FCS (DCM)- Artigos em Revistas Científicas Internacionais com Arbitragem Científica
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- Agrobacterium-mediated transformation of the terpenoid indole alkaloid-producing plant species Tabernaemontana pandacaquiPublication . Cardoso, M.I. Lopes; Meijer, A.H.; Hoge, J.H.C.Plants of the Apocynaceae family produce a wide range of terpenoid indole alkaloids (TIAs) which have important pharmaceutical applications. Studies of the molecular mechanisms controlling TIA biosynthesis may eventually provide possibilities to improve product yield by genetic modification of plants or cell cultures. However, these studies suffer from the lack of transformation/regeneration protocols for Apocynaceae plants. We chose to study the feasibility of Agrobacterium tumefaciens-mediated transformation of Tabernaemontana pandacaqui, because of the availability of an efficient regeneration procedure for this member of the Apocynaceae family. A procedure to produce transgenic T. pandacaqui plants was established, albeit with low efficiency. Transgenic expression was demonstrated of an intron-containing β-glucuronidase reporter gene and of a gene coding for the TIA biosynthetic enzyme strictosidine synthase from Catharanthus roseus, another Apocynaceae species.
- Alkaloid production by a Cinchona officinalis "Ledgeriana" hairy root culture containing constitutive expression constructs of tryptophan decarboxylase and strictosidine synthase cDNAs from Catharanthus roseusPublication . Geerlings, A.; Hallard, D.; Martinez Caballero, A.; Cardoso, I. Lopes; Heijden, R. Van Der; Verpoorte, R.Cinchona officinalis ‘Ledgeriana’, former called Cinchona ledgeriana, hairy roots were initiated containing constitutive-expression constructs of cDNAs encoding the enzymes tryptophan decarboxylase (TDC) and strictosidine synthase (STR) from Catharanthus roseus, two key enzymes in terpenoid indole and quinoline alkaloid biosynthesis. The successful integration of these genes and the reporter gene gus-int was demonstrated using Southern blotting and the polymerase chain reaction. The products of TDC and STR, tryptamine and strictosidine, were found in high amounts, 1200 and 1950 mg g–1 dry weight, respectively. Quinine and quinidine levels were found to rise up to 500 and 1000 mg g–1 dry weight, respectively. The results show that genetic engineering with multiple genes is well possible in hairy roots of C. officinalis. However, 1 year after analyzing the hairy roots for the first time, they had completely lost their capacity to accumulate alkaloids.
- An assay for secologanin in plant tissues based on enzymatic conversion into strictosidinePublication . Hallard, Didier; Heijden, Robert Van Der; Contin, Adriana; Tomaz Jiméréz, Emília M.; Snoeijer, Wim; Verpoorte, Robert; Jensen, Soren R.; Cardoso, M. Inês Lopes; Pasquali, Giancarlo; Memelink, Johan; Hoge, J. Harry C.The secoiridoid glucoside secologanin is the terpenoid building block in the biosynthesis of terpenoid indole alkaloids. A method for its determination in plant tissues and in cell suspension cultures has been developed. This assay is based on the condensation of secologanin with tryptamine, yielding strictosidine, in a reaction catalysed by the enzyme strictosidine synthase (STR; E.C. 4.3.3.2). Subsequently, the formation of strictosidine is quantified by high performance liquid chromatography (HPLC). STR was isolated from transgenic Nicotiana tabacum cells expressing a cDNA-derived gene coding for STR from Catharanthus roseus. The high specificity of STR for secologanin, in combination with a sensitive and selective HPLC system, allows a simple extraction of secologanin from plant tissue. The detection limit of this method is 15 ng secologanin. Using this assay, secologanin contents were determined in tissues of various plant species; Lonicera xylosteum hairy roots were found to contain 1% of secologanin on a dry weight basis. # 1998 John Wiley & Sons, Ltd.
- Apert syndrome and repercussions in Dental MedicinePublication . Paula, Lígia de; Cardoso, Inês LopesApert’s syndrome is a craniosynostosis syndrome caused by mutations in the gene coding for the fibroblast growthfactor receptor 2 (FGFR2), characterized by craniosynostosis, midface hypoplasia and syndactyly of the hands and feet. It has several oral manifestations, such as ogival palate, maxillary transverse and sagittal hypoplasia, dental crowding, eruptive delay and ectopic position of the teeth. The diagnosis of Apert’s syndrome is established in a proband with classic clinical characteristics, and genetic tests can also be performed. Patients with this syndrome often require craniofacial team care and dental, orthodontic and orthognathic surgical management because of their esthetic and functional problems such as Class III malocclusion and midface hypoplasia. The aim of this study is to present a literature review on oral manifestations of Apert’s syndrome and their impact on dental medicine.
- Assessment of Rhodopirellula rubra as a supplementary and nutritional food source to the microcrustacean Daphnia magnaPublication . Marinho, Maria da Conceição; Lage, Olga Maria; Sousa, Carla D.; Catita, José; Antunes, Sara C.The daily use of the planctomycete Rhodopirellula rubra as an alternative or supplementary food source for Daphnia magna and its feasibility in the nutrition of transgenerational populations were studied. The life history parameters, fatty acids (saturated, mono- and polyunsaturated; SFAs, MUFAs and PUFAs), glycogen and protein contents of organisms during feeding assays and of the first generation were analysed. An increase in the yields of D. magna with the increase of the cell concentration of R. rubra was evident, but overall, bacteria supplied as the only food source was nutritionally insufficient as observed for all the parameters analysed. However, when R. rubra was added as supplement to the microalgae Raphidocelis subcapitata a significant improvement in the life history parameters was observed namely in the reproductive output and the somatic growth rate. The identified SFAs, MUFAs and PUFAs were the fatty acids more abundant in daphniids, and the feed regimens influenced daphniids fatty acid profiles. Additionally, the mixed diet resulted in a larger number and size of offspring in the different F1 broods as also observed with the results of F0 generation. The pink colouration present in D. magna body and eggs confirmed that bacteria were absorbed, the pigment(s) retained and passed on to the next generation. Our results showed that R. rubra can play an essential role in D. magna diet as a nutritional supplement showing potential biotechnological applications.
- Bacterial contamination of gutta-percha points from different brands and the efficacy of a chairside disinfection protocolPublication . Bracciale, Francesca; Marino, Nicole; Noronha, Anariely; Manso, M. Conceição; Gavinha, Sandra; Cardoso, Inês Lopes; Pina, Cristina Maria San Román Gomes de; Teles, Ana MouraObjective: To evaluate the bacterial contamination of different brands of Gutta-Percha (GP) points routinely used in clinical practice and the efficacy of a chairside disinfection protocol with sodium hypochlorite. Methods: GP points (n=240), in sizes A, B, C, D, K15, K20, K25, K30, K35, K40, F1, F2, F3 (Dentsply®, Proclinic®, ProTaper® and R&S®), were randomly sampled from commercial packages already in use. These were added directly to Fluid Thioglycolate Medium (one GP point per tube) and incubated at 37ºC for 21 days. During this period, the presence/absence of turbidity was evaluated. To evaluate the efficacy of a chairside disinfection protocol, all detected contaminated GP points were immersed for 1 minute in 10 mL of 5.25% sodium hypochlorite, followed by 5 minutes in 10 mL of detergent solution (3% Tween 80 and 5% sodium thiosulfate) and a final rinse with 10 mL of sterile distilled water and incubated. The data was analysed using the chi-square test and differences between characteristics of dichotomic variables were performed using the binomial test. The significance level was set at P<0.05. Results: Bacterial growth was observed in 22.9% of the total study samples. Dentsply® and R&S® showed the highest level of contamination, 47.3% each, although without significant differences to the other commercial brands. The most contaminated GP point size was K30 (16.4%). The chairside disinfection protocol was effective in disinfection of 76.4% of GP points (P<0.001). Conclusion: A real small number of GP points in clinical use harboured bacteria, including after the Chairside Disinfection Protocol that, anyway, proved to be effective. No significant difference was observed between tested commercial brands.
- Biochemical mechanisms of the effect of environmental factors (diet, fluorine, alcohol and tobacco) on oral healthPublication . Cardoso, Inês Lopes; Leal, Fernanda; Santos, D.R.; Azevedo, W.S.Nowadays, oral diseases are prevalent chronic diseases present in worldwide population. Dental caries results from a complex interaction between acid-producing bacteria, fermentable carbohydrates and host factors, including teeth and saliva. Risk factors for caries development include physical, biological, environmental, behavioral factors such as high numbers of cariogenic bacteria, inadequate salivary flow, poor oral hygiene and poor eating habits. Moreover, due to its efficacy in caries prevention, fluorine has been added to toothpaste, supplements and water. However, despite of its beneficial effect, excessive intake can lead to acute or chronic toxic effects, that depend on ingestion period and ingested amount, age, cardiovascular or renal problems and genetic changes. The most common adverse effects of excessive fluorine intake are dental and skeletal fluorosis. Another oral health problem is oral cancer, one of the most common cancers in the world and a highly relevant problem of global public health, mainly affecting lips and oral cavity. It is a preventable disease, where tobacco and alcohol, considered major risk factors, are present with a synergic effect in 90% of cases. Alcohol can act as a risk factor both locally and systemically, being causally associated with oropharyngeal and larynx cancer, esophageal cancer, among others.
- Bisphenol A in Odontology: recent dataPublication . Vautherin, Thomas; Cardoso, Inês LopesBackground and Objective: Bisphenol A (BPA) is an organic compound of the aromatic family used mainly as a monomer, for more than 50 years in the production of polycarbonate plastics and epoxy resins. Nowadays, bisphenol A is a worrying substance that deserves special attention. The goal of this work is to focus on bisphenol A and its biological effects on the human body. Methods: A bibliographic search was performed in databases such as PubMed, b-On and Elsevier. The keywords used were: "Bisphenol A", "BPA", "Endocrin Disruptor Chemicals". Results: There are several known effects on the human body resulting from the exposure to BPA such as in the reproductive system, dentition, brain, behavior, cardiovascular and in immune systems. In the oral cavity, dental composites release bis-GMA and BPA immediately after placement but released doses are usually too low to lead to biological effects. Conclusion: There are other materials such as silorans, Ormocer® and ionomer glass cements that can be good alternatives. Clinical studies are still neaded to prove that these materials are better than the composites, being these ones still the most used materials in conservative dental medicine.
- C reactive protein and cardiovascular diseasePublication . Cardoso, Inês Lopes; Paulos, Ana TeresaCardiovascular Diseases (CVD) are the first cause of death in developed countries, therefore it is of interest to reduce this public health problem. The development of atherosclerosis is the main cause of CVD. This pathology results from the accumulation of lipids in the arterial wall, that leads to a complex inflammatory process. Inflammatory biomarkers are a valuable tool in the detection and monitorization of the evolution of this process, as well as in the choice of therapy to implement. C-Reactive Protein (CRP), determined by high sensitivity methods (hs-CRP), is the most studied biomarker and stands out among the others, being considered an important marker of inflammation. Its importance comes from the fact that its plasma level is not affected by large diurnal or seasonal variations, and for this reason is indicated as an important mediator of the atherosclerotic process. The goal of this work is to deepen knowledge concerning the importance of C reactive protein as a risk factor for the development of cardiovascular diseases and discuss how it can be used in primary as well as secondary prevention of CVD. The use of C reactive protein as a biomarker allows detection, monitorization and prevention of cardiovascular disease.
- Comparison of neutral lipid fatty acids composition in organisms from different trophic levelsPublication . Lage, Olga M.; Antunes, Sara C.; Marinho, Conceição; Catita, JoséThe profiles of total fatty acids (TFAs) and the neutral lipid fatty acids (NLFAs) were compared for the bacterium Rhodopirellula rubra and the alga Raphidocelis subcapitata (conventional food source for Daphnia magna). D. magna NLFAs were assessed when this crustacean was fed with bacterium and alga, individually or in combination. After NLFA extraction, the profiles of the various organisms were characterized by gas chromatography. Results evidenced the relevance of the different composition of the fatty acid (FAs) fractions in the different organisms, R. rubra and R. subcapitata. In these species, the NFLA analyses revealed high amounts of long chain FAs (C19). The FA profile of D. magna was influenced by the different diets provided although the preferred diet was the alga. D. magna showed the capacity to adapt to the available food resources as it defines its FA profile according to its needs, namely for the long chain FAs (C19).