Publications by authors named "Rúben Fernandes"

40 Publications

The Influence of miRNAs on Radiotherapy Treatment in Prostate Cancer - A Systematic Review.

Front Oncol 2021 3;11:704664. Epub 2021 Aug 3.

LaBMI - Laboratory of Medical & Industrial Biotechnology, Porto Research, Technology & Innovation Center (PORTIC), P.PORTO - Polytechnic Institute of Porto, Porto, Portugal.

In the last years, extensive investigation on miRNomics have shown to have great advantages in cancer personalized medicine regarding diagnosis, treatment and even clinical outcomes. Prostate cancer (PCa) is the second most common male cancer and about 50% of all PCa patients received radiotherapy (RT), despite some of them develop radioresistance. Here, we aim to provide an overview on the mechanisms of miRNA biogenesis and to discuss the functional impact of miRNAs on PCa under radiation response. As main findings, 23 miRNAs were already identified as being involved in genetic regulation of PCa cell response to RT. The mechanisms of radioresistance are still poorly understood, despite it has been suggested that miRNAs play an important role in cell signaling pathways. Identification of miRNAs panel can be thus considered an upcoming and potentially useful strategy in PCa diagnosis, given that radioresistance biomarkers, in both prognosis and therapy still remains a challenge.
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http://dx.doi.org/10.3389/fonc.2021.704664DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8369466PMC
August 2021

Cell-adhesion molecules as key mechanisms of tumor invasion: the case of breast cancer.

Curr Mol Med 2021 Aug 6. Epub 2021 Aug 6.

National Health Institute Ricardo Jorge, Porto. Portugal.

Cancer is a major health problem worldwide and the second leading cause of death only overcome by cardiovascular diseases. Breast cancer is the leading cause of mortality and morbidity among women and one of the most common malignant neoplasms prompt to metastatic disease. In the present review, the mechanisms of the major cell adhesion molecules involved in tumor invasion are discussed, focusing in the case of breast cancer. A non-systematic updated revision of the literature was performed in order to assemble information regarding the expression of the adhesion cell molecules associated with metastasis.
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http://dx.doi.org/10.2174/1566524021666210806155231DOI Listing
August 2021

Suitability for e-health of non-pharmacological interventions in connective tissue diseases: scoping review with a descriptive analysis.

RMD Open 2021 Jul;7(2)

Section for Outcomes Research, Medical University of Vienna, Vienna, Austria

Objective: Non-pharmacological interventions support patients with connective tissue diseases to better cope with and self-manage their diseases. This study aimed to map existing evidence on non-pharmacological interventions in patients with systemic lupus erythematosus (SLE), systemic sclerosis (SSc) and mixed connective tissue diseases regarding content, feasibility and potential suitability in an e-health setting.

Methods: A literature search was performed in eight different databases in July 2020. The intervention's content was extracted using the 'Better reporting of interventions: template for intervention description and replication (TIDieR) checklist and guide'. A Sankey diagram and descriptive statistics were used to analyse the data and illustrate the relationships between the interventions.

Results: Of 8198 identified records, 119 papers were eligible. One hundred and four of them (87.4%) were conducted between 2000 and 2020, mainly in the USA (SLE n=24 (21.2%), SSc n=16 (14.2%)), Brazil (SLE n=8 (7.1%), SSc n=5 (4.4%)) and Italy (SLE n=0 (0%), SSc n=12 (10.6%)). Fifty-two studies (SLE n=24 (21.2%), SSc n=28 (24.8%)) used multicomponent interventions. The single interventions were physical exercises (SLE n=16 (14.2%), SSc n=17 (15.0%)), coaching/counselling (SLE n=11 (18.0%), SSc n=0 (0%)) and education (SLE n=2 (1.8%), SSc n=3 (2.7%)). Primary outcomes focused on physical function (SLE n=1 (0.9%), SSc n=15 (13.3%)), mouth opening in SSc (n=4 (5.9%)) and physical capacity (SLE n=2 (1.8%), SSc n=1 (0.9%)). No interventions for mixed connective tissue disease were found.

Conclusion: There was a great variety in the intervention's content due to differences in body structure, activity limitations and participation restrictions in SLE and SSc. These results highlight the need for personalised, multicomponent, non-pharmacological interventions, which could be delivered as e-health interventions.
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http://dx.doi.org/10.1136/rmdopen-2021-001710DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8323400PMC
July 2021

Plastic Antibody of Polypyrrole/Multiwall Carbon Nanotubes on Screen-Printed Electrodes for Cystatin C Detection.

Biosensors (Basel) 2021 May 31;11(6). Epub 2021 May 31.

Biomedical Engineering Laboratory, Federal University of Pernambuco, Recife-PE 50670-901, Brazil.

This work reports the design of a novel plastic antibody for cystatin C (Cys-C), an acute kidney injury biomarker, and its application in point-of-care (PoC) testing. The synthetic antibody was obtained by tailoring a molecularly imprinted polymer (MIP) on a carbon screen-printed electrode (SPE). The MIP was obtained by electropolymerizing pyrrole (Py) with carboxylated Py (Py-COOH) in the presence of Cys-C and multiwall carbon nanotubes (MWCNTs). Cys-C was removed from the molecularly imprinted poly(Py) matrix (MPPy) by urea treatment. As a control, a non-imprinted poly(Py) matrix (NPPy) was obtained by the same procedure, but without Cys-C. The assembly of the MIP material was evaluated in situ by Raman spectroscopy and the binding ability of Cys-C was evaluated by the cyclic voltammetry (CV) and differential pulse voltammetry (DPV) electrochemical techniques. The MIP sensor responses were measured by the DPV anodic peaks obtained in the presence of ferro/ferricyanide. The peak currents decreased linearly from 0.5 to 20.0 ng/mL of Cys-C at each 20 min successive incubation and a limit of detection below 0.5 ng/mL was obtained at pH 6.0. The MPPy/SPE was used to analyze Cys-C in spiked serum samples, showing recoveries <3%. This device showed promising features in terms of simplicity, cost and sensitivity for acute kidney injury diagnosis at the point of care.
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http://dx.doi.org/10.3390/bios11060175DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8228410PMC
May 2021

The influence of gut microbiota in cardiovascular diseases-a brief review.

Porto Biomed J 2021 Jan-Feb;6(1):e106. Epub 2021 Jan 18.

LABMI-PORTIC, Laboratory of Medical & Industrial Biotechnology, Porto Research, Technology and Innovation Center, Porto Polytechnic.

Lately, the gut microbiota has emerged as an important mediator of the development and the outcomes of certain diseases. It's well known that the gut microbiota plays an important role in maintaining human health. Still far from being completely understood and analyzed is the complexity of this ecosystem, although a close relationship between the gut microbiota and cardiovascular diseases (CVD) has been established. A loss of diversity in the microbiota will lead to physiological changes, which can improve inflammatory or infection states like atherosclerosis and hypertension, the basic pathological process of CVD. Targeting the gut microbiota and its metabolites are new and promising strategies for the treatment and prognosis of CVD.
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http://dx.doi.org/10.1097/j.pbj.0000000000000106DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7817281PMC
January 2021

A state of the art review on the novel mediator asprosin in the metabolic syndrome.

Porto Biomed J 2020 Nov-Dec;5(6):e108. Epub 2020 Dec 10.

Department of Medicine, Faculty of Medicine, University of Porto.

Metabolic syndrome is a complex and heterogeneous pathology characterized by a cluster of biochemical, clinical, and metabolic factors that came together in raising the risk of cardiovascular diseases, type 2 diabetes mellitus, and all-cause mortality. Some of these features are well defined in this syndrome like: obesity, inflammation, hypertension, insulin resistance, atherosclerotic dyslipidemias, endothelial dysfunction, and inflammation. This circuit is intermediated by a complex network of hormones, cytokines, transcription factors, and adipokines, among others. Some like leptin, adiponectin, Plasminogen activator inhibitor-1, interleukin-6, Tumor necrosis factor, and their influence on the metabolic syndrome are well described in the literature and new players are described continuously. One novel player was described in 2016 by Romere et al as a fasting-induced glycogenic protein hormone named asprosin. In order to perform a state-of-the-art, nonsystematic review of asprosin, a study of the available literature was carried out in the main database (Pubmed) and the results were studied and correlated to better understand the mechanism of action of this hormone. Asprosin is not only associated with the metabolic syndrome features like glucose and lipid metabolism, insulin resistance, obesity and inflammation but also in other pathologies metabolic syndrome related like diabetic retinopathy, polycystic ovary syndrome and anorexia nervosa. A limited number of pathways were already unveiled although much more research is needed to better understand the therapeutical potential of asprosin in the metabolic syndrome.
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http://dx.doi.org/10.1097/j.pbj.0000000000000108DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7732265PMC
December 2020

Schistosomiasis and Infertility: What Do We Know?

Trends Parasitol 2019 12 14;35(12):964-971. Epub 2019 Oct 14.

Instituto de Investigação e Inovação em Saúde da Universidade do Porto (i3S), Porto, Portugal; Instituto Nacional de Saúde Dr Ricardo Jorge (INSA), Porto, Portugal. Electronic address:

There is increasing attention on the complex interactions occurring between schistosome parasites and their hosts. However, little is known about the occurrence, epidemiology, and mechanisms of schistosomiasis-associated infertility. In this article, we argue that an in-depth understanding of the interplay between parasites and the host endocrine system may significantly enhance current knowledge of infertility in infected individuals. We discuss the basic hormonal mechanisms that may lead to the discovery of entirely novel anthelmintic interventions against schistosomiasis.
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http://dx.doi.org/10.1016/j.pt.2019.09.001DOI Listing
December 2019

Vitamin A Enhances Macrophages Activity Against B16-F10 Malignant Melanocytes: A New Player for Cancer Immunotherapy?

Medicina (Kaunas) 2019 Sep 18;55(9). Epub 2019 Sep 18.

School of Health, Porto Polytechnic (ESS, P. Porto), 4200 Porto, Portugal.

: The incidence of cutaneous melanoma has been increasing. Melanoma is an aggressive form of skin cancer irresponsive to radiation and chemotherapy, rendering this cancer a disease with poor prognosis: In order to surpass some of the limitations addressed to melanoma treatment, alternatives like vitamins have been investigated. In the present study, we address this relationship and investigate the possible role of vitamin A. : We perform a co-culture assay using a macrophage cell model and RAW 264.7 from mouse, and also a murine melanoma cell line B16-F10. Macrophages were stimulated with both lipopolysaccharides (LPS) as control, and also with LPS plus vitamin A. : Using B16-F10 and RAW 264.7 cell lines, we were able to demonstrate that low concentrations of vitamin A increase cytotoxic activity of macrophages, whereas higher concentrations have the opposite effect. : These findings can constitute a new point of view related to immunostimulation by nutrients, which may be considered one major preventive strategy by enhancing the natural defense system of the body.
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http://dx.doi.org/10.3390/medicina55090604DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6780654PMC
September 2019

Pharmacogenomics, CYP2D6, and Tamoxifen: A Survey of the Reasons Sustaining European Clinical Practice Paradigms.

Medicina (Kaunas) 2019 Jul 5;55(7). Epub 2019 Jul 5.

School of Health, Porto Polytechnic (P. Porto), 4200-072 Porto, Portugal.

Tamoxifen is a drug that is often used in the clinical management of breast cancer. CYP2D6 is a key metabolizing enzyme that is involved in the conversion of tamoxifen to its active drug metabolites. CYP2D6 has several alleles that metabolize tamoxifen and other drugs at different rates that can alter therapeutic impact, a characteristic that renders it one of the most studied enzymes in the field of pharmacogenetics. : Portugal has no implemented measures based on pharmacogenomics analysis prior to therapy that might function as a cultural sample control when analyzing the individual and economic factors present in clinical practice paradigms. Therefore, we aim to investigate the impact of CYP2D6 genotyping of the tamoxifen metabolizing enzymes in the clinical management of breast cancer patients. : Qualitative/quantitative studies regarding the impact of pharmacogenomics in breast cancer; personal interviews in different Portuguese laboratories within hospital setting using a survey. Analysis of data through interviews to management board and/or decision makers from major oncological centers. : Reasons for common adoption of pharmacogenomics practice are contradictory and based both in economic factors and cultural/clinical bias. : This research study identifies specific cultural and/or clinical bias that act as obstacles to pharmacogenomic implementation and proposes viable courses of action that might bring about change in cultural/medical habits.
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http://dx.doi.org/10.3390/medicina55070344DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6681270PMC
July 2019

ESBL and AmpC β-Lactamases in Clinical Strains of from Serra da Estrela, Portugal.

Medicina (Kaunas) 2019 Jun 12;55(6). Epub 2019 Jun 12.

School of Health, Polytechnic of Porto, 4200 Porto, Portugal.

: Given the considerable spatial, temporal, and ecological factors, heterogeneity, which affects emergency response, persistence, and dissemination of genetic determinants that confer microorganisms their resistance to antibiotics, several authors claim that antibiotics' resistance must be perceived as an ecological problem. The aim of this study was to determine the prevalence of broad-spectrum genes, not only Extended-spectrum β-lactamases (ESBL) but also AmpC-types, in clinical strains of isolated from Portugal (in the highest region of the country, Serra da Estrela) to disclose susceptibility profiles among different genotypes, and to compare the distribution of genes expressing broad-spectrum enzymes. Clinical strains of presenting resistance to third generation (3G) cephalosporins and susceptibility to inhibition by clavulanic acid were studied by means of phenotypic and molecular profiling techniques for encoding β-lactamases genes. Strains were mainly isolated from hospital populations (97%). Molecular analysis enabled the detection of 49 genes, in which 55% (27/49) were identified as , 33% (16/49) as , 10% (5/49) as , and 2% (1/49) were identified as genes (AmpC). Among all detected, about 59% of strains expressed at least another gene. Co-production of β-lactamases was observed in 40% of strains, with the co-production of CTX-M group 1 and OXA-1- occurring as the most frequent. This is the first study using microorganisms isolated from native people from the highest Portuguese mountain regions, showing an unprecedent high prevalence of genes in this country.
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http://dx.doi.org/10.3390/medicina55060272DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6632026PMC
June 2019

Avoiding the Interference of Doxorubicin with MTT Measurements on the MCF-7 Breast Cancer Cell Line.

Methods Protoc 2019 Apr 12;2(2). Epub 2019 Apr 12.

School of Health, Polytechnic Institute of Porto, 4200 Porto, Portugal.

Doxorubicin (DOXO) is an adjuvant chemotherapy agent and is also commonly used in cell biology research. Cytotoxic assays in cell culture are frequently used in order to stablish drug concentrations that are useful for controlling cell proliferation. One common cytotoxic method used is 3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide (MTT). Our present research aims to support future studies in engaging MTT assay using DOXO that exhibits a strong red coloration and fluorescence, and so it is assumed that DOXO may interfere with commonly used colorimetric assays such as MTT. The interference of DOXO in the MTT determination was evaluated in a Breast Cancer cell line Michigan Cancer Foundation-7 (MCF-7). The interference was evaluated by means of spectroscopic methods in particular spectrophometry and fluorescence spectroscopy of MTT and DOXO. We postulate that the medium and the MTT reagent itself can interfere on the metabolic activity method, so in order to achieve better results, DMEM was replaced by a neutral buffer like Phosphate-buffered saline (PBS). This protocol may be extremely useful in future studies involving DOXO.
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http://dx.doi.org/10.3390/mps2020029DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6632110PMC
April 2019

Imbalance of Steroid Hormones in Hamsters Infected with .

Endocr Metab Immune Disord Drug Targets 2019 ;19(8):1122-1126

INSA, National Institute of Health Dr. Ricardo Jorge, Department of Health Promotion and Chronic Diseases, Porto, Portugal.

Objective: Schistosomiasis is a debilitating disease that affects 200 million people worldwide. Schistosoma haematobium and Schistosoma mansoni are the major causative agents of this disease. Cancer-association and infertility-association in Schistosoma haematobium infection have already been described and it is known that the parasite produces a catechol-estrogen molecule that induces a hormonal imbalance in the host.

Methods: In order to better understand the relation of hormonal imbalance in experimental Schistosoma mansoni infection, we investigated a serum panel of steroid hormones in Schistosoma mansoni infected hamsters.

Results: We found a decrease in the serum levels of Estradiol (E2), Testosterone and Progesterone in infected females and an increase of Testosterone and a decrease in Progesterone in infected males in comparison with controls.

Conclusion: These results indicate that S. mansoni alters the levels of steroid hormones in infected males and females and it will increase the repertoire of data about the host-parasite molecular interplay and its relation with the endocrine system.
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http://dx.doi.org/10.2174/1871530319666190529121204DOI Listing
April 2020

Prevalence of Antibiotic Resistance Genes in Multidrug-Resistant on Portuguese Livestock Manure.

Antibiotics (Basel) 2019 Mar 13;8(1). Epub 2019 Mar 13.

Environment Department, Research Centre for Natural Resources, Environment and Society (CERNAS), College of Agriculture, Polytechnic of Coimbra, 3045-601 Coimbra, Portugal.

The exposure of both crop fields and humans to antibiotic-resistant bacteria in animal excreta is an emergent concern of the One Health initiative. This study assessed the contamination of livestock manure from poultry, pig, dairy farms and slaughterhouses in Portugal with resistance determinants. The resistance profiles of 331 isolates to eight β-lactam (amoxicillin, cefoxitin, cefotaxime, cefpirome, aztreonam, ceftazidime, imipenem and meropenem) and to five non-β-lactam antibiotics (tetracycline (TET), trimethoprim/sulfamethoxazole (SXT), ciprofloxacin (CIP), chloramphenicol (CHL) and gentamicin) was investigated. Forty-nine integron and non-β-lactam resistance genes were also screened for. Rates of resistance to the 13 antibiotics ranged from 80.8% to 0.6%. Multidrug resistance (MDR) rates were highest in pig farm samples (79%). Thirty different integron and resistance genes were identified. These were mainly associated with resistance to CHL (I and II), CIP (mainly, S, B and ), TET (mainly (A) and (M)) and SXT (mostly Ia group and 3). In MDR isolates, integron presence and non-β-lactam resistance to TET, SXT and CHL were positively correlated. Overall, a high prevalence of MDR was found in livestock manure. The high gene diversity for antibiotic resistance identified in this study highlights the risk of MDR spread within the environment through manure use.
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http://dx.doi.org/10.3390/antibiotics8010023DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6466527PMC
March 2019

Warburg Effect Inversion: Adiposity shifts central primary metabolism in MCF-7 breast cancer cells.

Life Sci 2019 Apr 9;223:38-46. Epub 2019 Mar 9.

School of Health, Polytechnic of Porto (ESS/P.PORTO), Porto, Portugal; Instituto de Inovação e Investigação em Saúde (I3S), University of Porto, Portugal; Faculty of Medicine, University of Santiago de Compostela, Galiza, Spain. Electronic address:

Aims: Obesity is a complex health disorder and a trigger to many diseases like Diabetes mellitus (DM) and breast cancer (BrCa), both leading causes of morbidity and mortality worldwide. Also evidence demonstrates that abnormal glucose metabolism termed 'the Warburg effect' in cancer cell is closely associated with malignant phenotypes and promote the aggressiveness of several types of cancer, including BrCa. In this study, we evaluated the breast cancer cell metabolism in normoglycemia, hyperglycemia and in an obesity condition in order to clarify the potential underlined mechanisms that link these disorders.

Materials And Methods: MCF-7 cells were exposed to low and high glucose levels, the latter either in the presence of 3T3-L1 adipocyte conditioned medium (CM), thus mimicking the adiposity observed in obese patients. Cell viability, migration, proliferation, cytotoxicity and cell death assays were performed under the different culture conditions. Hormonal and lipid profile were also characterized by biochemical assays and primary metabolism was determined by Nuclear Magnetic Resonance (NMR)-based metabolomics.

Results: Our results show an increased aggressiveness in the condition mimicking diabetogenic obesity with an altered energy/lipid metabolism. Interestingly in the experimental obesity-mimicking status, lipids and amino acids were expended while glucose was produced by tumor cells from lactate. These findings reveal a shift on tumor cells metabolism that is opposite to 'the Warburg effect'.

Conclusions: Overall, this experimentally obesity-mimicking condition not only revealed an increased tumor proliferation and aggressiveness but also disclosed a new mechanism of cancer metabolism, the 'Warburg Effect Inversion'.
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http://dx.doi.org/10.1016/j.lfs.2019.03.016DOI Listing
April 2019

Adipocyte proteome and secretome influence inflammatory and hormone pathways in glioma.

Metab Brain Dis 2019 02 9;34(1):141-152. Epub 2018 Oct 9.

School of Health, Polytechnic of Porto, Porto, Portugal.

Gliomas 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.
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http://dx.doi.org/10.1007/s11011-018-0327-yDOI Listing
February 2019

Extract Induces Cell Death of Mammalian Cells.

Antiinfect Agents 2018 ;16(2):144-146

I3S, Instituto de Investigação e Inovação da Universidade do Porto, Portugal.

Background: Fascioliasis is a neglected tropical disease that affects poor people from poor and developing countries. In the world, it has been estimated that at least 2.6 million people are affected with this disease. The International agency for Research on Cancer, states that and , also liver flukes, are considered as definitive causes of cholangiocarcinoma. However, fascioliasis caused by has not been associated with cancer to date. There are not any known causative associations between this parasite and liver cancer (cholangiocarcinoma).

Methods: Chine Hamster Ovary (CHO) cells were treated with extracts and cell proliferation was assessed by using the indirect method for estimating cell number based on the mitochondrial activity with MTS cell proliferation reagent. We observed unexpected death of these cells when treated with extracts.

Results: We now hypothesize that this parasite could be used as a medically-important trematode pathogen in cancer therapy.
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http://dx.doi.org/10.2174/1570180815666180531102555DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6075840PMC
January 2018

Commentary: Parasites Secrete a Prolyl Isomerase to Maintain Host Leukocyte Transformation.

Front Med (Lausanne) 2018 27;5:120. Epub 2018 Apr 27.

I3S, Instituto de Investigação e Inovação da Universidade do Porto, Porto, Portugal.

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http://dx.doi.org/10.3389/fmed.2018.00120DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5934426PMC
April 2018

Sensing CA 15-3 in point-of-care by electropolymerizing O-phenylenediamine (oPDA) on Au-screen printed electrodes.

PLoS One 2018 1;13(5):e0196656. Epub 2018 May 1.

BioMark/ISEP, School of Engineering, Polytechnic Institute of Porto, Porto, Portugal.

This work presents an alternative device for cancer screening in liquid biopsies. It combines a biomimetic film (i) with electrochemical detection (ii). The biomimetic film (i) was obtained by electro-polymerizing amine-substituted benzene rings around a CA 15-3 target. This protein target was previously adsorbed on a gold (Au) support and incubated in charged monomers (4-Styrenesulfonate sodium and 3-Hydroxytyraminium chloride). The protein was further eliminated by enzymatic activity, leaving behind vacant sites for subsequent rebinding. Electrochemical detection (ii) was achieved on an Au working electrode, designed on commercial screen-printed electrodes. Raman spectroscopy, atomic force microscopy and ellipsometric readings were used to follow the chemical modification of the Au surface. The ability of the material to rebind CA15-3 was monitored by electrochemical techniques. The device displayed linear responses to CA15-3 ranging from 0.25 to 10.00 U/mL, with detection limits of 0.05 U/mL. Accurate results were obtained by applying the sensor to the analysis of CA15-3 in PBS buffer and in serum samples. This biosensing device displayed successful features for the detection of CA 15-3 and constitutes a promising tool for breast cancer screening procedures in point-of-care applications. Moreover, its scale-up seems feasible as it contains a plastic antibody assembled in situ, in less than 1 minute, and the analysis of serum takes less than 30 minutes.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0196656PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5929556PMC
July 2018

Estrogen Metabolism-Associated CYP2D6 and IL6-174G/C Polymorphisms in Schistosoma haematobium Infection.

Int J Mol Sci 2017 Nov 28;18(12). Epub 2017 Nov 28.

Department of Health Promotion and Chronic Diseases, National Institute of Health Dr. Ricardo Jorge (INSA), Rua Alexandre Herculano 321, 4000-055 Porto, Portugal.

is a human blood fluke causing a chronic infection called urogenital schistosomiasis. Squamous cell carcinoma of the urinary bladder (SCC) constitutes chronic sequelae of this infection, and infection is accounted as a risk factor for this type of cancer. This infection is considered a neglected tropical disease and is endemic in numerous countries in Africa and the Middle East. Schistosome eggs produce catechol-estrogens. These estrogenic molecules are metabolized to active quinones that induce modifications in DNA. The cytochrome P450 (CYP) enzymes are a superfamily of mono-oxygenases involved in estrogen biosynthesis and metabolism, the generation of DNA damaging procarcinogens, and the response to anti-estrogen therapies. IL6 Interleukin-6 (IL-6) is a pleiotropic cytokine expressed in various tissues. This cytokine is largely expressed in the female urogenital tract as well as reproductive organs. Very high or very low levels of IL-6 are associated with estrogen metabolism imbalance. In the present study, we investigated the polymorphic variants in the gene and the C-174G promoter polymorphism of the gene on -infected children patients from Guine Bissau. inactivated alleles (28.5%) and G-174C (13.3%) variants were frequent in -infected patients when compared to previously studied healthy populations (4.5% and 0.05%, respectively). Here we discuss our recent findings on these polymorphisms and whether they can be predictive markers of schistosome infection and/or represent potential biomarkers for urogenital schistosomiasis associated bladder cancer and infertility.
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http://dx.doi.org/10.3390/ijms18122560DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5751163PMC
November 2017

Angiogenesis in Schistosoma haematobium-associated urinary bladder cancer.

APMIS 2017 Dec 28;125(12):1056-1062. Epub 2017 Sep 28.

I3S, Instituto de Investigação e Inovação da Universidade do Porto, Porto, Portugal.

Schistosoma haematobium, a parasitic flatworm that infects more than 100 million people, mostly in the developing world, is the causative agent of urogenital schistosomiasis, and is associated with a high incidence of squamous cell carcinoma (SCC) of the bladder. During infection, eggs are deposited in the bladder causing an intense inflammatory reaction. Angiogenesis is defined as the formation of new blood vessels from preexisting ones and is recognized as a key event in cell proliferation and carcinogenesis and spread of malignant lesions. A growing amount of evidence points to angiogenesis playing a key role in schistosomiasis-associated bladder cancer. Thus, identifying biomarkers of this process plays an important role in the study of cancer. Here, we review recent findings on the role of angiogenesis in bladder cancer and the growth factors that induce and assist in their development, particularly SCC of the bladder associated to urogenital schistosomiasis.
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http://dx.doi.org/10.1111/apm.12756DOI Listing
December 2017

Adipocyte Secretome Increases Radioresistance of Malignant Melanocytes by Improving Cell Survival and Decreasing Oxidative Status.

Radiat Res 2017 05 31;187(5):581-588. Epub 2017 Mar 31.

a   Department of Biochemistry, Faculty of Medicine, University of Porto, Portugal.

Radiotherapy 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 phenotype 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.
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http://dx.doi.org/10.1667/RR14551.1DOI Listing
May 2017

Melanoma and obesity: Should antioxidant vitamins be addressed?

Life Sci 2016 Nov 23;165:83-90. Epub 2016 Sep 23.

Ciências Químicas e das Biomoléculas, Centro de Investigação em Saúde e Ambiente, Escola Superior de Saúde, Instituto Politécnico do Porto, Portugal; I3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal. Electronic address:

Melanoma is an aggressive form of skin cancer refractory to conventional therapies. Obesity has reached epidemic dimensions acting as a risk factor for several cancer types, such as melanoma. Several reactive species of oxygen are also involved in melanoma initiation and progression. Low levels of antioxidant content and/or activity in lightly pigmented cells could expose them to an extremely oxidative environment and rise the susceptibility to oxidative damage and consequently loss of cell homeostasis. Despite the knowledge about melanoma biology, pathogenesis and developed therapies, is extremely important to understand the antioxidant modulation of melanoma under an environment of obesity, especially the effect of some natural compounds of the diet, such as antioxidant vitamins A, C and E and selenium in order to establish alternatives to conventional therapies, which are known to be ineffective against melanoma.
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http://dx.doi.org/10.1016/j.lfs.2016.09.015DOI Listing
November 2016

3-Nitrotyrosine quantification methods: Current concepts and future challenges.

Biochimie 2016 Jun 26;125:1-11. Epub 2016 Feb 26.

Ciências Químicas e das Biomoléculas, Centro de Investigação em Saúde e Ambiente, Escola Superior de Tecnologia da Saúde do Porto, Instituto Politécnico do Porto, Portugal; I3S - Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Portugal. Electronic address:

Background: Measurement of 3-nitrotyrosine (3-NT) in biological samples can be used as a biomarker of nitrosative stress, since it is very stable and suitable for analysis. Increased 3-NT levels in biological samples have been associated with several physiological and pathological conditions. Different methods have been described for the detection and quantification of this molecule, such as (i) immunological methods; (ii) liquid chromatography, namely high-pressure liquid chromatography (HPLC)-based methods that use ultraviolet-visible (UV/VIS) absorption, electrochemical (ECD) and diode array (DAD) detection, liquid chromatography-mass spectrometry (LC-MS) and liquid chromatography-tandem mass spectrometry (LC-MS/MS); (iii) gas chromatography, such as gas chromatography-mass spectrometry (GC-MS) and gas chromatography-tandem mass spectrometry (GC-MS/MS).

Methods: A literature review on nitrosative stress, protein nitration, as well as 3-NT quantification methods was carried out.

Results: This review covers the different methods for analysis of 3-NT that have been developed during the last years as well as the latest advances in this field. Overall, all methods present positive and negative aspects, although it is clear that chromatography-based methods present good sensitivity and specificity. Regarding this, GC-based methods exhibit the highest sensibility in the quantification of 3-NT, although it requires a prior time consuming derivatization step. Conversely, HPLC does not require such derivatization step, despite being not as accurate as GC.

Conclusion: It becomes clear that all the methods described during this literature review, although accurate for 3-NT quantification, need to be improved regarding both sensitivity and specificity. Moreover, optimization of the protocols that have been described is clearly needed.
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http://dx.doi.org/10.1016/j.biochi.2016.02.011DOI Listing
June 2016

Novel and simple electrochemical biosensor monitoring attomolar levels of miRNA-155 in breast cancer.

Biosens Bioelectron 2016 Jun 14;80:621-630. Epub 2016 Feb 14.

BioMark-CINTESIS/ISEP, School of Engineering, Polytechnic Institute of Porto, Portugal. Electronic address:

This work, describes for the first time, a simple biosensing design to yield an ultrasensitive electrochemical biosensor for a cancer biomarker detection, miRNA-155, with linear response down to the attomolar range. MiRNA-155 was selected for being overexpressed in breast cancer. The biosensor was assembled in two stages: (1) the immobilization of the anti-miRNA-155 that was thiol modified on an Au-screen printed electrode (Au-SPE), followed by (2) blocking the areas of non-specific binding with mercaptosuccinic acid. Atomic force microscopy (AFM) and electrochemical techniques including cyclic voltammetry (CV), impedance spectroscopy (EIS) and square wave voltammetry (SWV) confirmed the surface modification of these devices and their ability to hybridize successfully and stably with miRNA-155. The final biosensor provided a sensitive detection of miRNA-155 from 10 aM to 1.0 nM with a low detection limit (LOD) of 5.7 aM in real human serum samples. Good results were obtained in terms of selectivity towards breast cancer antigen CA-15.3 and bovine serum albumin (BSA). Raw fluid extracts from cell-lines of melanoma did not affect the biosensor response (no significant change of the blank), while raw extracts from breast cancer yielded a positive signal against miRNA-155. This simple and sensitive strategy is a promising alternative for simultaneous quantitative analysis of multiple miRNA in physiological fluids for biomedical research and point-of-care (POC) diagnosis.
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http://dx.doi.org/10.1016/j.bios.2016.02.035DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6366556PMC
June 2016

Effect of Adipocyte Secretome in Melanoma Progression and Vasculogenic Mimicry.

J Cell Biochem 2016 07 15;117(7):1697-706. Epub 2016 Jan 15.

Department of Biochemistry, Faculty of Medicine, University of Porto, Porto, Portugal.

Obesity, favored by the modern lifestyle, acquired epidemic proportions nowadays. Obesity has been associated with various major causes of death and morbidity including malignant neoplasms. This increased prevalence has been accompanied by a worldwide increase in cutaneous melanoma incidence rates during the last decades. Obesity involvement in melanoma aetiology has been recognized, but the implicated mechanisms remain unclear. In the present study, we address this relationship and investigate the influence of adipocytes secretome on B16-F10 and MeWo melanoma cell lines. Using the 3T3-L1 adipocyte cell line, as well as ex vivo subcutaneous (SAT) and visceral (VAT) adipose tissue conditioned medium, we were able to show that adipocyte-released factors play a dual role in increasing melanoma cell overall survival, both by enhancing proliferation and decreasing apoptosis. B16-F10 cell migration and cell-cell and cell-matrix adhesion capacity were predominantly enhanced in the presence of SAT and VAT released factors. Melanocytes morphology and melanin content were also altered by exposure to adipocyte conditioned medium disclosing a more dedifferentiated phenotype of melanocytes. In addition, exposure to adipocyte-secreted molecules induced melanocytes to rearrange, on 3D cultures, into vessel-like structures, and generate characteristic vasculogenic mimicry patterns. These findings are corroborated by the released factors profile of 3T3-L1, SAT, and VAT assessed by microarrays, and led us to highlight the mechanisms by which adipose secretome from sub-cutaneous or visceral depots promote melanoma progression. J. Cell. Biochem. 117: 1697-1706, 2016. © 2015 Wiley Periodicals, Inc.
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http://dx.doi.org/10.1002/jcb.25463DOI Listing
July 2016

Adipocyte secreted factors enhance aggressiveness of prostate carcinoma cells.

PLoS One 2015 30;10(4):e0123217. Epub 2015 Apr 30.

Department of Anatomy, Unit for Multidisciplinary Research in Biomedicine (UMIB), Institute for Biomedical Sciences Abel Salazar (ICBAS), University of Porto, Porto, Portugal.

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.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0123217PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4415768PMC
December 2015

Antibiotic resistance in wastewater: occurrence and fate of Enterobacteriaceae producers of class A and class C β-lactamases.

J Environ Sci Health A Tox Hazard Subst Environ Eng 2015 ;50(1):26-39

a Departamento de Ambiente, CERNAS , Escola Superior Agraria do Instituto Politécnico de Coimbra , Coimbra , Portugal.

Antibiotics have been intensively used over the last decades in human and animal therapy and livestock, resulting in serious environmental and public health problems, namely due to the antibiotic residues concentration in wastewaters and to the development of antibiotic-resistant bacteria. This study aimed to access the contribution of some anthropological activities, namely urban household, hospital and a wastewater treatment plant, to the spread of antibiotic resistances in the treated wastewater released into the Mondego River, Coimbra, Portugal. Six sampling sites were selected in the wastewater network and in the river. The ampicillin-resistant Enterobacteriaceae of the water samples were enumerated, isolated and phenotypically characterized in relation to their resistance profile to 13 antibiotics. Some isolates were identified into species level and investigated for the presence of class A and class C -lactamases. Results revealed high frequency of resistance to the -lactam group, cefoxitin (53.5%), amoxicillin/clavulanic acid combination (43.5%), cefotaxime (22.7%), aztreonam (21.3) cefpirome (19.2%), ceftazidime (16.2%) and to the non--lactam group, trimethoprim/sulfamethoxazol (21.1%), tetracycline (18.2%), followed by ciprofloxacin (14.1%). The hospital effluent showed the higher rates of resistance to all antibiotic, except two (chloramphenicol and gentamicin). Similarly, higher resistance rates were detected in the wastewater treatment plant (WWTP) effluent compared with the untreated affluent. Regarding the multidrug resistance, the highest incidence was recorded in the hospital sewage and the lowest in the urban waste. The majority of the isolates altogether are potentially extended-spectrum -lactamases positive (ESBL(+)) (51.9%), followed by AmpC(+) (44.4%) and ESBL(+)/AmpC(+) (35.2%). The most prevalent genes among the potential ESBL producers were blaOXA (33.3%), blaTEM (24.1%) and blaCTX-M (5.6%) and among the AmpC producers were blaEBC (38.9%), blaFOX (1.9%) and blaCIT (1.9%). In conclusion, the hospital and the WWTP activities revealed to have the highest contribution to the spread of multidrug resistant bacteria in the study area. Such data is important for future management of the environmental and public health risk of these contaminants. This is the first embracing study in the water network of Coimbra region on the dissemination of antibiotic resistance determinants. Moreover, it is also the first report with the simultaneous detection of multiresistant bacteria producers of AmpC and ESBLs -lactamases in aquatic systems in Portugal.
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http://dx.doi.org/10.1080/10934529.2015.964602DOI Listing
April 2015

Quinoxaline, its derivatives and applications: A State of the Art review.

Eur J Med Chem 2015 Jun 1;97:664-72. Epub 2014 Jul 1.

Ciências Químicas e Biomoléculas, Centro de Investigação em Saúde e Ambiente (CISA), Escola Superior de Tecnologia da Saúde do Instituto Politécnico do Porto (ESTSP-IPP), Rua Valente Perfeito 322, 4400-330 Vila Nova de Gaia, Portugal; REQUIMTE/CQFB, Departamento de Química, FCT, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal. Electronic address:

Quinoxaline derivatives are an important class of heterocycle compounds, where N replaces some carbon atoms in the ring of naphthalene. Its molecular formula is C8H6N2, formed by the fusion of two aromatic rings, benzene and pyrazine. It is rare in natural state, but their synthesis is easy to perform. In this review the State of the Art will be presented, which includes a summary of the progress made over the past years in the knowledge of the structure and mechanism of the quinoxaline and quinoxaline derivatives, associated medical and biomedical value as well as industrial value. Modifying quinoxaline structure it is possible to obtain a wide variety of biomedical applications, namely antimicrobial activities and chronic and metabolic diseases treatment.
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http://dx.doi.org/10.1016/j.ejmech.2014.06.058DOI Listing
June 2015

Molecular characterization of ESBL-producing Enterobacteriaceae in northern Portugal.

ScientificWorldJournal 2014 13;2014:782897. Epub 2014 Feb 13.

Ciências Químicas e das Biomoléculas (CQB) e Centro de Investigação em Saúde e Ambiente (CISA), Escola Superior de Tecnologia da Saúde do Porto (ESTSP), Instituto Politécnico do Porto (IPP), 4400-330 Porto, Portugal ; CFBQ-Centro de Farmacologia e Biopatologia Química (U38-FCT), Faculdade de Medicina do Porto, Porto, Portugal ; Centro Hospitalar da Universidade de Coimbra (CHUC), Coimbra, Portugal.

Extended-spectrum β-lactamases (ESBLs) prevalence was studied in the north of Portugal, among 193 clinical isolates belonging to citizens in a district in the boundaries between this country and Spain from a total of 7529 clinical strains. In the present study we recovered some members of Enterobacteriaceae family, producing ESBL enzymes, including Escherichia coli (67.9%), Klebsiella pneumoniae (30.6%), Klebsiella oxytoca (0.5%), Enterobacter aerogenes (0.5%), and Citrobacter freundii (0.5%). β -lactamases genes blaTEM, blaSHV, and blaCTX-M were screened by polymerase chain reaction (PCR) and sequencing approaches. TEM enzymes were among the most prevalent types (40.9%) followed by CTX-M (37.3%) and SHV (23.3%). Among our sample of 193 ESBL-producing strains 99.0% were resistant to the fourth-generation cephalosporin cefepime. Of the 193 isolates 81.3% presented transferable plasmids harboring bla ESBL genes. Clonal studies were performed by PCR for the enterobacterial repetitive intragenic consensus (ERIC) sequences. This study reports a high diversity of genetic patterns. Ten clusters were found for E. coli isolates and five clusters for K. pneumoniae strains by means of ERIC analysis. In conclusion, in this country, the most prevalent type is still the TEM-type, but CTX-M is growing rapidly.
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http://dx.doi.org/10.1155/2014/782897DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3950362PMC
January 2015

Iron metabolism: from health to disease.

J Clin Lab Anal 2014 May 29;28(3):210-8. Epub 2014 Jan 29.

Ciências Químicas e das Biomoléculas e Unidade de Mecanismos Moleculares da Doença do Centro de Investigação em Saúde e Ambiente, Escola Superior de Tecnologia da Saúde do Porto, Instituto Politécnico do Porto, Portugal.

Background: Iron is vital for almost all living organisms by participating in a wide range of metabolic processes. However, iron concentration in body tissues must be tightly regulated since excessive iron may lead to microbial infections or cause tissue damage. Disorders of iron metabolism are among the most common human diseases and cover several conditions with varied clinical manifestations.

Methods: An extensive literature review on the basic aspects of iron metabolism was performed, and the most recent findings on this field were highlighted as well.

Results: New insights on iron metabolism have shed light into its real complexity, and its role in both healthy and pathological states has been recognized. Important discoveries about the iron regulatory machine and imbalances in its regulation have been made, which may lead in a near future to the development of new therapeutic strategies against iron disorders. Besides, the toxicity of free iron and its association with several pathologies has been addressed, although it requires further investigations.

Conclusion: This review will provide students in the fields of biochemistry and health sciences a brief and clear overview of iron physiology and toxicity, as well as imbalances in the iron homeostasis and associated pathological conditions.
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http://dx.doi.org/10.1002/jcla.21668DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6807557PMC
May 2014
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