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Research Interests: DNA damage, Medicine, Environmental Biotechnology, Humans, Toxicity, and 12 moreMutation, Coefficient of Variation, Micronucleus, Observer Variation, Occupational Exposure, Reproducibility of Results, Formaldehyde, Lymphocytes, Micronucleus Test, Micronucleus tests, Pharmacology and pharmaceutical sciences, and Wilcoxon Signed Rank Test
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Dietary habits are recognized to be an important factor influencing cancer risk and tumour behaviour. Diet can influence cancer development in several ways, as direct action of carcinogens in food can damage DNA, and some diet components... more
Dietary habits are recognized to be an important factor influencing cancer risk and tumour behaviour. Diet can influence cancer development in several ways, as direct action of carcinogens in food can damage DNA, and some diet components (macro or micronutrients) can block or induce enzymes involved in activation or deactivation of carcinogenic substances. Moreover, inadequate intake of some molecules involved in DNA synthesis, repair or methylation can influence mutation rate or changes in the gene expression mechanism. From a mechanistic view of carcinogenesis, food mutagens are classified as genotoxic and non-genotoxic. Genotoxic agents cause DNA damage resulting in gene point mutations, deletions, and insertions, recombination, rearrangements, and amplifications, as well as chromosomal aberrations. Non-genotoxic agents are less distinctively defined in terms of their modes of action, but they are presumed to indirectly affect cell proliferation as tumours promoters, with or without accompanying chronic cell damage. Another mechanism that diet can influence DNA mutation, and consequently cancer risk, is energy balance and growth rates since nutrition will influence hormone levels and growth factors that will influence the rate of cell division, cell cycling and consequently influence time for DNA repair and/or replication of DNA lesions. Nutritional genomics studies the functional interaction of food and its components, macro and micronutrients, with genome at molecular, cellular, and systemic level. One of the goals is to identify biomarkers that will provide better guidance on the relationship between nutrition and health. Also relevant are the implications of genetic polymorphisms and their role in the interaction between diet, environmental factors, lifestyles, and cancer risk. The recognition of the importance of adequate dietary levels of micronutrients in maintaining genomic stability is very significant because the latter is also affected by inadequate nutrient intakes, such as lack of vitamins A, D, E, folate, selenium, and others.info:eu-repo/semantics/publishedVersio
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Abstract Human exposure to mycotoxins can be determined by environmental and biological monitoring. Mycotoxins can assume carcinogenic, mutagenic, teratogenic, and immunotoxic properties, and biomonitoring instruments can be applied to... more
Abstract Human exposure to mycotoxins can be determined by environmental and biological monitoring. Mycotoxins can assume carcinogenic, mutagenic, teratogenic, and immunotoxic properties, and biomonitoring instruments can be applied to assess their effects. Cytokinesis-block micronucleus assay is a method that allows cytological scoring of nuclear abnormalities resulting from genotoxic insult. Its use is well established in in vitro genetic toxicology testing and has become an accepted standard method to assess the genotoxic hazard of chemicals, which has led to the development of the Organization for Economic Cooperation and Development. Generally, citrin, patulin, ochratoxin, and zearaleone genotoxicity are demonstrated by the increased presence of micronucleus. The comet assay, a standard method for assessing DNA damage, has been widely used to assess genotoxic effects, namely owing to oxidative damage, using formamidopyrimidine DNA glycosilase. Studies performed to evaluate genotoxicity in zearaleone, ochratoxin, aflatoxin, citrinin, and patulin demonstrate increased values in the parameters measured.
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ABSTRACT Cytokinesis blocked micronucleus (CBMN) assay and comet assay are the most promising short-term genotoxicity assays for human risk assessment and their combination is recommended to monitor populations chronically exposed to... more
ABSTRACT Cytokinesis blocked micronucleus (CBMN) assay and comet assay are the most promising short-term genotoxicity assays for human risk assessment and their combination is recommended to monitor populations chronically exposed to genotoxic agents. Nutrition is recognized to be an important lifestyle factor that influences cancer risk, and should be taken into account at an individual level. Laboratories are occupational settings where chemical agents are handled and workers are exposed. Formaldehyde and cytostatic drugs, in particular, are chemical agents handled in laboratories that are considered carcinogenic for humans and special protective measures should thus be adopted against them. The aim of this investigation is to contribute to the development of a biomonitoring programme that includes genotoxicity assessment related with genetic susceptibility biomarkers, and lifestyle factors, namely nutrition. The experimental planning used was a case-control blinded study. Four separated samples were formed comprising two samples of subjects exposed (n = 56 for formaldehyde; n = 46 for cytostatics), and two samples of non-exposed controls (n = 85 and n = 46, respectively). Participants filled-in a personal and a food frequency questionnaires. CBMN and comet assays were used to assess genotoxicity. Individual susceptibility was investigated by Real Time PCR. Measurements of serum vitamins A and E were performed by HPLC, and vitamin D by ELISA. The risk of genotoxicity in those exposed was then compared with the risk in the controls, allowing for the quantitative measurement of association between exposure and genotoxicity. In both occupational settings, the genotoxicity biomarkers were significantly higher in the exposed than in the non-exposed controls (p<0.05). In the formaldehyde occupational context, significant associations were found between XRCC3 genotypes and nuclear buds; but that was not the case either for ADH5 or VDR. Comet assay did not identify significant differences between those exposed to cytostatics and controls, and the same lack of association applies to the OGG1 genotypes. Micronutrients association to the genotoxicity biomarkers was controversial; we have found positive correlations for vitamin A and negative ones for vitamin E. Our findings emphasize the need for the implementation of a regular biomonitoring programme of personnel occupationally exposed to drugs like formaldehyde and cytostatics.
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Human biomonitoring consists, in one hand by the research and investigation of environmental hazards and, in other hand in risk estimating of developing cancer by exposure to those conditions. Since carcinogenesis is a process that takes... more
Human biomonitoring consists, in one hand by the research and investigation of environmental hazards and, in other hand in risk estimating of developing cancer by exposure to those conditions. Since carcinogenesis is a process that takes time, the biomarkers applied to recognize abnormal biological events have been developed in molecular epidemiological studies. The biomarkers allow the quantification and identification of the progression of normal to abnormal biological conditions at a molecular level. In general, biomarkers can be classified in exposure, effect and genetic susceptibility. Genotoxicity biomarkers are a sub-type of effect biomarkers and are widely used in the assessment of genomic effects caused by exposure – environmental or occupational, being considered predictors of carcinogenesis development. Taken together, and inserted in a biomonitoring program, probably is the most growing tool available nowadays for the prevention of health effects from occupational exposure to chemicals. Ethical considerations should always be taken into consideration in the planning and implementation of the biomarkers involved.info:eu-repo/semantics/publishedVersio
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Risk assessment of chemicals mainly relies on exposure to single chemicals and their hazardous effects, although the real scenario in occupational settings is more commonly characterized by exposure to chemi-cal mixtures. Chromium (Cr),... more
Risk assessment of chemicals mainly relies on exposure to single chemicals and their hazardous effects, although the real scenario in occupational settings is more commonly characterized by exposure to chemi-cal mixtures. Chromium (Cr), Nickel (Ni), and Polycyclic Aromatic Hydrocarbons (PAHs) are often pre-sent in occupational settings, such as aeronautic industries, where workers are exposed primarily through inhalation. Considering that those substances are recognized lung carcinogens acting mainly by genotoxic mechanisms, it is likely that interactive effects occur, indicating that the risk from occupational exposure to these chemicals should be assessed as a mixture. Such issue is being addressed in the HBM4EU Initiati-ve, where a real scenario of occupational exposure has been studied. In order to provide support to the hazard assessment of the referred mixture, the present work aimed to evaluate the combined toxicity of Cr(VI), Ni, and benzo(a)pyrene (BaP), using a human lung cell line (A549 cells). MTT assay was performed for each individual chemical and a dose-response curve was esta-blished, enabling the determination of the IC50. The combined toxicity of the Cr and Ni mixture as well as that of Cr, Ni and BaP were determined comparatively to the single chemicals’ toxicity to ascertain whet-her additive effects or deviations from additivity towards synergism or antagonism was obtained. The results will be presented and discussed and are expected to contribute to the overall mixture’s risk asses-sment.Funded by the ChemMix (IDI&CA, IPL, 2019) and HBM4EU (GA 733032) projects and by FCT/MCTES through national funds (UIDB/00009/2020; UIDP/00009/2020).N/
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Funded by the HBM4EU Initiative (GA 733032). ChemMix (IDI&CA, IPL, 2019). FCT_UIDB/00009/2020. FCT_UIDP/00009/2020.The environment within industrial settings is commonly characterized by the existence of a complex mixture of chemicals... more
Funded by the HBM4EU Initiative (GA 733032). ChemMix (IDI&CA, IPL, 2019). FCT_UIDB/00009/2020. FCT_UIDP/00009/2020.The environment within industrial settings is commonly characterized by the existence of a complex mixture of chemicals from different raw materials and transformation processes. Occupational co-exposure to chromium (Cr), Nickel (Ni), and Polycyclic Aromatic Hydrocarbons (PAHs) may occur in diverse workplaces, such as aeronautic and waste management, (e.g incineration) sectors. Such co-exposure raises concern in terms of occupational health, as these substances are recognized lung carcinogens and mainly act by genotoxic mechanisms, increasing the likelihood of interactive toxic effects. The fact that current regulatory practices are usually focused on single chemical substances, without integrating the possibility of combined or aggregated exposures and effects, may lead to a risk underestimation. This work, developed under the scope of HBM4EU Initiative (https://www.hbm4eu.eu), a literature-based mixture risk assessment (MRA) exercise for occupational exposure to metals and PAHs was performed. In addition, in vitro toxicity data was obtained for the same mixtures to provide support to its hazard assessment. Human biomonitoring (HBM) data on Cr(VI), Ni, and/or PAHs were extracted from occupational studies conducted in the European Union and searched in literature databases. Selected reference values were used to calculate risk quotients (RQ) for each substance based on the retrieved exposure data; the combined risk was given by the sum of the RQ, i.e., the Background Exposure Exceedance Score(BEES). In parallel, we evaluated the combined cyto- and genotoxicity of the same chemicals (assessed by the MTT and micronucleus assays) in the human alveolar A549 cell line. In most of the analyzed studies, we observed that BEES levels, estimated from the exposure to metals mixture or to metals and PAHs, exceeded RQ levels considered acceptable for the individual substances. Only two studies, conducted in hazard waste incinerator settings, presented urinary exposure levels for the three substances. They showed a value of BEES of concern (>1) for all exposure scenarios, even for workers performing activities considered of low exposure or no exposure, such as laboratory and administrative workers. In vitro assays supported that A549 cells' exposure to these substances resulted in interactive cytotoxic and genotoxic effects that may underlie health effects different from those predicted from single exposures. Our findings show the limitations of applying occupational exposure reference values defined on a single substance basis to workplaces where exposure to chemical mixtures occurs, highlighting the relevance of performing MRA as a more realistic approach to guide suitable risk management measures in occupational settings.info:eu-repo/semantics/publishedVersio
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Research Interests: Bioinformatics, Occupational Health, Biology, Life Sciences, Medicine, and 15 moreExposure Assessment, Biomedical Research, Occupational health and safety, Micronucleus, Occupational Exposure, Group, Health risk, In Vitro Studies, Formaldehyde, Occupational Safety and Health Knowledge, Carcinogen, Control Group, ENVIRONMENTAL SCIENCE AND MANAGEMENT, Cumulant, and Micronucleus Test
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Emerging contaminants such as nanoplastics (NPs), as well as manufacturing by-products such as plasticizers, have gained global attention and concern due to their limited biodegradability and their potential impact on human health, in... more
Emerging contaminants such as nanoplastics (NPs), as well as manufacturing by-products such as plasticizers, have gained global attention and concern due to their limited biodegradability and their potential impact on human health, in particular the effects on respiratory tissue. In parallel, in vitro cell culture techniques are key to the assessment and characterization of toxic effects and cellular mechanisms in different types of tissues and should provide relevant information to understand the hazardous potential of these emergent contaminants. This systematic review presents the main results on the current knowledge of the effects of NPs and plasticizers on lung cells, as assessed with the use of in vitro cell culture techniques. From the selected studies (n = 10), following the PRISMA approach, it was observed that cell viability was the most frequently assessed endpoint and that most studies focused on epithelial cells and exposures to polystyrene (PS). It was observed that e...
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The comet assay is a versatile method to detect nuclear DNA damage in individual eukaryotic cells, from yeast to human. The types of damage detected encompass DNA strand breaks and alkali-labile sites (e.g., apurinic/apyrimidinic sites),... more
The comet assay is a versatile method to detect nuclear DNA damage in individual eukaryotic cells, from yeast to human. The types of damage detected encompass DNA strand breaks and alkali-labile sites (e.g., apurinic/apyrimidinic sites), alkylated and oxidized nucleobases, DNA–DNA crosslinks, UV-induced cyclobutane pyrimidine dimers and some chemically induced DNA adducts. Depending on the specimen type, there are important modifications to the comet assay protocol to avoid the formation of additional DNA damage during the processing of samples and to ensure sufficient sensitivity to detect differences in damage levels between sample groups. Various applications of the comet assay have been validated by research groups in academia, industry and regulatory agencies, and its strengths are highlighted by the adoption of the comet assay as an in vivo test for genotoxicity in animal organs by the Organisation for Economic Co-operation and Development. The present document includes a series of consensus protocols that describe the application of the comet assay to a wide variety of cell types, species and types of DNA damage, thereby demonstrating its versatility.
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Background Calculating the disease burden due to injury is complex, as it requires many methodological choices. Until now, an overview of the methodological design choices that have been made in burden of disease (BoD) studies in injury... more
Background Calculating the disease burden due to injury is complex, as it requires many methodological choices. Until now, an overview of the methodological design choices that have been made in burden of disease (BoD) studies in injury populations is not available. The aim of this systematic literature review was to identify existing injury BoD studies undertaken across Europe and to comprehensively review the methodological design choices and assumption parameters that have been made to calculate years of life lost (YLL) and years lived with disability (YLD) in these studies. Methods We searched EMBASE, MEDLINE, Cochrane Central, Google Scholar, and Web of Science, and the grey literature supplemented by handsearching, for BoD studies. We included injury BoD studies that quantified the BoD expressed in YLL, YLD, and disability-adjusted life years (DALY) in countries within the European Region between early-1990 and mid-2021. Results We retrieved 2,914 results of which 48 performed...
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Hexavalent chromium (Cr(VI)), nickel (Ni) and polycyclic aromatic hydrocarbons (PAHs) are genotoxic co-occurring lung carcinogens whose occupational health risk is still understudied. This study, conducted within the European Human... more
Hexavalent chromium (Cr(VI)), nickel (Ni) and polycyclic aromatic hydrocarbons (PAHs) are genotoxic co-occurring lung carcinogens whose occupational health risk is still understudied. This study, conducted within the European Human Biomonitoring Initiative (HBM4EU), aimed at performing a mixtures risk assessment (MRA) based on published human biomonitoring (HBM) data from Cr(VI), Ni and/or PAHs occupational co-exposure in Europe. After data extraction, Risk Quotient (RQ) and Sum of Risk Quotients (SRQ) were calculated for binary and ternary mixtures to characterise the risk. Most selected articles measured urinary levels of Cr and Ni and a SRQ > 1 was obtained for co-exposure levels in welding activities, showing that there is concern regarding co-exposure to these substances. Similarly, co-exposure to mixtures of Cr(VI), Ni and PAHs in waste incineration settings resulted in SRQ > 1. In some studies, a low risk was estimated based on the single substances’ exposure level (RQ ...
Research Interests: Portuguese, Environmental Health, Environmental Chemistry, Risk assessment, Medicine, and 15 moreHealth Risk Assessment, Genotoxicity, Biomonitoring, Workers, Comet Assay, Micronucleus, Occupational Exposure, Hexavalent Chromium, Risk Assessment, Toxics, Human Biomonitoring, Air Samples, alkaline comet assay, Peripheral blood, and Hazard quotient
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Background Assessment of disability-adjusted life years (DALYs) resulting from non-communicable diseases (NCDs) requires specific calculation methods and input data. The aims of this study were to (i) identify existing NCD burden of... more
Background Assessment of disability-adjusted life years (DALYs) resulting from non-communicable diseases (NCDs) requires specific calculation methods and input data. The aims of this study were to (i) identify existing NCD burden of disease (BoD) activities in Europe; (ii) collate information on data sources for mortality and morbidity; and (iii) provide an overview of NCD-specific methods for calculating NCD DALYs. Methods NCD BoD studies were systematically searched in international electronic literature databases and in grey literature. We included all BoD studies that used the DALY metric to quantify the health impact of one or more NCDs in countries belonging to the European Region. Results A total of 163 BoD studies were retained: 96 (59%) were single-country or sub-national studies and 67 (41%) considered more than one country. Of the single-country studies, 29 (30%) consisted of secondary analyses using existing Global Burden of Disease (GBD) results. Mortality data were mai...
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Introdução: O carcinoma da mama é a forma de cancro mais comum na mulher e as taxas de incidência têm vindo a subir gradualmente. No passado recente, diversas estratégias têm vindo a ser aplicadas para um combate mais eficiente a esta... more
Introdução: O carcinoma da mama é a forma de cancro mais comum na mulher e as taxas de incidência têm vindo a subir gradualmente. No passado recente, diversas estratégias têm vindo a ser aplicadas para um combate mais eficiente a esta doença, nomeadamente a terapêutica por trastuzumab® que possui como alvo a glicoproteína transmembranar de 185 kD codificada pelo proto-oncogene HER-2. Verificou-se que em 15% a 30% dos casos de carcinoma da mama, o gene HER-2 se encontra amplificado, o que conduz à produção de proteína HER-2 em redundância e consequentemente torna os mecanismos de estimulação do crescimento tumoral demasiado exuberantes. A identificação da expressão de HER-2 no carcinoma da mama por Imunocitoquímica (ICQ) é indispensável quando se considera a elegibilidade dos doentes para a terapêutica com o trastuzumab®. Objectivo: Este trabalho tem como principal objectivo descrever o papel da ICQ na qualificação do status HER-2 e apresentar a consequente abordagem terapêutica pelo trastuzumab®.info:eu-repo/semantics/publishedVersio
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Funded by the HBM4EU Initiative (GA 733032). ChemMix (IDI&CA, IPL, 2019). FCT_UIDB/00009/2020. FCT_UIDP/00009/2020.The environment within industrial settings is commonly characterized by the existence of a complex mixture of chemicals... more
Funded by the HBM4EU Initiative (GA 733032). ChemMix (IDI&CA, IPL, 2019). FCT_UIDB/00009/2020. FCT_UIDP/00009/2020.The environment within industrial settings is commonly characterized by the existence of a complex mixture of chemicals from different raw materials and transformation processes. Occupational co-exposure to chromium (Cr), Nickel (Ni), and Polycyclic Aromatic Hydrocarbons (PAHs) may occur in diverse workplaces, such as aeronautic and waste management, (e.g incineration) sectors. Such co-exposure raises concern in terms of occupational health, as these substances are recognized lung carcinogens and mainly act by genotoxic mechanisms, increasing the likelihood of interactive toxic effects. The fact that current regulatory practices are usually focused on single chemical substances, without integrating the possibility of combined or aggregated exposures and effects, may lead to a risk underestimation. This work, developed under the scope of HBM4EU Initiative (https://www.hbm4eu.eu), a literature-based mixture risk assessment (MRA) exercise for occupational exposure to metals and PAHs was performed. In addition, in vitro toxicity data was obtained for the same mixtures to provide support to its hazard assessment. Human biomonitoring (HBM) data on Cr(VI), Ni, and/or PAHs were extracted from occupational studies conducted in the European Union and searched in literature databases. Selected reference values were used to calculate risk quotients (RQ) for each substance based on the retrieved exposure data; the combined risk was given by the sum of the RQ, i.e., the Background Exposure Exceedance Score(BEES). In parallel, we evaluated the combined cyto- and genotoxicity of the same chemicals (assessed by the MTT and micronucleus assays) in the human alveolar A549 cell line. In most of the analyzed studies, we observed that BEES levels, estimated from the exposure to metals mixture or to metals and PAHs, exceeded RQ levels considered acceptable for the individual substances. Only two studies, conducted in hazard waste incinerator settings, presented urinary exposure levels for the three substances. They showed a value of BEES of concern (>1) for all exposure scenarios, even for workers performing activities considered of low exposure or no exposure, such as laboratory and administrative workers. In vitro assays supported that A549 cells' exposure to these substances resulted in interactive cytotoxic and genotoxic effects that may underlie health effects different from those predicted from single exposures. Our findings show the limitations of applying occupational exposure reference values defined on a single substance basis to workplaces where exposure to chemical mixtures occurs, highlighting the relevance of performing MRA as a more realistic approach to guide suitable risk management measures in occupational settings.info:eu-repo/semantics/publishedVersio
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Electromagnetic fields (EMF) are classified as "possibly carcinogenic" by the International Agency for Research on Cancer (IARC). Some publications have reported associations between EMF exposure and DNA damage, but many other... more
Electromagnetic fields (EMF) are classified as "possibly carcinogenic" by the International Agency for Research on Cancer (IARC). Some publications have reported associations between EMF exposure and DNA damage, but many other studies contradict such findings. Cytomorphological changes, such as micronuclei (MN), indicative of genomic damage, are biomarkers of genotoxicity. To test whether mobile phone-associated EMF exposure affects the MN frequency in exfoliated buccal cells, we obtained cells smears from the left and right inner cheeks of healthy mobile phone users, aged 18-30 (n=86), who also completed a characterization survey. MN frequencies were tested for potential confounding factors and for duration of phone use and preferential side of mobile phone use. No relationship was observed between MN frequency and duration of mobile phone use in daily calls. Cells ipsilateral to mobile phone use did not present a statistically significantly higher MN frequency, compared ...
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It is relevant to develop new monitoring techniques of carcinogenic risk associated to environmental exposition to genotoxic chemicals. The conventional biomonitoring techniques are based on laborious, expensive methods as the ones... more
It is relevant to develop new monitoring techniques of carcinogenic risk associated to environmental exposition to genotoxic chemicals. The conventional biomonitoring techniques are based on laborious, expensive methods as the ones requiring isolation of lymphocytes from peripheral blood, in vitro cell culture, followed by e.g. cytokinesis-block assay and microscope observation of chromosomal abnormalities. The present work evaluated an infrared spectroscopy method, based on a simple, more economic and high-throughput procedure of analysis of whole blood processed with methanol. It was possible to identify ratios of spectral bands that are statistically different between hospital professionals occupationally exposed to antineoplastic drugs, such as 5-fluorouracil and non-hospital professionals without this exposure. It was also identified ratios of spectral bands which are statistically different between participants presenting lymphocytes with chromosomal abnormalities (as micronucleus, nuclear buds and nucleoplasmatic bridges) and participants not presenting these abnormalities. The infrared spectroscopy-based method presents therefore appealing characteristics to be applied in more intensive and/or large-scale studies of monitoring genotoxic risks.
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Biomarcadores: alterações celulares, bioquímicas ou moleculares mensuráveis em meios biológicos, tais como tecidos, células ou fluídos; determinantes quantificáveis de eventos biológicos que permitem a discriminação entre condições... more
Biomarcadores: alterações celulares, bioquímicas ou moleculares mensuráveis em meios biológicos, tais como tecidos, células ou fluídos; determinantes quantificáveis de eventos biológicos que permitem a discriminação entre condições biológicas normais e anormais; idealmente devem ser acessíveis, não invasivos, não destrutivos e fáceis de quantificar e pouco dispendiosos. Biomarcadores tumorais: proteínas e proteínas conjugadas ( hormonas, fragmentos de proteínas), péptidos ou hidratos de carbono; substâncias bioquímicas associadas com a carcinogénese; podem ser produzidos pelas células cancerígenas ou pelo indivíduo em resposta ao cancro; existem 3 tipos principais de marcadores tumorais que são libertados na circulação e medidos, normalmente em amostras de sangue.info:eu-repo/semantics/publishedVersio
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O estudo baseou-se na alteração do processo de desidratação de rotina, em que foi suprimida a concentração intermédia (96%) de etanol, com o intuito de tentar diminuir o tempo do processamento histológico sem provocar alterações nos... more
O estudo baseou-se na alteração do processo de desidratação de rotina, em que foi suprimida a concentração intermédia (96%) de etanol, com o intuito de tentar diminuir o tempo do processamento histológico sem provocar alterações nos tecidos. Objectivo do estudo: verificar se existem diferenças significativas a nível tecidular e celular em diferentes tipos de tecidos entre o método de desidratação de rotina e o alternativo