéquico, foram os compostos fenólicos maioritários identificados no extrato, estáveis durante 5 dias. Durante o ensaio experimental, com duração de 33 dias, foi registado o consumo de água, e as massas corporais individuais e consumo de comida. No final do ensaio todos os animais foram sacrificados, tendo sido recolhido sangue, para estudo bioquímico e danos oxidativos pelo ensaio dos cometas. Observou-se que o extrato de tília não provocou alterações no aspeto dos animais nem na média da massa corporal. O consumo médio de alimento foi idêntico entre grupos, os murganhos K14HPV16 consumiram mais água. Os parâmetros bioquímicos avaliados (creatinina, ureia, alanina aminotransferase e aspartato aminotransferase) não apresentaram diferenças entre grupos. Através do ensaio dos cometas não foram registados danos oxidativos entre grupos. No geral, conclui-se que a concentração de tília utilizada não apresentou efeitos adversos neste modelo animal. Os autores agradecem a Fundação para a Ciência e a Tecnologia (FCT, Portugal) e ao FEDER no ambito do programa PT2020 pelo apoio financeiro ao CIMO (UID/AGR/00690/2013) e ao programa FEDER-Interreg Espafia-Portugal pelo apoio financeiro atrav6s do projeto 0377 lberphenoi_6_E. info:eu-repo/semantics/publishedVersion