Study of Tissue-Specific Reactive Oxygen Species Formation by Cell Membrane Microarrays for the Characterization of Bioactive Compounds

The production of reactive oxygen species (ROS) increases considerably in situations of cellular stress, inducing lipid peroxidation and multiple alterations in proteins and nucleic acids. However, sensitivity to oxidative damage varies between organs and tissues depending on the triggering process. Certain drugs used in the treatment of diverse diseases such as malaria have side effects similar to those produced by oxidative damage, although no specific study has been conducted. For this purpose, cell membrane microarrays were developed and the superoxide production evoked by the mitochondrial activity was assayed in the presence of specific inhibitors: rotenone, antimycin A and azide. Once the protocol was set up on cell membrane isolated from rat brain areas, the effect of six antimalarial drugs (atovaquone, quinidine, doxycycline, mefloquine, artemisinin, and tafenoquine) and two essential oils (Rosmarinus officinalis and Origanum majoricum) were evaluated in multiple human samples. The basal activity was different depending on the type of tissue, the liver, jejunum and adrenal gland being the ones with the highest amount of superoxide. The antimalarial drugs studied showed specific behavior according to the type of human tissue analyzed, with atovaquone and quinidine producing the highest percentage of superoxide formation, and doxycycline the lowest. In conclusion, the analysis of superoxide production evaluated in cell membranes of a collection of human tissues allowed for the characterization of the safety profile of these antimalarial drugs against toxicity mediated by oxidative stressThis work has been partially supported by grant Ministry of Economy and Competitiveness PID2019-106222RB-C31/SRA (State Research Agency, 10.13039/501100011033), the Basque Government Department of Economic Development, Sustainability and Environment (PUE21-03 and Bikaintek program: 48-AF-W2-2019-7 and 007-B2/2020) and University of the Basque Country (price to the best Master Thesis in collaboration with Institutions and Business)

Anti-Trypanosomatidae Activity of Essential Oils and Their Main Components from Selected Medicinal Plants

Kinetoplastida is a group of flagellated protozoa characterized by the presence of a kinetoplast, a structure which is part of a large mitochondria and contains DNA. Parasites of this group include genera such as Leishmania, that cause disease in humans and animals, and Phytomonas, that are capable of infecting plants. Due to the lack of treatments, the low efficacy, or the high toxicity of the employed therapeutic agents there is a need to seek potential alternative treatments. In the present work, the antiparasitic activity on Leishmania infantum and Phytomonas davidi of 23 essential oils (EOs) from plants of the Lamiaceae and Asteraceae families, extracted by hydrodistillation (HD) at laboratory scale and steam distillation (SD) in a pilot plant, were evaluated. The chemical compositions of the EOs were determined by gas chromatography-mass spectrometry. Additionally, the cytotoxic activity on mammalian cells of the major components from the most active EOs was evaluated, and their anti-Phytomonas and anti-Leishmania effects analyzed. L. infantum was more sensitive to the EOs than P. davidi. The EOs with the best anti-kinetoplastid activity were S. montana, T. vulgaris, M. suaveolens, and L. luisieri. Steam distillation increased the linalyl acetate, β-caryophyllene, and trans-α-necrodyl acetate contents of the EOs, and decreased the amount of borneol and 1,8 cineol. The major active components of the EOs were tested, with thymol being the strongest anti-Phytomonas compound followed by carvacrol. Our study identified potential treatments against kinetoplastids.Publishe

Characterization of the Antitumor Potential of Extracts of Cannabis sativa Strains with High CBD Content in Human Neuroblastoma

Cannabis has been used for decades as a palliative therapy in the treatment of cancer. This is because of its beneficial effects on the pain and nausea that patients can experience as a result of chemo/radiotherapy. Tetrahydrocannabinol and cannabidiol are the main compounds present in Cannabis sativa, and both exert their actions through a receptor-mediated mechanism and through a non-receptor-mediated mechanism, which modulates the formation of reactive oxygen species. These oxidative stress conditions might trigger lipidic changes, which would compromise cell membrane stability and viability. In this sense, numerous pieces of evidence describe a potential antitumor effect of cannabinoid compounds in different types of cancer, although controversial results limit their implementation. In order to further investigate the possible mechanism involved in the antitumoral effects of cannabinoids, three extracts isolated from Cannabis sativa strains with high cannabidiol content were analyzed. Cell mortality, cytochrome c oxidase activity and the lipid composition of SH-SY5Y cells were determined in the absence and presence of specific cannabinoid ligands, with and without antioxidant pre-treatment. The cell mortality induced by the extracts in this study appeared to be related to the inhibition of the cytochrome c oxidase activity and to the THC concentration. This effect on cell viability was similar to that observed with the cannabinoid agonist WIN55,212-2. The effect was partially blocked by the selective CB1 antagonist AM281, and the antioxidant α-tocopherol. Moreover, certain membrane lipids were affected by the extracts, which demonstrated the importance of oxidative stress in the potential antitumoral effects of cannabinoids.This work has been partially supported by a grant from the Ministry of Economy and Competitiveness (DIN2019-010902 and DIN2020-011349) and the Basque Government Department of Economic Development, Sustainability and Environment (Bikaintek program: 005-B2/2021)

Konexio zilindrikodun hortz inplanteak: torlojuen aurreteinkaketaren eta erresonantzia maiztasunen arteko erlazioaren azterketa

[EU]Gradu Amaierako Lan honetan konexio zilindrikodun hortzetako inplanteetako torlojuen aurreteinkaketaren zeharkako kalkulua burutu nahi da. Hortzetako inplanteen egonkortasuna torlojuaren aurre-kargak definitzen du eta aldi berean, inplante hori aurreteinkatuta egon behar da, apurketarik edota lekualdaketarik jasan ez dezan eta ondorioz erosotasun eta aho osasun egokia ahalbidetzeko. Horretarako torlojuaren aurreteinkaketaren eta maiztasun naturalen arteko erlazioa ikertuko da. Egun, ahoaren neurria kontuan izanik, ez da metodo erosorik ezagutzen aurre-teinkaketaren kalkulua burutzeko. Hori dela eta, ikerketa gai izango da aurrerago aipatutako metodoa zeinetan kitzikadura harmoniko edo mikroimpaktoen bidez, sistemaren erresonantzia maiztasunak neurtuz eta aurreteinkaketarekin duen erlazioa erabiliz, torlojuaren aurreteinkaketa balioa lortuko da. Azaldutako lana aurrera eramateko, entsegu bankuan dagoen inplante baten analisia Elementu Finituen metodoa erabiliz gauzatuko da; metodoa baliogarria den ala ez ondorioztatuko da.[ES]Mediante este Trabajo de Fin de Grado, se pretende realizar el cálculo indirecto de la precarga de un tornillo de un implante dental de conexión cilíndrica. La estabilidad de un implante viene definida por la precarga de su tornillo y, en lo que concierne a la salud bucal y a la comodidad de un implante, es imprescindible que dicho implante permanezca precargado para evitar su movimiento o, en su defecto, su ruptura. Para ello, se realizará el estudio de un método en el que se relacionarán la precarga del tornillo y las frecuencias naturales del sistema con el fin de calcular la precarga. Por el momento, no se conoce ningún método relativamente cómodo para el cálculo de la precarga. Por ello, será objeto de estudio el método mencionado anteriormente en el cual mediante excitaciones harmónicas o microimpactos se medirán las frecuencias naturales del sistema y, utilizando la relación entre la precarga y las frecuencias naturales, se obtendrá la precarga del tornillo. Para llevar a cabo el trabajo, se realizará el análisis de un implante dental sobre un banco de pruebas utilizando el Método de Elementos Finitos y se concluirá con la aprobación o desaprobación del método en estudio.[EN]In this End of Grade work, the preload of a cylindrical connection dental implant`s screw will be indirectly calculated. The stability of an implant is defined by the preload of the screw. So, the preload is essential to prevent implant’s movement or break which define the oral health and comfort. Therefore, a method will be studied in which the bolt preload and the natural frequencies of the system will be related. At the moment, a comfortable preload calculating method is not known. For this reason, the method mentioned above will be studied in which by a harmonic excitations or microimpacts the system natural frequencies will be obtained and, using the relationship between preload and natural frequencies, bolt preload will be calculated. To carry out the work, dental implant will be analyzed in the testing bench using the Finite Element Method and to conclude with the approval or disapproval of the method under study

Cultivation of Lavandula luisieri for the production of bioactive extracts (póster)

Lavandula luisieriPublishe

Screening System of <i>Cannabis sativa</i> Extracts Based on Their Mitochondrial Safety Profile Using Cytochrome c Oxidase Activity as a Biomarker

The development of Cannabis sativa strains with high cannabidiol (CBD) and low tetrahydrocannabinol (THC) content is a growing field of research, both for medical and recreational use. However, the mechanisms behind clinical actions of cannabinoids are still under investigation, although there is growing evidence that mitochondria play an important role in many of them. Numerous studies have described that cannabinoids modulate mitochondrial activity both through activation of mitochondrial cannabinoid receptors and through direct action on other proteins such as mitochondrial complexes involved in cellular respiration. Thus, the aim of this study was to determine the actions of a panel of extracts, isolated from high-CBD varieties of Cannabis sativa, on the activity of the mitochondrial electron transport chain complex IV, cytochrome c oxidase (CCO), in order to select those with a safer profile. After demonstrating that Cannabis sativa strains could be identified by cannabinoids content, concentration–response curves were performed with a collection of extracts from strains with high-CBD and low-THC content using bovine CCO. The CCO rate was clearly modified by specific extracts of Cannabis sativa plants compared to others. Half maximal inhibitory concentrations (IC50) of extracts and the inhibitory effects evoked at 1 × 10−4 g/mL displayed a significant correlation with the THC. Therefore, the screening of extracts based on CCO activity provides a powerful and rapid methodology to identify those plants with higher mitochondrial toxicity or even mito-protective actions