Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A toxoplasmose é uma zoonose amplamente distribuída que afeta mais de um terço da população mundial e de grande importância na saúde pública. A maioria das infecções em humanos por Toxoplasma gondii é assintomática. A toxoplasmose é amplamente investigada visto que se apresenta como uma doença grave em pessoas imunodeprimidas (portadores da síndrome da imunodeficiência adquirida (SIDA), não tratados, indivíduos transplantados, paciente em tratamento quimioterápico ou em uso de drogas supressoras e gestantes). A toxoplasmose congênita frequentemente pode levar ao aborto espontâneo ou até mesmo resultar na formação de crianças com algum grau de atraso no desenvolvimento mental e/ou físicos, deste modo, a transmissão congênita pode ser muito mais importante do que se pensava, pois os parasitos encontrados na circulação sanguinea são capazes de infectar as células endoteliais dos vasos e os tecidos circunjacentes, podendo resultar no encistamento do T. gondii. Atualmente a toxoplasmose vem sendo investigada devido a sua associação a inúmeras outras doenças, assim, estudos sobre a evolução da infecção por T. gondii em diferentes tipos de células hospedeiras se fazem necessários para uma abordagem terapêutica adequada. Ao invadir a célula hospedeira o parasito possui a capacidade de recrutar as mitocôndrias promovendo mudanças na organização mitocondrial ao longo da progressão da infecção, garantindo um ambiente favorável a sua multiplicação. Diante disso, investigamos se o parasito possui a capacidade de interferir no metabolismo mitocondrial e na resposta apoptótica da célula endotelial. O presente trabalho teve como objetivo analisar o metabolismo mitocondrial através da respirometria de alta-resolução e da resposta apoptótica através do western blotting das células endoteliais da veia umbilical humana (HUVEC) infectadas por 2, 6 e 20 horas por taquizoítos de T. gondii. A respirometria de alta-resolução revelou que o parasito interfere no metabolismo energético da célula hospedeira. A análise do conteúdo de proteínas da família Bcl-2 por western blotting revelou maior estímulo apoptótico no tempo inicial de infecção, quando comparado aos demais tempos. Os resultados dos conteúdos de caspase 3, proteína efetora da apoptose, não demonstrou diferença nos tempos iniciais de infecção Entretanto, em tempos mais tardios, o conteúdo de caspase 3 mostrou-se significativamente aumentado quando comparado às HUVEC não infectadas. A dinâmica de replicação do parasito foi observada através do monitoramento pelo sistema Time-Lapse Nikon BioStation IMQ em tempo real das células infectadas por T.gondii. Portanto, nossos resultados sugerem que o protozoário ao recrutar as mitocôndrias da célula hospedeira interfere no metabolismo mitocondrial e na modulação da apoptose para garantir um ambiente favorável a sua multiplicação. / Toxoplasmosis is a widespread zoonosis that affects more than a third of the world population and of great public health importance. Most human infections with Toxoplasma gondii are asymptomatic. Toxoplasmosis is widely investigated since it presents itself as a serious disease in immunocompromised persons (holders of acquired immunodeficiency syndrome (AIDS), untreated, transplant recipients, patients undergoing chemotherapy or suppressing drugs and pregnant). Congenital toxoplasmosis can often lead to miscarriage or even result in the formation of children with some degree of developmental delay mental and / or physical, thus congenital transmission may be much more important than previously thought, because the parasites found In the bloodstream are able to infect endothelial cells of blood vessels and surrounding tissues, which may result in encystment T. gondii. Currently toxoplasmosis has been investigated because of their association with other diseases, so, studies of the evolution of T.gondii infection in different types of host cells are necessary for an adequate therapeutic approach. To invade the host cell, the parasite has the ability to recruit mitochondria promoting changes in mitochondrial organization along the progression of infection, ensuring a favorable environment for their multiplication. Therefore, we investigated whether the parasite has the ability to interfere with mitochondrial metabolism and apoptotic response of endothelial cells. This study aimed to analyze the mitochondrial metabolism by high-resolution respirometry and apoptotic response by western blotting of endothelial cells of human umbilical vein (HUVEC) infected for 2, 6 and 20 hours per tachyzoites of T. gondii. The high-resolution respirometry revealed that the parasite interferes with the energy metabolism of the host cell. The analysis of the family protein content of Bcl-2 by western blotting revealed higher apoptotic stimulus at the initial time of infection, as compared to other times. The results of the contents of caspase 3 protein effector of apoptosis, showed no difference in the initial days of infection However, in more recent times, the content of caspase 3 was significantly increased when compared to non-infected HUVEC. The dynamics of parasite replication was observed by monitoring the system Time-Lapse Nikon BioStation IMQ in real time from infected cells by T. gondii. Therefore, our findings suggest that mitochondria in recruiting protozoan host cell interfere with mitochondrial metabolism and in the modulation of apoptosis to ensure a favorable environment for multiplication.

Le NAD+ apparaît comme un régulateur majeur du fonctionnement mitochondrial. En effet, ce cofacteur régule non seulement l’activité de nombreuses enzymes impliqués dans le métabolisme énergétique (enzymes de la β-oxydation des acides gras, du cycle de Krebs) mais joue également un rôle dans la production d’espèces réactives de l’oxygène (ROS). Le NAD+ est aussi le cofacteur des sirtuines, des enzymes déacétylases régulatrices notamment du métabolisme mitochondrial. De plus, la mitochondrie est l’organite au sein duquel la concentration en NAD+ est la plus élevée (jusqu’à 70% du NAD cellulaire). Le complexe I, qui possède une activité NADH déshydrogénase, pourrait être l’un des régulateurs majeurs du rapport NADH/NAD+ mitochondrial. L’objectif de ce travail de thèse a été d’étudier le rôle du rapport NADH/NAD+ mitochondrial dans le métabolisme énergétique et l’implication du complexe I dans les pathologies mitochondriales. Nous avons mis en évidence qu’une modulation du rapport NADH/NAD+ mitochondrial (augmentation par un activateur pharmacologique ou diminution consécutive à une mutation touchant une sous-unité du complexe I, modifie de manière drastique le métabolisme énergétique notamment en activant ou inhibant la protéine SIRT3, isoforme mitochondriale des sirtuines. Le complexe I semble jouer un rôle majeur dans cette modulation. Le resveratrol, ciblant le complexe I, ainsi que le NMN, un précurseur du NAD+, permettent de restaurer ce rapport et d’améliorer ainsi le métabolisme mitochondrial. Nos résultats suggèrent donc que le rapport NADH/NAD+ pourrait être une cible thérapeutique particulièrement intéressante dans les déficits du complexe I. / NAD+ appears as a main regulator of the mitochondrial function. Indeed, this compound not only regulates the enzymatic activity of enzymes involved in energetic metabolism (fatty acid oxidation, tricarboxylic acid cycle) but is also involved in ROS production. NAD+ is also the cofactor of sirtuins, deacetylase enzymes, in particular regulating the mitochondrial function. Moreover, mitochondria sequester most of the cellular NAD+ (up to 70 %). The complex I, which possesses an NADH dehydrogenase activity, is thought to be the most important regualtor of the mitochondrial NADH/NAD+ ratio. The work presented here aimed at studying the role of the mitochondrial NADH/NAD+ ratio in mitochondrial metabolism and to test the involvement of the complex I in mitochondrial disorders. We show that a modulation of the mitochondrial NADH/NAD+ ratio (increase by a pharmacological agent or decrease in complex-I mutated fibroplasts) severely affects the mitochondrial energetic function especially by interacting with SIRT3 a mitochondrial sirtuin isoform. The NADH/NAD+ ratio is highly regulated by complex I activity. Resveratrol, which targets the complex I, as well as NMN, a NAD+ precursor, improves the mitochondrial NADH/NAD+ ratio and consequently increases the mitochondrial metabolism. Our results strongly suggest that the mitochondrial NADH/NAD+ ratio could be an interesting therapeutic target especially in complex I- deficient patients.

Yes / Aims/hypothesis Hypoglycaemia is a major barrier to good glucose control in type 1 diabetes. Frequent hypoglycaemic episodes impair awareness of subsequent hypoglycaemic bouts. Neural changes underpinning awareness of hypoglycaemia are poorly defined and molecular mechanisms by which glial cells contribute to hypoglycaemia sensing and glucose counterregulation require further investigation. The aim of the current study was to examine whether, and by what mechanism, human primary astrocyte (HPA) function was altered by acute and recurrent low glucose (RLG). Methods To test whether glia, specifically astrocytes, could detect changes in glucose, we utilised HPA and U373 astrocytoma cells and exposed them to RLG in vitro. This allowed measurement, with high specificity and sensitivity, of RLG-associated changes in cellular metabolism. We examined changes in protein phosphorylation/expression using western blotting. Metabolic function was assessed using a Seahorse extracellular flux analyser. Immunofluorescent imaging was used to examine cell morphology and enzymatic assays were used to measure lactate release, glycogen content, intracellular ATP and nucleotide ratios. Results AMP-activated protein kinase (AMPK) was activated over a pathophysiologically relevant glucose concentration range. RLG produced an increased dependency on fatty acid oxidation for basal mitochondrial metabolism and exhibited hallmarks of mitochondrial stress, including increased proton leak and reduced coupling efficiency. Relative to glucose availability, lactate release increased during low glucose but this was not modified by RLG. Basal glucose uptake was not modified by RLG and glycogen levels were similar in control and RLG-treated cells. Mitochondrial adaptations to RLG were partially recovered by maintaining euglycaemic levels of glucose following RLG exposure. Conclusions/interpretation Taken together, these data indicate that HPA mitochondria are altered following RLG, with a metabolic switch towards increased fatty acid oxidation, suggesting glial adaptations to RLG involve altered mitochondrial metabolism that could contribute to defective glucose counterregulation to hypoglycaemia in diabetes. / Diabetes UK (RD Lawrence Fellowship to CB; 13/0004647); the Medical Research Council (MR/N012763/1) to KLJE, ADR and CB; and a Mary Kinross Charitable Trust PhD studentship to CB, ADR and RW to support PGWP. Additional support for this work came from awards from the British Society for Neuroendocrinology (to CB and KLJE), the Society for Endocrinology (CB), Tenovus Scotland (CB) and the University of Exeter Medical School (CB and KLJE). AR was also supported by a Royal Society Industry Fellowship.

Uma estreita relação entre a ação dos hormônios tireoidianos (HT) no sistema cardiovascular sendo mediada por componentes do sistema renina-angiotensina (SRA) tem sido descrita na literatura. Já foi demonstrado que o processo de isquemia/reperfusão (I/R) promove diminuição na expressão do receptor de angiotensina II do tipo 2 (AT2) no miocárdio, com consequente prejuízo funcional, enquanto o aumento de sua expressão melhorou a recuperação da função cardíaca. A hipótese do presente estudo é que o papel cardioprotetor exercido pelos HT ocorre com a participação do receptor AT2. Esta hipótese foi testada utilizando o modelo de I/R com a perfusão de coração isolado de camundongos selvagens e nocautes para o AT2, submetidos a tratamento por 14 dias com T3. Ainda foi avaliado o SRA mitocondrial, assim como o papel do óxido nítrico (NO) na recuperação pós-isquêmica. Os resultados apontam que a cardioproteção induzida pelo T3 é mediada pelo AT2, com consequente aumento na produção de NO e modulação de parâmetros do metabolismo mitocondrial. / Some authors have shown a close relationship between the action of thyroid hormone (TH) on cardiovascular system and renin angiotensin system (RAS) activation. Have been shown that ischemia/reperfusion (I/R) promotes decrease on angiotensin II type 2 receptor (AT2) expression in the myocardium, with functional worsening, on the other hand the AT2R increased improves the cardiac function after ischemia episodes. So, we have hypothesized that the cardioprotective effect of TH in I/R model may occur with the participation of AT2. This hypothesis was tested using I/R model in isolated hearts AT2 knockout and wild-type mice submitted to high levels of T3 by 14 days. The mitochondrial RAS was evaluated, as well as the nitric oxide (NO) role in post-ischemic recovery. The results show that TH induces cardioprotection through AT2 receptor, some mitochondrial metabolism parameters were modulated by TH and the NO production was increased.

Pancreatic ß-cells are heterogeneous in their secretory responsiveness, glucose sensitivity and metabolic rate. A diminished and delayed first-phase insulin release is an early sign of failing ß-cells in diabetes. Mechanisms controlling functional characteristics, such as lag time for insulin release or magnitude of the response in each individual cell are unknown. To find out whether the heterogeneity represents a random phenomenon in ß-cell or is a manifestation of reproducible characteristics, we compared parameters of Ca2+ response in Fura-2 labelled ob/ob mouse ß-cells during two consecutive stimulations with glucose. Lag times, as well as peak heights and nadirs of initial lowering showed a strong correlation between the first and second stimulation. Thus, timing and magnitude of the early Ca2+ response were specific for each cell. ß-Cells from lean mice, diabetic db/db mice and rats also showed cell-specific responses characteristics. This indicates that a cell-specific Ca2+ response to glucose is common in rodent ß-cells, both normal and diabetic. Another question was whether aggregated ß-cells show cell-specific responses. Using the same protocol as for dispersed ß-cells, we analysed Ca2+ responses in clusters of different size and in intact islets from ob/ob and lean mice. Correlations were found between the first and second stimulation for timing and magnitude of [Ca2+]i rise, and for the initial lowering. Next, we tested if the ß-cell response is cell-specific, when induced at different steps of the stimulus-secretion coupling. The glycolytic intermediate glyceraldehyde, the mitochondrial substrate KIC, the KATP-channel blocker tolbutamide and arginine were used as tools. [Ca2+]i changes were studied in dispersed ß-cells from lean, ob/ob and db/db mice. NADH responses to glucose and KIC were analyzed as a measure of metabolic flux. The correlation between Ca2+ and insulin response from individual ß-cells was tested using Fluo-3 and Fluozin-3. Both timing and magnitude of calcium responses were cell-specific in lean mouse ß-cells with all tested secretagogues. ß-Cells from ob/ob and db/db mice showed cell-specific timing of Ca2+ responses to glyceraldehyde but not to KIC, tolbutamide or arginine. However, ob/ob mouse ß-cells within intact islets showed cell-specific timing of tolbutamide-induced response. NADH responses to glucose were cell-specific in all three mouse models, but the timing of NADH responses to KIC was cell-specific only in lean mice. Thus, a cell-specific response can be induced in normal ß-cells at several steps of stimulus-secretion coupling for nutrient-stimulated insulin release. Cell-specific properties of ß-cell ion channels and the mitochondrial metabolism are affected in db/db and ob/ob mice. The relation between mitochondrial mass and parameters of Ca2+ responses were investigated in Mitotracker Red and Fluo-3 labelled ß-cells using confocal microscopy. Data show that ß-cell mitochondrial state may play an important role in determining the timing of [Ca2+]i changes. In summary, the early Ca2+ response pattern in ß-cells, including the lag time, the nadir of initial lowering and the height of the first peak response is cell-specific. Isolated and functionally coupled ß-cells show cell-specific timing of Ca2+ responses when stimulated with metabolic and non-metabolic agents. This may be a robust mechanism of importance for the adequate function of ß-cells and a basis for the pacemaker function of some cells. A disturbed cell specificity of the mitochondrial metabolism and ion channel function appears to be a marker of ß-cell dysfunction in hyperglycemia and diabetes and may explain the delayed insulin release in ß-cells from diabetic subjects.

Le cerveau adulte des mammifères conserve sa capacité à générer de nouvelles cellules cérébrales à partir de cellules souches neurales (CSNs), principalement localisées dans deux régions cérébrales spécifiques, l'hippocampe et la zone sous-ventriculaire (SVZ). Ce processus, appelé neurogenèse, permet la formation de nouveaux neurones et de nouvelles cellules gliales (astrocytes et oligodendrocytes). Différents signaux contrôlent la prolifération et la différenciation des CSNs. Parmi ces signaux, les hormones thyroïdiennes (HTs) sont impliquées dans la prolifération des CSNs de la SVZ et dans la différenciation neuronale. À l’inverse des cellules différenciées, telles que les neurones ou les glies, les CSNs ont un fonctionnement – ou métabolisme – principalement basé sur la glycolyse et sur une faible respiration mitochondriale. Or l'évolution du métabolisme des CSNs peut influencer leur choix de destin cellulaire. Les HTs jouant un rôle important dans l'activation du métabolisme mitochondrial, j'ai testé l'hypothèse selon laquelle le choix du destin des CSNs de la SVZ adulte se ferait grâce à l'influence de la signalisation thyroïdienne sur l'activité mitochondriale. J'ai tout d'abord montré in vivo et in vitro que les HTs permettent la détermination des CSNs en précurseurs neuronaux dans la SVZ, tandis qu'une période d'hypothyroïdisme favorise la détermination gliale. La transthyrétine, protéine de liaison des HTs, est spécifiquement présente dans les cellules de la SVZ ayant un destin neuronal, alors que la désiodase de type 3, inactivatrice des HTs, est exprimée par les précurseurs oligodendrocytaires (OPCs), indiquant une activationdifférentielle de la signalisation thyroïdienne dans les deux lignages cellulaires. Par ailleurs j'ai pu observer que les cellules s'engageant vers un destin neuronal possèdent une plus grande activité mitochondriale que les OPCs. La présence d'HTs favorise de plus la respiration mitochondriale, ainsi que la production de dérivés réactifs de l'oxygène (ROS) issus de l'activité des mitochondries, dans les cellules de la SVZ. Un blocage des protéines de la chaîne respiratoire empêche les HTs de promouvoir la détermination neuronale, montrant la nécessité de l'activation mitochondriale pour l'engagement des CSNs en précurseurs neuronaux. On sait d'autre part que les modifications morphologiques (ou dynamiques) mitochondriales sont nécessaires à l'augmentation de la respiration. La division (ou fission) des mitochondries est en particulier essentielle à une bonne répartition intracellulaire de la production de l'énergie issue de la respiration, ainsi qu'à la migration cellulaire. Dans les cellules de la SVZ, j'ai montré que l'action des HTs permet l'activation de la protéine DRP1, médiatrice de la fission mitochondriale, et ce principalement dans les cellules du lignage neuronal. Les HTs favorisent donc la détermination des CSNs de la SVZ vers un destin neuronal grâce à l'activation de la respiration et de la fission mitochondriales. / The adult mammalian brain maintains its capacity to generate new cells from neural stem cells (NSCs), mainly localized in two specific brain regions, the hippocampus and the sub-ventricular zone (SVZ). This process, named neurogenesis, results in the production of new neurons and new glial cells (astrocytes and oligodendrocytes). Several signals control NSCs proliferation and differentiation. Among those, thyroid hormones (THs) are involved in NSCs proliferation in the SVZ and in neuronal differentiation. NSC metabolism relies mainly on glycolysis associated with a low mitochondrial activity, whereas mature cells, like neurons and glia, preferentially use oxidative phosphorylation. Changes in NSC metabolism can impact cell fate. As THs play an important part in activating mitochondrial metabolism, I hypothesized that the influence of TH signaling on mitochondrial activity triggers NSC fate choice in the adult SVZ. First, I showed in vivo and in vitro that THs allow NSC determination in neuronal precursors, whereas a short hypothyroidism favors glial determination. Transthyretine, a TH binding protein, is specifically present in the SVZ cells having a neuronal fate, while type 3 deiodinase, a TH inhibitor, is expressed by oligodendrocyte precursor cells (OPCs). These results indicate that THs signaling isdifferentially activated in neuronal and glial cell lineages. I observed that cells adopting a neuronal fate display a greater mitochondrial activity when compared to OPCs, and that TH signaling favors mitochondrial respiration and ROS production in the SVZ cells. Inhibiting the mitochondrial respiratory chain prevents TH-mediated promotion of neuronal determination, proving the need of mitochondrial activation for NSC commitment toward a neuronal phenotype. Besides, it is also known that modifications of mitochondrial morphology (or mitochondrial dynamics) are required for the respiration to increase. Among mitochondrial dynamics, fission is crucial for a good intracellular repartition of energy production, and for cell migration. In the SVZ cells, I showed that, DRP1, the main inducer of mitochondrial fission, is activated by THs mainly in cells adopting a neuronal fate. Thus, THs favor NSC fate choice toward a neuronal phenotype through the activation of mitochondrial metabolism and mitochondrial fission in the adult mouse SVZ.

<p>According to the Global Cancer Incidence, Mortality, and Prevention (GLOBOCAN) study for 2018, 2,089,000 women will have been diagnosed with breast cancer worldwide, with 627,000 breast cancer-related mortalities. It is estimated that between 15 – 20 % of breast cancer diagnoses are of the triple-negative subtype. Triple-negative breast cancers (TNBCs) do not express the receptors for estrogen, progesterone, and human epidermal growth factor 2, and hence cannot be treated using hormone receptor-targeted therapy. </p> <p>TNBCs are commonly of the basal-like phenotype, with high expression levels of proteins involved in epithelial-mesenchymal transition, extracellular-matrix (ECM) remodeling, cell cycle progression, survival and drug resistance, invasion, and metastasis. 5-year survival rates are significantly lower for TNBC patients, and the disease is characterized by poorer grade at the time of diagnosis as well as higher 5-year distant relapse rates, with a greater chance of lung and CNS metastases. Current treatments for TNBC take the form of aggressive cytotoxic chemotherapy regimens with multiple adverse side-effects. An important goal of on-going studies is to identify new compounds with significant TNBC-specificity, in order to improve patient survival outcomes while preserving a high quality of life during treatment.</p> <p> For several decades, compounds originally isolated from bioactive natural extracts, such as the taxanes and vinca<i> </i>alkaloids, have been at the forefront of chemotherapy. However, due to their non -specific mechanisms of action, treatment with these compounds eventually leads to significant toxicity to normal cells and tissues. Modern transcriptomics, metabolomics, and proteomics tools have greatly improved our understanding of the mechanisms governing cancer initiation and progression, and revealed the considerable heterogeneity of tumor cells. This has allowed for the identification of potential vulnerabilities in multiple cancers, including TNBCs. By leveraging these new technologies and insights with the tremendous diversity of bioactive compounds from organisms that remain unstudied, new classes of onco-drugs targeting pathways specific to TNBC cells could be identified in the near future.</p> <p>Here, we describe the cytotoxic effects of extracts from <i>Lippia origanoides </i>- a species of medicinal shrub native to Central and South America - on TNBC cells. We report that these extracts induce rapid, sustained, and irreversible apoptosis in TNBC cells <i>in vitro</i>, with significantly reduced cytotoxicity against normal mammary epithelial cells. The <i>L. origanoides </i>extracts LOE and L42 exploited two TNBC-specific characteristics to induce apoptosis in these cells: i) inhibiting the constitutively active survival and inflammatory NF-kB signaling pathway, and ii) significantly dysregulating the expression levels of mitochondrial enzymes required to maintain the TCA cycle and oxidative phosphorylation; metabolic pathways that are required for the maintenance of TNBC cell growth and proliferation.</p> <p>Finally, to lay the foundations for future studies on the abilities of these extracts to prevent tumor initiation and inhibit tumor growth <i>in vivo</i>, we also show that the <i>L. origanoides </i>extract, L42, is non-toxic<i> </i>to immunocompetent C57BL/6 mice, and have developed an <i>in vivo </i>model of human TNBC in athymic <i>nu/nu</i> mice. </p> <p>Collectively, our studies are the first to identify the anti-TNBC-specific properties of bioactive extracts from the <i>Lippia </i>species, and reveal that targeting NF-kB signaling and mitochondrial metabolism are potential avenues to new therapeutics against this subtype of breast cancer. Future work in our lab will focus on identifying the bioactive components (BACs) of the extract mediating its apoptotic effects, and shedding light on their protein binding partners within the cell.</p>

Recent progress in fundamental biological sciences and medicine has considerably increased the quantity ofdata that can be studied and processed. The main limitation now is not retrieving data, but rather extractinguseful biological insights from the large datasets accumulated. More recent advances have provided detailedhigh-density data regarding metabolism (metabolomics) and protein expression (proteomics). Clearly, no single analytic methods, can provide a comprehensive understanding. Rather, the ability to link available datatogether in a coherent manner is required to obtain a complete view. The improving application of MachineLearning (ML) techniques provides the means to make continuous progress in processing complex data sets.A brief discussion is offered on the advantages of ML, the state-of-the-art in Deep Learning (DL) for proteinpredictions and the importance of ML in biological data processing. Noise stemming from incorrect classification or arbitrary/ambiguous labelling of data may arise when ML techniques are applied to large datasets. Furthermore, the stochasticity of biological systems needs to be considered for correctly evaluating theoutputs. Here we show the potential of a workflow to respond biological questions taking into consideration aperturbation of the biological data.  For controlling the applicability of models and maximizing the predictivity, in silico filtering schemescan usefully be applied as an “Ockham’s razor” before using any ML technique. After reviewing differentDL approaches for protein prediction purposes, this work shows that a computational approach in filteringsteps is a valuable tool for proteins classification when biological features are not fully annotated or reviewed.The in silico approach has identified putative proline transporters in fungi and plants as well as carotenoidbiosynthetic gene products in the plant family Brassicaceae. The proposed method is suitable for extractingfeatures of classification and then maximizing the use of a DL approach.

No description available.