Rodrigues, C.F.

PharmD and Researcher

Recent Works

In vitro study of antifungal drug effect in a tissue conditioner

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Created: Friday, 30 June 2017 14:33
Assunção B; Henriques, M; Rodrigues, CF; Vaz P; Figueiral H; Braga AC; Fernandes S.
IV International Conference on Biodental Engineering, Porto, Portugal, June, 2016- ORAL PRESENTATION
Tissue conditioners are generally used in acrylic removable dentures to improve the fit of the prosthesis and to allow support tissues recovery. It is mostly by the elastic property of the conditioners, which translates the performance of the "cushion" effect by the forces exerted during mastication, that is obtained the tissue recovery. However, these tissue conditioners are also well known for its poor physical properties - easy sensitivity degradation and colonization by microorganisms. Thus, their characteristics favor the accumulation of plaque and colonization by species of Candida, which can irritate the oral tissues and promote the development of denture stomatitis, threatening the oral health of patients with removable dental prosthesis. It was shown that these same acrylic resin when placed at the base of the prosthesis, could act as a reservoir for microorganisms and have the potential for biofilm formation.
 

Candida glabrata’s biofilms response to amphotericin B

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Created: Friday, 30 June 2017 14:27
Rodrigues, CF; Silva, SC; Salgado JM; Abrunhosa L; Venâncio A; Azeredo, J; Henriques, M
FEMS 2015 - 6th Congress of European Microbiologists, Maastricht, The Netherlands, The Netherlands, 2015 - ORAL PRESENTATION
Candida species are responsible for recurrent human infections, mostly in immunocompromised patients, due to their high vulnerability. Candida glabrata has been showing to have a major role in these infections being the second most prevalent species involved in human fungemia. Amphotericin B (AmB), a common antifungal drug, is a hospital-environment exclusive polyene, normally being efficient when used to fight candidiasis. 
Objectives: To infer about the influence of AmB in Candida glabrata biofilms formation and its effect on matrix composition and ERG genes expression.
AmB reduces biofilms of C. glabrata.
ERG genes seem to be less involved than the matrix composition in C. glabrata biofilms response to AmB. 
C. glabrata biofilms matrices respond with an increase of carbohydrates, particularly β-1,3 glucans, and with a decrease of total proteins. 
New matrix artifact component was found: ergosterol.
The present work supports the theory of multifaceted mechanisms developed by C. glabrata biofilms as response to the presence of AmB.
 

Synergistic Antimicrobial Interaction Of Honey And Bacteriophage In Escherichia coli Biofilms

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Created: Friday, 30 June 2017 13:52
Ribeiro H; Oliveira A; Silva AC; Silva AD; Rodrigues, CF; Melo LDR; Henriques A; Sillankorva S.
X Encontro Nacional de Estudantes de Bioquímica (X ENEBIOQ), Braga, Portugal, April, 2017
Wound colonization by biofilms-forming bacteria is one of the main obstacles to the treatment of chronic wounds, causing a number of biological and financial problems. Biofilms are structured communities of bacterial cells enclosed in a self-produced polymeric matrix, adhered to inert or living surfaces, blocking antibiotics and patient’s immune cells from reaching bacteria. Bacterio(phages), viruses infecting exclusively bacteria, and honey are being considered as valuable alternatives to treat a variety of infections (Table 1). Phages are harmless to mammalian cells, specific for target bacteria therefore not affecting commensal microflora, have the ability to selfreplicate as long as the host is present, and are effective against antibiotic resistant bacteria. Honey is a complex substance with broad spectrum antimicrobial activity, essentially attributed to the high sugar content, low pH, the presence of hydrogen peroxide and methylglyoxal that reacts with important biological molecules (RNA, DNA and proteins). Honey had yet the potential to promote tissue regeneration, cicatrization, decrease inflammation, improving wound healing. In this work the combination of those two antimicrobial agents was considered in the control E. coli biofilms.
The Portuguese honey PF2 50% (w/v) showed great antibiofilm effect with 6-log cell reduction in 24 h-old biofilms. 
A synergistic effect was observed with PF2 25% (w/v) and EC3a in 24 and 48 h-old biofilms. 
48 h-old biofilms were more difficult to combat than 24 h-old biofilms. 
This is a promising strategy for biofilm control that will be further tested with other pathogens and using ex vivo models.
 
 

Fluconazole vs Voriconazole: Candida glabrata’s biofilms response to different azoles

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Created: Friday, 30 June 2017 13:28
Rodrigues, CF; Gonçalves B; Rodrigues ME; Silva, SC; Azeredo, J; Henriques, M
TIMM 2015 - 7th Congress on Trends in Medical Mycology, Portugal, October, 2015.
Candida glabrata is the second most prevalent yeast in fungal infections, especially in immunocompromised and/or hospitalized patients. The azole resistance within this species is very well-known and results in a low therapeutic response of C. glabrata infections, particularly when associated with biofilms.
Objective - To understand the different efficacies of two azoles against C. glabrata biofilms: - fluconazole (Flu), a long time used drug - voriconazole (Vcz), a latest drug used only in hospitals.
The pattern of C. glabrata biofilm resistance to Flu and Vcz is effectively different;
The cell genetic alterations in genes encoding for the ergosterol biosynthesis exists but is still unable to   explain the different resistance profiles obtained for these two agents;
There were alterations in the matrix composition in the presence of the azoles, but there was not found a direct correlation with Flu and Vcz efficacy; 
The diffusion of the drugs within the biofilms is azole’s dependent, with Vcz having a better diffusion and a better activity against C. glabrata biofilms than Flu, explaining its enhanced performance. 
 

The chemical structure of the biofilm matrices of Candida glabrata induces resistance to antifungal drugs

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Created: Friday, 30 June 2017 13:37
Rodrigues, CF; Rodrigues ME; Silva, SC; Azeredo, J; Henriques, M
BIOFILMS'7- Internation Congress of Biofilms, Porto, Portugal, June, 2016.
Candida infections are often associated to biofilms and consequently to high resistance to the most common drugs. These resistance mechanisms are not only associated with the biofilm yeast physiology, but also with the presence of a barrier imposed by the biofilm matrix. However, the biochemical role of the biofilm components remains very unclear. Therefore, this work intends to further enlighten the effect of antifungal agents on C. glabrata biofilm resistance. As a good biofilm former, Candida glabrata ATCC2001 was selected to this study. Several antifungal drugs, belonging to different groups, were used in this work, namely fluconazole, amphotericin B, caspofungin and micafungin and their effect on biofilm matrix was assessed. Biofilm matrix chemical composition and structure was evaluated by analytical methods. As expected, C. glabrata biofilms were resistant to the antifungals used in an agent-dependent manner. The results showed significant differences in polysaccharides and proteins contents, in the matrix of biofilms formed in the absence and in the presence of the drugs. Moreover, the diffusion through the matrix was evaluated demonstrating that different agents, even belonging to the same group, present very different profiles, explaining the different tolerance registered. So, with this study we confirmed that C. glabrata biofilm’s resistance to antifungal drugs is a very complex mechanism, where the matrix plays a major role.
The biofilm matrices of C. glabrata ATCC2001 strongly changes when in contact to antifungal drugs: protein profile changes and β-1,3 glucans  production increases.
It is known that β-1,3 glucans are linked to the antifungal resistance, by making it difficult to the drugs to diffuse through the biofilm matrices and reach the yeast cells. This change in the matrix composition may be an attempt of the cells to obtain a denser matrix, unflexible and more protective for their environment.
 

How do Candida glabrata's biofilms respond to antifungal drugs?

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Created: Friday, 30 June 2017 13:24
Rodrigues, CF; Rodrigues ME; Silva S; Henriques M.
TIMM 2015 - 7th Congress on Trends in Medical Mycology,, Lisboa, Portugal, 2015.
Candida species are responsible for recurrent human infections, mostly in immunocompromised patients, due to their high vulnerability.  Candida glabrata has been shown to have a major role in these infections being the second most prevalent species involved in human fungemia.
Objective: To understand the effect of three different antifungal agents – Fluconazole (Flu), Amphotericin B (AmB) and Caspofungin (Csf) - in C. glabrata’s biofilm formation, specially their role on matrix composition.
 
The pattern of C. glabrata biofilm response to Flu, AmB and Csf is clearly different;
The three agents had diverse effects on C. glabrata’s biofilm formation;
Matrices´ composition display variations when exposed to different antifungal agents, and these differences depend on the drug is used;
AmB, and especially Csf, were confirmed in this study, to be the most effective pharmacoterapies for eradication of C. glabrata infections associated to biofilms.
 

"Soy Proteins" - Encyclopedia of Biomedical Polymers and Polymeric Biomaterials

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Created: Tuesday, 10 December 2013 22:55
Editor: Dr. Munmaya Mishra. Taylor & Francis, New York, USA
ISBN-10: 1439898553, (1800 pages), 1st edition (2015).  ISBN-10: 1439898790, (8000 pages), 1st edition (2015)
Rodrigues, CF; Andrade, JC.
Soy protein products such as flour, concentrates and isolates are used in the food industry because of their functionality, nutritional value and low cost. This chapter presents an overview current knowledge of soy proteins, manufacture processes and modifications. The health benefits attributed to soy proteins are also discussed. In addition the biomedical applications are summarized.

"Pectin" - Encyclopedia of Biomedical Polymers and Polymeric Biomaterials

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Created: Tuesday, 10 December 2013 22:50
Editor: Dr. Munmaya Mishra. Taylor & Francis, New York, USA
ISBN-10: 1439898553, (1800 pages), 1st edition (2015).  ISBN-10: 1439898790, (8000 pages), 1st edition (2015)
Rodrigues, CF; Faria, L; Andrade, JC.
Pectin is a family of complex polysaccharides that occur as structural materials in all land-growing plants. Its main utilization is on the food industry mainly due to his gelling properties. Recently, pectin has been exploited for different biomedical applications namely for drug delivery and for tissue engineering. This chapter will discuss the chemistry and gelling formation mechanism and properties of pectin. Furthermore health aspects of pectin are overviewed as well as diverse current and emerging applications in the biomedical field.
 

"Gellan" - Encyclopedia of Biomedical Polymers and Polymeric Biomaterials

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Created: Tuesday, 10 December 2013 22:53
Editor: Dr. Munmaya Mishra. Taylor & Francis, New York, USA
ISBN-10: 1439898553, (1800 pages), 1st edition (2015).  ISBN-10: 1439898790, (8000 pages), 1st edition (2015)
Rodrigues, CF; Andrade, JC.
Gellan gum is a natural anionic extracellular polysaccharide obtained by fermentation of the bacteria Sphingomonas elodea. It has the distinctive property of temperature-dependent and cation-induced gelation. This chapter summarizes the current knowledge on production, structure and properties of gellan gum. In addition, an overview of the current and potential biomedical applications of gellan is given.

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"Milk Proteins" - Encyclopedia of Biomedical Polymers and Polymeric Biomaterials

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Created: Tuesday, 10 December 2013 22:59
Editor: Dr. Munmaya Mishra. Taylor & Francis, New York, USA
ISBN-10: 1439898553, (1800 pages), 1st edition (2015).  ISBN-10: 1439898790, (8000 pages), 1st edition (2015)
Rodrigues, CF; Andrade, JC.

Milk contains many types of proteins which are classified in two general categories – caseins and whey proteins. Due to their nutritional and functional properties milk proteins products are widely used in the food industry. This chapter summarizes the current knowledge on milk protein fraction and their biological properties. The production of the mains protein-enriched products is described. In addition an overview of the current and potential biomedical applications of milk proteins is given.

"Analysis of biofilm metabolites"

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Created: Tuesday, 22 July 2014 12:33
Clinical Biofilms – Current Concepts and Advanced Techniques
ISBN: 978-989-97478-4-5, Universidade do Minho – DEB, Braga (Portugal), 2014, pp: 173-192,
Almeida,C; Rodrigues CF; Jorge P; Costa AR.
Quorum Sensing (QS) involves the production, release, detection, and response to secondary metabolites that act as small hormones and are termed auto-inducers. These metabolites enable microorganisms to ‘sense’ cell density and alter their behavior on a population-wide scale in response to environmental factors, such as nutrient availability, toxic compounds and host–immune responses. N-acyl homoserine lactones (HSLs) are part of a basic QS circuit that has been firstly described for Vibrio fischeri, the LuxI/LuxR system, and that is very similar throughout Gram-negative bacteria. HSLs secreted by bacteria usually differ in the length of the acyl-chain moiety, saturation and substitution on the third carbon by either oxo or hydroxyl group. For instance, in P. aeruginosa the two main HSLs produced are the short-chain molecule N-butanoyl-l-homoserine lactone (C4-HSL) and the long-chain molecule N-(3-oxododecanoyl)-l-homoserine lactone (3-oxo-C12-HSL), which are involved in coordinately regulating virulence and secondary metabolite production. While no gram-positive HSL producers have been reported yet, it is known that some HSL can interact and modulate the behavior of gram-positive species. For instance, long-chain AHLs, such as 3-oxo-C12-HSL produced by P. aeruginosa, are capable of interacting with the S. aureus, reducing exotoxin production and down-regulating cell surface colonization proteins. In this practical class we will focus on the production of HSLs on mixed biofilm populations.  As a case study, a mixed biofilm of P. fluorescens and Staphyloccocus epidermidis, will be evaluated. Crude extracts of the mixed biofilm supernatants will be evaluated for the presence of HSL and compared with those of single species biofilms. First, a fractionation procedure, that will provide a partial purification of the biofilm supernatant, will be presented. Then, screening procedures for HSL and their isolation by High Pressure Liquid Chromatography (HPLC) will be addressed.