ESS - CQB - Ciências Químicas e das Biomoléculas
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- 11 - Collagen-like materials for tissue regeneration and repairPublication . Teixeira, Cátia; Ferraz, Ricardo; Prudêncio, Cristina; Gomes, PaulaPeptides can be designed to recreate the higher-order assembly (triple helix) and biological function of natural collagens. Collagen triple-helical domain contains Gly-Xaa-Yaa triplets where Xaa and Yaa are frequently proline and hydroxyproline, respectively. Glycine must be in every third position forming a hydrogen bond to an adjacent proline to stabilize collagen structure. Hydroxyproline residues are important in the thermal stability of the collagen triple-helix conformation. Although it is difficult to produce a stable triple helix, collagen-like peptides can be used to produce scaffolds that mimic natural tissue networks for biomedical engineering, without the liabilities of nonhuman natural collagens such us immunogenicity and pathogen transmission. This chapter describes the major approaches towards synthesis of collagen-like peptides, essentially aimed at production of artificial biocompatible and biofunctional frameworks for regenerative medicine.
- 4,9-Diaminoacridines and 4-Aminoacridines as Dual-Stage Antiplasmodial HitsPublication . Fonte, Mélanie; Tassi, Natália; Fontinha, Diana; Bóuzon-Arnáiz, Inês; Ferraz, Ricardo; Araújo, Maria J.; Fèrnandez-Busquets, Xavier; Prudêncio, Miguel; Gomes, Paula; Teixeira, CátiaMulti‐stage drugs have been prioritized in antimalarial drug discovery, as targeting more than one process in the Plasmodium life cycle is likely to increase efficiency, while decreasing the chances of emergence of resistance by the parasite. Herein, we disclose two novel acridine‐based families of compounds that combine the structural features of primaquine and chloroquine. Compounds prepared and studied thus far retained the in vitro activity displayed by the parent drugs against the erythrocytic stages of chloroquine‐sensitive and ‐resistant Plasmodium falciparum strains, and against the hepatic stages of Plasmodium berghei, hence acting as dual‐stage antiplasmodial hits.
- Absence of negative allelopathic effects of cylindrospermopsin and microcystin-LR on selected marine and freshwater phytoplankton speciesPublication . Pinheiro, Carlos; Azevedo, Joana; Campos, Alexandre; Loureiro, Susana; Vasconcelos, VítorCyanobacterial toxins have been regarded by some researchers as allelopathic substances that could modulate the growth of competitors. Nevertheless, often the concentrations of toxins used are too high to be considered ecologically relevant. In this work we tested the hypothesis that microcystin-LR (MC-LR) and cylindrospermopsin (CYN) at ecologically relevant concentrations have no allelopathic effects on some species of phytoplankton. Extracts containing the toxins as well as pure MC-LR and CYN toxins were used to assess their effects on the growth rates of Nannochloropsis sp., Chlamydomonas reinhardtii, and Chlorella vulgaris. Cyanobacterial crude extracts induced more pronounced effects on growth rates than pure toxins. Microcystis aeruginosa and Aphanizomenon ovalisporum crude extracts containing MC-LR and CYN at 0.025–2.5 mg l−1 stimulated growth rates of microalgae, whereas A. ovalisporum crude extracts containing 2.5 mg l−1 of CYN strongly inhibited growth rates of microalgae after 4 and 7 days of exposure. MC-LR and CYN at environmentally occurring concentrations were unable to affect negatively the growth of microalgae, and therefore these molecules may play roles other than allelopathy in natural ecosystems.
- Actinobacteria as new probiotics for preventing fish diseases in aquaculturePublication . Santos, Matilde; Girão, Mariana; Oliveira, Rui S.; Carvalho, FátimaIn order to meet the food needs of a constantly growing world population, aquaculture emerges as the fastest growing food production sector. However, disease outbreaks due to opportunistic bacterial pathogens cause large losses. Antibacterial chemicals used indiscriminately have resulted in resistant bacteria and antibiotic resistance genes. Ecological solutions such as probiotics are needed. Probiotics are microorganisms introduced into another organism that can have beneficial effects on the host as well as improve the external environment. Actinobacteria are an important biotechnological resource, as they produce a great diversity of secondary metabolites with a high range of bioactivities, including antimicrobial. The aim of this study was to explore the probiotic potential of some marine actinobacteria to prevent aquaculture fish diseases. For this, a screening of antimicrobial activity was performed using agar-based disk diffusion and Minimum Inhibitory Concentration (MIC) assays, against three Gram-negative bacterial fish pathogens: Aeromonas hydrophila, Edwardsiella tarda and Pseudomonas anguilliseptica. A total of 54 out of 103 actinobacterial extracts tested, presented bioactivity against these microorganisms, so that the growth of all three Gram-negative pathogens used was inhibited. The diameter of the inhibition halos varied between 8 and 20 mm. Tested extracts presented MIC values of 1000 µg/ml, except for Isoptericola chiayiensis (500 µg/ml). Microbispora bryophytorum, Isoptericola chiayiensis and Nocardia nova were some of the bacteria with the most promising results. Overall, actinobacteria associated with seaweed proved to be a potential source of probiotics to inhibit pathogens in aquaculture fish.
- Adipocyte proteome and secretome influence inflammatory and hormone pathways in gliomaPublication . Almeida, Joana; Costa, J.; Coelho, Pedro; Cea, V.; Galesio, M.; Noronha, J. P.; Diniz, M. S.; Prudêncio, Cristina; Soares, R.; Sala, C.; Fernandes, RúbenGliomas represent the most common primary malignant brain tumors in adults, with an extremely poor prognosis. Among several risk factors, lifestyle was also recently identified as a major risk factor for the development of primary glioma. In the present study, we explore the relationship between obesity and glioma in a cellular model. Thus, we have study the influence of adipocytes secretome on glioma cell line GL261. Using the 3T3-L1 adipocyte cell line, and its conditioned medium (adipokines-enriched medium), we showed that adipocyte-released factors relate with glioma angiogenic, growth, hormones and metabolic behavior by MALDI-TOF-MS and proteomic array analysis. In a first view, STI1, hnRNPs and PGK1 are under expressed on CGl. Similarly, both carbonic anhydrase and aldose reductase are even suppressed in glioma cells that grown under adipokines-enriched environment. Contrariwise, RFC1, KIF5C, ANXA2, N-RAP and RACK1 are overexpressed in GL261 cell the in the presence of the adipokines-enriched medium. We further identified the factors that are released by adipocyte cells, and revealed that several pro-inflammatory and angiogenic factors, such as IL-6, IL-11, LIF, PAI-1, TNF-α, endocan, HGF, VEGF IGF-I, were secreted to the medium into a high extent, whereas TIMP-1 and SerpinE1 were under expressed on CGl. This study discloses an interesting in vitro model for the study of glioma biology under a "obesity" environment, that can be explored for the understanding of cancer cells biology, for the search of biomarkers, prognostic markers and therapeutic approaches.
- Adipocyte Secreted Factors Enhance Aggressiveness of Prostate Carcinoma CellsPublication . Moreira, Ângela; Pereira, Sofia S.; Costa, Madalena; Morais, Tiago; Pinto, Ana; Fernandes, Rúben; Monteiro, Mariana P.Obesity has been associated with increased incidence and risk of mortality of prostate cancer. One of the proposed mechanisms underlying this risk association is the change in adipokines expression that could promote the development and progression of the prostate tumor cells. The main goal of this study was to evaluate the effect of preadipocyte and adipocyte secretome in the proliferation, migration and invasion of androgen independent prostate carcinoma cells (RM1) and to assess cell proliferation in the presence of the adiposity signals leptin and insulin. RM1 cells were co-cultured in with preadipocytes, adipocytes or cultured in their respective conditioned medium. Cell proliferation was assessed by flow cytometry and XTT viability test. Cell migration was evaluated using a wound healing injury assay of RM1 cells cultured with conditioned media. Cellular invasion of RM1 cells co-cultured with adipocytes and preadipocytes was assessed using matrigel membranes. Preadipocyte conditioned medium was associated with a small increase in RM1 proliferation, while adipocytes conditioned media significantly increased RM1 cell proliferation (p<0.01). Adipocytes also significantly increased the RM1 cells proliferation in co-culture (p <0.01). Cell migration was higher in RM1 cells cultured with preadipocyte and adipocyte conditioned medium. RM1 cell invasion was significantly increased after co-culture with preadipocytes and adipocytes (p <0.05). Insulin also increased significantly the cell proliferation in contrast to leptin, which showed no effect. In conclusion, prostate carcinoma cells seem to be influenced by factors secreted by adipocytes that are able to increase their ability to proliferate, migrate and invade.
- Adipocyte secretome increases radioresistance of malignant melanocytes by improving cell survival and decreasing oxidative statusPublication . Coelho, Pedro; Silva, Liliana; Faria, Isabel; Vieira, Mónica; Monteiro, Armanda; Pinto, Gabriela; Prudêncio, Cristina; Fernandes, Rúben; Soares, RaquelRadiotherapy is a treatment option for the majority of malignancies. However, because melanoma is known to be radioresistant, the use of ionizing radiation as an adjuvant therapy in cutaneous melanoma patients is ineffective. Obesity has now been recognized as a risk factor for melanoma. High adiposity is generally associated with a more pro-oxidative status. Oxidative stress is a major player in radiation therapy and also a common link between obesity and cancer. Several adipocyte-released proteins are known to have a role in controlling cellular growth and pro-survival signaling. For that reason, we investigated the influence of 3T3-L1 mature adipocyte secretome in B16-F10 malignant melanocyte radiosensitivity. We evaluated B16-F10 cell survival and redox homeostasis when exposed to four daily doses of ionizing radiation (2 Gy per day) up to a total of 8 Gy in a medical linear accelerator. B16-F10 melanocytes exhibited slight alterations in survival, catalase activity, nitrative stress and total oxidant concentration after the first 2 Gy irradiation. The motility of the melanocytes was also delayed by ionizing radiation. Subsequent irradiations of the malignant melanocytes led to more prominent reductions in overall survival. Remarkably, 3T3-L1 adipocyte-secreted molecules were able to increase the viability and migration of melanocytes, as well as lessen the pro-oxidant burden induced by both the single and cumulative X-ray doses. In vitro adipocyte-released factors protected B16-F10 malignant melanocytes from both oxidative stress and loss of viability triggered by radiation, enhancing the radioresistant phenoyype of these cells with a concomitant activation of the AKT signaling pathway These results both help to elucidate how obesity influences melanoma radioresistance and support the usage of conventional medical linear accelerators as a valid model for the in vitro radiobiological study of tumor cell lines.
- Adipocyte-released factors enhance melanocyte’s proliferation and motilityPublication . Fernandes, Rúben; Coelho, Pedro; Almeida, Joana; Prudêncio, Cristina; Soares, RaquelObesity, favored by the modern lifestyle, acquired epidemic proportions nowadays. Obesity has been associated with various major causes of death and morbidity including malignant neoplasms. Cutaneous melanoma incidence rates have also been increasing uring the last four decades in several countries. Obesity involvement in melanoma etiology has been recognized, but the implicated mechanisms remain unclear. We propose to address the above relationship and investigate the mechanism interplaying between obesity and an increased risk of melanoma onset.
- Affinity coefficient for clustering autoregressive moving average modelsPublication . Nascimento, Ana Paula; Oliveira, Alexandra; Faria, Brígida Mónica; Pimenta, Rui; Vieira, Mónica; Prudêncio, Cristina; Nicolau, Helena BacelarIn various fields, such as economics, finance, bioinformatics, geology, and medicine, namely, in the cases of electroencephalogram, electrocardiogram, and biotechnology, cluster analysis of time series is necessary. The first step in cluster applications is to establish a similarity/dissimilarity coefficient between time series. This article introduces an extension of the affinity coefficient for the autoregressive expansions of the invertible autoregressive moving average models to measure their similarity between them. An application of the affinity coefficient between time series was developed and implemented in R. Cluster analysis is performed with the corresponding distance for the estimated simulated autoregressive moving average of order one. The primary findings indicate that processes with similar forecast functions are grouped (in the same cluster) as expected concerning the affinity coefficient. It was also possible to conclude that this affinity coefficient is very sensitive to the behavior changes of the forecast functions: processes with small different forecast functions appear to be well separated in different clusters. Moreover, if the two processes have at least an infinite number of π- weights with a symmetric signal, the affinity value is also symmetric.
- Alterações anatómicas induzidas pela idade na coluna vertebral: intervenção para melhoria da saúde da comunidadePublication . Ribas, Jorge; Ribas, C.; Prudêncio, CristinaO aumento do envelhecimento da população implica uma crescente importância na implementação da política de cuidados de saúde primários. O envelhecimento leva a profundas alterações na morfologia da coluna vertebral do indivíduo. Estas alterações não estão ainda devidamente caracterizadas pelo que é necessário estudar e tipificar quais as mudanças induzidas pela idade. Desta forma será possível avaliar de que forma o patológico se poderá enquadrar num padrão normal de envelhecimento.