CQB-Day 2015
September 15th
Faculdade de Ciências – Universidade de Lisboa
ORGANISING COMMITTEE
ANA PAULA CARVALHO
OLINDA C. MONTEIRO
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WELCOME TO CQB-DAY 2015
Centro de Química e Bioquímica (CQB) was founded in 2001 aiming at creating an environment
oriented toward fruitful collaborations between chemistry and biochemistry groups. In 2015, the
Strategic Project for 2015-2020 (PEst 2015-2020) was approved by FCT and initiated. The
research areas were organized in two thematic lines, as discussed in CQB-Day 2014, which had
the format of a small conference:
1. Chemistry and Biochemistry for a Clean Environment.
2. Chemistry and Biochemistry for a Healthy Aging.
All the groups had the opportunity of presenting orally their contributions towards these topics.
Very recently, a restructuring of the project was proposed owing to the low level of funding
assigned to the unit, compared to the initial objectives and deliveries. The second thematic line
was renamed Human health: molecular interventions and regulation mechanisms.
In CQB-Day 2015, there will be four oral presentations addressing collaborative work and one
plenary lecture for each thematic line. A large number of poster presentations should contribute
to increase the degree of cross-fertilization between the twelve groups and their sixty three
integrated members and approximately one hundred collaborators. Two new members of the
External Advisory Committee (EAC) will be present and we hope that they will get to know
CQB and help to achieve the proposed objectives.
We count on all of you to make this CQB Day successful and provide further opportunities to
discuss how to make our Strategic Project successful, by strengthening collaborations between
groups supported by novel scientific synergies, find new common grounds for research, answer
funding calls and participate in existing initiatives.
Maria José Calhorda
CQB coordinator
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SCIENTIFIC PROGRAM
09:15-09:30
Opening Session
09:30-10:15
Invited Communication
Wastewater - charcteristics and monitorization: Challanges in the preent to
solutions in the future
Doutora Eugénia Cardoso (Coordenadora do Departamento de Águas Residuais
da Direção de Controlo de Qualidade da Água; Águas de Portugal: Águas de
Lisboa e Vale do Tejo-EPAL)
10:15-10:45
Science at CQB – Oral Communications
O1. Magnetic Fe nanoparticles as support in catalysis
C.I. Fernandes, C. Bravo, O.C. Monteiro, C.D. Nunes
O2. Influence of activated carbons porous structure on iopamidol adsorption
A.S. Mestre, M. Machuqueiro, M. Silva, R. Freire, I.M. Fonseca, M.S.C.S.
Santos, M.J. Calhorda, A. P. Carvalho
10:45-11:45
Coffee-break/ Poster communications
11:45-12:15
Science at CQB – Oral Communications
O3. Self-assembly of multifunctional Fe(III) magnetic switches
P.N. Martinho, A.I. Vicente, A.A. Joseph, S.P. Shannon, A.J. Fitzpatrick, L.P.
Ferreira, M.D. Carvalho, V. Rodrigues, M.E. Minas da Piedade, G.G. Morgan, G.
Redmond, M.J. Calhorda
O4. Bar adsorptive microextraction (BAµE) coated with mixed sorbents phases –
Determination of non-steroidal anti-inflammatory drugs in real matrices in
combination with capillary electrophoresis
S.M. Ahmad, C. Almeida, N.R. Neng, J.M.F. Nogueira
12:15-13:45
Lunch
13:45-14:45
Science at CQB – Poster Communications
14:45-15:45
Invited Communication
Lead Generation in Pharmaceutical Research
Professor Hans P. Wessel (Professor Catedrático Convidado da Universidade de
Aveiro)
15:45-16:45
Science at CQB – Oral Communications
O5. Influence of herbal infusions on essential amino acids absorption. Can
infusions in nanoparticles avoid this effect?
J. Peres, A. Silva, C. Ferreira, J. Marquês, P. Falé, A.S. Viana, R. Pacheco, M. L.
Serralheiro
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O6. Alkyl deoxy glycosides as antimicrobial agents: synthesis, surface properties
and mechanism of action
C. Dias, J. P. Pais, P. Serra, A. Almeida, R. F. M. de Almeida, A. Martins, M. S.
Santos, A.S. Viana, R. Almeida, A. Pelerito, A. P Rauter
O7. The centrosomal protein TBCCD1 regulates the dynamics of a microtubule
subpopulation involved in the nucleus-centrosome
C. Camelo, C. Peneda, A.I. Câmara, B. Carmona, H.S. Marinho, H. Soares
O8. Tracking new psychoactive substances in Portugal
H. Gaspar, C. Leal, C. Gonçalves, S. Ciríaco, A. Matias, J. Rodrigues, S. Santos
16:45-17:00
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Conclusions and Closing Session
INDEX
PLENARY AND ORAL COMMUNICATIONS
WASTEWATER-CHARACTERISTICS AND MONITORIZATION CHALLENGES IN THE
PRESENT TO SOLUTIONS IN THE FUTURE
E. Cardoso……………………………………………………………………………………...…… 3
LEAD GENERATION IN PHARMACEUTICAL RESEARCH
H.P. Wessel………………………………………………………………………..………...4
O1
MAGNETIC Fe NANOPARTICLES AS SUPPORT IN CATALYSIS
C. I. Fernandes, C. Bravo, O. C. Monteiro, C. D. Nunes.………………….………………………..5
O2
INFLUENCE OF ACTIVATED CARBONS POROUS STRUCTURE ON IOPAMIDOL
ADSORPTION
A. S. Mestre, M. Machuqueiro, M. Silva, R. Freire, I. M. Fonseca, M. S .C. S. Santos,
M. J. Calhorda, A. P. Carvalho………………………..………………………………………….....6
O3
SELF-ASSEMBLY OF MULTIFUNCTIONAL FE(III) MAGNETIC SWITCHES
P. N. Martinho, A. I. Vicente, A. A. Joseph, S. P. Shannon, A. J. Fitzpatrick, L. P. Ferreira,
M. D. Carvalho, V. Rodrigues, M. E. Minas da Piedade, G. G. Morgan, G. Redmond,
M .J. Calhorda……………………………………….……………….……………..…..…..7
O4
BAR ADSORPTIVE MICROEXTRACTION (BAµE) COATED WITH MIXED
SORBENT PHASES –DETERMINATION OF NON-STEROIDAL ANTIINFLAMMATORY DRUGS IN REAL MATRICES IN COMBINATION WITH
CAPILLARY ELECTROPHORESIS
S. M. Ahmad, C. Almeida, N. R. Neng, J. M.F.Nogueira...................................................................8
O5
INFLUENCE OF HERBAL INFUSIONS ON ESSENTIAL AMINO ACIDS ABSORPTION.
CAN INFUSIONS IN NANOPARTICLES AVOID THIS EFFECT?
J. Peres, A. Silva, C. Ferreira, J. Marquês, P. Falé, A. S.Viana, R. Pacheco, M. L. Serralheiro ........ 9
O6
ALKYL DEOXY GLYCOSIDES AS ANTIMICROBIAL AGENTS: SYNTHESIS,
SURFACE PROPERTIES AND MECHANISM OF ACTION
C. Dias, J. P. Pais, P. Serra, A. Almeida, R .F. M. de Almeida, A. Martins, M.S.C.S. Santos,
A. S. Viana, A. Pelerito, A. P. Rauter ............................................................................................... 10
O7
THE CENTROSOMAL PROTEIN TBCCD1 REGULATES THE DYNAMICS OF A
MICROTUBULE SUBPOPULATION INVOLVED IN THE NUCLEUS-CENTROSOME
CONNECTION
C. Camelo, C. Peneda, A. I. Câmara, B. Carmona, H. S. Marinho, H. Soares ................................ 11
O8
TRACKING NEW PSYCHOACTIVE SUBSTANCES IN PORTUGAL
H. Gaspar, C. Leal, C. Gonçalves, S. Ciríaco, A. Matias, J. Rodrigues, S. Santos ........................... 12
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POSTER COMMUNICATIONS
P1
NON-RELEASING BIOCIDAL COATINGS: A NEW STRATEGY TO PREVENT
BIOFOULING
E. R. Silva, O. Ferreira. J. C. M. Bordado, M. J. Calhorda…………………………….............…..15
P2
ZNO@TIO2 CORE-SHELL NANOSTRUCTURES FOR SOLAR CELLS
APPLICATIONS
T. Frade, K. Lobato, A. Gomes .........................................................................................................16
P3
A MEMBRANE BIOPHYSICS APPROACH TO THE MECHANISM OF ACTION OF
NEW ACTIVE RUTHENIUM ANTICANCER COMPOUNDS
A. D. de Sousa, T.C. Santos, F.C. Santos, M.S.C.S. Santos, M.L. Corvo, M.H. Garcia, A.I. Tomaz,
R.F.M. de Almeida ............................................................................................................................17
P4
MoO3 NANOPARTICLES A HIGHLY SELECTIVE CATALYST
C. I. Fernandes, S. C. Capelli, P. D. Vaz, C.D. Nunes ......................................................................18
P5
COMPOUNDS FROM HERBAL INFUSIONS AND FRUITS IN THE PREVENTION OF
AGE-RELATED DISEASES
E. Brito, J. Peres, M. Salas, F. Ferreira, S. Barata, A. Ressaissi, A. Maia, N. Marques, P. Lopes, A.
Graça, A. Silva, R. Pacheco, M. H. Florencio, P. L. Falé, M. L. Serralheiro ...................................19
P6
SYNTHESIS AND MAGNETIC CHARACTERIZATION OF MANGANESE(III)
COMPLEXES
G. Santos, P. N. Martinho, S. Realista, A. Fitzpatrick, L. P. Ferreira, S. Barroso, L. C. J. Pereira, M.
J. Calhorda.........................................................................................................................................20
P7
IN VIVO REAL-TIME RESPONSE OF SACCHAROMYCES CEREVISIAE TO A MILDSTRESS INDUCED BY HYDROGEN PEROXIDE
R. N. Bento, C. E. S. Bernardes, M. E. Minas da Piedade, F. Antunes ............................................21
P8
MODULATION OF THE BIOPHYSICAL PROPERTIES AND PERMEABILITY OF
MODEL MEMBRANES BY SPHINGOSINE
A. C. Carreira, R. F. M. de Almeida, L. C. Silva ..............................................................................22
P9
BIOMIMETIC SUPPORTED LIPID BILAYERS FOR IMMUNOSENSING
I. Almeida, J. T. Marquês, T.M.O. Paiva, Y. Niu, W. Li, G. Jin, R. F.M. de Almeida,
A. S.Viana .........................................................................................................................................23
P10 SPHINGOLIPID-DEPENDENT PLASMA MEMBRANE BIOPHYSICAL PROPERTIES
IN ANTIFUNGAL DRUG ACTION AND RESISTANCE
F. C. Santos, C. A. C. Antunes, J. T. Marquês, A. S. Fernandes, A. S. Viana, A. Videira,
H. S. Marinho, R.F.M. de Almeida ...................................................................................................24
P11 H2O2 CONSUMPTION RATE AND REDOX SIGNALING IN ENDOTHELIAL CELLS IS
MODULATED BY EXTRACELLULAR MATRIX
A. Bagulho, F. Vilas-Boas, A. Pena, C. Peneda, F. C. Santos, A. Jerónimo, R. F .M. de Almeida,
C. Real ...............................................................................................................................................25
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P12 CORE@SHELL NANOSTRUCTURES FOR MAGNETIC HYPERTHERMIA
APPLICATIONS
S. G. Mendo, A. F. Alves, L. P. Ferreira, M. M. Cruz, M. H. Mendonça, M. Godinho,
M. D. Carvalho.................................................................................................................................. 26
P13 LUMINESCENT HYDROPHOBIC SURFACES FOR SMART WINDOWS
J. B. Aldeias, S. Realista, M. J. Calhorda, P. N. Martinho................................................................ 27
P14 MODIFIED NANOSTRUCTURED ZnO PHOTOANODES SENSITIZED WITH N719 IN
ETHANOL AND ETHANOL/WATER
D. Siopa, S. Sério, M. E. Melo Jorge, F. Martins, M. S. C. S. Santos, K. Lobato, A. Gomes .......... 28
P15 pH-DEPENDENT INSERTION OF PHLIP PEPTIDE INTO BILAYERS: INSIGHTS
FROM COMPUTATIONAL SIMULATIONS
D. Vila-Viçosa, V. H. Teixeira, M. Machuqueiro ............................................................................. 29
P16 MANGANESE(II) COMPLEXES WITH PYRAZOLE LIGANDS: SYNTHESIS AND
OXIDATIVE CATALYSIS
T. A. G. Duarte, L. M. D. R. S. Martins, A. P. Carvalho, A. J. L. Pombeiro ................................... 30
P17 A TIME-RESOLVED PHOTOACOUSTIC CALORIMETRY STUDY OF
CYCLOHEXYL, 1,4-DIOXYL, AND TETRAHYDROFURAN-2-YL PEROXY
RADICALS
E. M. Gonçalves, F. Agapito, R. M. Borges dos Santos, J. A. Martinho Simões ............................. 31
P18 INTRAMOLECULAR ALLYLIC AMINATION CATALYZED BY Pd(II): A
COMPUTATIONAL STUDY
M. J. Calhorda, F. J. S. Duarte, M. M. Lorion, G. Poli ..................................................................... 32
P19 SYNTHESIS AND PHOTOCATALYTIC ACTIVITY OF COBALT DOPED TITANATE
NANOTUBES WITH DIFFERENT COBALT POSITION IN THE STRUCTURAL
B. Barrocas, A. J. Silvestre, C. D. Nunes, O. C. Monteiro ............................................................... 33
P20 BIOSYNTHESIS OF METAL SULPHIDE NANOPARTICLES AND THEIR
PHOTOCATALYTIC ACTIVITY IN DEGRADATION OF EMERGING POLLUTANTS
B. Vieira, G. Vitor, J. P. Lourenço, O. C. Monteiro, M. C. Costa .................................................... 34
P21 ADSORPTION OF PHARMACEUTICAL COMPOUNDS ON CARBONS OF
DIFFERENT MORPHOLOGIES
S. Marques, J. Marcuzzo, A. S. Mestre, R. Dias, C. O. Ania, A. P. Carvalho .................................. 35
P22 NOVEL TECHNOLOGIES FOR SAMPLE PREPARATION IN GREEN ANALYTICAL
CHEMISTRY
A. H. Ide, S. M. Ahmad, N. R. Neng, J. M. F. Nogueira .................................................................. 36
P23 EXPLORING THE CATALYTIC BEHAVIOUR OF HIERARCHICAL MCM-22
ZEOLITE IN LOW TEMPERATURE FRIEDEL-CRAFTS ACYLATION OF
HETEROAROMATICS
R. Aleixo, N. Nunes, R. Leitão, F. Martins, A. P. Carvalho, A. Brigas, A. Martins ........................ 37
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P24 HUMAN RED BLOOD CELLS EXOVESICLES: A STUDENTS’ APPROACH TO
PLASMA MEMBRANE DYNAMICS
C. P. Sabino, A. G. Flor, T.M.O. Paiva, R. F. M. de Almeida ..........................................................38
P25 HOW INERT ARE DRUG EXCIPIENTS? A BIOPHYSICAL STUDY OF M-CRESOLBIOMEMBRANE INTERACTIONS IN MODEL SYSTEMS AND NEURAL CELLS
T. M. O. Paiva, A. E. P. Bastos, J. M. Marquês, P. A. Lima, A. S. Viana, R. F. M. de Almeida .....39
P26 DEHYDRATION OF A ROBUST AND YET METASTABLE HEMIHYDRATE OF 4HYDROXYNICOTINIC ACID: THERMODYNAMICS, KINETICS AND MECHANISM
A. Joseph, C. E. S. Bernardes, A. S. Viana, M. F. M. Piedade, M. E.Minas da Piedade ..................40
P27 MOO3 NANOPARTICLES IN OXIDATION CATALYSIS
C. D. Nunes, A. Sanches, A. Bento, P. D. Vaz .................................................................................41
P28 POLLUTANTS REMOVAL USING NEW MAGNETIC-BASED PHOTOCATALYSTS
C. Bravo, C. D. Nunes, O. C. Monteiro ............................................................................................42
P29 ENZYME MODIFIED POLYDOPAMINE FILMS WITH HIGH CATALYTIC
PERFORMANCE
L. C. Almeida, A. C. Carreira, J. P. Correia, A. S. Viana .................................................................43
P30 THE AMBIGUOUS CASE OF POLYMORPHISM IN SIMVASTATIN: A SINGLE
CRYSTAL X-RAY DIFFRACTION, THERMODYNAMIC, AND MD SIMULATION
STUDY
R. G. Simões, C. E. S. Bernardes, M. F. M. Piedade, H. P. Diogo, M. E. Minas da Piedade ...........44
P31 SYNTHESIS, CHARACTERIZATION AND ANTI-CANCER EVALUATION OF NOVEL
RUTHENIUM (II) COMPLEXES OF THIOSEMICARBAZONE
S. Assis, A. C. Poeta, A. Valente, T. S. Morais, F. Marques, M. P. Robalo, S. Santos, A. I. Tomaz,
M. H. Garcia ......................................................................................................................................45
P32 SHAPE AND CURVATURE OF SURFACE TENSION ISOTHERMS FOR LIQUID
MIXTURES
M. S. C. S. Santos, J. C. R. Reis........................................................................................................46
P33 FORMULATION OPTIMIZATION AND STABILITY EVALUATION OF A
BIOACTIVE CATIONIC NANOEMULSION CONTAINING QUERCETIN
M. F. Dario, M. S. C. S. Santos, N. A. Bou-Chacra, M. E. Minas da Piedade, A. R. Baby,
M. V. R. Velasco ..............................................................................................................................47
P34 BIOPHYSICAL PROPERTIES OF STEAROYL-PHYTOCERAMIDE MIXED WITH A FLUID
PHOSPHOLIPID. ROLE OF SPHINGOID BASE HYDROXYLATION IN MEMBRANE
COMPARTMENT ORGANIZATION
J. T. Marquês, A. M. Cordeiro, A. S. Viana, A. Herrmann, H. S. Marinho, R. F.M. de Almeida ....48
P35 ADDING EXPLICIT -HOLES IN THE GROMOS FORCE FIELD: TOWARDS DRUG
DESIGN APPLICATIONS
D. Vila-Viçosa, M. Machuqueiro, P. J. Costa ...................................................................................49
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P36 DESIGN OF UREA-BASED ANION RECEPTORS WITH ANTICANCER PROPERTIES
J. M. Caio, M. J. Martins, T. Esteves, O. Cruz e Silva, V. Félix, C. Moiteiro.................................. 50
P37 SEPARATION OF THE ETHANE/ETHYLENE MIXTURE IN UIO-66 MOFS
J. Fernandes, M. Pinto, J. Pires ......................................................................................................... 51
P38 METABOLIC PROFILE OF LEISHMANIA INFANTUM PROMASTIGOTES AND
AMASTIGOTES BY FT-ICR-MS ANALYSIS
A. P. Marques, M. Maia, A. E. N. Ferreira, A. Figueiredo, F. Monteiro, M. Sebastiana,
S. Carvalho, A. M. Tomás, A. Ponces Freire, C. Cordeiro, M. Sousa Silva ..................................... 52
P39 DATEBASE OF PHENOLIC COMPOUNDS BY MASS SPECTROMETRY
C. Sánchez, M. H. Florêncio, M. L. Serralheiro, A. P. Marques ...................................................... 53
P40 ANTIOXIDANT PROPERTIES AND EVALUATION OF TANNINS IN DIFFERENT
CASTES OF GRAPES
M.E.M. Araújo, B. Oliveira .............................................................................................................. 54
P41 PHYTOCHEMICAL SCREENING OF MEDICINAL PLANTS FROM TUNISIA
(MARRUBIUM VULGARE AND GLOBULARIA ALYPUM)
M.E.M. Araújo, M. Rezgui, S.Bellili, L. Bettaieb Ben-Kaab, W. Mnif ........................................... 55
P42 TRACKING NEW PSYCHOACTIVE SUBSTANCES IN PORTUGAL
H. Gaspar, C. Leal, C. Gonçalves, S. Ciríaco, A. Matias, J. Rodrigues, S. Santos ........................... 56
P43 SYNTHESIS OF HYACINTH FRAGRANCE: CATALITIC BEHAVIOR OF
ACTIVATED CARBONS MADE FROM RAPESEED SOLID RESIDUE OF BIODIESEL
PRODUCTION
M. Batista, I. Fonseca, I. Matos, J. Vital, A. Mestre, A. P. Carvalho ............................................... 57
P44 MOB1 AN ACTIVE PLAYER IN MORPHOGENESIS, CYTOKINESIS AND CELL
PROLIFERATION CONTROL IN PROTOZOA ORGANISMS
A. Tavares, I. Delgado, S. Francisco, J. Coelho, A. Leitão, H. Soares, S. Nolasco .......................... 58
P45 MOLYBDENUM COMPLEXES WITH 2,2’-DIPYRIDYLAMINE DERIVATIVES AS
CATALYSTS IN OXIDATION REACTIONS
M. S. Saraiva, M. J. Calhorda ........................................................................................................... 59
P46 FUNCTIONAL NANOPARTICLES OF FOOD PROTEINS FOR REDUCTION OF
CARDIOVASCULAR DISEASES RISK
A. Silva, J. T. Marquês, A. S. Viana, R. Pacheco, M. L. M. Serralheiro .......................................... 60
P47 SYNTHESIS AND APPLICATION OF NEW TITANATE NANOTUBES SENSITIZED
WITH SILVER NANOPARTICLES FOR POLLUTANTS PHOTODEGRADATION
B. Barrocas, O. C. Monteiro ............................................................................................................. 61
P48 PKA VALUES OF TITRABLE AMINO ACIDS AT THE WATER/MEMBRANE
INTERFACE
V. H. Teixeira, D. Vila-Viçosa, P. B. P. S. Reis, M. Machuqueiro .................................................. 62
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P49 METHOD'S DEVELOPMENT FOR DRUG ANALYSIS BY MASS SPECTROMETRY
AND OTHER ANALYTICAL TECHNIQUES
R. A. Osawa, M. H. Florêncio, A. P. Carvalho, M. R. Bronze .........................................................63
P50 RECOVERY OF PALLADIUM FROM A SPENT INDUSTRIAL CATALYST BY
HYDROMETALLURGY
O. Ortet, A. P. Paiva, C. Nogueira ....................................................................................................64
P51 TRANSITION METAL COMPLEXES FOR CO2 REDUCTION
S. Realista, A. I. Melato, P. M. Martinho, M. J. Calhorda ................................................................65
P52 THE CENTROSOMAL PROTEIN TBCCD1 REGULATES THE DYNAMICS OF A
MICROTUBULE SUBPOPULATION INVOLVED IN THE NUCLEUS-CENTROSOME
CONNECTION
C. Camelo, C. Peneda, A. I. Câmara, B. Carmona, H. S. Marinho, H. Soares .................................66
P53 THEORETICAL STUDY ON THE INTERCALATION OF PHENANTHROLINE IN
DNA: WHEN DISPERSION FORCES ARE IMPORTANT BUT THE ELECTROSTATIC
CONTRIBUTION BECOMES CRUCIAL
A. Gil, V. Branchadell, M. J. Calhorda .............................................................................................67
P54 TARGETING BUTYRYLCHOLINESTERASE FOR NEW TREATMENTS IN
ALZHEIMER’S DISEASE AND CANCER
V. Cachatra, S. Schwarz, M. C. Oliveira, L. Gano, A. Paulo, A. P. Rauter..................................... 68
P55 THE PHENOL O–H BOND DISSOCIATION ENTHALPY
F. Agapito, R. M. Borges dos Santos, J. A. Martinho Simões ..........................................................60
P56 CHEMICAL STABILITY AND NEUROPROTECTIVE ACTIVITY EVALUATION OF
ERICA AUSTRALIS L. EXTRACTS
P. Dias, A. Martins, A. P. Rauter, P. L. Falé …………………..…………………………………70
P57 SELECTIVE RECOVERY OF SILVER FROM DILUTE SOLUTIONS BY ELECTROLESS
PRECIPITATION USING POLYANILINE FILMS
I. J. Pereira, J. P. Correia……………..…………………………………………………………….71
P58 NEW SUGAR-BASED SURFACTANTS TARGETTING LIPID BILAYERS: SYNTHESIS
AND MOLECULAR DYNAMICS STUDY
R. Nunes, J. Pais, D. Vila-Viçosa, M. Machuqueiro, A. P. Rauter……………………………….. 72
P59 NOVEL TRIAZOLE, PURINE AND SULFONOHYDRAZIDE GLYCOCONJUGATES
FOR THE TARGETING OF NUCLEOTIDE-DEPENDENT ENZYMES: SYNTHESIS
AND BIOACTIVITY EVALUATION
N. M. Xavier, S. D. Lucas, R. Jorda, S. Schwarz, A. Loesche, R. Csuk..........................................73
P60 NOVEL N-GLYCOSYLSULFONAMIDES DERIVED FROM D-RIBOSE AND
D-GLUCURONAMIDE: SYNTHESIS AND CONFORMATIONAL STUDIES
A. Fortuna, Paulo J. Costa, N. M. Xavier......................................................................................... 74
xvi
P61 SYNTHESIS AND BIOACTIVITY EVALUATION OF PURINE AND PYRIMIDINE
6´-ISONUCLEOSIDES
D. Batista, R. Jorda, S. Schwarz, A. Loesche, R. Csuk, N. M. Xavier............................................ 75
P62 BIOMASS-DERIVED ACTIVATED CARBONS: PROMISING GREEN MATERIALS
FOR ENVIRONMENTAL REMEDIATION
M. A. Andrade, A. S. Mestre, C. O. Ania, A. P. Carvalho....................................................................................... 76
P63 C-GLUCOSYLATION REACTIONS IN THE SYNTHESIS OF NEW SUGAR-LINKED
MOLECULES WITH POTENTIAL ANTI-AMYLOID ACTIVITY
A. M. Matos, A. P. Rauter............................................................................................................... 77
P64 ALKYL DEOXY GLYCOSIDES AS ANTIMICROBIAL AGENTS: SYNTHESIS,
SURFACE PROPERTIES AND MECHANISM OF ACTION
C. Dias, J. P. Pais, P. Serra, A. Almeida, R. F. M. Almeida, A. Martins, M.S.C.S. Santos, A.S.
Viana, A. Pelerito, A. P Rauter ......................................................................................................... 78
P65 GLYCOLIPIDS: ANTIBACTERIAL ACTIVITY AND INSIGHTS INTO MECHANISM
OF ACTION VIA GENETIC AND METABOLIC APPROACHES
J. Pais, A. S. Viana, A. P. Rauter, L. Sobral, R. Dias, R. Tenreiro.................................................. 79
P66 SYNTHESIS AND IN VITRO STUDY OF C-GLUCOSYL DIHYDROCHALCONES AS
POTENTIAL SODIUM-GLUCOSE CO-TRANSPORTER (SGLT) INHIBITORS FOR
THE TREATMENT OF DIABETES
A. R. Jesus, T. Dore, A. P. Rauter.................................................................................................... 80
P67 PREPARATION OF SPHERICAL NANOPOROUS CARBONS BY K2CO3
ACTIVATION OF BIOMASS ACID LIQUORS DERIVED CHARS
F. Hesse, A.S. Mestre, C. Freire, C.O. Ania, A.P. Carvalho............................................................. 81
P68 HIERARCHICAL ZEOLITES: A GREEN ALTERNATIVE TO CONVENTIONAL
FRIEDEL-CRAFTS ACYLATION OF HETEROAROMATICS
R. Aleixo, N. Nunes, R. Leitão, F. Martins, A.P. Carvalho, A. Brigas, A. Martins .......................... 82
P69 REMOVAL OF MICROCYSTIN-LR BY CORK-BASED AND COMMERCIAL ACTIVATED
CARBONS
C. Menezes, S. José, P. Pereira, S. P. Silva, A. P. Carvalho, A. S. Mestre .................................................... 83
P70 CYSTOSEIRA TAMARISCIFOLIA AS A POTENTIAL BIOMEDICAL SOURCE FOR
ANTIOXIDANT AND ANTI-HEPATOCARCINOMA COMPOUNDS
C. Vizetto-Duarte, L. Custódio, J. H. G. Lago, T. R. Morais, C. B. de Sousa, K. N. Gangadhar, L.
Barreira, F. Albericio, A. P. Rauter, J. Varela………………………………………………………………………….84
P71 BIOMASS CONVERSION INTO CARBON BIOSORBENTS
P. S. de Velasco, A. S. Mestre, V. Hernández, A. P. Carvalho............................................................................. 85
xvii
Science at CQB
Plenary and Oral Communications
CQB-Day 2015
WASTEWATER-CHARACTERISTICS AND MONITORIZATION
CHALLENGES IN THE PRESENT TO SOLUTIONS IN THE FUTURE
Eugénia Cardoso
Águas de Portugal: Águas de Lisboa e Vale do Tejo - EPAL
The role of wastewater treatment plants in the environment is presented along with the enterprise Águas
de Lisboa e Vale do Tejo, SA (EPAL-LVT) and is responsibility in the collection, treatment and disposal
of the waters from 85 municipalities in Portugal, correspondent to 3.278.500 inhabitants from River
Zêzere and Côa, to Tagus basin. Analytical monitorization needs and resources are reviewed in present
and future perspective.
Critical points connections are established considering that wastewater treatment plants are assumed as an
important source of organic and inorganic contaminants to the aquatic environment, that becomes to the
system from different origins: industrial effluents, hospital-clinical effluents with pharmaceutical
compounds, consumption of ilicitt drugs, that are sent to public wastewater treatment plants (WWTP),
along with domestic sewage, either in specific points discharge or from disperse, diffuse sources. Short
overview of WWTP concerning the need of new, fast, reliable forms to control processes and discharges
in environment, as well as reclaimed water and sludge are discussed by comparison with classical
analytical resources. Respirometric tool with direct evaluation of effects in biological sludge: Strathox is
presented as a case study that aimed to study an integrated and multidisciplinary tool to control potential
toxic influents and optimization energetic efficiency of biological process. Other projects concerning the
identification of pharmaceutical and illicit drugs in the sewer, as well as studies intended to improve
efficiency of biological treatment process are presented as examples of the role and importance of
chemistry and biochemistry to a clean environment.
3
CQB-Day 2015
LEAD GENERATION IN DRUG RESEARCH
Hans Peter Wessel
University of Aveiro, Portugal
Lead Generation is the important stage in the long drug research and development path from concept to
market in which the basic chemical structures of a project will be determined. While historically drug
candidates were solely derived from natural product isolates, modern drug research relies largely on
screening of compound libraries of different origin. Various routes to chemical and virtual library
constructions as well as screening techniques including DNA-encoded library technology will be
discussed, garnished with practical research examples from experience at F. Hoffmann-La Roche, Basel,
Switzerland. On the other hand, mimetics approaches have been successful as exemplified by the
development of agents against viral infections.
4
CQB-Day 2015
O1
MAGNETIC Fe NANOPARTICLES AS SUPPORT IN CATALYSIS
C. I. Fernandes, C. Bravo, O. C. Monteiro, C. D. Nunes
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, Ed. C8, 1749-016 Lisboa,
Portugal, ([email protected])
Nowadays nanoparticles (NPs) research is a very dynamic field that ranges across many scientific areas.
In the particular case of catalysis many studies have been addressed showing that the investment in this
kind of systems is beneficial, since the high surface/volume ratio presented by them yields, generally,
high performance. The catalytic properties in many cases can be successfully combined with other
properties (such as magnetism) allowing an important added value for the use of multi functionalized NPs
in such processes.
Following this research line, in this work the organometallic fragment [MoI2(CO)3] was coordinated to
magnetic iron oxide nanoparticles which have been previously coated with a silica shell and grafted with
a pyridine derivative ligand.
The synthesis of MNP, possessing magnetic properties, had the aim of favoring and simplifying the
separation process after a catalytic process.
Preparation of these organometallic decorated magnetic nanoparticles was further confirmed by evidence
obtained from extensive characterization by powder XRD, SEM/TEM analysis, as well as FTIR
spectroscopy.
The catalytic activity of the prepared hybrid materials were evaluated in several (photo)catalytic
processes, including pollutants photodegradation and olefin epoxidation reactions.
The epoxidation catalytic studies show that the catalysts yield selectively the desired epoxides of a series
of olefins. In addition, these catalysts are found to work under a wide temperature range and over several
catalytic cycles without notorious performance loss in most cases.
The photocatalytic performance of these hybrid materials was also studied using distinct organic
molecules: terephthalic acid (as probe for hydroxyl radical production study), phenol and Rhodamine B.
All the nanomaterials showed good catalytic behavior. However the results obtained indicates that the
stability of the hybrid material is dependent on the experimental conditions used mainly (photo)catalysis
media.
Figure 1. TEM images of MNP and MNP-Si
Acknowledgements The authors are grateful to FCT for financial support project UID/MULTI/00612/2013.
References
[1] C.I. Fernandes, M.D. Carvalho, L.P. Ferreira, C.D. Nunes, P.D. Vaz (2014) J. Organomet. Chem. 760: 2-10.
5
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CQB-Day 2015
INFLUENCE OF ACTIVATED CARBONS POROUS STRUCTURE ON IOPAMIDOL ADSORPTION
A. S. Mestre,a M. Machuqueiro,a M. Silva,a,b R. Freire,a,b I. M. Fonseca,b M. S. C. S. Santos,a
M. J. Calhorda,a A. P. Carvalhoa
a Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal b Departamento
de Química, REQUIMTE CQFB, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, Quinta da Torre, 2829-516
Caparica, Portugal ([email protected])
In the present study the adsorption of iopamidol (a pharmaceutical compound used in soft tissues
diagnostic imaging based on X-ray radiography) onto activated carbons was assessed, since this is an
effective technology to remove pharmaceutical compounds that fail to be degraded in conventional
wastewater treatments.
Three sisal-based carbons prepared by chemical activation with KOH, and two commercial carbons, were
selected in order to understand the role of the porous structure in the removal of iopamidol from aqueous
phase. The kinetic and equilibrium adsorption results indicate that iopamidol is adsorbed in mesopores
and also in larger micropores (supermicropores). The adsorption isotherms reflect a complex mechanism
originating unusual two-step isotherms, which highlights the influence of the porous structure. As proven
using conductivity measurements and computational calculations, iopamidol can be adsorbed as a single
molecule or in the form of aggregates, filling either small or much larger pores in a discontinuous way
(Fig. 1).
The adsorption capacity of the carbons appears to be related with the volume of larger micropores and
mesopores. However, the micropore size distribution must also be taken into account to explain the
iopamidol adsorption process, which in some cases can originate unusual two-step isotherms. A
multidisciplinary approach, combining computational studies and conductivity measurements, showed the
stability of progressively larger iopamidol species (dimer and trimer). Comparing the molecular
dimensions of these iopamidol aggregates with the micropore size distribution of the samples, it could be
concluded that the appearance of two-step isotherms is linked with micropore size distributions without
pores in the range between 1.2 and 2.0 nm.1
Figure 1. Schematic drawing of iopamidol species (monomer, dimer and trimer) adsorption onto activated carbon
pore structure.
Acknowledgements The authors thank the Fundação para a Ciência e Tecnologia, Portugal, for financial support
(Project PEst-OE/QUI/UI0100/ 2014) and a fellowship to ASM (SFRH/BPD/86693/2012). Cordex, Quimitejo, and
Hovione are also acknowledged for providing, respectively, the sisal wastes, the commercial carbon samples, and
iopamidol.
References
1. A. S. Mestre, M. Machuqueiro, M. Silva, R. Freire, I. M. Fonseca, M. S. C. S. Santos, M. J. Calhorda, A, P.
Carvalho, Carbon, 77 (2014) 607
6
CQB-Day 2015
O3
SELF-ASSEMBLY OF MULTIFUNCTIONAL FE(III) MAGNETIC SWITCHES
P. N. Martinhoa, A. I. Vicentea, A. A. Josepha, S. P. Shannonb, A. J. Fitzpatrickb, L. P. Ferreirac,d, M. .D.
Carvalhoa, V. Rodriguesd, M. E. Minas da Piedadea, G. G. Morganb, G. Redmondb, M. J. Calhordaa
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected]).
bSchool of Chemistry & Chemical Biology, University College Dublin, Belfield, Dublin 4, Ireland
cBioISI, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, 1749-016 Lisboa, Portugal
dDepartamento de Física, Faculdade de Ciências e Tecnologia, Universidade de Coimbra, 3004-516 Coimbra, Portugal
Spin crossover (SCO) complexes have long been considered as promising candidates for the next
generation of data storage devices [1,2]. SCO candidate compounds can be found among a limited group
of 3d4–3d7 transition metal ions, the most common being Fe(II), Fe(III) and Co(II). Fe(III), with its
advantageous redox stability, is a good candidate for fabrication of SCO materials, an area towards which
research has been moving [3]. Much effort has been expended in recent years to develop the materials
assembly of SCO complexes and impressive results have been achieved in stabilising and isolating
monodisperse nanoparticles, nanocrystals, nanowires, thin films, micro- and nanopatterned media,
Langmuir- Blodgett surface mono- and multilayers and hysteretic soft media assemblies.
We recently synthesised a mononuclear Fe(III) compound displaying a wide hysteresis window centred at
room temperature. We also found that this compound undergoes a phase transition coupled with the
thermosalient effect resulting in crystal fragmentation with no loss of both SCO and hysteresis. Here we
present the synthesis and characterisation of a [Fe(SalEen)2]+ derivative and its self-assembly into
nanoparticles and nanowires using template-free fabrication techniques. Variable temperature Raman and
AFM are used to investigate both the SCO and thermosalient effect of the new materials.
Acknowledgements We thank FCT for financial support UID/MULTI/00612/2013 and UID/MULTI/04046/2013.
PNM thanks FCT for financial support (SFRH/BPD/73345/2010).
References
1. O. Kahn and J. P. Launay, Chemtronics 3, 140-144, (1988).
2. O. Kahn, J. Krober and C. Jay, Adv. Mater. 4, 718-728, (1992).
3. M. A. Halcrow, Spin-Crossover Materials: Properties and Applications (John Wiley & Sons Ltd, 2013).
7
O4
CQB-Day 2015
BAR ADSORPTIVE MICROEXTRACTION (BAµE) COATED WITH MIXED SORBENT PHASES –
DETERMINATION OF NON-STEROIDAL ANTI-INFLAMMATORY DRUGS IN REAL MATRICES IN
COMBINATION WITH CAPILLARY ELECTROPHORESIS
S. M. Ahmad, C. Almeida, N. R. Neng, J. M. F. Nogueira
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
Non-steroidal anti-inflammatory drugs (NSAIDs), such as acetylsalicylic acid, diclofenac and naproxen,
are the most widely prescribed medications worldwide [1]. These compounds are used to reduce pain and
inflammation, because of their antipyretic, analgesic and anti-inflammatory properties. In spite of these
therapeutic effects, toxicity and side effects such as gastro-intestinal effects or renal problems, can occur
[2]. For these reasons, their determination in biological fluids, such as urine and plasma, is very
important. This information is required for assessing the safety, mechanisms of action and therapeutic
effect of these drugs, having a strong impact on areas such as forensic and clinical toxicology.
Furthermore, NSAIDs have been also evidenced as potential contaminants of aquatic resources such as
surface, estuary, river, sea and drinking waters, having a possible toxicological effect in the environment
[3].
In this contribution, the development, optimization, validation and application of a novel method, based
on the “Floating Sampling Technology” [4], for the determination of NSAIDs in urine and environmental
water matrices are discussed. Thus, bar adsorptive micro-extraction technique [5] was used in
combination with mixed polymeric sorbent phases, followed by liquid desorption and analysis by
capillary electrophoresis with diode array detection (BaμE(PMIX)-LD/CE-DAD). Under optimized
experimental conditions, it was possible to obtain average efficiencies of 86.6 and 104.1 % for all the
NSAIDs under study.
Acknowledgements
The authors wish to thank Foundation for Science and Technology for funding (UID/MULTI/00612/2013), Nuno R.
Neng Post-Doc grant (SFRH/BPD/86071/2012) and C. Almeida PhD grant (SFRH/BD/69951/2010).
References
[1] A. Inotai, B. Hanko, A. Meszaros, Pharmacoepidem. Dr. S., 19 (2010) 183-190.
[2] A. Pilotto, D. Sancarlo, F. Addante, C. Scarcelli, M. Franceschi, Surg. Oncol., 19 (2010) 167-172.
[3] M. Gonzalez-Rey, M.J. Bebianno, Aquat. Toxicol., 148 (2014) 221-230.
[4] J.M.F. Nogueira, Anal. Chim. Acta, 757 (2012) 1–10.
[5] N.R. Neng, A.R.M. Silva, J.M.F. Nogueira, J. Chromatogr. A, 1217 (2010) 7303–7310.
8
CQB-Day 2015
O5
INFLUENCE OF HERBAL INFUSIONS ON ESSENTIAL AMINO ACIDS ABSORPTION. CAN INFUSIONS
IN NANOPARTICLES AVOID THIS EFFECT?
J. Peresa, A.Silvaa, C. Ferreiraa, J.Marquêsa, P. Faléa,b, A.S. Vianaa, R. Pachecoa,c, M. L. Serralheiroa,d
a.Centro de Química e Bioquímica, Faculdade de Ciências Universidade de lisboa; b Institute of Pharmaceutical Science.
Faculty of Life Sciences & Medicine. King's College London. Franklin-Wilkins Building. 150 Stamford Street. Waterloo. London
SE1 9NH, UK.; cÁrea Departamental de Engenharia Química, Instituto Superior de Engenharia de Lisboa, Av. Conselheiro
Emídio Navarro, 1959-007 Lisboa, Portugal; dDepartamento de Quimica e Bioquimica, FCUL, Campo Grande 1749-016
Lisboa ([email protected])
Essential amino acids are an important part of the human diet. The consumption of herbal infusions
during a meal may pose some problems in amino acid absorption. Several infusions previously analysed
for dietary cholesterol reduction [1,2] were used to study the permeation of amino acids. The system used
was Caco-2 cell lines, which underwent differenciation in the appropriate transwell system developing
microvillosities to simulate the intestinal absorption system. The results indicated that all plant infusions
under evaluation could diminish the absorption of essential amino acids present in the cell culture
medium. These results seem to indicate that the effect depends on the amino acid transporter system
present on the apical cell membrane. These results pave the way to a new line of studies. In order to avoid
this effect and to take some benefit of the decrease in the diet cholesterol absorption previously detected,
encapsulation of herbal infusions in protein nanoparticles are being tested at present. The effect of these
infusions on the nanoparticles morphology was evaluated using AFM. Besides hiding the phenolic
compounds inside the protein nanostructures, the system under development can also function as a new
source of proteins, adding nutritional value to the herbal phenolic compounds.
Figure 1: Plants studied and amino acids
Acknowledgements: PTDC/QUI/BIQ/13477/2009; UID/MULTI/00612/2013
References
1. P.L. Falé, C. Ferreira, A.M. Rodrigues, F.N. Frazão, M.L.Serralheiro, J. Med Plant Res 8 (2014) 9.
2. P.L.Falé, C. Ferreira, F. Maruzzella, M.H. Florencio, F.N. Frazão, M.L. Serralheiro, J. Ethnopharmacol 150
(2013) 718.
9
O6
CQB-Day 2015
ALKYL DEOXY GLYCOSIDES AS ANTIMICROBIAL AGENTS: SYNTHESIS, SURFACE PROPERTIES
AND MECHANISM OF ACTION
C. Dias,a J. P. Pais,a P. Serra,a A. Almeida,a R. F. M. Almeida,a A. Martins,a M.S.C.S. Santos,a
A.S. Viana,a A. Pelerito,b A. P Rautera
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected]). bInstituto Nacional de Saúde Doutor Ricardo Jorge, Avenida Padre Cruz, 1749-016 Lisboa, Portugal
The search for new antimicrobial drugs is currently one of the major ongoing research areas due to the
spread of multidrug-resistance, making research on new antibacterial agents with new mechanisms of
action highly relevant. Another matter of concern is the fact that infections in elderly populations are
known to be not only more frequent but also more severe, being this susceptibility often related to
neurodegenerative diseases such as dementia and Alzheimer’s [1]. Alkyl 2-deoxy/2,6-dideoxy-arabinohexopyranosides with a potent antimicrobial activity in several Bacillus species have been previously
described by our research group [2, 3]. Moreover, promising preliminary results arising from interaction
studies of some of these 2,6-dideoxyglycosides with cystatin B amyloid fibrils, assessed by NMR
spectroscopy, show their potential for neurodegenerative diseases as well.
We present now the synthesis and the antimicrobial activity of related glycosides differing in the glycon
structure (D- and L-series, hexo- and pentopyranosides, arabino and threo configuration) and in the
lipophilic chain (chain size, fluorinated chain, S-linked chain, amide containing chain), for the recognition
of the structural features that determine the selectivity for Bacillus species. The synthetic approach is
based on the reaction of glycals with alcohols catalysed by triphenylphosphane hydrobromide. Moreover,
aiming at a better understanding of the importance of the deoxygenation pattern, synthetic methodologies
towards new alkyl 3-deoxy, 4-deoxy and 6-deoxy glycosides were also investigated.
The surface properties of the most active compounds, in terms of adsorption and aggregation parameters,
were assessed and suggested that surface activity is required for the bioactivity. However, one of the most
promising surface active glycosides was not active. In order to have insights into the relationship between
surface activity and bioactivity of this family of compounds expected to target bacterial membranes,
collaborative work based on biophysical methods for the study of the interaction of these molecules with
lipossomes was encouraged.
This work clearly demonstrates the uniqueness of carbohydrates which stereochemistry and chemical
structure can tune the bioactivity exhibited by stereoisomers.
Acknowledgements This work was supported by FEDER-QREN-SI I&DT, Co-Promotion Project nr. 21457. The
authors thank the FCT for financial support (PEST UID/MULTI/00612/2013), and FCT and Cipan are also
gratefully acknowledged for the Ph.D. grants of C. Dias (SFRH/BDE/51998/2012) and J. P. Pais
(SFRH/BDE/51957/2012).
References
1. G. Gavazzia, K.-H. Krause. Lancet Infec. Dis. 2 (11) (2002) 659.
2. A. P. Rauter, S. Lucas, T. Almeida, D. Sacoto, V. Ribeiro, J. Justino, A. Neves, F. V. M. Silva, M. C.
Oliveira, M. J. Ferreira, M. S. Santos, E. Barbosa. Carbohydr. Res. 340 (2005) 191.
3. F. Silva, M. Goulart, J. Justino, A. Neves, F. Santos, J. Caio, S. Lucas, A. Newton, D. Sacoto, E. Barbosa, M. S.
Santos, A. P. Rauter. Bioorg. Med. Chem. 16 (2008) 4083.
10
CQB-Day 2015
O7
THE CENTROSOMAL PROTEIN TBCCD1 REGULATES THE DYNAMICS OF A MICROTUBULE
SUBPOPULATION INVOLVED IN THE NUCLEUS-CENTROSOME CONNECTION
C. Cameloa, *, C. Penedaa, *, A.I. Câmaraa, B. Carmonaa,b, H. S. Marinhoa H. Soaresa,b#
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa; bEscola Superior de
Tecnologia da Saúde de Lisboa, 1990-096 Lisboa, Portugal; #([email protected])
*These authors have contributed equally for this work.
The centrosome is a major microtubule-organizing center (MTOC) in animal cells, usually localized at
the cell center in close association with the nucleus. It has been reported to have an important role in
several cell processes such as organelle positioning, cell migration, adhesion, polarity, spindle pole
formation and assemble cilia axoneme. The TBCC domain-containing human protein (TBCCD1) is a
centrosomal protein previously characterized by our group 1. TBCCD1 contains two identified domains:
a TBCC domain, which is associated with the tubulin folding pathway, and a CARP domain, that is
thought to be associated with actin filaments. We identified two different splicing transcripts that encode
TBCCD1 proteins with cytoplasmic localization. The overexpression of each of the proteins seems to
regulate the endogenous expression of the other. Furthermore, TBCCD1 knockdown in RPE-1 cells
causes a pronounced increase in the centrosome-nucleus distance, a cell cycle delay, GA disorganization,
lower efficiency to assemble primary cilia and delayed wound healing close accompanied by collective
cell migration alterations. Recent data suggests that TBCCD1 may be involved in the control of
centrosome-nucleus connection by regulating the dynamics of a microtubule subpopulation surrounding
the nucleus. We hope to soon elucidate this role by the identification of the TBCCD1 and TBCCD1
splicing variant partners using the BioID system.
Acknowledgements: This work was funded by CQB`s PEST UID/MULTI/00612/2013
References
1.
J. Gonçalves, S. Nolasco, R. Nascimento, M Lopez Fanarraga, JC Zabala, H. Soares, EMBO Reports 11
(2010) 194.
11
O8
CQB-Day 2015
TRACKING NEW PSYCHOACTIVE SUBSTANCES IN PORTUGAL
H. Gaspara, C. Leala,b,C. Gonçalvesa, S. Ciríacoa, A. Matiasa,b, J. Rodriguesb and S. Santosa
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, 1749-016 Lisboa,
Portugal([email protected]). bLaboratório de Polícia Cientifica da Polícia Judiciária, Novo edifício-sede da
Polícia Judiciária, Rua Gomes Freire, 169-007 Lisboa, Portugal ([email protected])
In the last decade, there has been an uprising of New Psychoactive Substances (NPS) available in
“smartshops” and over the Internet. NPS are defined as “new narcotic or psychotropic drugs that are not
listed in the Single Convention on Narcotic Drugs of 1961 or the Convention on Psychotropic Substances
of 1971, but which may pose a public health threat comparable to that posed by substances listed in those
conventions”. These compounds have been traditionally sold as innocuous products, becoming a legal
alternative to illicit drugs, such as cocaine, ecstasy or cannabis. The products have been advertised as
being herbal incenses, plant feeders or bath salts, with the caveat of not being for human consumption. Up
to 2014, more than 450 NPS have been reported in the European Union (EU). The two most reported
classes of NPS are synthetic cathinones and synthetic cannabinoids and, just in 2014, these two groups of
substances accounted for approximately 50% of the total NPS reported in the EU [1].
Recently, a new decree has been implemented in Portugal (Dec-Lei 54/2013) [2], which forbids the
production and commercialisation of about 159 NPS, being liable to fast updates, in order to keep up with
the everyday appearance of new substances. The rapidly growing problem of NPS makes the time
management for international control a real challenge, with the traditional detection methods becoming
increasingly inadequate. In order to tackle this issue, a collaboration was established in May 2014 with
the Laboratório de Polícia Científica da Polícia Judiciária (LPC), in which the role of our group is to
supply NPS standards to forensic laboratories, obtained either by synthesis or by isolation from LPC
samples, and to develop adequate analytical methodologies for the identification and quantification of
NPS in different matrices.
With the decree, several products previously sold in 8 Portuguese smartshops were delivered to LPC.
Within the scope of this collaboration, we have already studied 169 of those products. This led to the
identification of 26 NPS, through NMR and/or GC hyphenated techniques (MS and FID). The isolation
and unequivocal structural characterization of NPS permits their use as secondary standards for routine
analyses in forensic laboratories and allowed the creation of an in-house library with 21 NPS: 12
synthetic cathinones, 8 synthetic cannabinoids and ethylphenidate. The developed methodology was
applied in 4 seized samples, allowing the identification of two novel NPS: SGT-25, a synthetic
cannabinoid recently reported in the EU; and 4F-PBP (Figure 1), a new synthetic cathinone in the EU
reported by us during this project [3].
The application of the developed methodology in seized products allows a faster and easier tackling of the
rapid appearance of NPS in the market, illustrating the importance of the scientific knowledge in the
resolution of current public health threats.
Figure 1. 4F-PBP, a novel NPS
Acknowledgements The authors thank Fundação para a Ciência e a Tecnologia (FCT) for financial support through
project UID/MULTI/00612/2013 and the toxicology sector at LPC/PJ.
References
1. European Monitoring Centre for Drugs and Drug Addiction, European Drug Report 2015 - Trends and
Developments, Publications Office of the European Union, Luxembourg, 2015.
2. Decreto-Lei 54/2013, Diário da República, 1.ª série, No. 75, 17 de abril de 2013.
3. H. Gaspar, S. Bronze, S. Ciríaco, C.R. Queirós, A. Matias, J. Rodrigues, C. Oliveira, C. Cordeiro, S. Santos, For.
Sci. Int. 215 (2015) 168.
12
Science at CQB
Poster Communications
CQB-Day 2015
P1
NON-RELEASING BIOCIDAL COATINGS: A NEW STRATEGY TO PREVENT BIOFOULING
E. R. Silva,a, b O. Ferreira,b J. C. M. Bordado,b M. J. Calhordaa
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal ([email protected],
[email protected]), bCentro de Recursos Naturais e Ambiente, Instituto Superior Técnico, Universidade de Lisboa, 1049-001
Lisboa, Portugal ([email protected], [email protected])
The adhesion of microorganisms on submerged surfaces forms the so-called biofilms. Later on, fouling
organisms (e.g. hard foulers, such algae) will attach over time forming the macrofouling1). This biofouling
constitutes a global problem in water management systems for several industrial activities. It is
responsible for serious environmental and economic consequences. In the Marine Transport Industry in
particular, its adhesion and further accumulation on ships´ hulls is not only associated to hulls
deterioration (biocorrosion), but can also lead to drag friction increases up to 40 % and subsequent power
penalties of up to 86% at cruising speed2). Chemical control is nowadays the most effective strategy to
control this biofouling adhesion. It is based on the employment of coatings which act by a controllable
biocide releasing mechanism. However the continued releasing of toxic agents to the environment has led
to serious side effects on ecosystems, and rigid international regulations have been issued (EU
Regulation, 2012). More are expected to come in a near future, and may compromise permission to use
biocides currently in use. Non-toxic technologies are therefore sought. The most recent coatings’
technologies for biofouling surfaces protection (e.g. foul-releasing or self-polishing) still evidence
environmental and/or technical limitations. They are based on materials with a low surface energy, such
as silicone based coatings, which lead to minimal adhesive smooth surfaces (non-stick properties)3).
However, and in order to remove the weak attached organisms, these coatings with typically 5 to 10 years
lifetime are only effective for non-static applications, for examples for fast moving vessels (> 15 knot) 4).
New alternatives are needed to overcome the existent limitations.
This work aims to develop synergistic non-releasing coating systems by providing isocyanate functional
proved bioactive biocides suitable to be tethered to the most effective marine silicone based coatings.
This non-releasing biocide strategy was achieved by an effective biocide (Econea and Irgarol)
immobilisation. The binding of the product from reaction of the biocides with a conventional polyol
coating component was proved by FTIR analysis. Promising coatings’ antifouling performance (up to 23
weeks or about 6 months) were found on submerged coated surfaces under static marine conditions. Tests
are still on progress to assess coatings antifouling behaviour for longer immersion periods (a year).
Figure 1. Silicone based coated prototypes exposed for 23 weeks in Atlantic seawater (Peniche, Portugal): (left) 2.5 wt.%
of biocides (Econea + Irgarol), and (right) the control without biocide
Acknowledgements: The authors thank the financial support provided by the FP7 FOUL-X-SPEL project by
European Commission. E. R. Silva is also grateful for a research grant awarded by Fundação para a Ciência e a
Tecnologia (FCT) (SFRH / BPD / 88135 / 2012). E. R. Silva and M.J. Calhorda thank FCT (Pest
UID/MULTI/00612/2013). The authors also thank the support of HEMPEL A/S and ENP SA for the coating
providing and field tests facilities.
References
1. E. Almeida, T.C. Diamantino, O. Sousa, Prog. in Organic Coatings 59 (2007) 2.
2. E. R. Silva, O. Ferreira, J. C. M. Bordado, M. J. Calhorda, Boletim da Sociedade Portuguesa de Química 134
(2014) 43.
3. M. Lejars, A. Margaillan, C. Bressy, Chem. Reviews 112 (2012) 4347.
4. C. Stevens, D.E. Powell, P. Mäkelä, C. Karman, Mar. Pollution Bulletin 42 (2001) 536.
15
P2
CQB-Day 2015
ZNO@TIO2 CORE-SHELL NANOSTRUCTURES FOR SOLAR CELLS APPLICATIONS
T. Frade,a K. Lobato,b A. Gomesa
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected]; [email protected]). bInstituto Dom Luiz, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa,
Portugal ([email protected]).
Recently, increases in world energy demand and climate change have focused research attention on
renewable and clean energy sources, for which photovoltaic devices (PV) are promising examples.
Concerning photovoltaic applications, several nanostructured semiconductors with a wide band gap such
as TiO2, SnO2, Nb2O5 and ZnO have been employed as ordered electron acceptors1. Among these
materials, ZnO has attracted much attention in due to its excellent properties such as low cost, direct band
gap (3,37 eV), high exciton energy (60 m.eV), high electron mobility (200 cm2.V-1.s-1), and non toxicity2.
There are several publications describing 1D ZnO nanostructures such as nanorods, nanowires or
nanotubes that provide direct and continuous pathways for charge transport. ZnO nanorods can be easily
grown on different substrates by different physical and chemical methods, such as chemical vapor
deposition, magnetron sputtering, hydrothermal and electrochemical deposition methods. Electrochemical
deposition is probably the most promising one for such purposes because it is easy to scale-up, low-cost
and ecofriendly3.
Core-shell materials have attracted a great deal of interest due to their versatile structures and properties
for electronics, magnetism, optics, catalysis, mechanics, and other4. The idea of applying an oxide shell is
to suppress recombination by (i) introducing an energy barrier that increases the physical separation
between photoinjected electrons and the oxidized redox species in the electrolyte, (ii) forming a tunneling
barrier that keeps electrons within the conducting cores, or (iii) passivating recombination centers on the
oxide surface5.
In this work we present1D ZnO nanostructures prepared by pulsed electrochemical deposition, coated
with TiO2 obtained for the first time by solvothermal method (Figure 1). The TiO2 was tested because of
its similar band gap (3,2 eV) and its proved performance on photovoltaic devices. The films were
characterized by their morphological, structural, optical and photoelectrochemical properties.
Figure 1. SEM image of ZnO@TiO2 nanostructures after 4 h of solvothermal treatment at 150 °C.
Acknowledgements The authors acknowledge financial support from Fundação para a Ciência e Tecnologia
(Portugal), under PhD grant SFRH/BD/84669/2012, and CQB’s PEST UID/MULTI/00612/2013.
References
1. Q. F. Zhang, T. P. Chou, B. Russo, S. A. Jenekhe, G. Z. Cao, Adv. Funct. Mater. 18 (2008) 1654.
2. M. Mehrabian, H. Afarideh, K. Mirabbaszadeh, L. Lianshan, T. Zhiyong, J. Opt. Soc. Korea 18 (2014) 307.
3. C. Z. Yao, B. H. Wei, P. Zhang, X. H. Lu, P. Liu, Y. X. Yong, J. Rare Earths 29 (2011) 133.
4. Y. H. Deng, W. D. Qi, C. H. Deng, X. M. Zhang, D. Y. Zhao, J. Am. Chem. Soc. 130 (2008) 28.
5. M. Law, L. E. Greene, A. Radenovic, T. Kuykendall, J. Liphardt, P. Yang, J. Phys. Chem. B 110 (2006) 22652.
16
CQB-Day 2015
P3
A MEMBRANE BIOPHYSICS APPROACH TO THE MECHANISM OF ACTION OF NEW ACTIVE
RUTHENIUM ANTICANCER COMPOUNDS
A. D. de Sousaa,b, T. C. Santosa,b, F. C. Santosa, M.S.C.S. Santosa, M. L. Corvoc, M. H. Garciab, A. I.
Tomazb, R. F. M. de Almeidaa
aCentro
de Química e Bioquímica and bCentro de Química Estrutural, Faculdade de Ciências, Universidade de Lisboa, 1749016 Lisboa, Portugal; cFARM-ID, Departamento de Farmácia Galénica e Tecnologia Farmacêutica, Faculdade de Farmácia,
Universidade de Lisboa ([email protected])
Membrane lipid domains, such as the so-called lipid rafts, with their unique composition and biophysical
properties, have crucial roles in cell signaling and sorting. Changes in those properties may have vast
implications in defining cellular fate, and therefore are intimately related with cancer conditions. In fact,
it is known that lipid composition and organization is markedly different in cancer cells versus non-cancer
counterparts [1]. Thus, membrane lipids are emerging as key targets in the action of novel therapeutics
which can be designed to change membrane biophysical properties, either directly or through alterations
in lipid metabolism [1,2].
In cancer chemotherapy, platinum-based agents are the only metallodrugs approved for use in the Clinic
worldwide. However, their use presents some severe limitations given their high toxicity, which has
prompted research in the field to pursue better alternatives. Some of these limitations have been exceeded
by the use of ruthenium complexes. The mechanisms underlying the action of ruthenium complexes are
not well known. Thus, in this work we studied the possible role of the membrane on the molecular
mechanism of action of three selected ruthenium complexes, which we have previously shown to be
active against human tumor cells [3]. Four model membranes were used. One-component models
represent the two extremes of disorder (dioleoylphosphatodylcholine, DOPC), fluid phase, and order
(dipalmitoylphosphatidylcholine, DPPC), gel phase. The more complex models, containing
phosphatidylcholine, phosphatidylethanolamine, sphingomyelin and cholesterol mimic either the
“canonical” composition of mammalian plasma membrane (1:1:1:1 mole ratio) or have the composition
of U-118 glioma cell membrane (36:24:7:33 mole ratio) [1,2].
The three complexes have moderate to high affinity for the fluid lipid bilayer, as quantified from the
membrane/water partition coefficients, Kp. Comparing the Kp values and the changes on photophysical
parameters upon interaction with DOPC liposomes of complexes and ligands it was possible to assess
which ligand was promoting the interaction. Importantly, all the complexes increased the leakiness of the
membrane without compromising its integrity.
Changes in the biophysical properties of the membrane studied by fluorescence spectroscopy were
observed in the presence of the complexes, in all types of membrane studied. The presence of the
complexes promoted an increased water penetration into the lipid bilayer. To better understand the effect
of complexes in cancer cells, membrane dipole potential was also measured. The dipole potential affects
the activity of many membrane proteins and is strongly dependent on cholesterol-sphingolipid
interactions, wich as mentioned above are changed in human tumor cells. The results show an increased
dipole potential of the U-118 glioma cell membrane model in the presence of two of the complexes, while
no effect was detected on the mammalian plasma membrane model.
These results highlight the importance that the cell membrane biophysical properties may have on the
mode of action of these Ruthenium complexes, motivating further research on the changes induced by
active Ru complexes in both lipids and proteins involved in membrane domain organization.
Acknowledgements This work was financed through the Portuguese Foundation for Science and Technology within the scope of
projects UID/QUI/00100/ 2013, UID/MULTI/00612/2013, and the Investigador FCT 2012/2013 Initiatives (POPH, FSE).
References
[1] G. Barceló-Coblijn, M.L. Martin, R.F.M. de Almeida, M.A. Noguera-Salvà, A. Marcilla-Etxenike, F. Guardiola-Serrano, A. Lüth, B. Kleuser,
J.E. Halver, P.V. Escribá. Proc. Natl. Acad. Sci. USA 108 (2011) 19569.
[2] A. Khemelinskaia, M. Ibarguren, R.F.M. Almeida, D. J. López, V. Paixão, H. Ahayayauch, F. M. Goñi, P. Escribá. Langmuir 30 (2014) 2117.
[3] C.P. Matos, A. Valente, F. Marques, P. Adão, M.P. Robalo, R.F.M. de Almeida, J.C. Pessoa, I. Santos, M.H. Garcia, A.I. Tomaz, Inorg Chim.
Acta 394 (2013) 616.
17
P4
CQB-Day 2015
MOO3 NANOPARTICLES A HIGHLY SELECTIVE CATALYST
C. I. Fernandes1, S. C. Capelli2, P. D. Vaz1,2, C. D. Nunes1
1 CQB, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, Ed. C8, 1749-016 Lisboa, Portugal 2 ISIS Neutron &
Muon Source, Rutherford Appleton Laboratory, Chilton, Didcot, Oxfordshire, OX11 0QX, United Kingdom*
[email protected]
-MoO3 nanoparticles can be synthesized by different methods, being the hydrothermal synthesis the
most popular one. This is a low temperature process which yields various nanostructures with controlled
size, stoichiometry and shape [1]. -MoO3 nanoparticles were prepared with an average size below 100
nm by solvothermal synthesis of MoO2 and subsequent thermal oxidative annealing [2]. The MoO3
nanoparticles were used as catalysts and revealed to be very active in olefin epoxidation of a number of
olefins using tert-butylhydroperoxide (tbhp) as oxygen source. Catalytic tests show that the catalyst
conducts selectively to the desired epoxides.
Figure 1. TEM image of synthesized nanoparticles.
Acknowledgements The authors are grateful to FCT for financial support (projects UID/MULTI/00612/2013 and
EXPL/QEQ-QIN/1137/2013). CIF also thanks FCT for a grant (SFRH/BD/81029/2011).
References
[1] W.-S. Kim, H.-C. Kim, S.-H. Hong, J. Nanopart. Res. 12 (2010) 1889.
[2] C.I. Fernandes, S.C. Capelli, P.D. Vaz, C.D. Nunes, Appl. Catal. A: Gen. (2015) http://dx.doi.org/
10.1016/j.apcata.2015.02.027
18
CQB-Day 2015
P5
COMPOUNDS FROM HERBAL INFUSIONS AND FRUITS IN THE PREVENTION OF AGE-RELATED
DISEASES
E. Brito a, J. Peres a, M. Salas, F. Ferreira a, S. Barata a, A. Ressaissi a, A. Maia a, N. Marques a, P. Lopes a,
A. Graça a, A. Silva a, R. Pacheco a,b, M. H. Florêncioa, P. L.Falé a, c, , M. L. Serralheiro a
a.
Centro de Química e Bioquímica, Faculdade de Ciências da Universidade de Lisboa. Campo Grande 1749-016, Lisboa.
Portugal; b. Área Departamental de Engenharia Química, Instituto Superior de Engenharia de Lisboa, Av. Conselheiro Emídio
Navarro, 1959-007 Lisboa, Portugal; c. Institute of Pharmaceutical Science. Faculty of Life Sciences & Medicine. King's College
London. Franklin-Wilkins Building. 150 Stamford Street. Waterloo. London SE1 9NH, UK.
Aqueous extracts from plants have been consumed throughout history for the prevention or amelioration
of several diseases. Nowadays part of the molecular mechanism underlying these effects have been
disclosed. The effect of infusions or decoctions containing mainly rosmarinic acid and chlorogenic acid
were analysed on what concerns their effect on the prevention of age-related diseases like Alzheimer
disease (AD), and atherosclerosis. As these diseases are multifactorial-caused the main selected topics
will be the inhibitory activity of these compounds or extracts on enzymes involved in AD treatment like
acetylcholinesterase and the enzymes of the digestive tract catalysing the hydrolyses of hydrocarbons
present in the diet, alpha-amylase, alpha-glucosidase. The effect of these caffeic acid derivatives on the
absorption of cholesterol from the diet at membrane-transporter proteins will be envisaged, together with
the effect on the permeation of aminoacids under the influence of infusions. Several infusion from
different herbs either collected in Portugal or bought in the supermarket, as well as the peel of several
fruits, were analysed. In vitro studies are being developed to simulate the in vivo environment in a more
realistic way (under collaboration with Parvez Haris, from Montfort University, Leicester. UK)
Digestion
Plant
infusions
Alzheimer
Objective
Fruits
Cholestero
l
Biological
activities
Protein
Nutrition
Future
Functional Foods
development
Acknowledgements:.PTDC/QUI/BIQ/113477/2009; UID/MULTI/00612/2013
19
P6
CQB-Day 2015
SYNTHESIS AND MAGNETIC CHARACTERIZATION OF MANGANESE(III) COMPLEXES
G. Santos,a P. N. Martinho,a S. Realista,a A. Fitzpatrick,b L. P. Ferreira,c,d S. Barroso,e L. C. J. Pereira,f
M. J. Calhordaa
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal, bSchool of
Chemistry and Chemical Biology, University College Dublin, Dublin 4, Ireland, cBioISI, Faculdade de Ciências, Universidade
de Lisboa, Campo Grande, 1749-016 Lisboa, Portugal, dDepartamento de Física, Faculdade de Ciências e Tecnologia,
Universidade de Coimbra, 3004-516 Coimbra, Portugal, eCentro de Química Estrutural, Instituto Superior Técnico,
Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisboa, Portugal, fSolid State Group, UCQR, IST/CTN, Universidade de
Lisboa, Instituto Superior Técnico, 2695-066 Bobadela, Loures, Portugal.
([email protected])
Manganese(III) is a d4 metal ion which in an octahedral coordination environment displays Jahn-Teller
distortion. This makes Mn(III) an interesting ion to both study its magnetic properties, namely spin
crossover1 and single ion magnet2 and to investigate the effect of the Jahn-Teller distortion on these
properties. Here we report the synthesis of seven new Mn(III) complexes with tridentate Schiff-base
ligands. All compounds were characterised by FTIR, elemental analysis, single crystal X-ray
crystallography and SQUID magnetometry. Potential single ion magnets were further characterised by
PPMS magnetometry. Our focus was on the engineering of the ligand and the coordination geometry
around the metal. SQUID magnetometry showed that all seven compounds stabilise in the high-spin state
with one pair of bond lengths (Mn-Namine) considerably longer than the others, Figure 1. AC susceptibility
measurements carried out using a MagLab2000 system showed that one of the new compounds displays
single ion magnet behaviour, thus making it a good candidate for further investigation and possible
application in data storage.
Figure 1. Octahedral geometry distortion around the Mn(III) centre.
Acknowledgements We thank FCT for financial support UID/MULTI/00612/2013 and UID/MULTI/04046/2013.
PNM thanks FCT for financial support (SFRH/BPD/73345/2010).
References
1. P. N. Martinho, B. Gildea, M. M. Harris, T. Lemma, A. D. Naik, H. Müller-Bunz, T. E. Keyes, Y. Garcia, G. G.
Morgan, Angew. Chem. Int. Ed. 2012, 51, 11995.
2. G. A. Craig, M. Murrie, Chem. Soc. Rev. 2015, 44, 2135.
20
CQB-Day 2015
P7
IN VIVO REAL-TIME RESPONSE OF SACCHAROMYCES VEREVISIAE TO A MILD-STRESS INDUCED BY
HYDROGEN PEROXIDASE
R. N. Bento, C. E. S. Bernardes, M. E. Minas da Piedade, F. Antunes
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
Microcalorimetry, a probe-free method, has proved to be a very sensitive technique to follow cell
metabolism in real time. The results of those experiments are normally given as heat-flow rate versus time
(-t) or power versus time (P-t) curves. The shape of these curves strongly depends on the
microorganism nature and on the physiological conditions in the calorimetric vessel (e.g. composition of
the growth medium or the oxygen concentration). Signatures of specific biological events (e.g. diauxic
shift) and of cell responses to the addition of extraneous substances to the growing medium are also
possible to detect. The latter feature, has, for example, been used to assess antibiotics effectiveness.
Hydrogen peroxide (H2O2) is a ubiquitous biological molecule that has intracellular signaling functions at
low doses, but causes oxidative stress at high doses. It is, however, known that cells develop a response
against H2O2, even at low doses. This type of response was investigated in this work for Saccharomyces
cerevisiae (Sc) by using heat-conduction flow calorimetry. Based on the typical growth profile followed
by calorimetry and by optical density (Figure 1), the main findings were: (i) metabolism is most active
during the lag-phase of growth; (ii) exposure to three 150 M consecutive H2O2 bolus additions, over a
period of 80 min, decreases cell division be near 6 % and decreases the heat dissipated associated to
glucose consumption by 19 %, implying major metabolic changes; (iii) exposure to a 150 M H2O2
steady-state adaptive dose, obtained by delivering approximately 600 M H2O2 during 90 min, does not
change the heat dissipated associated to glucose consumption. Thus, response to H2O2 involves different
mechanisms, depending on the method of H2O2 delivery, with the bolus addition causing a larger
response from the energetic point of view.
Figure 1. Calorimetric apparatus and typical growth curve followed by calorimetry and optical density.
Acknowledgements This work was supported by Fundação para a Ciência e a Tecnologia (FCT), Portugal, through
project UID/MULTI/00612/2013) and a Post-Doctoral (SFRH/BPD/101505/2014) grant awarded to C. E. S.
Bernardes.
21
P8
CQB-Day 2015
MODULATION OF THE BIOPHYSICAL PROPERTIES AND PERMEABILITY OF MODEL MEMBRANES
BY SPHINGOSINE
A .C. Carreira,1,2 R. F. M. de Almeida2, L. C. Silva1
–Research Institute for Medicines, Faculdade de Farmácia, Universidade de Lisboa,1649-003 Lisboa, Portugal
([email protected], [email protected]). 2Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa,
Campo Grande, 1749-016 Lisboa, Portugal ([email protected])
1iMed.ULisboa
Sphingosine is a common sphingoid base in mammalians and duo to its bioactive action in the cell, it can
interfere with important biological processes1,2. This lipid is generated inside the lysosome and in some
situations it abnormally accumulate inside the acidic organelle, leading to Niemann Pick type C1, one of
the most complex storage diseases in terms of molecular phenotype3. At the moment little is known about
the interaction of sphingosine with other membrane lipids. It is our aim to characterize the effect of
sphingosine on the biophysical properties of model membranes composed of different lipids, so that new
insights into its mode of action might arise. Using an established fluorescence spectroscopy approach we
evaluated the effect of sphingosine in membrane organization and permeability of different 1-palmitoyl2-oleoyl-sn-glycero-3-phosphocholine/Sphingomyelin/Cholesterol raft-mimicking mixtures. Our results
showed that sphingosine accumulation leads to the formation of sphingosine-enriched gel domains in raft
mimicking mixtures. These domains are more easily formed at neutral pH mimicking the plasma
membrane environment as compared to the acidic lysosomal membrane environment, where higher
sphingosine concentrations are required. The determination of ζ-potential further revealed that in plasma
membrane-raft models sphingosine is mainly neutral, whereas in lysosomal membrane-raft models the
positive charge of sphingosine leads to electrostatic repulsion, reducing sphingosine ability to form gel
domains. It was also shown that the external addition of sphingosine affects the membrane permeability,
being the effect more noticeable in acidic environment. These results support the hypothesis that Sph
biological actions, including those related to Niemann Pick type C1, could be exerted through biophysical
changes in cellular membranes.
Acknowledgements: Fundação para a Ciência e Tecnologia (FCT), Portugal: PTDC/BBB-BQB/0506/2012,
SFRH/BD/88194/2012 to ACC and Investigador FCT 2012 and 2014 to RFMA and LCS, respectively (POPH,
FSE). CQB´s PEST UID/MULTI/00612/2013
References
1. S. T. Pruett, A. Bushnev, K. Hagedorn, M. Adiga, C. A. Haynes, M. C. Sullards, D. C. Liotta, A. H Merrill. J.
Lipid Res. 49 (2008) 1621.
2. W. Zheng, J. Kollmeyer, H. Symolon, A. Momin, E. Munter, E. Wang, S. Kelly, J. C. Allegood, Y. Liu, Q.
Peng, et al. Biochim. Biophys. Acta. 1758 (2006) 1864.
3. E. Lloyd-Evans, A. J. Morgan, X. He, D. A. Smith, E. Elliot-Smith, D. J. Sillence, G. C. Churchill, E. H.
Schuchman, A. Galione, F. M. Platt. Nat. Med. 14 (2008) 1247.
22
CQB-Day 2015
P9
BIOMIMETIC SUPPORTED LIPID BILAYERS FOR IMMUNOSENSING
I. Almeidaa, J. T. Marquêsa, T.M.O. Paivaa, Y. Niub, W. Lib, G. Jinb, R. F.M. de Almeidaa, A. S.Vianaa
a Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, 1749-016 Lisboa, Portugal
b Institute of Mechanics, Chinese Academy of Sciences, Beijing, 100190, China ([email protected])
The development of new and enhanced biosensing devices is challenging [1], and the use of lipid-based
platforms is a highly promising approach since they provide a biomembane-like environment. To this
end, the lipids should be carefully chosen to tailor the biophysical properties of the membranes and to
form a planar, uniform and continuous supported bilayer on suitable transduction surfaces, namely gold
[2,3].
In this work we describe a novel approach that takes full advantage of both the biophysical and the
chemical properties of lipids in order to build a simple and efficient immunosensing platform. The new
methodology presented is based on a binary lipid system incorporating a low percentage of decanethiol,
which will enable the ready formation, from vesicles, of a robust bilayer on gold with thiol linkages. The
platform displays important requirements for immunosensing assays: allows easy covalent attachment of
antibodies, is able to inhibit protein non-specific adsorption and presents mechanical stability in a flow
cell during real-time immunoassays. In addition, this configuration avoids the use of specially synthesized
thiolipids and preserves the natural fluidity and dynamics of the lipids. The sensitive detection of specific
antibody/antigen interaction was monitored by surface plasmon resonance and confirmed by ellipsometric
measurements. This lipid-based biosensing platform is versatile and can be adapted to the biorecognition
reaction of interest.
Acknowledgements
Financial
support
by
FCT:
SFRH/BD/64442/2009,
SFRH/BD/70673/2010,
UID/Multi/00612/2013, IF2012/13 (FSE, POPH) and 7th Sino-Portugal Scientific and Technological Cooperation.
References
1. Y. Niu, A. I. Matos, L. M. Abrantes, A.S. Viana, G. Jin, Langmuir 28 (2012) 17718.
2. J.T. Marquês, R.F.M. de Almeida, A.S. Viana, Soft Matter, 8 (2013) 2007.
3. J.T. Marquês, A.S. Viana, R.F.M. de Almeida, Langmuir 30 (2014) 12627.
23
P10
CQB-Day 2015
SPHINGOLIPID-DEPENDENT PLASMA MEMBRANE BIOPHYSICAL PROPERTIES IN ANTIFUNGAL
DRUG ACTION AND RESISTANCE
F. C. Santosa, C. A. C. Antunesa, J. T. Marquêsa, A. S. Fernandesb,c, A. S. Vianaa, A. Videirab,c,
H. S. Marinhoa, R. F .M. de Almeidaa
aCentro
de Química e Bioquímica - Faculdade de Ciências, Universidade de Lisboa, Lisboa, Portugal ([email protected];
[email protected]; [email protected]; [email protected]; [email protected]; [email protected]). bIBMC Instituto de Biologia Molecular e Celular, Universidade do Porto, Porto, Portugal. cICBAS - Instituto de Ciências Biomédicas de
Abel Salazar, Universidade do Porto, Porto, Portugal. ([email protected]; [email protected]).
Fungal drug resistance is becoming a pressing concern. In recent years, evidence has emerged linking
sphingolipids to antifungal resistance. Sphingolipids actively regulate membrane biophysical properties,
ranging from lipid domain organization, to membrane fluidity, hydration and dipole potential. Therefore,
we hypothesize that membrane biophysical properties regulated by sphingolipids are involved in the
mechanisms of resistance to antifungal agents. Our goal is, thus, to obtain solid evidence supporting this
hypothesis.
In this work we used two biological models of fungal drug resistance involving sphingolipid-associated
phenomena, where a role for ergosterol could be clearly ruled-out. Saccharomyces cerevisiae ipt1Δ cells
do not synthesize the sphingolipid mannosyl-diinositolphosphorylceramide, M(IP)2C. Despite having
ergosterol content similar to the wild-type (wt), they exhibit higher resistance towards Nystatin and
Miconazole. Thus, we evaluated how the absence of M(IP)2C affects membrane organization in S.
cerevisiae wt and ipt1Δ strains, in living cells in mid-exponential phase using state-of-the-art fluorescence
spectroscopy approaches, and in supported lipid bilayers reconstituted from lipid extracts employing
high-resolution imaging by atomic force microscopy. Neurospora crassa conidia are naturally devoid of
ergosterol, meaning that membrane microdomains are due to sphingolipids. Upon stimuli with the
antifungal drug Staurosporine, the multidrug-resistance protein ABC-3, analogous to a human protein
intimately involved in sphingolipid transport, is overexpressed >100×. To understand the organization of
sphingolipid domains in the plasma membrane (PM) of N. crassa and their role in Staurosporine
resistance, we studied conidial suspensions by advanced fluorescence spectroscopic methods.
In budding yeast S. cerevisiae, the absence of M(IP)2C in ipt1Δ cells does not affect the rigidity of
sphingolipid domains, but decreases their fraction when compared to the wt. Moreover, it decreases
membrane fluidity, and thickens the disordered regions of the membrane. These results suggest that
M(IP)2C not only affects sphingolipid domain abundance, but also modulates the properties of the
remaining membrane.
According to our results, filamentous fungus N. crassa possesses a highly ordered PM, probably with
several types of sphingolipid domains. In slime, a cell wall-less strain, Staurosporine induces PM
reorganization, leading to increased global fluidity and less packed sphingolipid domains, whereas in the
wt, the abundance of ordered domains increases. These observations, strongly suggest that in N. crassa
sphingolipid domains are intimately involved in the response to Staurosporine challenge. We also found
that Staurosporine affects the PM lipids hydration in different manners in the wt and slime.
In two fungal species, clear relations between membrane biophysical properties involving sphingolipids
and differences in drug resistance were found, strongly supporting our initial hypothesis. Therefore, this
class of lipids represents a potentially important target to overcome the emerging health problem of
fungal drug resistance.
Acknowledgements This work was financed by Portuguese national funds through Fundação para a Ciência e a
Tecnologia (F.C.T., I.P.): Ph.D. fellowship SFRH/BD/64442/2009; research grant PEst 2015–2020
(UID/Multi/00612/2013), and IF2012/2013 initiative (POPH, Fundo Social Europeu).
24
CQB-Day 2015
P11
H2O2 CONSUMPTION RATE AND REDOX SIGNALING IN ENDOTHELIAL CELLS IS MODULATED BY
EXTRACELLULAR MATRIX
A. Bagulhoaδ, F. Vilas-Boasaδ, A. Penaa, C. Penedaa, F. C. Santosa, A. Jerónimoa, R. F. M. de Almeidaa,
C. Reala
δ These authors contributed equally to this work
a Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
The molecular processes that are crucial for cell function, such as proliferation, migration and survival,
are regulated by hydrogen peroxide (H2O2)1. Although environmental cues, such as growth factors,
regulate redox signaling, it was still unknown whether the ECM, a component of the cell
microenvironment, had a function in this process.
Here, we showed that the ECM differently regulated H2O2 consumption by endothelial cells and that this
effect was not general for all types of cells. The analysis of biophysical properties of the endothelial cell
membrane suggested that this modification in H2O2 consumption rates was not due to altered membrane
permeability. Instead, we found that the ECM regulated GPx activity, a known H2O2 scavenger. Finally,
we showed that the extent of PTEN oxidation was dependent on the ECM, indicating that it was able to
modulate H2O2-dependent protein oxidation.
The microenvironment surrounding a particular cell influences cell fate through the activation of
particular signaling pathways2. Our results show that the ECM is an important microenvironment
component that modifies the kinetics of H2O2 consumption and the redox signaling outcome. The ability
of the ECM to modulate H2O2 consumption might be important for endothelial cell function during
angiogenesis, for endothelial cell migration and tube formation, and in the quiescent state, for endothelial
cell survival and homeostasis regulation3. Our results, therefore, demonstrate the existence of a crosstalk
between ECM-dependent signaling and redox signaling in order to direct endothelial cell behavior.
Figure 1. The ECM regulates GPx activity, which is responsible for H 2O2 degradation. Adhesion to different matrices
changes PTEN oxidation levels induced by extracellular H2O2, indicating that the ECM is able to contribute to redox
signaling in endothelial cells.
Acknowledgements This work was supported by Fundação para a Ciência e a Tecnologia (FCT), grants
PTDC/BIA-PRO/101624/2008, PEst-OE/QUI/UI0612/2013 and UID/Multi/00612/2013. Filipe Vilas-Boas is
supported by FCT, individual fellowship SFRH/BPD/74715/2010. Rodrigo F.M. de Almeida acknowledges IF2012
Program (FCT, FSE, POPH). Carla Real acknowledges Ciência2008 Program (FCT).
References
1. H.S. Marinho, C. Real, L. Cyrne, H. Soares, F. Antunes, Redox Biol. 2 (2014) 535–562.
2. R.O. Hynes, J. Thromb. Haemost. 5 Suppl 1 (2007) 32–40.
3. R.H. Adams, K. Alitalo, Nat. Rev. Mol. Cell Biol. 8 (2007) 464–478.
25
P12
CQB-Day 2015
CORE @SHELL NANOSTRUCTURES FOR MAGNETIC HYPERTHERMIA APPLICATIONS
S. G. Mendoa, A. F. Alvesa, L. P. Ferreirab,c, M. M. Cruzb, M. H. Mendonçaa, M. Godinhob,
M. D. Carvalhoa
aCQB,
Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal ([email protected]);bBioISI,
Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal; cDepartment of Physics, University of Coimbra,
300-516 Coimbra, Portugal)
Core@shell structures with magnetic nanomaterials have been studied in the last years for their
potential interest in biomedical applications, including for magnetic hyperthermia . The magnetic
properties of these nanostructures depend on the composition of the core and of the shell and also on the
organization at the interface. The combination of different magnetic phases like Fe3O4, CoFe2O4 and
MnFe2O4 can provide unique magnetic characteristics enhancing the hyperthermia performance, as
reported previously 1,2.
The most used method to synthesize this type of core@shell nanostructures has been the thermal
decomposition. This method has already showed its versatility in allowing the combination of different
phases and a precise control of the particle size and size distribution. However, the use of organic solvents
and hazardous reactants, the difficulty of the method itself and the cost of the reagents are drawbacks
which should be avoided. Therefore, new, easier, cheaper and more environmentally friendly synthesis
methods are needed.
In this work an alternative route to synthesize core@shell nanoparticles is presented. The core was
prepared by an established procedure in gelatin medium3, and the core@shell nanostructures were
obtained by the hydrothermal method performed in an oven equipped with a rotative system. Different
combinations of core and shell Fe3O4, CoFe2O4 and MnFe2O4 were investigated.
To evaluate the structure and the morphology of the synthesized core@shell nanoparticles X-ray
diffraction (XRD) and scanning and transmission electron microscopy (SEM and TEM) were performed.
The magnetic characterization was investigated by SQUID magnetometry and 57Fe Mössbauer
spectroscopy. In order to assess their potential interest for magnetic hyperthermia applications, induction
heating measurements under an alternating magnetic field were performed in optimized adiabatic
conditions4 and the specific loss power (SLP) parameter was determined. The results show an
enhancement of the SLP values for the core@shell nanostructures when compared with the individual
phases.
Acknowledgements This work was funded by the Portuguese Foundation for Science and Technology (FCT) under
the project PTDC/CTM-BIO/2102/2012 (including a grant for Sofia G. Mendo and André F. Alves),
UID/MULTI/00612/2013 and UID/MULTI/04046/2013.
References
1. J.H. Lee, J.-T. Jang, J.S Choi, S. H. Moon, S.H. Noh, J.W. Kim, J.G. Kim, I.S. Kim, K. I. Park, K. I.
Cheon, J. Nat. Nanotechnol. 6 (2011) 418.
2. M. Angelakeris, Z.A. Li, M. Hilgendorff, K. Simeonidis, D. Sakellari, M. Filippousi, H. Tian, G.V.
Tendeloo, M. Spasova, M. Acet, M.Farle, J. Magn. Magn. Mater. 381 (2015) 179.
3. A. F. Alves, S. G. Mendo, L. P. Ferreira, M. H. Mendonça, P. Ferreira, M. Godinho, M. M. Cruz, M. D.
Carvalho, (2015) (submitted).
4. S. G. Mendo, A. F. Alves, L. P. Ferreira, M. M. Cruz, M. H. Mendonça, M. Godinho, M. D. Carvalho,
New J. Chem. (2015) (DOI:10.1039/C5NJ00009b).
26
CQB-Day 2015
P13
LUMINESCENT HYDROPHOBIC SURFACES FOR SMART WINDOWS
J. B. Aldeias, S. Realista, M. J. Calhorda, P. N. Martinho
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected]).
The general properties of a material and its wettability may have a huge impact on its usage and
application in everyday life. Recently hydrophobic surfaces have been a hot topic of research and teams
have put some effort to combine wettability/hydrophobicity with other interesting properties.
Here we report the synthesis of new Zn(II) complexes with both hydrophobic and luminescent properties
and their deposition on glass surfaces by the low-cost drop-casting method. The surfaces were prepared
by dissolving the new Zn(II) Schiff-base complexes in three different solvents (chloroform, toluene and
tetrahydrofuran) using different evaporation rates. Several parameters were taken into account:
concentration of the Zn(II) complexes, solvent, temperature, number of depositions and surface
wettability. The tecniques used to characterise both complexes and surfaces were 1H and COSY NMR
spectroscopy, FTIR spectroscopy, UV-vis spectroscopy, elemental analysis and contact angle
microscopy. We have found that the best solvent to combine hydrophobicity, luminescence and
transparency with our Zn(II) Schiff-base compounds was toluene.
Figure 1. Zn(II) Schiff-base hydrophobic luminescent surfaces.
Acknowledgements We thank FCT for financial support UID/MULTI/00612/2013. PNM thanks FCT for financial
support (SFRH/BPD/73345/2010).
References
1 Q. Wei, C. Schlaich, S. Prévost, A. Schulz, C. Böttcher, M. Gradzielski, Z. Qi, R. Haag and C. a. Schalley, Adv.
Mater., 26 (2014) 7358–7364.
2 S. Subhash Latthe, J. Surf. Eng. Mater. Adv. Technol., 02 (2012) 76–94.
3 L. Y. Liu Y, Zhao X, Cai B, Pei T, Tong Y, Tang Q, Pub.Med., (2014).
27
P14
CQB-Day 2015
MODIFIED NANOSTRUCTURED ZnO PHOTOANODES SENSITIZED WITH N719 IN ETHANOL AND
ETHANOL/WATER
D. Siopaa, S. Sériob, M. E. Melo Jorgea, F. Martinsa, M. S. C. S. Santosa, K. Lobatoc, A. Gomesa
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected], [email protected], [email protected], [email protected] and [email protected]) bCEFITEC – Departamento
de Física, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal
([email protected]) cInstituto Dom Luiz, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected])
The modification of transparent conductive fluor tin oxide (FTO) substrates by the deposition of a zinc
oxide (ZnO) thin film using DC reactive magnetron sputtering allowed the formation of a seed layer
suitable for electrochemical growth of vertically oriented 1D nanorod structures with diameters of about
40 nm.
The characterization of these sputtered seed layers by scanning electron microscopy (SEM) and X-ray
diffraction (XRD) was used to correlate its morphological and structural properties with the
electrodeposition process conditions (Figure 1). Electrochemical impedance spectroscopy (EIS) was
carried out to construct Mott-Schottky plots for the determination of the carrier density (ND) of the ZnO
thin films sputtered with different deposition parameters.
Electrodeposition step was performed under potentiostatic control using an aqueous nitrate electrolyte
bath at relatively low temperature [1].
The nanostructured ZnO thin films were sensitized with ruthenium organometallic dye N719 in ethanol
and ethanol/water mixtures. Their performance on a test dye solar cell was analyzed and compared.
Interactions between dye and solvent were explored through a systematic spectroscopic study in solution,
using solvatochromic probes.
Figure 1. Electrodeposited ZnO nanorods (left) and voltage-current curves (right) for photoanodes sensitized in etanol and
etanol/water.
Acknowledgements The authors acknowledge financial support from Fundação para a Ciência e Tecnologia under
CQB’s PEST UID/MULTI/00612/2013.
References
1. D. Siopa, A. Gomes, J. Electrochem. Soc. (2013) 160, D476
28
CQB-Day 2015
P15
pH-DEPENDENT INSERTION OF PHLIP PEPTIDE INTO BILAYERS: INSIGHTS FROM
COMPUTATIONAL SIMULATIONS
D. Vila-Viçosa, V. H. Teixeira, M. Machuqueiro
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected]).
The pH (low) insertion peptide (pHLIP)1-3 is a family of peptides that are able to insert into a lipid bilayer
at acidic pH. These peptides are based on a transmembrane sequence of bacteriorhodopsin that is
unstructured in solution (Stage I), interacts with lipid bilayers remaining unstructured at neutral pH (Stage
II) and inserts into the bilayer with a significant α-helical content at acidic pH (Stage III). This family of
peptides have already been used to target tumor cells in vivo since acidosis is a hallmark of these tissues.
The relation between the pK of insertion of pHLIP peptides and the pKa of some key residues is yet to be
clarified. In this work, we used MD simulations and Poisson-Boltzmann / Monte Carlo calculations in a
linear response approximation (LRA) to determine the pKa of these residues. Four different pHLIP
variants were used to study the importance of the Asp positions and its mutation to Glu, and the results
were compared with the available experimental data. Finally, we also propose the addition of a His
residue to the sequence, which we expect to turn off the pHLIP peptide insertion (Stage III) at too low pH
values.
Stages of pHLIP insertion in lipid bilayers
Acknowledgements
The authors acknowledge financial support from Fundação para a Ciência e Tecnologia, Portugal, through project
grants UID/MULTI/00612/2013 and PTDC/QUI-BIQ/113721/2009 and fellowship SFRH/BD/81017/2011.
References
1. J. F. Hunt, T. N. Earnest, O. Bousché, K. Kalghatgi, K. Reilly, C. Horváth, K. J. Rothschild, D. M. Engelman,
Biochemistry 36 (1997) 15156
2. M. Musial-Siwek, A. Karabadzhak, O. A. Andreev, Y. K. Reshetnyak, D. M. Engelman, Biochimica et
Biophysica Acta 1798 (2010) 1041
3. J. Fendos, F. N. Barrera, D. M. Engelman, Biochemistry 52 (2013) 4595
29
P16
CQB-Day 2015
MANGANESE(II) COMPLEXES WITH PYRAZOLE LIGANDS: SYNTHESIS AND OXIDATIVE
CATALYSIS
T. A. G. Duartea, L. M. D. R. S. Martinsa,b, A. P. Carvalhoc, A. J. L. Pombeiroa
a Centro de Química Estrutural, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001 Lisboa,
Portugal ([email protected]) b Chemical Engineering Department, Instituto Superior de Engenharia de
Lisboa, Instituto Politécnico de Lisboa, R. Conselheiro Emídio Navarro, 1959-007 Lisboa, Portugal c Centro de Química e
Bioquímica, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, 1749-016 Lisboa, Portugal
Oxidation reactions catalyzed by transition metal complexes are quantitatively the most important
homogeneously catalyzed reactions in chemical industry. The use of metallic complexes with Chomoscorpionate poly(pyrazol-1-yl)methane ligands as oxidation catalysts is experiencing significant
development, mainly due to their coordinative versatility that allows the electronic and steric features at a
metal center to be tuned. [1,2]
Herein we report the design of manganese(II) complexes containing the C-homoscorpionate
tris(pyrazol-1-yl)methane ligand, HC(pz)3 (pz = pyrazolyl), or pyrazole (Hpz), formed by the reactions
depicted in Scheme 1, for application as catalysts for the partial oxidation of saturated hydrocarbons, a
field of great interest due to the considerable inertness of the substrates.
The effects of some catalytic parameters on the oxidation reactions are also discussed.
Scheme 1
Acknowledgements This work has been partially supported by the Foundation for Science and Technology (FCT),
Portugal (UID/QUI/00100/2013 and UID/MULTI/00612/2013). T. A. G. Duarte is thankful to FCT for his PhD
fellowship (PD/BD/105993/2014). The authors acknowledge the Portuguese NMR Network (IST-UTL Centre) for
access to the NMR facility, and the IST Node of the Portuguese Network of mass-spectrometry for the ESI-MS
measurements.
References
[1] Silva, T. F. S., Martins, L. M. D. R. S., M. Guedes da Silva, M. F. C., Kuznetsov, M. L., Fernandes, A. R., Silva,
A., Pan, C.-J., Lee, J.-F, Hwang, B.-J., Pombeiro, A. J. L., Chem. Asian J. 9 (2014), 1132.
[2] Martins, L. M. D. R. S., Pombeiro, A. J. L., Coord. Chem. Rev. 265 (2014) 74.
30
CQB-Day 2015
P17
A TIME-RESOLVED PHOTOACOUSTIC CALORIMETRY STUDY OF
CYCLOHEXYL, 1,4-DIOXYL, AND TETRAHYDROFURAN-2-YL PEROXY RADICALS
E. M. Gonçalves,a F. Agapito,a R. M. Borges dos Santos,a,b, José A. Martinho Simõesa
a
Centro de Química e Bioquímica,Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal; emails:
{emdgoncalves; agapito; rmsantos; jams}@fc.ul.pt. b Center for Biomedical Research, Universidade do Algarve, Campus de
Gambelas, 8005-139 Faro, Portugal
The main focus of research into automotive-class engines is on low-temperature combustion (LTC)
techniques, that can reduce emissions in-cylinder without sacrificing engine efficiency and fuel
consumption. Under these conditions, peroxy radicals play a crucial role in the overall combustion
process. For the development of both fuels and LTC engines, accurate thermodynamic data on peroxy
radicals is thus essential.1
Unfortunately, peroxy radicals have been among the most difficult radicals to study experimentally,2
mainly due to their manifolds way of reacting, which makes it very difficult to account for the complete
kinetic mechanism of the reaction under study. Our time-resolved photoacoustic calorimetry (TR-PAC)
technique, previously established for the study of carbon radicals, was recently adapted for the study of
peroxy radicals. TR-PAC allows the simultaneous determination of the enthalpy and rate constants of
only the initial steps of radical reactions, when the desired radicals are formed, dramatically simplifying
the interpretation of the results.3
We used our updated TR-PAC technique to obtain the C‒H bond dissociation enthalpy (BDE) of the
model compounds cyclohexane, 1,4-dioxane, and tetrahydrofuran, and the C‒OO● BDE of the
corresponding peroxyl radicals cyclohexyl, 1,4-dioxyl, and tetrahydrofuran-2-yl (Fig. 1). A fundamental
question related to the latter determinations is the solvation enthalpy of the peroxy radicals, a largely
unstudied subject. We addressed this issue by using a new experimental TR-PAC methodology and
quantum chemistry calculations.
Figure 1. The structures of the cyclohexyl (a), 1,4-dioxyl (b), and tetrahydrofuran-2-yl (c) peroxy radicals.
Acknowledgements This work was supported by Fundação para a Ciência e a Tecnologia (FCT), Portugal
(PTDC/QUI-QUI/110542/2009 and UID/MULTI/00612/2013). E.M.G. and F.A. thank FCT for post-doctoral grants
(SFRH/BPD/88607/2012 and SFRH/BPD/74195/2010, respectively).
References
1. M. Mehl, W. J. Pitz, C. K. Westbrook, H. J. Curran, Proc. Combust. Inst. 33 (2011) 193.
2. Y.-R Luo, Comprehensive Handbook of Chemical Bond Energies. CRC Press: Boca Raton, 2007.
3. R. C. Santos, F. Agapito, E. M. Gonçalves, J. A. Martinho Simões, R. M. Borges dos Santos, J. Chem.
Thermodyn. 61 (2013) 83.
31
P18
CQB-Day 2015
INTRAMOLECULAR ALLYLIC AMINATION CATALYZED BY PD(II): A COMPUTATIONAL STUDY
M. J. Calhorda,*a F. J. S. Duarte,a M. M. Lorionb, G. Polib
a Centro de Química e Bioquímica, DQB, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal.
bSorbonne Universités, UPMC Univ Paris 06, UMR 8232, Institut Parisien de Chimie Moléculaire, F-75005, Paris, France
In a previous computational study of the full mechanism of the intramolecular allylic amination reaction
of but-3-enyl-N-tosylcarbamate catalyzed by Pd(II),1,2 it was shown that the C-H activation was the ratedetermining step (RDS). We now extend the DFT methodology to rationalize the effect of modifying the
unsaturated N-tosyl carbamate and the pH of the solution.3
In acidic conditions and when Y = CH2, the aminopalladation pathway is favored over the C-H activation.
This change results from the smaller steric effects in the -elimination transition state (see Figure). The
C-H activation pathway, involving an intramolecular hydrogen atom abstraction, is not affected by the Y
group, but by the length of the carbon chain and the double bond position when other substrates are
considered. The coordination and C-H activation steps depend particularly on the double bond position.
The transition state (TS) energy may increase by ~20 kJ/mol as result of steric interactions between the
carbon chain and the Pd ligands, when the double bond is no longer terminal. For certain carbamates, the
TS for the metal reoxidation process has an energy similar to the energy of the TS for C-H activation,
becoming the new RDS.
Figure 1. Schematic energy profile for aminopalladation and C-H activation
Acknowledgements MJC and FJSD are grateful to FCT (SFRH/BPD/76878/2011 and PEst-OE/QUI/UI0612/20132014). GP thanks CNRS, UPMC for financial support. We thank CMST COST Action CM1205 (CARISMA).
References
1. F. Nahra, F. Liron, G. Prestat, C. Mealli, A. Messaoudi, G. Poli Chem. Eur. J. 15 (2009) 11078.
2. F. J. S. Duarte, G. Poli, M. J. Calhorda (submitted).
3. J. Rajabi, M.M. Lorion, V. L. Ly, F. Liron, J. Oble, G. Prestat, G. Poli Chem. Eur. J. 20 (2014) 1539.
32
CQB-Day 2015
P19
SYNTHESIS AND PHOTOCATALYTIC ACTIVITY OF COBALT DOPED TITANATE NANOTUBES WITH
DIFFERENT COBALT POSITION IN THE STRUCTURAL
B. Barrocasa, A. J. Silvestreb, C. D. Nunesa, O. C. Monteiroa
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected]).
bÁrea Departamental de Física e CeFEMA, Instituto Superior de Engenharia de Lisboa, IPL, 1959-007 Lisboa, Portugal.
Over the past years, the nanostructured materials have become one of the most important research topics
and have established a remarkable development in several scientific areas. In particular, the use of
nanocrystalline semiconductors on the photocatalytic treatment of wastewaters has been generating great
interest. Titanate nanotubes (TNTs) are very interesting since they combine the properties and
applications of conventional TiO2 nanoparticles, with the properties of layered titanates, such as
photocatalytic activity and ion-exchange ability, respectively. However their high charge recombination
rate and wide band gap limit their practical application. Therefore, the synthesis of TNT-based materials
with either a broader range of light absorption or a lower charge recombination rate would be an
important achievement toward the development of successful photoactive catalytic materials. For
instance, the modification of TNTs with a transition metal (e.g. Co), could extend their optical absorption
and catalytic performance of the pristine material.
Figure 1. TEM images of the cobalt doped TNTs.
Nanocrystalline TNTs materials modified by cobalt doping (CoTNTs) and by ion-exchange (TNTsCo)
were successful prepared, by hydrothermal method, and characterized using XRD, TEM, DRS and ICPOES. The influence of the Co content on the optical and photocatalytic activity of the samples was
investigated using the terephthalic acid as the probe molecule to study the catalytic production of
hydroxyl radical (•OH). The photocatalytic degradation of model pollutant molecules (anionic and
cationic organic molecules) using the modified TNTs samples was also investigated. The results show
that the Co modified TNTs samples (CoTNTs and TNTsCo) are better catalysts than the unmodified
TNTs, being the photocatalytic performance dependent on the cobalt position in the TNTs crystalline
structure.
Acknowledgements
The authors thank FCT for financial support (PTDC/CTM-NAN/113021/2009, UID/MULTI/00612/2013 and
SFRH/BD/101220/2014).
References
1. E. K.Ylhäinen, M. R. Nunes, A. J. Silvestre, O. C. Monteiro, J. Mater. Sci. 47 (2012) 4305.
2. V. Bem, M. C. Neves, M. R. Nunes, A. J. Silvestre, O. C. Monteiro, J. Phtochem and Photobiol. A: Chemistry
232 (2012) 50.
3. M. R. Nunes, O. C. Monteiro, A. L. Castro, D. A. Vasconcelos, A. J. Silvestre, Eur. J. Inorg. Chem. 28 (2008)
961.
33
P20
CQB-Day 2015
BIOSYNTHESIS OF METAL SULPHIDE NANOPARTICLES AND THEIR PHOTOCATALYTIC ACTIVITY
IN DEGRADATION OF EMERGING POLLUTANTS
B. Vieiraa, G. Vitorb, J. P. Lourençob,c, O. C. Monteirod, M. C. Costaa
a
Centro de Ciências do Mar, CCMAR, Faculdade de Ciências e Tecnologia, Universidade do Algarve, Campus de Gambelas,
8005-139 Faro, Portugal.
b Faculdade de Ciências e Tecnologia, Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal.
c Centro de Investigação em Química do Algarve, CIQA, Faculdade de Ciências e Tecnologia, Universidade do Algarve, Campus
de Gambelas, 8005-139 Faro, Portugal.
d Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal.
The production of nanomaterials, using byproducts of an environmental remediation process, is a
welcome approach to convert potentially harmful wastes into useful products for innovative applications.
In this sense, this work describes the production of nanosized crystalline materials, as a result of sulphatereducing bioremediation process for the treatment of acid mine drainage (AMD), by precipitating metal
ions into the corresponding metal sulphides. Different nanoparticles of metal sulphides were successfully
synthesized, such as CdS, CuS and the respective TiO2 nanocomposites.
These nanomaterials were morphologically and structurally characterized using different techniques, like
X-ray powder diffraction, transmission electron microscopy, diffuse reflectance spectroscopy and
elemental analysis. Nanoparticles of metal sulphides might possess interesting optical properties, and
studies concerning their potential use in environmentally-friendly applications, such as additives to solar
panels and as photoactive materials for catalytic photodegradation of emerging pollutants, are now in
progress.
Pollutants removal by photodegradation, using semiconductors as catalysts, results in fast oxidation of the
pollutants, which makes a promising and greener technology for the elimination of toxic compounds from
wastewaters. In this work, each sample photocatalytic performance was first evaluated through the
production of the hydroxyl radical, using therephthalic acid as probe molecule. The samples with the
highest photocatalytic ability for the production of this oxidizing species were then tested for the removal
of pollutants, such as chloramphenicol and paracetamol.
TEM images of the CdS nanoparticles (A) and CdS/TiO2 nanocomposites (B). (Insets: SAED of the prepared
powders).
Acknowledgements Funding by Fundação para a Ciência e a Tecnologia (FCT) through project PTDC/AAGTEC/2721/2012 and PEST UID/MULI/00612/2013
34
CQB-Day 2015
P21
ADSORPTION OF PHARMACEUTICAL COMPOUNDS ON CARBONS OF DIFFERENT MORPHOLOGIES
S. Marques, a,b J. Marcuzzo, c A. S. Mestre, a R. Dias, b C. O. Ania, d A. P. Carvalhoa
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal. bMicrobiology
and Biotechnology Lab, Center for Biodiversity, Functional & Integrative Genomics, Edificio ICAT, Campo Grande 1749-016
Lisboa, Portugal. cFaculdade de Tecnologia do Estado de São Paulo FATEC- Campus São José dos Campos, 12247-014 São
José dos Campos SP,Brasil. dADPOR Group, Instituto Nacional del Carbón, INCAR-CSIC33080, Oviedo, Espanha.
([email protected])
One of the main environmental problems affecting public health is aquatic environment contamination.
Pharmaceutical compounds are considered emergent pollutants that have been detected in all sorts of
water streams all over the world. Acetaminophen and clofibric acid are among the most detected
compounds since both are highly consumed medicines continuously introduced in the aquatic media, due
to the lack of effectiveness of the conventional water treatment technologies. Advanced wastewater
treatments processes, namely those based on adsorption onto activated carbons, can be used to solve this
problem.1 Although these adsorbents are normally employed in granular or powder morphologies, other
forms of activated carbons have been developed based on the carbonization and activation of cloths from
different polymeric materials, called activated carbon cloths.2
The aim of this work was to study the adsorption properties of four activated carbons with different
morphologies (powder, granular and cloth) for the removal acetaminophen and clofibric acid from
aqueous solution by kinetic and equilibrium assays. Regeneration cycles at 400 and 600C were also
performed.
The results show that, regarding the adsorption process, the activated carbon cloth performance is
comparable only to the powder sample that has a developed mesopore network, pointing out the
importance of the accessible microporosity present in the cloth. Additionally, cloth morphology has the
advantage of, as in the case of granular carbon, allowing thermal regeneration. After three re-use cycles
the activated carbon cloth retains ~45% of the adsorption capacity obtained with the fresh sample, while
powder sample retains less than 20% of the initial value.
Acknowledgements To FCT (Project UID/MULTI/00612/2013) and fellowships to SM (SFRH/BD/91767/2012)
and ASM (SFRH/BPD/86693/2012).
References
1. N. Bolong, A. Ismail, M. Salim, T. Matsuura, Desalination 239 (2009) 229-246.
2. A. Cukierman, ISRN Chem. Eng (2013) - DOI: 10.1155/2013/261523.
.
35
P22
CQB-Day 2015
NOVEL TECHNOLOGIES FOR SAMPLE PREPARATION IN GREEN ANALYTICAL CHEMISTRY
A. H. Ide, S. M. Ahmad, N. R. Neng, J. M. F. Nogueira
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
Trace analysis generally requires a sample preparation step before quantification, especially when
working with a complex matrix. Therefore, analytical methods must be developed for sample enrichment.
New approaches aim miniaturized and easy to use techniques, in agreement with the green chemistry
concepts, which demand small volumes of sample and reduce the use of toxic organic solvents.
With the development of novel miniaturized sorption-based methods, a new microextraction technique,
based on a static floating sampling mode, bar adsorptive microextraction (BAµE) was proposed recently.
Floating sampling technology is a new enrichment concept based on the use of an analytical device, light
in weight, simultaneously with a conventional Teflon magnetic stirring bar at the bottom of a sampling
flask. When the sample matrix is rapidly spinning around due to centripetal force promoted by the
magnetic bar, the analytical device is left under free-floating motion just below the centre of the vortex.
During a static process, the analytes migrate by diffusion from the bulk sample and then are retained in a
convenient sorbent phase, where the microextraction takes place. Several coating phases have been tested
with remarkable performance, including many types of activated carbons and polymers. This novel
technology has been applied in combination with suitable chromatographic and hyphenated systems (e.g.
GC, HPLC, GC-MS or LC-MS), mainly for trace or ultra-level analysis.
Applications can be performed in several types of samples and matrices, such as environmental
(pharmaceutical and personal care products, pesticides, endocrine disruptors), forensic (drugs of abuse,
legal highs, steroid hormones) and food (disinfection by-products of water, flavonoids, polyphenols).
BAµE presents several advantages, such as the use of convenient sorbents with remarkable selectivity and
sensitivity for a broad range of priority compounds, excellent performance for trace and ultra-level
analysis in several scientific areas. It is also a cost-effective analytical approach, ideally for combination
with instrumental separation systems and the use of small volumes of organic solvents based on the
concepts of green chemistry. The present contribution is an overview on the advantages of floating
sampling technology, as a novel emerging concept for static microextraction analysis.
Figure 1 – Bar adsorptive microextraction operating under the floating sampling technology.
Acknowledgements
The authors thank “Coordenação de Aperfeiçoamento de Pessoal de Nível Superior” (Brazil) for the Ph.D. grant
(CAPES BEX 0394-14-9), “Fundação para a Ciência e Tecnologia” (Portugal) for the Post-Doc grant
(SFRH/BPD/86071/2012) and funding (UID/MULTI/00612/2013).
References
1. J. M. F. Nogueira, Anal. Chim. Acta 757 (2012) 1.
2 N. R. Neng, A. R. M. Silva, J. M. F. Nogueira, J. Chromatogr. A 1217 (2010) 7303.
3 J. M. F. Nogueira, TrAC (2015). doi:10.1016/j.trac.2015.05.002
36
CQB-Day 2015
P23
EXPLORING THE CATALYTIC BEHAVIOUR OF HIERARCHICAL MCM-22 ZEOLITE IN LOW
TEMPERATURE FRIEDEL-CRAFTS ACYLATION OF HETEROAROMATICS
R. Aleixo,a N. Nunes,a,b R. Leitão,a,b F. Martins,b A. P. Carvalho,b A. Brigas,c A. Martinsa,b
a Área
Departamental de Engenharia Química, ISEL, IPL, R. Conselheiro Emídio Navarro, 1959-007 Lisboa, Portugal. b Centro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016, Lisboa, Portugal. c Departamento de
Química Farmacêutica, UAlg, Campus da Penha, Estrada da Penha, 8005-139, Faro, Portugal. ([email protected],
[email protected])
MCM-22 is a synthetic zeolite (Mobil 1990) with a peculiar structure, presenting three independent pore
systems, two of them internal and the last one located at the external surface of the crystals. Even though
this structure presents internal supercages, with a high concentration of acidic active sites, they are only
accessed through narrow windows. Thus, the modification of the textural properties of this zeolite,
through the creation of a micro + mesopore hierarchical structure, allows the improvement of molecular
diffusion and access to the active sites, especially when larger molecules are involved. In this study
hierarchical MCM-22 was prepared by performing post-synthesis basic or a combination of basic + acid
treatments. The experimental procedures as well as the discussion of structural, textural and acidic
characterization are reported elsewhere.1 Friedel-Crafts acylation of heteroaromatics is an important
industrial reaction that traditionally uses homogenous catalysts, such as AlCl 3 and FeCl3 that are harmful
to the environment. The use of commercial zeolites as environmentally friendly catalysts has been
reported. However, the use of hierarchical zeolites, and especially hierarchical MCM-22, has never been
explored. In this study the catalytic behaviour was investigated in the acylation of simple heteroaromatics
such as furan, anisole or pyrrole, by acetic anhydride (Scheme 1) using a molar ratio of 1:5 and 150 mg of
zeolite, at 60 ºC. Samples of the reaction mixture were analysed periodically by GC to follow conversion
and yields as a function of time. Langmuir-Hinshelwood model was used to calculate kinetic parameters
as well as turnover frequencies (TOF). The analysis of the results shows an improved catalytic behaviour
for hierarchical MCM-22 modified through alkaline + acid treatment when compared to parent MCM-22,
presenting enhanced mass transport and access to the active sites, making hierarchical MCM-22 a
promising catalyst for Friedel – Crafts acylation of larger heteroaromatic molecules.
Scheme 1: General scheme for the acylation of heteroaromatic compounds.
Acknowledgements: We thank FCT for financial support under PEst UID/MULTI/00612/2013.
References
1. V. Machado, J. Rocha , A.P. Carvalho, A. Martins App. Catal. A: Gen. 329 (2012) 445.
37
P24
CQB-Day 2015
HUMAN RED BLOOD CELLS EXOVESICLES: A STUDENTS’ APPROACH TO PLASMA MEMBRANE
DYNAMICS
C. P. Sabino, A. G. Flor. T.M.O. Paiva, R. F. M. de Almeida
Centro de Química e Bioquímica, Faculdade de Ci~encias, Unviersidade de Lisboa, 1740-016 Lisboa, Portugal
Under certain physiologic stimuli erythrocytes produce exovesicles, whose main biological functions are
still controversial, but seem to play an important role in intercellular communication. In our laboratory,
we used human blood from healthy donors kindly provided by the Portuguese Institute of Blood and
Transfusions (IPST, IP). By means of ionophore-assisted calcium influx mimicking the physiologic
response to prostaglandin E2, we were able to induce exovesiculation1. The presence of exovesicles was
confirmed by atomic force microscopy, and their size is consistent with previous reports 1 and with the
expected for mammalian membrane microdomains2. Several biochemical features were characterized on
the exovesicles and compared to the whole plasma membrane (erythrocyte ghosts). Exovesicles have a
specific acetylcholinesterase (AchE, E.C. 3.1.1.7) activity at least 40% higher than that of the whole
membrane. AChE is a GPI-anchored protein, and as such, typical of membrane microdomains (lipid
rafts). Furthermore, we managed to identify the main components of exovesicles, as phosphatidylcholine,
sphingomyelin and cholesterol, using Thin Layer Chromatography. Since the only analytical method used
that detects glycosphingolipids was gravimetry, we cannot exclude an enrichment of the vesicles in these
lipids. In conclusion, the analysis of exovesicles from the human red blood cells suggests that they are
formed from microdomains present on the plasma membrane, and that they can be used to further
elucidate membrane structure and dynamics, in health and disease.
Figure 1. The formation of erythrocyte exovesicles and their relation with plasma membrane microdomains.
Acknowledgements
UID/MULTI/00612/2013; Structure and Dynamics of Biomembranes alumni, 2013/2014 and 2014/2015.
References
1 U. Salzer, P. Hinterdorfer, U. Hunger, C. Borken, R. Prohaska. Blood 99 (2002) 2569.
2 A.E.P. Bastos, S. Scolari, M. Stöckl, R.F.M. de Almeida. Methods Enzymol. 504 (2012) 57.
38
CQB-Day 2015
P25
HOW INERT ARE DRUG EXCIPIENTS? A BIOPHYSICAL STUDY OF M-CRESOL-BIOMEMBRANE
INTERACTIONS IN MODEL SYSTEMS AND NEURAL CELLS
T. M. O. Paiva, A. E. P. Bastos, J. M. Marquês, P. A. Lima, A. S. Viana, R. F. M. de Almeida
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected])
m-Cresol, a molecule with ability to interact with artificial lipids 1, is a preservative used in numerous
pharmaceutical formulations, including parenteral insulin and vaccines2-4. Humulin® is a form of
injectable insulin, and one example of those formulations that contain high levels of m-cresol. Therefore,
we studied the effects of m-cresol in a range of concentrations from 10 nM to 3 mM on membrane model
systems mimicking lipid-rafts and living neural-cells, in order to understand if this compound when
present in pharmaceutical formulations can have dangerous effects on human cell membranes.
To that end, the intrinsic fluorescence of m-cresol was first studied. Both its fluorescence lifetime and
anisotropy increased in the presence of liposomes, indicating reduced mobility of the molecule. This
interaction was, however, strongly dependent on membrane lipid composition. To elucidate this,
liposomes were labeled with several membrane probes spanning a range of in-depth locations and with
preference for distinct lipid domains. The probes located in the bilayer core reported no effect, even for an
m-cresol concentration of 300 M, whereas for the more superficial probes, ≥30 M m-cresol induced a
significant fluorescence lifetime decrease, with a compositional dependence similar to m-cresol intrinsic
fluorescence.
Atomic force microscopy was performed on ternary supported lipid bilayers containing raft-like liquid
ordered domains (lo). It was observed that upon addition of m-cresol in the M range, a small reduction of
the lo phase fraction occurs without changing their thickness. For higher m-cresol concentrations, raft-like
domains are not detected at all. All the changes observed occur in less than one hour.
Whole-cell voltage-clamp recordings from pyramidal-neurons isolated from the CA1 region of rat
hippocampus (p21-p29) and from N1E-115 neuroblastoma cells were also performed. m-Cresol was
applied during constant superfusion, and a read-out for the leak-current shows an increase for m-cresol
≥100 M.
As a whole, we show that m-cresol interacts with the membrane surface, affecting lipid raft organization
and neural-cell leak current. Furthermore, m-cresol contained in Humulin® has qualitatively similar
effects, but much less pronounced, possibly because its interaction with other formulation components
decreases the amount of m-cresol available to interact directly with membrane lipids.
Acknowledgements
F.C.T. for grant UID/MULTI/00612/2013; Ph.D. fellowships: SFRH/BD/64442/2009 and SFRH/BD/88199/2012;
Investigador FCT Initiatives (POPH, FSE) 2012 (RFMdeA), 2013 (ASV).
References
1Fujisawa, S. et al. J. Biomed. Mat. Res. 21 (1987) 89.
2 Gualandi-Signorini, A.M. Eur. Rev. Med. Pharma. Sci. 5 (2001) 73.
3 Elder, D.P. et al. Am.Pharm. Rev. 38885 (2012).
4 Patent EP 2129394 B1 (Papillomavirus vaccine composition).
39
P26
CQB-Day 2015
DEHYDRATION OF A ROBUST AND YET METASTABLE HEMIHYDRATE OF 4-HYDROXYNICOTINIC
ACID: THERMODYNAMICS, KINETICS AND MECHANISM
A. Joseph,a C. E. S. Bernardes,a,b A. S. Viana,a M. F. M. Piedade,a,b M. E. Minas da Piedade,a
aCentro
de Química e Bioquímica e Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa,
1749-016 Lisboa, Portugal bCentro de Química Estrutural, Instituto Superior Técnico, Universidade de Lisboa,1049-001 Lisboa,
Portugal.
Hydrates are the most common type of solvates and certainly the most important ones for industries such
as pharmaceuticals which strongly rely on the development, production, and marketing of organic
molecular solids. A survey of the European Pharmacopeia indicates, for example, that 29% of the listed
compounds are known to form hydrates. The stability of hydrates has, therefore, received increasing
attention over the last decades, due to the potential impact of their formation on the development,
processing and performance of solid dosage forms. Particularly important within this context, is the
thermodynamic and kinetic characterization of hydration/dehydration reactions, if tight control over the
production and processing of crystalline organic products is to be achieved. Also relevant, from a more
fundamental point of view, is to understand the mechanism of the dehydration process. In spite of the
important efforts made to systematize the mechanisms of solid state dehydrations, the topic remains a
challenge, particularly when there is no obvious relationship between the crystal lattices of the hydrate
and the anhydrate product (no topotatic relationship), and no evidence of reversibility.
Nicotinic acid and its hydroxyl derivatives have received considerable attention in recent years. This
interest has been mostly driven by their biological significance and ample industrial applications, namely
as additives in food, forage or cosmetics and, particularly, as active pharmaceutical ingredients (API). A
recent study indicated that, in contrast with thermodynamic predictions, a new hemihydrate of 4hydroxynicotinic acid (4HNA∙0.5H2O) did not undergo facile spontaneous dehydration at ambient
temperature and pressure. The origin of this robustness and the mechanism of dehydration were
investigated in this work, through a combined approach which involved kinetic studies by
thermogravimetry (TGA), calorimetric measurements, crystal packing analysis based on X-ray diffraction
data, and microscopic observations by hot stage microscopy (HSM), scanning electron microscopy
(SEM), and atomic force microscopy (AFM).
The TGA experiments indicated that water loss was indeed hindered by the presence of an activation
barrier, which varied in the range 85 - 133 kJ·mol-1, depending on the particle size. It also was found that
the dehydration reaction conformed to the Avarami Erofeev A2 model, which assumes a nucleation and
growth mechanism. Crystal packing analysis suggested that the dehydration process was not topotatic.
Microscopic observations (hot stage microscopy, scanning electron microscopy and atomic force
microscopy) evidenced structural motifs that are compatible with one dimensional random nucleation and
growth of the anhydrous phase which is rarely observed.2 Finally, a reaction Gibbs energy profile for the
dehydration process (Figure 1) could be determined based on the obtained thermodynamic and kinetic
data.
Figure 1. Some structural and energetic features of 4HNA·0.5H 2O dehydration.
Acknowledgements This work was supported by FCT, Portugal (Project UID/MULTI/00612/ 2013), PhD
(SFRH/BD/90386/2012), and Post-Doctoral (SFRH/BPD/101505/2014) grants from FCT are also gratefully
acknowledged by A. Joseph and C. E. S. Bernardes, respectively.
References
1. E.P. Matias, C.E.S. Bernardes, M.F.M Piedade, M.E. Minas da Piedade, Cryst Growth Des. 11 (2011) 2803-2810
2. A. Joseph, C.E.S. Bernardes, A.S. Viana, M.F.M. Piedade, M.E. Minas da Piedade, Cryst Growth Des. 15 (2015) 3511-3524.
40
CQB-Day 2015
P27
MOO3 NANOPARTICLES IN OXIDATION CATALYSIS
C. D. Nunes1, A. Sanches1, A. Bento1, P. D. Vaz1,2
1Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, Ed. C8, 1749-016 Lisboa,
Portugal 2 ISIS Neutron & Muon Source, Rutherford Appleton Laboratory, Chilton, Didcot, Oxfordshire, OX11 0QX, United
Kingdom [email protected]
Nanoscale materials with different morphologies have stimulated great interest due to unique properties
and wide range of potential applications[1] in contrast to their bulky counterparts. MoO2 nanoparticles
were prepared by a hydrothermal method using ethylenediamine as reducing and hydroquinone or iron
oxide (Fe2O3) as assisting agents, respectively. [2,3] The successful preparation of the MoO 2
nanoparticles is evidenced by an extensive characterization process using powder XRD, SEM/TEM
analysis, and FTIR. The resulting material was tested as catalytic precursor in the epoxidation of ciscyclooctene, styrene, R-(+)-limonene and trans-hex-2-en-1-ol, using tert-butylhydroperoxide (tbhp) as
oxygen source. The catalytic studies show that the synthesized material yields selectively the desired
epoxides of the tested substrates with very good results, especially at higher temperatures and using
toluene as solvent.
Figure 1: SEM image of MoO2 nanoparticles.
Acknowledgements The authors are grateful to FCT for financial support (projects UID/MULTI/00612/2013 and
EXPL/QEQ-QIN/1137/2013).
References
[1] C. A. Ellefson, O. Marin-Flores, S. Ha, M. Grant Norton, J. Mater. Sci. 47 (2012), 2057–2071.
[2] L.C. Yang, Q.S. Gao, Y.H. Zhang, Y. Tang, Y.P. Wu, Electrochem. Commun. 10 (2008) 118-122.
[3] A. Bento, A. Sanches, E. Medina, C. D. Nunes, P. D. Vaz, Appl. Catal A: Gen. DOI:
http://dx.doi.org/10.1016/j.apcata.2015.03.024
41
P28
CQB-Day 2015
POLLUTANTS REMOVAL USING NEW MAGNETIC-BASED PHOTOCATALYSTS
C. Bravo, C. D. Nunes, O. C. Monteiro
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, Ed. C8, 1749-016 Lisboa,
Portugal, [email protected]
The presence of organic pollutants in the aquatic environments is one of today's major environmental
issues. Therefore, it is urgent to research and develop new methodologies using promising functional
materials that provide high pollutants removal efficiency at a rational cost. In this work, iron oxide
magnetic nanoparticles (MNP) were synthetized and then covered with a silica layer (MNP-Si), and
afterwards were functionalized with the ligand isonicotinoyl chloride (MNP-Si-inic) and finally reacted
with an organometallic precursor of Mo(II) (MNP-Si-inic-Mo)1. The synthesis of MNP, which have
magnetic properties, had the aim of favoring and simplifying the separation process after a photocatalytic
process. The photocatalytic ability of MNP, MNP-Si, MMP-Si-inic and MNP-Si-inic-Mo was tested with
three different organic pollutants: terephthalic acid, phenol and Rhodamine B. Overall, all nanomaterials
showed good photocatalytic behavior.
Figure 1: Scheme of the synthesis
Acknowledgements The authors are grateful to FCT for financial support project UID/MULTI/00612/2013.
References
[1] C.I. Fernandes, M.D. Carvalho, L.P. Ferreira, C.D. Nunes, P.D. Vaz (2014) J. Organomet. Chem. 760: 2-10.
42
CQB-Day 2015
P29
ENZYME MODIFIED POLYDOPAMINE FILMS WITH HIGH CATALYTIC PERFORMANCE
L. C. Almeida, A. C. Carreira, J. P. Correia, A. S. Viana
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, 1749-016 Lisboa, Portugal
([email protected])
In recent years, polydopamine (PDA) films have been extensively studied due to their great
biocompatibility and adhesive properties [1]. They are highly functional films enriched with quinone
groups which are able to covalently bind target biomolecules, such as enzymes, through a Schiff base
reaction [2]. In addition, their easy preparation on a variety of surfaces make them good alternatives to
more common electrode modification approaches, e.g. self-assembled monolayers [3], used for biosensors
and biofuel cell applications.
The objective of this work is to exploit the potential of PDA films in the immobilisation of Laccase and
Glucose Oxidase on graphitic surfaces, in a way they preserve or even improve their catalytic ability for
the molecular oxygen reduction and glucose oxidation, respectively. To fulfil this purpose, several PDA
films with different thicknesses were grown on glassy carbon and their electrochemical properties were
assessed by cyclic voltammetry. Ellipsometry has been used to evaluate the increase of the film thickness
with time of deposition, whereas morphological information was provided by AFM onto highly oriented
pyrolytic graphite. Contact angle measurements disclose the hydrophilicity of the films that was
independent from the film thickness. The powerful combination of these techniques allowed to select the
most suitable PDA film that maximizes the catalytic performance of commercial graphite bioelectrodes,
achieved by cyclic voltammetric and chronoamperometric measurements, with promising currents values
for miniaturized electronic devices applications.
Acknowledgements Financial Support by FCT: CQB’s PEST UID/MULTI/00612/2013 and IF 2013 (POPH, Fundo
Social Europeu).
References
1. Y Lui, K Ai and L Lu, Chemical Reviews 114 (2014) 5077.
2. E. Faurea, C. Falentin-Daudréa, C. Jérômea, J. Lyskawa, D. Fournier, P. Woisel, C. Detrembleur, Progress in
Polymer Science 38 (2013) 236.
3. I. Ameida, V.C. Ferreira, M.F. Montemor, L.M. Abrantes, A.S. Viana, Electrochimica Acta 30 (2012) 311.
43
P30
CQB-Day 2015
THE AMBIGUOUS CASE OF POLYMORPHISM IN SIMVASTATIN: A SINGLE CRYSTAL
X-RAY DIFFRACTION, THERMODYNAMIC, AND MD SIMULATION STUDY
R. G. Simões,a C .E. S. Bernardes,a M. F. M. Piedade,b,c H. P. Diogo,b M. E. Minas da Piedadea
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, 1749-016 Lisboa, Portugal.
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, 1749-016 Lisboa,
Portugal. cCentro de Química Estrutural, Instituto Superior Técnico, Universidade de Lisboa, 1049-001 Lisboa, Portugal.
bDepartamento
Simvastatin (Figure 1) is one of the most widely used active pharmaceutical ingredients (API) for the
treatment of hyperlipidemias. Because the compound is employed as a solid in drug formulations,
particular attention should be given to the characterization of different polymorphs, their stability
domains and the nature of the phase transitions that relate them. Indeed, because modifications of crystal
packing may be accompanied by significant changes in physical properties, the lack of control over
polymorphism may create serious problems for the reproducible production and safe use of an API. In
this work, the crystal structures of three previously reported simvastatin phases 1 were experimentally
revisited based solely on single crystal X-ray diffraction experiments. A key aim was to examine if there
is really polymorphism in simvastatin in the classical sense of McCrone’s definition, which implies the
existence of at least two phases with different arrangements of the molecules in the solid state and rules
out differences from changes in shape such as those involving dynamic isomerism or tautomerism.
Another goal was to evaluate the nature and reversibility of the solid-solid phase transitions relating forms
I, II, and III simvastatin.
Preliminary results indicated that all phases correspond to an orthorhombic structure, space group
P212121, with Z’/Z = 1/4. This is in accordance with the previously reported studies, except for phase III,
which had been assigned as monoclinic, space group P21, Z’/Z = 2/4.1 Differential scanning calorimetry
experiments evidenced two reversible phase transitions: III  II at 235.9±0.1 K, with ΔtrsHm = 0.95±0.06
kJ·mol-1; II  I at 275.2±0.2 K, with ΔtrsHm = 3.3±0.1 kJ·mol-1. No other phase transitions were found
above 275.2 K, up to the melting point, which occurred at 412.2±0.2 K with ΔfusHm = 30.4±0.2 kJ·mol-1.2
These results, in conjunction with quantum chemistry calculations and molecular dynamics simulations,
suggest that the structural differences between the three simvastatin solid phases are essentially related to
a progressive increase of internal rotation freedom of the “ester tail” (Figure 1) as the temperature
increases and not to different arrangements of the molecules in the crystal lattice. Free rotation is
essentially achieved on entering the liquid domain.2 Thus, overall, the obtained results point to an
“ambiguous” type of polymorphism, which does not conform to McCrone’s definition.
The fact that the two transitions were found to be fast and reversible with very small hysteresis, suggests
that polymorphism in unlikely to be a problem for pharmaceutical formulations employing crystalline
simvastatin because, if present, the III and II phases will readily convert to phase I, which is the most
stable form at ambient temperature.
Figure 1. Molecular structure of simvastatin.
Acknowledgements This work was supported by projects UID/MULTI/00612/2013, PTDC/QUI-QUI/098216/2008, and
PTDC/QUI-QUI/116847/2010 and grants (SFRH/BD/48410/2008) (SFRH/BPD/101505/2014) from FCT, Portugal.
References
1. Čejka, J.; Kratochvíl, B.; Císařová, I.; Jegorov, A., Acta Crystallogr. C59 (2003) O428-O430; Hušák, M.; Kratochvíl, B.;
Jegorov, A.; Brus, J.; Maixner, J.; Rohlíček, J., Struct. Chem. 21 (2010) 511-518.
2. R.G. Simões, C.E.S. Bernardes, H.P. Diogo, F. Agapito, M.E. Minas da Piedade, Mol. Pharmaceutics 10 (2013) 2713-2722.
44
CQB-Day 2015
P31
SYNTHESIS, CHARACTERIZATION AND ANTI-CANCER EVALUATION OF NOVEL RUTHENIUM (II)
COMPLEXES OF THIOSEMICARBAZONES
S. Assis,a,b A. C. Poeta,a,b A. Valente,a T. S. Morais,a F. Marques,c M. P. Robalo,d S. Santos,b*
A. I. Tomaz,a M. H. Garciaa
aCentro
de Química Estrutural, Faculdade de Ciências, Universidade de Lisboa, 1749-016, Lisboa, Portugal. bCentro de
Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected]) . cCentro de Ciências e Tecnologias Nucleares, Instituto Superior Técnico, Universidade de
Lisboa, 2695-066 Bobadela LRS, Portugal. dCentro de Química Estrutural, Instituto Superior Técnico, Universidade de Lisboa,
1049-001 Lisboa, Portugal.
Cancer is the second largest cause of death in developed countries. Two in every five people born today
will be diagnosed with cancer at some time during their life, and one of them will effectively perish from
that condition. According to the World Health Organization, cancer mortality is projected to rise to over
13.1 million people in 20301. Cisplatin and analogues are to date the only metal-based chemotherapeutics
approved worldwide for clinical use. Although highly effective, dose-limiting side effects and the
development of resistance to treatment severely limit their clinical value, and ruthenium compounds are a
proven effective alternative2. Ruthenium-based compounds have consistently shown excellent anti-cancer
properties both in vitro and in vivo, and are quite promising in cancer chemotherapy, typically offering a
wide spectrum of activity and the potential to overcome tumor platinum-resistance, as well as different
mechanisms of action and a general lower toxicity2].
Research in this field has been quite extensive and several families of compounds based on Ru(II) and
Ru(III) showing structural diversity have been developed. In the search for complexes with suitable
stability and solubility in aqueous media, octahedral Ru(NN)2L complexes (NN being a polypyridyl-type
ligand) have shown interesting profile, their therapeutic activity being modulated to some extent by the
bidentate ligand L, especially if it exhibits biological activity itself. In this context, thiosemicarbazones
(TSCs), particularly those including heterocyclic moieties, possess a broad range of biological activity,
such as antimalarial, antimicrobial, antifungal and antitumor properties, and can be used as the ligand L.
In this work five new aromatic thiosemicarbazones (TSC) were synthesized and used as ligands in the
preparation of novel ruthenium (II) complexes of the type [(NN)2Ru(TSC)][X] (Fig.1). TSC were chosen
as ligands due to their ability to chelate with several metals, including ruthenium, and proved
pharmacological properties as antibacterial, antifungal, and antitumor activities3.
All ligands and complexes were fully characterized by NMR, IR, UV-Vis, ESI-MS, Elemental Analysis
and Cyclic Voltammetry. Their anti-cancer activity was evaluated in vitro against ovarian
adenocarcinoma and triple-negative breast.
Fig.1. Structure of the ligands and complexes synthesized in this study.
Acknowledgements This work was financed by FCT within the scope of projects PTDC/QUI-QUI/118077/2010,
UID/QUI/00100/2013, UID/QUI/UI0612/2013 and the Investigador FCT Initiative IF/01179/2013 and IF/01302/2013.
References
1. The World Health Organization, http://www.who.int/mediacentre/factsheets/fs297/en/index.html accessed on 9th June (2015).
2. A. Levina, A. Mitra, P. A. Lay, Metallomics 1 (2009) 458.
3. F. A. Beckford, M. Shaloski Jr., G. Leblanc et. al, Dalton Trans. 48 (2009) 10757.
45
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SHAPE AND CURVATURE OF SURFACE TENSION ISOTHERMS FOR LIQUID MIXTURES
M. S. C. S. Santosa, J. C. R. Reisb
a Centro
de Química e Bioquímica, Faculdade de Ciências,Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected]) bCentro de Química Estrutural, Faculdade de Ciências,Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected]).
The different shapes of surface tension isotherms consistent with Butler’s ideal model for the planar
surface phase of binary liquid mixtures are comprehensively worked out using the ratio of pure
constituent molar surface areas, r  A A , and a parameter named surface factor, E      A RT ,which depends
AB

A

B
B
*
A
*
B
*
B
only on pure-constituent properties. Butler’s equation1 is expressed and solved as a function of these two
independent parameters and the geometrical features for the plots of reduced surface pressure against the
mole fraction of component with lower surface tension are established for every possible combination of
those two parameters. Three main shape types are found, namely: concave down (negative curvature);
concave down passing through an inflexion point followed by concave up; and concave up (positive
curvature). The range of parameter values leading to each shape type is obtained and shown graphically
and demonstrated that Butler’s model excludes the possibility of an ideal system presenting a shape
comprising an initial negative curvature, an inflexion point and continuing with positive curvature.
Figure 1. Domains of values for the surface factor of B, E , and the molar surface area ratio, r , leading to different types
B
A B
of Butler’s reduced surface pressure isotherms.
Acknowledgements We thank FCT for supporting this work under projects UID/MULTI/00612/2013 and
UID/QUI/00100/2013.
References
1. J.A.V. Butler, Proc. R. Soc. London A 135 (1932) 1348–1375.
46
CQB-Day 2015
P33
FORMULATION OPTIMIZATION AND STABILITY EVALUATION OF A BIOACTIVE CATIONIC
NANOEMULSION CONTAINING QUERCETIN
M. F. Darioa,b, M. S. C. S. Santosb, N. A. Bou-Chacraa, M. E. Minas da Piedadeb, A. R. Babya,
M. V. R. Velascoa
aDepartamento
de Farmácia, Faculdade de Ciências Farmacêuticas, Universidade de São Paulo, 05508-000 São Paulo,
Brasil,bCentro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
Colloidal dispersions containing very small particles have numerous advantages over macroemulsions
namely increased stability towards aggregation and gravitational separation, useful rheological properties,
and the ability to increase the bioavailability of incorporated bioactive lipophilic substances. This work
describes the preparation of a cationic nanoemulsion containing the antioxidant quercetin by the sub-PIT
method. The procedure consists of heating all the components, under constant stirring, until minimum
turbidity (clearing boundary temperature – Tcb) is achieved, followed by fast cooling. The ideal
temperature (Tmáx) to achieve the nanoemulsion was determined monitoring conductivity and turbidity
over heating cycles (45.0 <  < 80.0 ºC). The oil droplets average diameter and polydispersity index were
determined by dynamic light scattering (DLS) and droplets morphology was characterized by Cryo-TEM.
Long term stability assays were performed and Ostwald ripening constants were calculated.
The nanoemulsion is formed upon heating above 75 ºC, but the best parameters are observed at 80 ºC
(Fig. 1, 2). The nanoemulsion presents spherical droplets with very low diameter (201nm). Ostwald
ripening destabilization phenomena associated with individual spreading of oil molecules between
adjacent droplets (ω3) occurred at high temperature (45.0 ± 2.0 ºC). Disruption was not observed at low
or room temperature, where the nanoemulsion remained stable over 90 days. This methodology was
effective to attain a bioactive cationic nanoemulsion comprising 20 nm diameter spherical droplets
containing 0.5% (w/w) of the bioactive lipophilic antioxidant quercetin, stable at room temperature for at
least 90 days.
Figure 1. Relationship between turbidity, conductivity and temperature of bioactive cationic nanoemulsion. Legend: (♦)
Turbidity; () Conductivity
10000
1
0.5
PDI
D / nm
100
1
0
60
65 70 heating
75 80
Maximum
…
Figure 2. Average droplet diameter (D) and polydispersity index (PDI) of nanoemulsion stir-quenched from different initial
temperatures to  =25º C.
Acknowledgements The support of FCT through project UID/MULTI/00612/2013, and a FASESP grant to M.
Dario is acknowledged.
47
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CQB-Day 2015
BIOPHYSICAL PROPERTIES OF STEAROYL-PHYTOCERAMIDE MIXED WITH A FLUID PHOSPHOLIPID.
ROLE OF SPHINGOID BASE HYDROXYLATION IN MEMBRANE COMPARTMENT ORGANIZATION
J. T. Marquêsa, A. M. Cordeiroa, A. S. Vianaa, A. Herrmanna, H. S. Marinhoa, R. F. M. de Almeidaa
a
Centro de Química e Bioquímica, Departamento de Química e Bioquímica, Faculdade de Ciências da Universidade de Lisboa,
Ed. C8, Campo Grande, 1749-016, Lisboa, Portugal
b Department of Biology, Molecular Biophysics, Humboldt University, Berlin, Germany
Phytoceramide is the backbone of major sphingolipids in fungi and plants and is essential in several
tissues of animal organisms, such as human skin. Its sphingoid base, phytosphingosine, differs from that
usually found in mammals by the addition of a hydroxyl group to the 4-ene, which may be a crucial factor
for the different properties of membrane microdomains among those organisms and tissues. Recently,
sphingolipids hydroxylation in animal cells emerged as a key feature in several physiopathological
processes. Hence, the study of the biophysical properties of phytosphingolipids is relevant also in that
context, since it helps to understand the effects of sphingolipid hydroxylation. In this work, binary
mixtures of N-stearoyl-phytoceramide (PhyCer) with palmitoyloleoylphosphatidylcholine (POPC) were
studied. Steady-state and time-resolved fluorescence of different membrane probes, X-ray diffraction,
atomic force microscopy and confocal microscopy were employed. As for other saturated ceramides,
highly rigid gel domains start to form with just ~5 mol% PhyCer at 24ºC. However, PhyCer gel-enriched
domains in coexistence with POPC-enriched fluid present additional complexity, since their properties
(maximal order, shape and thickness) change at specific POPC:PhyCer molar ratios, suggesting the
formation of highly stable stoichiometric complexes with their own properties, distinct from both POPC
and PhyCer. A POPC/PhyCer binary phase diagram (Figure 1), supported by the different experimental
approaches employed, is proposed with complexes of 3:1 and 1:2 stoichiometries, stable at least from ~15
C to ~55 C, providing mechanisms for in vivo formation of sphingolipid-enriched gel domains, that
may account for stable membrane compartments and diffusion barriers in eukaryotic cell membranes.
Figure 1. Proposed phase diagram for the binary mixture POPC:PhyCer.
Acknowledgements Fundação para a Ciência e a Tecnologia (FCT), Portugal is acknowledged for PTDC/QUIBIQ/104311/2008, PEst 2015-2020 (UID/Multi/00612/2013), and IF2012/2013 initiatives (POPH, Fundo Social
Europeu). J.T.M. acknowledges a PhD. scholarship SFRH/BD/64442/2009.
48
CQB-Day 2015
P35
ADDING EXPLICIT -HOLES IN THE GROMOS FORCE FIELD: TOWARDS DRUG DESIGN
APPLICATIONS
D. Vila-Viçosa, M. Machuqueiro, P. J. Costa
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal ([email protected])
Halogen atoms are commonly used in drug design to fill hydrophobic cavities in protein binding sites,
improve blood–brain barrier crossing or facilitate membrane permeability. Additionally, halogens are
capable of establishing a directional, non-covalent interaction, known as halogen bond, consisting on an
R­X∙∙∙B interaction, where X = Cl, Br, I and B = Lewis base. These bonds are explained by the existence
of a positive region at the tip of X, called -hole, which interacts with Lewis bases.
Halogen bonds play an important role in several protein-drug interactions and several structures deposited
in the RCSB Protein Data Bank show this type of interaction, thus showing its potential for rational drug
design. In spite of this importance, the implementation of halogen bonding in biomolecular force fields is
rare and recent relying on the use of massless points of charge to emulate the -hole, but depend solely on
the nature of the halogen. In the commonly used GROMOS force field, the implementation is nonexistent and the halogen atom is described by an erroneous punctual negative charge with large LennardJones parameters.
Herein, we try to overcome the limitations of the current implementations by presenting an approach that
takes into account not only the halogen, but also its substituents, known to influence the halogen donor
capability. Additionally, we broaden the application of our approach in biomolecular simulations by
implementing the halogen boding terms in the GROMOS 54A7 force field. Therefore, in this work we
will present a way to parametrize drug molecules containing halogen atoms, taking this interaction into
account which latter could be used in drug design applications. The introduction of the massless points of
charge is critical for the proper description of the halogen bonding interaction (Figure 1).
Figure 1 – Iodobenzene – Ammonia dimer after energy minimization with a molecular dynamics force field without hole (a) and with -hole (b).
Acknowledgements The Authors acknowledge financial support from Fundação para a Ciência e Tecnologia,
Portugal, through project grant UID/MULTI/00612/2013 and fellowship SFRH/BD/81017/2011. P.J. Costa
acknowledges FCT for the Investigador FCT Programme (IF/00069/2014).
49
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CQB-Day 2015
DESIGN OF UREA-BASED ANION RECEPTORS WITH ANTICANCER PROPERTIES
J. M. Caio,a,b M. J. Martins,a T. Esteves,a O. Cruz e Silva,b V. Félix,b C. Moiteiroa
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected]). biBiMED and CICECO, Universidade de Aveiro, 3810-193 Aveiro, Portugal ([email protected]).
Channelopathies are diseases caused by genetic mutations of the proteins responsible for the regulation of
the ion transport in cells.1 For instance, a chloride transport deficiency is expressed in cystic fibrosis (CF),
which is the most common life-threatening genetic disease in humans.2 In addition there are several
studies that infer a link between anion transport and anticancer activity, for instance, it is known that the
activation of K+ and Cl- efflux channels originate a cell volume decrease, inducing apoptosis of cancer
cells.3
On the other hand, Ataluren (TranslarnaTM, Figure 1) is a small molecule incorporating 1,2,4-oxadiazole
ring, and is currently in III clinical trial in the USA and recommended for approval in the EU for CF. 4–7
Inspired in this drug-like molecule, we have devised two series of small anion receptors composed of urea
binding moiety linked by the ethylenediamine to an amide binding unit decorated with 1,3,4- or 1,2,4oxa-diazole rings (Figure 1, 1 and 2, respectively). Additionally, a third series of derivatives 3,4disubstituted-1,2,5-oxadiazole with urea or amide groups were also prepared (Figure 1, 3). Herein, we
report the synthesis and binding affinity of these receptors for biological anions (chloride and
bicarbonate) together with their transmembrane anion transport and cytotoxicity properties against human
cancer cell lines.
Figure 1. Ataluren and the tree series of urea-based synthetic anion receptors.
Acknowledgements This work was financially supported by National Funds through FCT under projects
PTDC/QUI-QUI/101022/2008 and PEst-UID/MULTI/00612/2013. J.M. Caio also thanks FCT for his PhD
fellowship SFRH/BD/66789/2009.
References
1. R. S. Kass, JCI 115 (2005) 1986.
2. F. M. Ashcroft, Ion Channels and Disease: Channelopathies. Academic Press, San Diego, 2000.
3. a) P. A. Gale, et al. Angew. Chem. Int. Ed. 52 (2013) 1374-1382; b) P. A. Gale, et al. Chem. Sci. 3 (2012) 25012509; c) R. Quesada et al., Accounts Chem. Res. 46 (2013) 2801-2813; d) P. A. Gale, et al. J. Am. Chem. Soc. 133
(2011) 14136-14148.
4. E. Kerem et al. Lancet Respir. Med. 2 (2014) 539.
5. E. Kerem, et al. Lancet 372 (2008) 719.
6. E. Dolgin Nat. Med. 17 (2011) 396.
7. M. Wilschanski, et al. Eur. Respir. J. 38 (2011) 59–69.
50
CQB-Day 2015
P37
SEPARATION OF THE ETHANE/ETHYLENE MIXTURE IN UIO-66 MOFS
J. Fernandesa, M. Pintob, J. Piresa
aCentro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected])., bCERENA, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, n° 1, 1049-001
Lisboa, Portugal ([email protected] ).
The separation of ethylene from ethane, is one of the most energy-intensive single distillations
practiced. This separation could be alternatively made by an adsorption process if the adsorbent would
preferentially adsorb ethane over ethylene, reducing energy consumption and, therefore, contribute to a
cleaner environment.
In this work we studied metal-organic frameworks (MOFs) more precisely the MOFs UiO-66 and it
variant with CF3 substituent group (UiO-66-CF3) and showed that both materials are ethane selective
adsorbents (that is, both adsorb higher amounts of ethane than ethylene) at 25 and 46 ºC for pressures up
to 10 bar.
Figure 1. Adsorption isotherms of ethane and ethylene in the MOF UiO-66 at 25 ºC.
Selectivity values are shown also.
Acknowledgements Thanks are due to FCT by PEST UID/MULTI/00612/2013 (CQB) and UID/ECI/04028/2013
(CERENA)
References
1. J. Pires, M. L. Pinto, V. K. Saini, ACS Appl. Mater. Interfaces 6 (2014) 12093−12099.
51
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CQB-Day 2015
METABOLIC PROFILE OF LEISHMANIA INFANTUM PROMASTIGOTES AND
AMASTIGOTES BY FT-ICR-MS ANALYSIS
A. P. Marquesa, M. Maiaa, b, A. E. N. Ferreiraa, A. Figueiredob, F. Monteirob, M. Sebastianab,
S. Carvalho c, A. M. Tomás c,d, A. Ponces Freirea, C. Cordeiroa, M. Sousa Silvaa*
a
Centro de Química e Bioquímica, Departamento de Química e Bioquímica, Faculdade de Ciências da Universidade de Lisboa,
1749-016 Lisboa, Portugal; *email [email protected]. b Centro de Biodiversidade, Genómica Integrativa e Funcional, Faculdade
de Ciências da Universidade de Lisboa, 1749-016 Lisboa, Portugal. c IBMC, Instituto de Biologia Molecular e Celular,
Universidade do Porto, Porto, Portugal. d ICBAS, Instituto de Ciências Biomédicas Abel Salazar, Universidade do Porto, Porto,
Portugal
Leishmaniasis is caused by the protozoan Leishmania parasites which are transmitted by the bite of
infected sandflies. The major risks factors associated with this disease are malnutrition, population
displacement, poor housing, a weak immune system and lack of resource. An estimated 1.3 million new
cases and 20000 to 30000 deaths occur annually [1]. There are several forms of this disease, but the most
severe and fatal without treatment is the Leishmania infantum which occurs mainly in 6 countries such as
Bangladesh, Brazil, Ethiopia, India, South Sudan and Sudan. The infected dogs, domestic and/or wild, are
the principal reservoir for Leishmania infantum and can be a threat to public health. Leishmania species
can exist in two main forms depending on the stage of their lifecycle: promastigote and amastigote that
can be found in invertebrate (sandfly) and vertebrate (mammalian) hosts, respectively. To understand
better Leishmania infantum and to search new treatments it is important to know the differences between
these two life forms.
Metabolomics is a new and growing research area which involves “… a comprehensive analysis in which
all the metabolites of a biological system are identified and quantified...” according to Oliver Fiehn [2]. In
the quest for whole metabolome identification it is essential to achieve maximum coverage, sensitivity
and specificity. These three requirements can only be simultaneously reached by high-resolution and
high-mass accuracy mass spectrometry. In this work, we developed a metabolite extraction method from
Leishmania parasites using different solvents such as chloroform, water, methanol and acetonitrile. All
the fractions were analysed by direct infusion in a Fourier Transform Ion Cyclotron Resonance Mass
Spectrometer
(FT-ICR-MS) in positive and negative ionization modes. The data collected in the form of an m/z values
list was submitted to SimMet (version 1.5, by Premier Biosoft), a software for mass spectrometry
metabolite data analysis.
Acknowledgements This work was supported by projects UID/MULTI/00612/2013 and REDE/1501/REM/2005
from Fundação para a Ciência e Tecnologia.
References
1. World Health Organization, http://www.who.int/topics/en/ (updated February 2015).
2. O. Fiehn, Plant Mol. Biol. 48 (2002) 155-171.
52
CQB-Day 2015
P39
DATEBASE OF PHENOLIC COMPOUNDS BY MASS SPECTROMETRY
C. Sáncheza, M. H. Florêncio a, M. L. Serralheiroa, A. P. Marquesa*
a Centro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal; *email
[email protected]
Mass spectrometry is an analytical technique with high sensitivity, low detection limit and short analysis
time. This technique is very versatile and can be applied in various fields, such as chemistry, biology,
pharmacy, environmental and forensic sciences. Mass spectrometry provides information on the structure
of molecules, allowing to identify unknown compounds and quantify known compounds [1]. Liquid
chromatography coupled to mass spectrometry (LC-MS) is also an important analytical technique, since it
combines the power of chromatography separation with the capacity of mass spectrometric identification.
Phenolic compounds like flavonoids, caffeic acid derivatives and others are present in several foods
(e.g. fruits and vegetables) and drinks (e.g. coffee, tea and infusions). The identification of these
compounds in this type of products allows to establish a relationship between the biological activity
attributed to these food products with the chemical compounds existing in them. This can therefore
contribute to a better understanding of the beneficial effects of a Mediterranean diet characterized by the
elevated consumption of fruits and vegetables [2].
In this work, we built a database of phenolic compounds through the injection of different standards by
LC-MS and direct infusion in positive and negative ionization modes. The apparatus used was a HPLC
Surveyor Plus coupled to a LCQ Duo ion trap mass spectrometer equipped with an ESI source from
Thermo Scientific. The standards analyzed are divided in two series: caffeic acid, chlorogenic acid,
rosmarinic acid and syringic acid (first series), apigenin, fisetin, kaempherol, luteolin, rutin and taxifolin
(second series). These standards were chosen because they are present in many samples studied in Grupo
de Espectrometria de Massa Ambiental e Biológica (Departamento de Química e Bioquímica,
Universidade de Lisboa, Portugal). All the results were acquired and processed by Xcalibur (version 1.2
from Thermo Scientific).
Acknowledgements This work was supported by projects UID/MULTI/00612/2013 and REDE/1501/REM/2005
from Fundação para a Ciência e Tecnologia.
References
1. J. K. Prasain (Ed.), Tandem Mass Spectrometry Applications and Principles (2012). InTech, Croatia.
2. A. S. Dontas, N. S. Zerefos, D. B. Panagiotakos and D. A. Valis. Clin. Interv. Aging. 2 (2007) 109-115.
53
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CQB-Day 2015
ANTIOXIDANT PROPERTIES AND EVALUATION OF TANNINS IN DIFFERENT CASTES OF GRAPES
M.E.M. Araújo, B. Oliveira
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected]).
Wine is a product well known and their antioxidant property is very well studied as well. Wine is
commonly commercialized as a mixture obtained of various types of grapes, and therefore the antioxidant
values presented by the majority of the samples are usually very similar. The goal of this project is to
study the variation of the antioxidant properties of three types of grapes generated under the same
conditions and what might be the influence of the presence of tannins in different quantities in the grapes
antioxidant variation.
The antioxidant properties were studied using the DPPH and the β-carotene bleaching tests and both tests
showed strong differences between different castes.
Tannins in three Portuguese wines are also under study using the ATR-FTIR technique.
Several solutions of tannic acid were prepared and a plot of tannic concentration against the KubelkaMunk values was obtained using the intensities of two bands present in all classes of tannins, 1207 cm-1
and 1328 cm-1 (Figure 1). Good correlations were obtained which indicate that the content of tannins in
wines can be evaluated, as tannic acid equivalents, using this technique, first in liquid samples and then
using dried samples of wine.
a)
b)
𝜈 =1207 cm
-1
𝜈 =1328 cm
-1
Figure 1. Plot of the intensities of two bands of tannic acid (ATR-FTIR) and the concentration of the solution
Acknowledgements CQB’s PEST UID/MULTI/00612/2013
References
L. Falcão, M.E.M. Araújo, Vibrational Spectroscopy 74 (2014) 98-103
54
CQB-Day 2015
P41
PHYTOCHEMICAL SCREENING OF MEDICINAL PLANTS FROM TUNISIA
(MARRUBIUM VUL GARE AND GLOBULARIA ALYPUM)
M.E.M. Araújo,a M. Rezgui,b S. Bellili,c L. Bettaieb Ben-Kaab, b W. Mnif c
[a] Centro de Química e Bioquímica, Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade de
Lisboa, Campo Grande Ed. C8, 1749-016 Lisboa, Portugal. [email protected]. [b] Faculty of Sciences of Tunisia, University
Tunis El Manar, Tunis, Tunisia. [c] 1LR11-ES31 Biotechnologie et Valorisation des Bio-Géo Ressources, Institut Supérieur de
Biotechnologie de Sidi Thabet, BiotechPole de Sidi Thabet, 2020, Université de la Manouba, Tunisie.
Tunisia has a rich flora including numerous medicinal and aromatic plants. More than 500 species out of 2103
(approximately 25% of the total flora) are used in therapeutic [1]. In Tunisia, salinity affects about 10% of the whole
territory Crops are more and more exposed to this problem accentuated by increasing climate aridity [2]. Plants
subjected to high salinity levels undergo various physiological and biochemical changes. First investigation
concerned Marrubium vulgare (Lamiaceae) to study its degree of resistance to salt stress. Plants were watered with
five levels of NaCl concentration: 0, 25, 50, 100 and 150 mM during 5 months. The first results showed that salt
treatment reduced significantly total phenol content of methanolic extracts, nevertheless those treated with NaCl 100
mM showed the highest capacity to scavenge the DPPH radical (IC50=199.86 µg∙mL-1), and were quite potent in the
β-carotene bleaching assay (40%). Similarly, extracts reducing power ability increased with increasing salinity. As
marrubiin is the major constituent of M. vulgare, the effect of salt stress on the production of this compound was
investigated and quantified by NMR spectroscopy. The above results showed that marrubiin content decreases with
salt stress which was confirmed using the intensities of peaks at 6.29 and 7.25 ppm. Hence, Marrubium vulgare can
be considered as salt sensitive. Second investigation concerned Globularia alypum (Globulariaceae) which is a wild
perennial shrub found throughout the Mediterranean area, Europe and northeastern Africa [3]. The present study
aimed at assessing the phenolic content, flavonoids and antioxidant activities (β-carotene, ferric reducing power).
Also, the effectiveness of α-glucosidase inhibition was evaluated to access the antidiabetic effects of Globularia.
Infusion, methanolic and aqueous extracts were prepared for the (leaves, flowers, steams and roots) separately, the
results showed that the highest total phenolic content was in leaves (8.62 mg EAG/g DW), the greatest total
flavonoid content was also revealed in leaves (1.75 mg Rut E/ gDW). Concerning the alpha-glucosidase inhibition,
the study has been performed on the leaves methanolic extract at two different concentrations (1mg/mL and 0.5
mg/mL) and showed that plants exhibited respectively 98% and 80 % inhibitory effect. The rest of analysis for
differents extracts and tests are under investigation.
Figure 1. localisation of Marrubium vulgare (1) and Globularia alypum (2)
Acknowledgements The work of Maria Eduarda M. Araújo was funded by Fundação para a Ciência e a Tecnologia (FCT), Portugal (PEstOE/QUI UI0612/2013).
References [1] E. Le Floc’h. (1983). Contribution à une étude ethnobotanique de la flore Tunsienne. Programme Flore et Végétation tunisienne,
Ministère de l’Enseignement supérieur et de la Recherche scientifique, Tunis, pp 113–114.
[2]
M. Hachicha, OJ. Job, A. Mtimet. (1994). Sols salés et la salinisation en Tunisie. Sols de Tunisie, 5, 271–341.
[3]
N. Es-Safi, S. Khlifi, L. Kerhoas, A. Kollmann, A. El Abbouyi, PH Ducrot. (2005). Antioxidant constituents of the aerial parts of Globularia
alypum growing in Morocco. J. Nat. Prod 68: 1293–1296.
55
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TRACKING NEW PSYCHOACTIVE SUBSTANCES IN PORTUGAL
H. Gaspara, C. Leala,b,C. Gonçalvesa, S. Ciríacoa, A. Matiasa,b, J. Rodriguesb, S. Santosa
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, 1749-016 Lisboa,
Portugal([email protected]). bLaboratório de Polícia Cientifica da Polícia Judiciária, Novo edifício-sede da
Polícia Judiciária, Rua Gomes Freire, 169-007 Lisboa, Portugal ([email protected])
In the last decade, there has been an uprising of New Psychoactive Substances (NPS) available in
“smartshops” and over the Internet. NPS are defined as “new narcotic or psychotropic drugs that are not
listed in the Single Convention on Narcotic Drugs of 1961 or the Convention on Psychotropic Substances
of 1971, but which may pose a public health threat comparable to that posed by substances listed in those
conventions”. These compounds have been traditionally sold as innocuous products, becoming a legal
alternative to illicit drugs, such as cocaine, ecstasy or cannabis. The products have been advertised as
being herbal incenses, plant feeders or bath salts, with the caveat of not being for human consumption. Up
to 2014, more than 450 NPS have been reported in the European Union (EU). The two most reported
classes of NPS are synthetic cathinones and synthetic cannabinoids and, just in 2014, these two groups of
substances accounted for approximately 50% of the total NPS reported in the EU [1].
Recently, a new decree has been implemented in Portugal (Dec-Lei 54/2013) [2], which forbids the
production and commercialisation of about 159 NPS, being liable to fast updates, in order to keep up with
the everyday appearance of new substances. The rapidly growing problem of NPS makes the time
management for international control a real challenge, with the traditional detection methods becoming
increasingly inadequate. In order to tackle this issue, a collaboration was established in May 2014 with
the Laboratório de Polícia Científica da Polícia Judiciária (LPC), in which the role of our group is to
supply NPS standards to forensic laboratories, obtained either by synthesis or by isolation from LPC
samples, and to develop adequate analytical methodologies for the identification and quantification of
NPS in different matrices.
With the decree, several products previously sold in 8 Portuguese smartshops were delivered to LPC.
Within the scope of this collaboration, we have already studied 169 of those products. This led to the
identification of 26 NPS, through NMR and/or GC hyphenated techniques (MS and FID). The isolation
and unequivocal structural characterization of NPS permits their use as secondary standards for routine
analyses in forensic laboratories and allowed the creation of an in-house library with 21 NPS: 12
synthetic cathinones, 8 synthetic cannabinoids and ethylphenidate. The developed methodology was
applied in 4 seized samples, allowing the identification of two novel NPS: SGT-25, a synthetic
cannabinoid recently reported in the EU; and 4F-PBP (Figure 1), a new synthetic cathinone in the EU
reported by us during this project [3].
The application of the developed methodology in seized products allows a faster and easier tackling of
the rapid appearance of NPS in the market, illustrating the importance of the scientific knowledge in the
resolution of current public health threats.
Figure 1. 4F-PBP, a novel NPS
Acknowledgements The authors thank Fundação para a Ciência e a Tecnologia (FCT) for financial support through project
UID/MULTI/00612/2013 and the toxicology sector at LPC/PJ.
References
1. European Monitoring Centre for Drugs and Drug Addiction, European Drug Report 2015 - Trends and Developments,
Publications Office of the European Union, Luxembourg, 2015.
2. Decreto-Lei 54/2013, Diário da República, 1.ª série, No. 75, 17 de abril de 2013.
3. H. Gaspar, S. Bronze, S. Ciríaco, C.R. Queirós, A. Matias, J. Rodrigues, C. Oliveira, C. Cordeiro, S. Santos, For. Sci. Int. 215
(2015) 168.
56
CQB-Day 2015
P43
SYNTHESIS OF HYACINTH FRAGRANCE: CATALYTIC BEHAVIOR OF ACTIVATED CARBONS MADE
FROM RAPESEED SOLID RESIDUE OF BIODIESEL PRODUCTION
M. Batista,a,b I. Fonseca,b I. Matos,b J. Vital,b A. Mestre,a A. P. Carvalhoa
aDepartamento
de Química e Bioquímica and CQB, Faculdade de Ciências da Universidade de Lisboa, Ed C8, Campo Grande,
1749-016 Lisboa, Portugal bDepartamento de Química REQUIMTE, CQFB, Faculdade de Ciências e Tecnologia,Universidade
Nova de Lisboa, Quinta da Torre, 2829-516 Caparica, Portugal ([email protected])
The use of biomass as reagent or base material is of utmost importance for the optimization of available
resources and the minimization of residues. This study not only presents the production of activated
carbon from a biomass waste, but also it’s use as catalyst in the transformation of glycerol which is a
secondary product of biodiesel production and may became a residue itself. The rapeseed-based activated
carbons were prepared by chemical activation with K2CO3 rapeseed was mixed with ground K2CO3. The
textural properties of the materials were evaluated through N2 and CO2 adsorption assays. Activated
carbons with apparent surface area of around 1000m2g-1 were obtained.[1]The synthesis of
phenylacetaldehyde glycerol acetals, 2-benzyl-4-hydroxymethyl-1,3-dioxolane (1), 2-benzyl-5-hydroxy1,3-dioxane (2) which are flavoring compounds, have been carried out successfully by acetalization of
phenylacetaldehyde with glycerol using toluene as solvent and rapeseed activated carbon as catalyst,
according to the scheme in Figure 1.[2,3] The effect of various reaction parameters such as temperature,
molar ratio of reagent to glycerol were studied.
OH
HO
O
O
O
+
OH + HO
O
1
HO
O
2
Fig.1. Reaction Scheme of the formation of 2-benzyl-4-hydroxymethyl-1,3-dioxolane (1), 2-benzyl-5-hydroxy-1,3dioxane (2).
Acknowledgments The authors thank IBEROL- Sociedade Ibérica de Oleaginosas by provided the solid residues
used in this work. Mary Batista and Ana S. Mestre, thank the financial support of FCT for a
(SFRH/PBD/84542/2012) and (SFRH/PBD/86693/2012) Post-Doc grants, respectively).
References
1. M. Batista, I. Fonseca, A. Mestre, A. Carvalho, Poster, XII GEC, Madrid, 2013.
2. M. Climent, A. Corma, A. Velty, Applied Catalysis A: General 263 (2004) 155.
3. T. Ceia, A. Silva, C. Ribeiro, J. Pinto, M. Casimiro, A. Ramos, J. Vital, Catalysis Today 236 (2014) 98.
57
P44
CQB-Day 2015
MOB1 AN ACTIVE PLAYER IN MORPHOGENESIS, CYTOKINESIS AND CELL PROLIFERATION
CONTROL IN PROTOZOA ORGANISMS
A. Tavaresa,c,e, I. Delgadoc, S. Franciscoc, J. Coelhoc, A. Leitãod, H. Soaresa,b,e S. Nolascob,c,e
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa; bEscola Superior de
Tecnologia da Saúde de Lisboa, 1990-096 Lisboa, Portugal;c Centro de Investigação Interdisciplinar em Sanidade Animal,
Faculdade de Medicina Veterinária, Universidade de Lisboa, Avenida da Universidade Técnica, 1300-477 Lisboa, Portugal,
dInstituto de Investigação Científica Tropical, CVZ, CIISA, Avenida da Universidade Técnica, 1300-477 Lisboa, Portugal,
eInstituto Gulbenkian de Ciência, Apartado 14, 2781-901 Oeiras, Portugal ([email protected]);
Intrinsic cell polarity relies in elaborate intracellular organization allowing cells to define, for example,
anterior-posterior, dorsal-ventral and left-right axis. Also, accuracy of cell division depends on the correct
division plane placement, which is dependent on the polarity axis.
Mob1 is a core protein of the Mitotic Exit Network and of the Hippo pathway, fundamental signaling
cascades for the correct metaphase to anaphase transition and for the proper balance between cell
proliferation and death. In metazoan, Mob1 is also involved in cell proliferation and organ size control
1. We may wonder if this Mob1 role was also early established during evolution and is already present
in single-cell organisms. If true, Mob1 will be an excellent target to study the control of protozoan
parasite replication, a key matter in parasite/host interaction. To explore this hypothesis we have
investigated the role of Mob1 in Toxoplasma gondii. T. gondii is an obligate intracellular protozoan
parasite (phylum Apicomplexa) that infects warm-blooded vertebrates. This parasite is an important lifethreatening opportunistic pathogen in AIDS patients and other immunosuppressed individuals. T. gondii
presents a polarized cell structure with an anterior-posterior organization and is able to form tissue cysts
that can be seen as supra cellular structures organizations with a characteristic size inside the vertebrate
host organism. Thus, like tissue growth and correct size maintenance in multicellular organisms, the
number of parasites inside the structure of the cyst has also to be tightly controlled.
Our studies show that also in T. gondii Mob1 has a polarized localization in the antero-posterior axis. This
cellular localization resembles that of the ciliate protozoa Tetrahymena thermophila, in which Mob1
accumulates in the posterior pole basal bodies, creating a gradient through the anterior-posterior axis of
the cell that is required to correct cytokinesis and therefore proper cell division 2. Strikingly, the
Toxoplasma mob1 gene is down-regulated when parasites start to replicate inside of the host cell.
Additionally, we have constructed a transgenic parasite strain that overexpresses Mob1 and these
parasites show a significant delay in the replication process. Our results strongly support that Mob1 is not
only a core factor that links cytokinesis to morphogenesis but is also a regulator in cell proliferation in
unicellular organisms, which parallels with the role of Mob1 in the metazoans Hippo pathway.
Acknowledgements: This
UID/MULTI/00612/2013
work
was
funded
by
EXPL/C
VT-EPI/1945/2013
and
References
1 Hergovich A. Cell Signal. 23 (2011)1433.
2. Tavares A, Gonçalves J, Florindo C, Tavares AA, Soares H. 125 (2012) J Cell Sci. 516.
58
CQB`s
PEST
CQB-Day 2015
P45
MOLYBDENUM COMPLEXES WITH 2,2’-DIPYRIDYLAMINE DERIVATIVES AS CATALYSTS IN
OXIDATION REACTIONS
M. S. Saraiva, M. J. Calhorda
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected]).
New molybdenum complexes from the family [MoBr(ƞ3-C3H5)(CO)2(L)] were synthesised by reaction of
the precursor [MoBr(ƞ3-C3H5)(CO)2(CH3CN)2] with a family of bidentate ligands derived from 2,2’dipyridylamine, as shown in Figure 1. All new compounds prepared were characterized by FTIR, NMR
of 1H e 13C as well as elemental analysis.
Figure 1.Molybdenum complexes from 2,2’-ddipyridylamine derivatives.
All complexes were tested as homogeneous pre-catalysts in the oxidation of olefins and sulfides, such as
cis-cyclooctene, styrene, cis-3-hexen-1-ol, trans-2-hexen-1-ol, geraniol, methylphenylsulfide and
diphenylsulfide, using hydrogen peroxide, tert-butyl hydroperoxide or cumene hydroperoxide as the
oxidant. These studies were carried out in order to determine the influence of the length of the alkyl
chains of the ligands on the complexes catalytic activity.
Acknowledgements: We thank Fundação para a Ciência e Tecnologia for financial support (PEST UID
/MULTI/00612/2013). MSS thanks FCT for Grant SFRH / BPD / 88082 / 2012.
References
1. M.S. Saraiva, S. Quintal, F.M.C. Portugal, T.A. Lopes, V. Félix, J.M.F. Nogueira, M. Meireles, M.G.B.
Drew, M.J. Calhorda, Journal of Organometallic Chemistry 693 (2008) 3411–3418.
2. M.J. Rauterkus, S. Fakih, C. Mock, I. Puscasu, B. Krebs, Inorganica Chimica Acta 350 (2003) 355-365.
59
P46
CQB-Day 2015
FUNCTIONAL NANOPARTICLES OF FOOD PROTEINS FOR REDUCTION OF CARDIOVASCULAR
DISEASES RISK
A. Silvaa, b, J.T. Marquêsa, A.S. Vianaa, R. Pacheco a, b, M.L.M. Serralheiroa
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected]; [email protected])
bArea Departamental de Engenharia Química, Instituto Superior de Engenharia de Lisboa, Av. Conselheiro Emídio Navarro,
1959-007 Lisboa ([email protected]);
Nowadays with the problem of human population ageing there is increasingly high risk of cardiovascular
diseases. These are among the highest cause of death in EU and also in Portugal [1]. One of the risk
factors of these diseases is the high cholesterol level in the blood. People having high levels of cholesterol
are often medicated with drugs and, the drugs currently prescribed are reported to present various
secondary effects. The group members expertise [2, 3] in the use of plant infusions and decoctions, often
named as herbal teas, containing flavonoids and other polyphenols, for reducing serum cholesterol levels,
suggested the main idea for this work: the use of nanotechnology to obtain a biological product by
encapsulation of polyphenols from the plant decoctions in food proteins and to study the application of
these functional foods in hypercholesterolemia treatment. One of the objectives of the encapsulation of
the plants infusion compounds into nanopreparations is to increase the low polyphenols uptake by the
cells [3], which is expected to have an effect of cholesterol reduction. Another of the objectives is to use
as carries food proteins that are considered safe and have additional nutritional value, hence the biological
products obtained can be applied as functional foods.
At the present state of work, different methodologies and conditions were tested for the synthesis of
nanoparticles using bovine serum albumin loaded with various sources of flavonoids, such as quercetin
and rutin, as standards, and Annona cherimola leaves decoctions. The obtained nanoparticles (NPs) were
characterized by Atomic Force Microscopy demonstrating nanoscale size. Toxicity of the NPs were tested
using human colorectal carcinoma lines (Caco2 ATCC#HTB37) and also the stability of the preparations
at the digestive action using in vitro simulation of gastric and pancreatic juices. Bioavailability studies in
Caco2 cells, an epithelial cell line normally used to simulate the intestinal barrier, were conducted. The
results obtained in these studies will be presented.
10 nm
Figure 1. Rutin-BSA nanoparticles
Acknowledgements UID/MULTI/00612/2013
References
1. ESC/EAS Guidelines for the management of dyslipidaemias, European Heart Journal 32 (2011) 1769
2. P. L. Falé, C. Ferreira, ,A. M. Rodrigues, F. N. Frazão, M. L. M. Serralheiro, Journal of Medicinal Plants
Research, 8 (2014) 917.
3. P. L. Falé , C. Ferreira , F. Maruzzella, M. H. Florêncio , F.N. Frazão, M.L. Serralheiro. Ethnopharmacol. 25
(2013) 71823.
60
CQB-Day 2015
P47
SYNTHESIS AND APPLICATION OF NEW TITANATE NANOTUBES SENSITIZED WITH SILVER
NANOPARTICLES FOR POLLUTANTS PHOTODEGRADATION
B. Barrocas, O. C. Monteiro
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected]).
The disposal, after use, of pharmaceuticals and personal care products (PPCPs) has become very
problematic nowadays. They are extremely resistant to conventional treatments and even at very low
concentrations, they may impose toxicity at all biological hierarchy levels. Several approaches have been
proposed to overcome this dramatic environmental problem but with unsatisfactory results. New solutions
with clear effectiveness are mandatory. Titanate nanotubes (TNTs) are a semiconductor material that
combines the properties and applications of conventional TiO2 nanoparticles (including photocatalysis)
with the properties of layered titanates. In addition, the intrinsic properties of these 1D materials, mainly
surface area, physical and chemical adsorption ability, optical and photocatalytic properties, can be
tailored and adjusted to a specific application/interaction.
a
b
Figure 1. TEM images of TNTs before (a) and after (b) sensitization with nanocrystalline Ag particles.
The synthesis, using a new methodology, of TNTs modified with Ag nanocrystalline particles was
successful achieved. The best conditions for the production of nanomaterials with different TNT/Ag ratio
were established. The nanomaterials were morphologically and structurally characterized using X-ray
powder diffraction, transmission electron microscopy. Optical and electronic properties were also
evaluated. The application of these new photoactive materials for emergent pollutants catalytic
photodegradation was studied and the results will be present and discussed.
Acknowledgements
The authors thank FCT for financial support (UID/MULTI/00612/2013 and SFRH/BD/101220/2014).
References
1. E. K.Ylhäinen, M. R. Nunes, A. J. Silvestre, O. C. Monteiro, J. Mater. Sci. 47 (2012) 4305.
2. V. Bem, M. C. Neves, M. R. Nunes, A. J. Silvestre, O. C. Monteiro, J. Phtochem and Photobiol. A: Chemistry
232 (2012) 50.
3. M. R. Nunes, O. C. Monteiro, A. L. Castro, D. A. Vasconcelos, A. J. Silvestre, Eur. J. Inorg. Chem. 28 (2008)
961.
61
P48
CQB-Day 2015
PKA VALUES OF TITRABLE AMINO ACIDS AT THE WATER/MEMBRANE INTERFACE
V. H. Teixeira, D. Vila-Viçosa, P. B. P. S. Reis, Miguel Machuqueiro
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected])
pH is a crucial physicochemical property that affects most biomolecules. Changes in protonation
equilibrium of susceptible sites will modify the electrostatic environment and, consequently, have an
effect on the molecular structure, stability and catalysis. 1 The pKa values of the typical titrable amino
acids can be significantly influenced by changes in the solvent mixture or due to insertion into a lipid
bilayer.2-3
The main objective of this work is a comprehensive study of how a membrane environment can shift the
pKa values of common pH-sensitive amino acids (Asp, Glu, His, Lys, Cys, Tyr, and the N- and Ctermini). For this, we used our recently developed CpHMD-L methodology4 with a DMPC membrane and
the model Ala-based pentapeptides that have already been well characterized in water by Pace and coworkers.5
With
this
approach,
we
intend
to
capture
the
coupling
between
conformation/configuration/insertion and protonation at the membrane interface, taking into consideration
that if complete insertion occurs, the peptides will probably no longer be able to exchange protons with
the solvent.
Figure 1. The Ala2-Glu-Ala2 pentapeptide at the water/membrane interface.
Acknowledgements: We acknowledge financial support from FCT through projects PTDC/QUI-BIQ/113721/2009,
UID/MULTI/00612/2013, Incentivo/QUI/UI0612/2014 and grant SFRH/BD/81017/2011.
References
1. J. Matthew, Annu. Rev. Biophys. Biophys. Chem. 14 (1985) 387.
2. J. F. Hunt, P. Rath, K. J. Rothschild, D. M. Engelman, Biochemistry 36 (1997) 15177.
3. P. R. Magalhães, M. Machuqueiro, A. M. Baptista, Biophys. J. 108 (2015) 2282.
4. D. Vila-Viçosa, V. H. Teixeira, A. M. Baptista, M. Machuqueiro, J. Chem. Theory Comput. 11 (2015) 2367.
5. R. L. Thurlkill, G. R. Grimsley, J. M. Scholtz, C. N. Pace, Protein Sci. 15 (2006) 1214.
62
CQB-Day 2015
P49
METHOD'S DEVELOPMENT FOR DRUG ANALYSIS BY MASS SPECTROMETRY AND OTHER
ANALYTICAL TECHNIQUES
R. A. Osawaa, M. H. Florêncioa, A. P. Carvalhoa, M. R. Bronzeb
a
b
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa Portugal
Faculdade de Farmacia, Universidade de Lisboa, 1649-019 Lisboa Portugal
Pharmaceutical drugs are considered emerging contaminants, which by definition are chemical
compounds that are not monitored because they have unknown toxicological effects or have no regulatory
legislation. Emerging compounds may be of synthetic or natural origin, found in products consumed by a
large population and end up in natural ecosystems through the discharge of effluents in superficial
waters1. Many of these drugs have unknown effects on biota and humans. These compounds are found in
superficial waters, underground waters and sewage effluents2,3.
The study aimed to optimize chromatographic parameters for the analysis of drugs: amitriptyline (AMI),
bupropion (BUP), venlafaxine (VEN), citalopram (CIT), trazadone (TRA) and duloxetine (DUL) group of
antidepressants and methotrexate (TEM), ifosfamide (IF), cyclophosphamide (CYP) and 5-fluorouracil
(5-FU) group of anticancer, verify the recovery of these compounds in filters and test their removal in
distilled water using activated carbon (granulometry between 0.42 and 0.29 µm).
LC-MS/MS (Triple Quadrupole) in MRM (Multiple Reaction Mode) was used for the detection of drugs.
The compounds obtained low detection limits, between 0.15 and 1.5 µg L-1 and quantification limits
between 0.5 and 5 µg L-1 and showed good selectivity. In the future, samples will be collected at
environmental matrices with volumes of 250 ml to be extracted in SPE (Solid Phase Extraction) with final
volume of 1 ml, the sample will have a concentration factor of 250. The quantification limits will result
between 2 to 20 ng L-1.
The nylon showed the best result among the compounds (Figure 1) using syringe filters (83-144%) and
the carbon adsorption experiment (temperature at 30 °C for 5 and 17 hour), all drugs were removed in
concentrations of 10 µg L-1.
The method developed using LC-MS/MS for the determination of pharmaceutical compounds is adequate
for application in environmental matrices. For future studies it is intended to collect sample of effluent
treatment plants to determine the removal of these compounds. In the carbon adsorption experiment, the
removal was 100% in the reported conditions. For future experiments it is intended to change the
concentrations of the contaminants and the adsorption time to determine the adsorption kinetics.
Cellulose Acetate
Cellulose Regenerated
PTFE
Nylon
Glass Fiber + Cellulose Acetate
160
140
Relative Response (%)
120
100
80
60
40
20
0
IF
LU
5F
YP
C
M
ET
C
IT
TR
A
L
U
VE
N
AM
I
D
BU
P
-20
Figure 1 - Adsorption experiments for different filter
Acknowledgements CAPES – n° 0845/14-0; CQB’s PEST UID/MULTI/00612/2013
References
1. M. Stuart, D. Lapworth, E. Crane, A. Hart. The Science of the Total Environment 416 (2012) p.1–21.
2. J. Giebułtowicz, J. Grzegorznałęcz Ecotoxicology and Environmental Safety 104 (2014) p.103–109.
3. O. Golovko, V. Kumar, G. Fedorova, T. Randak, R. Grabic. Chemosphere 111 (2014) p.418–426.
63
P50
CQB-Day 2015
RECOVERY OF PALLADIUM FROM A SPENT INDUSTRIAL CATALYST BY HYDROMETALLURGY
O. Ortet,a,b A. P. Paiva,a C. Nogueirac
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected]). bDepartamento de Ciência e Tecnologia, Universidade de Cabo Verde, Praia, Cabo Verde. cLaboratório
Nacional de Energia e Geologia (LNEG), I.P., Estrada do Paço do Lumiar, 22, 1649-038 Lisboa, Portugal.
Precious metals such as silver, platinum, gold, palladium have similar chemical properties, and are
extremely important from the commercial point of view. These metals are used in industrial and/or
automobile catalytic converters, as well as in the production of jewelry, electronics, etc. [1]. Focusing on
palladium, which is one of the metals belonging to the class of the platinum-group metals (PGMs), as it is
one PGM actually rare in the earth crust, there is an increasing tendency to its recovery from different Pdcontaining spent materials. Palladium is used to catalyze the synthesis of various organic compounds in
the chemical industry. The recovery of palladium from these catalysts is an important economic aspect;
most catalysts are usually supported on alumina/silica, with varying percentages and different proportions
of Pd, with concentration ranges from 2.5 to 20% [2]. Several researchers [2-5] have adapted
pyrometallurgy and hydrometallurgy for recovery of palladium from catalysts by application of leaching
processes.
In this communication, the experimental development of a leaching process of a spent industrial catalyst,
from a petrochemical industry in Portugal, is presented. After leaching, the process of solvent extraction
followed, in order to recover the palladium content from the leached solution. The investigated catalyst is
composed by metals like aluminum, palladium and chromium. Several parameters were controlled under
the leaching process, such as the temperature (27, 40, 60, 80 and 90ºC), the reaction time, the
hydrochloric acid concentration of the leaching solution (2, 4 and 6 M HCl), the concentration of the
oxidizing agents (1 M HNO3, 1 M H2O2), the liquid-solid ratio (L/S), and particle size. Leaching with 2 M
HCl and 1 M H2O2, at 27ºC for 10 minutes, with an L/S=2 and an average particle size of 176 µm, was
adequate to solubilize more than 95% of palladium and less than 2% and 1% of aluminum and chromium,
respectively. The “best” liquors obtained in the leaching process have been tested for preliminary
extraction studies, aiming to the selective recovery of palladium by solvent extraction using 0.03M Ncyclohexyl-N-methyloctanthioamide in toluene as the organic solvent [6].
Acknowledgements The financial support for this work has been kindly provided by FCT “Fundação para a Ciência
e a Tecnologia” (Lisbon, Portugal), under the project UID/MULTI/00612/2013, and through the PhD grant
SFRH/BD/78289/2011, offered to O. Ortet.
References
1. A. Cieszynska, M. Wisniewski, Sep. and Purif. Technol. 80 (2011) 385.
2. B. Singh, Journal of Hazardous Materials 167 (2009) 24.
3. M. A. Barakat, M. H. H. Mahmoud, Y. S. Mahrous, Applied Catalyst A: General 301 (2006) 182.
4. M. K. Jha, J. Lee, M. Kim, J. Jeong, B. Kim, V. Kumar, Hydrometallurgy 133 (2013) 23.
5. A. Das, R. Ruhela, A. K. Singh, R. C. Hubli, Sep. and Purif. Technol. 125 (2014) 151.
6. O. Ortet, A. P. Paiva, Hydrometallurgy 151 (2015) 33.
64
CQB-Day 2015
P51
TRANSITION METAL COMPLEXES FOR CO2 REDUCTION
S. Realistaa, A. I. Melatoa, P. M. Martinhoa, M. J. Calhordaa
a Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected])
Carbon dioxide is the main greenhouse gas resulting from everyday human activities.[1] The increasing
concentration of CO2 in the atmosphere affects Earth’s energy balance. Changes in the greenhouse effect,
in the energy reaching Earth’s surface and alterations in its atmosphere and surface reflectivity are some
of the known consequences. For these reasons it is urgent to find efficient pathways for carbon utilisation
and recycling to form products with significant commercial value. Recent reports of reaction of
pressurised CO2 with epoxides to form polycarbonates[2] or cyclic carbonates[3] have attracted both
academic and industrial interest. While recycling of CO2 to produce high value products have mainly
used Fe, Ni, Zn and Co based catalysts,[4] reports on Cu catalysts for efficient CO2 reduction are scarcely
known. We describe the synthesis of several mononuclear and binuclear Schiff base complexes and its
performance for homogeneous and heterogeneous CO2 reduction. The synthesis and characterisation of
new asymmetric Cu(II)/Cu(II) and Ni(II)/Ni(II) binuclear Schiff base complexes obtained via a one-pot
reaction introducing different aldehyde building blocks and overcoming solubility issues is also
presented.[5] Both binuclear complexes and their mononuclear parent salphen subunits were used for CO2
reduction in homogeneous (figure 1) and heterogeneous media. Several studies such as controlled
potential electrolysis and cyclic voltammetry were performed to highlight a route for CO 2 recycling
catalysts.
Figure 1
Acknowledgements: S. Realista thanks FCT for the scholarship SFRH/BD/52368/2013 – CATSUS doctoral
program. The authors also thank to Fundação Calouste Gulbenkian (Programa Estímulo à Investigação 2013) and to
CQB’s PEST UID/MULTI/00612/2013.
References
1. D. T. Whipple, P. J. A. Kenis, J. Phys. Chem. Lett., 1, (2010), 3451.
2. G. W. Coates, D. R. Moore, Angew. Chem. Int. Ed., 43, (2004), 6618.
3. J. Meléndez, M. North, R. Pasquale, Eur. J. Inor. Chem., 21, (2007), 3323.
4. J. Qiao, Yuyu Liu, F. Hong, J. Zhang, Chem. Soc. Rev., 43, (2014), 631.
5. S. Realista, A. S. Viana, B. de P. Cardoso, A. M. Botelho do Rego, P.D. Vaz, A. I. Melato, P.N. Martinho, M. J.
Calhorda, RSC Adv., 5, (2015), 39495.
65
P52
CQB-Day 2015
THE CENTROSOMAL PROTEIN TBCCD1 REGULATES THE DYNAMICS OF A MICROTUBULE
SUBPOPULATION INVOLVED IN THE NUCLEUS-CENTROSOME CONNECTION
C. Cameloa, *, C. Penedaa, *, A. I. Câmaraa, B. Carmonaa,b, H. S. Marinhoa H. Soaresa,b
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa; bEscola Superior de
Tecnologia da Saúde de Lisboa, 1990-096 Lisboa, Portugal; ([email protected])
*These authors have contributed equally for this work.
The centrosome is a major microtubule-organizing center (MTOC) in animal cells, usually localized at
the cell center in close association with the nucleus. It has been reported to have an important role in
several cell processes such as organelle positioning, cell migration, adhesion, polarity, spindle pole
formation and assemble cilia axoneme. The TBCC domain-containing human protein (TBCCD1) is a
centrosomal protein previously characterized by our group 1. TBCCD1 contains two identified domains:
a TBCC domain, which is associated with the tubulin folding pathway, and a CARP domain, that is
thought to be associated with actin filaments. We identified two different splicing transcripts that encode
TBCCD1 proteins with cytoplasmic localization. The overexpression of each of the proteins seems to
regulate the endogenous expression of the other. Furthermore, TBCCD1 knockdown in RPE-1 cells
causes a pronounced increase in the centrosome-nucleus distance, a cell cycle delay, GA disorganization,
lower efficiency to assemble primary cilia and delayed wound healing close accompanied by collective
cell migration alterations. Recent data suggests that TBCCD1 may be involved in the control of
centrosome-nucleus connection by regulating the dynamics of a microtubule subpopulation surrounding
the nucleus. We hope to soon elucidate this role by the identification of the TBCCD1 and TBCCD1
splicing variant partners using the BioID system.
Acknowledgements: This work was funded by CQB`s PEST UID/MULTI/00612/2013
References
1. J. Gonçalves, S. Nolasco, R. Nascimento, M Lopez Fanarraga, JC Zabala, H. Soares, EMBO Reports 11
(2010)194.
66
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THEORETICAL STUDY ON THE INTERCALATION OF PHENANTHROLINE IN DNA: WHEN
DISPERSION FORCES ARE IMPORTANT BUT THE ELECTROSTATIC CONTRIBUTION BECOMES
CRUCIAL
A. Gil,*a V. Branchadell,b M. J. Calhorda*a
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected]; [email protected] ). bDepartament de Química, Universitat Autònoma de Barcelona, 08193 Bellaterra,
Barcelona, Spain
The effects of phenanthroline (phen) intercalation in the structure, energetics and bonding of AdenineThymine and Guanine–Cytosine tetramers (A-T/T-A and G-C/C-G) were studied through Density
Functional Theory (DFT) using functionals that were recently improved considering the effect of
dispersion forces. Our results given by the Energy Decomposition Analysis show that dispersion
contribution, Edisp, is the most important contribution to the interaction energy, Eint. However, it is not
enough to compensate the Pauli repulsion term, EPauli, and the role of the orbital contribution, Eorb, and
specially the role of the electrostatic contribution, Eelstat, become crucial for the stabilization of the
structures in the intercalation process. Moreover, formation energies are higher when intercalation takes
place from Major Groove (MG) in G-C/C-G systems but no appreciable differences are found for A-T/TA systems. For G-C/C-G systems hydrogen bonding (HB) interactions are more important than stacking
(S) interactions, whereas for A-T/T-A systems, HB and S become competitive. On the other hand,
intercalation produces important changes not only in the hydrogen bonds of base pairs because S and HB
are deeply connected, but also in other characteristic geometric parameters of the base pairs. Interactions
and bond properties are analyzed in terms of dipole moments, polarizability, molecular electrostatic
potential maps, electronic density, charge transfer, and frontier orbitals.
Acknowledgements This research was financially supported by the Fundação para a Ciência e a Tecnologia (FCT)
by means of the postdoctoral grant SFRH/BPD/89722/2012 to A. G. and the grant PEst-OE/QUI/UI0612/2013).
67
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CQB-Day 2015
TARGETING BUTYRYLCHOLINESTERASE FOR NEW TREATMENTS IN ALZHEIMER’S DISEASE AND
CANCER
V. Cachatra,a S. Schwarz,b M. C. Oliveira,c L. Gano,c A.Paulo,c A. P. Rautera
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal. bBereich
Organische Chemie, Martin-Luther-Universität Halle-Wittenberg, Kurt-Mothes-Str. 2, D-06120 Halle (Saale), Germany. cCentro
de Ciências e Tecnologias Nucleares, IST, Universidade de Lisboa, Estrada Nacional 10, 2695-066 Bobadela LRS, Portugal
Worldwide, there is a growing concern in providing a healthy ageing mainly due to the increase in
average life expectancy. Butyrylchlolinesterase (BChE) is an enzyme that is responsible for the
degradation of acetylcholine in the brain of patients in latter stages of Alzheimer’s disease (AD). While
some of the approved treatments are based in single inhibition of acetylcholinesterase or dual inhibition of
both enzymes, research focusing on selective BChE inhibition to access a better understanding of BChE
role in AD is mandatory.[1]
Cancer is also one of the main challenges for researchers dedicated to the search for new treatments
aiming to provide a healthy and active ageing. Not only new therapies with fewer side-effects are required
but also more effective imaging agents for detection of malignant cells at earlier stages are needed. BChE
levels have been used as a biochemical marker in the management of head, neck and cervical cancer.[2]
Selective inhibitors of BChE has been a major research area of our group. We have found a new family of
nucleosides embodying an unusual bicyclic sugar moiety (type I) that demonstrated potent and selective
inhibition of BChE.[3] With the goal of simplifying the structure of these nucleosides, new BChE
inhibitors with general structure type II and III were synthesized exhibiting also potent activity.[4] To
assess their biokinetics and potential interest as radioprobes for imaging BChE activity in AD or cancer
patients, the most promising compounds were successfully radioiodinated with 125I and biologically
evaluated. The results will be presented and discussed.
Figure 1 - General structures of nucleosides
Acknowledgements The authors thank Fundação para a Ciência e a Tecnologia for Vasco Cachatra PhD grant
(SFRH/BD/90359/2012) and for the support of the project UID/MULTI/00612/2013.
References
[1]
M. J. Ehret, K. W. Chamberlin, Clinical Therapeutics, 2015, in press.
K. Prabhu, D. Naik, S. Ray, Vadiraj, A. Rao, A. Kamath, Australas. Med. J., 2011, 4(7), 374.
[3]
F. Marcelo, F. M. V. Silva, M. Goulart, J. Justino, P. Sinaÿ, Y. Blériot, A. P. Rauter, Bioorg. & Med. Chem.,
2009, 17(14), 5106.
[4]
S. Schwarz, R. Csuk, A. P. Rauter, Org. Biomol. Chem., 2014, 12, 2446.
[2]
68
CQB-Day 2015
P55
THE PHENOL O–H BOND DISSOCIATION ENTHALPY
F. Agapito,a R. M. Borges dos Santos,a,b J. A. Martinho Simõesa
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal ({agapito,
rmsantos, jams}@fc.ul.pt), bInstitute for Biotechnology and Bioengineering Center for Molecular and Structural Biomedicine,
Universidade do Algarve, Campus de Gambelas, 8005-139 Faro, Portugal
Phenolic compounds, from both natural and synthetic sources, are ubiquitously present in human
nutrition.1 These compounds play a vital biological role as antioxidants (e.g. vitamin E), and several of
them possess anticarcinogenic activity.2 One key thermochemical property in the study of phenolic
compounds is their O–H bond dissociation enthalpy (BDE). The O–H BDE of phenol is particularly
relevant since the BDEs of more complex compounds are often anchored to this value.3 Over the years
this value has been the subject of lively debate, fueled by competing experimental and theoretical
predictions. While some sources recommend a value of ca. 371 kJ/mol for this BDE, 3 others posit a
significantly lower value of ca. 363 kJ/mol.4 In the present work we take on the task of reevaluating this
critical thermochemical property using state of the art high-accuracy quantum chemical methods.
Acknowledgements This work has been supported by Fundação para a Ciência e a Tecnologia (FCT), Portugal,
through project UID/MULTI/00612/2013 and a post doctoral grant awarded to F. Agapito
(SFRH/BPD/74195/2010).
References
1. M.-T. Huang, T. Ferraro Phenolic Compounds in Food and Cancer Prevention in Phenolic Compounds in Food
and Their Effects on Health II, M.-T. Huang, C.-T. Ho, C. Y. Lee, Am. Chem. Symp. Ser. No. 507. Washington,
DC, 1992.
2. F. M. F. Roleira, E. J. Tavares da Silva, C. L. Varela, S. C. Costa, T. Silva, J. Garrido, F. Borges Food Chem.
183 (2015) 235.
3. R.M. Borges dos Santos, J.A. Martinho Simões, J. Phys. Chem. Ref. Data 27, 707 (1998).
4. P. Mulder, H. Korth, D. Pratt, G. DiLabio, L. Valgimigli, G. Pedulli, K. Ingold J. Phys. Chem. A 109 (2005)
2647.
69
P56
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CHEMICAL STABILITYAND NEUROPROTECTIVE ACTIVITY EVALAUATION OF
ERICA AUSTRALIS L. EXTRACTS
P. Dias, A. Martins, A. P. Rauter, P. L. Falé,
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected]).
Erica australis L. (Ericaceae) is used in traditional medicine to treat many free-radical related ailments. In
the present work, the stability and biological activity of the plant aqueous extracts submitted to an in vitro
digestive process were investigated. Infusion and decoction chemical stability was monitored by HPLCDAD and LC-MS/MS, while their bioactivities were evaluated through the inhibition of AChE and DPPH
radical scavenging activity. Both extracts, whose main components were flavonol glycosides, inhibited
AChE, showing IC50 values lower than the ones found in the literature for many other plant aqueous
extracts [1] and several species used as Portuguese food spices [2]. Significant radical scavenging
activities were also presented by the infusion and by the decoction. After submission to gastric and
pancreatic juices, no remarkable alterations in the composition or in the bioactivities were observed,
suggesting that the extracts may pass through the gastrointestinal tract, keeping their composition and
therefore their biological properties. Our results support the potential of E. australis as a source of
functional ingredients with antioxidant and neuroprotective properties.
Acknowledgements This work was supported by Fundação para a Ciência e a Tecnologia
(UID/MULTI/00612/2013). The authors also thank the European Commission for approval of the
INOVAFUNAGEING commitment and the support of the project “PERsonalised ICT supported Service for
Independent Living and Active Ageing”, FP7-ICT-2013-10, Project Nr. 610359, 2013-2016
References
1. P. L. Falé, F. Amaral, P. J. A. Madeira, M. S. Silva, M. H. Florencio, F. N. Frazão, M. L. M. Serralheiro, Food
Chem Toxicol 50 (2012) 2656.
2. A. T. Mata, C. Proença, A. R. Ferreira, M. L. M. Serralheiro, J. M. F. Nogueira, M. E. M. Araújo, Food Chem
103 (2007) 778.
70
CQB-Day 2015
P57
SELECTIVE RECOVERY OF SILVER FROM DILUTE SOLUTIONS BY ELECTROLESS PRECIPITATION
USING POLYANILINE FILMS
I. J. Pereira, J. P. Correia
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected])
It is well known that the fully oxidized state of polyaniline (pernigraniline form) reduces to produce the
protonated emeraldine in acid medium at open circuit potential. It is so expected that in the presence of
metallic ions that can act as oxidizing species, spontaneous deprotonation occur with simultaneous
reduction of the metal ion - eventually to zero oxidation state (scheme 1) – continuing the process while
the polymer is exposed to the solution [1]. The high electrochemical potentials of noble metals make them
suitable for this electroless precipitation process in polyaniline (PAni) films [2]. This spontaneous,
selective (to noble metals) and sustained reduction of metal ions is of particular importance in the field of
extractive metallurgy [3].
In this work the process of electroless precipitation of silver from acidic dilute solutions of silver ions is
investigated. Thin PAni films were electrochemically synthesized on vitreous carbon and stainless steel
electrodes and exposed to 1 mM silver solutions for different periods at ambient temperature. The amount
of reduced metal in each experiment was assessed by atomic absorption spectroscopy. The effect of film
thickness and immersion time in the silver extraction efficiency was evaluated by optical microscopy and
electrochemical characterization of the pristine films and after exposure to the silver containing solutions.
It was observed that the amount of reduced silver increases with polymer thickening and the metal
presence doesn’t affect the electroactivity of the polyaniline film. The selectivity of the electroless
precipitation methodology to reduce noble metal ions was also evaluated by adding significant amounts
of engineering metal ions to the silver solutions.
Scheme 1
Acknowledgements This work is funded by FCT (Fundação para a Ciência e a Tecnologia) through the project
UID/MULTI/00612/2013
References
1. L.M. Abrantes, J.P. Correia, Mat. Sci. Forum 191 (1995) 235.
2. V. Lyutov, V. Tsakova, J. Solid State Electrochem. 15 (2011) 2553-2561
3. Y.P. Ting, K.H. Neoh, E.T. Kang, K.L. Tan, J. Chem. Tech. Biotechnol. 59 (1994) 31-36
71
P58
CQB-Day 2015
NEW SUGAR-BASED SURFACTANTS TARGETTING LIPID BILAYERS: SYNTHESIS AND MOLECULAR
DYNAMICS STUDY
R. Nunes, J. Pais, D. Vila-Viçosa, M. Machuqueiro, A. P. Rauter
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal [email protected])
The search for new antibiotics with innovative mechanisms of action represents a foremost challenge to
the scientific community owing to the increasing spread of bacterial resistance to the available
therapeutics. In this context, our research group is particularly interested in exploiting the potential
usefulness of sugar-based surfactants as antibacterial agents. These molecules are known for their
biocompatibility properties and feature low toxicity, making them suitable for several industrial and
medicinal applications. In previous work (1), we have reported a series of alkyl 2-deoxy/2,6-dideoxyarabino-hexopyranosides exhibiting antimicrobial activity against several pathogens including Bacillus
anthracis, which is considered a bioterrorism agent. Their synthesis has been efficiently accomplished by
reaction of a variety of alcohols with protected glycals in the presence of triphenylphosphane
hydrobromide, a procedure that stereoselectively delivers the bioactive α-anomers in high yields. The
antimicrobial activity is modulated by the deoxygenation pattern of the sugar moiety, and preliminary
studies indicate that these glycosides act through destabilization of bacterial cell membranes.
In this communication we present the synthesis and biological screening of a small library of alkyl
glycosides structurally related to the most active compound, including 2-deoxyglycosides derived from
pentoses, 2-fluorinated analogues and glycosides deoxygenated at 6-position of the sugar, which were
accessed by distinct methodologies starting from either glycals or glycosyl trichloroacetimidates. This
study led to several promising molecular entities, providing important insights into the key structural
features required for tuning their antimicrobial properties. Aiming at further contributing to the
elucidation of the mechanism of action for this family of compounds, molecular dynamics (MD)
simulations were performed in order to study their effect on model membranes. In particular, we studied
the partitioning of glycosides from micelles in solution at the interface of a phospholipid bilayer and
subsequently analyzed the structural properties of the bilayer and the molecular interactions involved in
lipid phase reorganization. These results will also be highlighted and discussed.
n
O
HO
R
O
HO
MD
X
Acknowledgements: The authors thank QREN-COMPETE program for financial support through project FACIB
(21547). FCT is gratefully acknowledged for funding CQB research unit (strategic project
UID/MULTI/00612/2013), as well as for the PhD grants SFRH/BDE/51957/2012 and SFRH/BD/81017/2011.
Thanks are also due to COST for funding through action CMST/COST-Action/CM1102.
References
1. A. Martins, M. S. Santos, C. Dias, P. Serra, V. Cachatra, J. Pais, J. Caio, V. H. Teixeira, M. Machuqueiro, M. S.
Silva, A. Pelerito, J. Justino, M. Goulart, F. V. Silva, A. P. Rauter, Eur. J. Org. Chem. 8 (2013) 1448.
72
MD
MD
CQB-Day 2015
P59
NOVEL TRIAZOLE, PURINE AND SULFONOHYDRAZIDE GLYCOCONJUGATES FOR THE TARGETING
OF NUCLEOTIDE-DEPENDENT ENZYMES: SYNTHESIS AND BIOACTIVITY EVALUATION
N. M. Xavier,a S. D. Lucas,b R. Jorda,c S. Schwarz,d A. Loesche,d R. Csukd
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, Ed. C8, 2º Piso, Campo Grande, 1749-016
Lisboa,: bInstituto de Investigaç ão do Medicamento, Faculdade de Farmácia, Universidade de Lisboa, Av. Prof. Gama Pinto,
1649-003 Lisboa,: cLaboratory of Growth Regulators, Palacký University & Institute of Experimental Botany ASCR, 78371
Olomouc, Czech Republic:;Bereich Organische Chemie, Martin-Luther-Universität Halle-Wittenberg, Kurt-Mothes-Str. 2, D06120 Halle (Saale), Germany ([email protected])
Nucleotide-dependent enzymes encompass a variety of enzymes which play crucial roles in fundamental
biological processes such as cell metabolism, cell division or nucleic acid synthesis, among others.
Enzymes of these type are frequently deregulated in diseases such as cancer or viral infections and hence
the development of mimetics of their nucleotide substrates may be a liable therapeutic strategy. 1 Among
these enzymes, cyclin-dependent kinases, which are ATP-dependent, regulate the cell cycle. Their
overexpression/overactivity play essential roles in tumor development, which turned them promising
therapeutic targets for cancer.2 Most CDK inhibitors are purine derivatives, which do not mimic
important ATP-CDK interactions, particularly those arising from the phosphate moiety.
In this communication we report on the synthesis of new nucleotide mimetics comprising an aromatic or a
N-heteroaromatic unit, such as a triazole unit, a sugar moiety and a sulfonohydrazide functionality. These
molecules were aimed to target CDK-2, whose inhibition potentially blocks tumor cell proliferation with
no effect on normal cells.3 In addition, a new structural framework for potential mimetics of nucleoside
diphosphate sugars, which are substrates for enzymes such as glycosyltransferases,4 was designed. The
synthesis of an analog of diphosphate-linked disaccharides, containing an uncharged surrogate of a
diphosphate system, and that of a purine nucleoside derivative, was carried out.
The synthetic work and the results of the molecular docking studies, which contributed to identify
promising CDK-2 inhibitors, are revealed. Results on the evaluation of the biological profile of the new
compounds, namely their antiproliferative activity on tumour cells and their inhibitory effects on CDK-2
as well as on other enzymes of therapeutic relevance, such as cholinesterases and carbonic anhydrase-II,
will also be presented.
Acknowledgements: FCT is acknowledged for funding through the FCT Investigator Program to N.M. Xavier and
to S.D. Lucas (projects IF/01488/2013 and IF/00472/2014) and for financial support of the strategic projects
UID/MULTI/00612/2013 and Pest-OE/SAL/UI4013/2014.
References
1. L. P. Jordheim, D. Durantel, F. Zoulim, C. Dumontet, Nat. Rev. Drug. Discov. 12 (2013) 447-64.
2. S. Lapenna, A. Giordano, Nat. Rev. Drug Discov. 8 (2009) 547-566
3. G. I. Shapiro, J. Clin. Oncol. 24 (2006) 1770-1783.
4. L.L. Lairson, B. Henrissat, G.J. Davies, S.G. Withers., Annu. Rev. Biochem. 77 (2008) 521-55
73
P60
CQB-Day 2015
NOVEL N-GLYCOSYLSULFONAMIDES DERIVED FROM D-RIBOSE AND D-GLUCURONAMIDE:
SYNTHESIS AND CONFORMATIONAL STUDIES
A. Fortuna, P. J. Costa, N. M. Xavier
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, Campo Grande,1749-016 Lisboa, Portugal
([email protected])
The sulfonamide moiety is of particular relevance in medicinal chemistry since it is present in many
bioactive molecules, some of them exhibiting antitumor, anti-inflammatory and antibacterial activities.1
In particular, sugar derivatives comprising sulfonamide functions, namely anomeric sulfonamides, have
shown interesting biological properties, such as antitumor effects.2,3 N-Glycosylsulfonamides constitute
rather stable N-glycosyl analogs of O-glycosides when compared with the majority of glycosylamines,
whose stability is normally low.
These aspects prompted us to focus on the synthesis of novel N-glycofuranosyl and N-glycopyranosyl
sulfonamides (Figure 1), containing ribosyl and glucuronamide-derived moieties, to study the
stereochemical and conformational outcome of the reactions involved in their synthesis and the possible
pyranose/furanose equilibration in ribosyl derivatives.
Conformational analyses were performed by a combination of Molecular Dynamics and DFT calculations
in order to understand the effect of the structure of the sugar moiety on the conformation adopted by the
target molecules. The synthetic work and the insights obtained by the conformational analysis studies will
be presented.
Figure 1.General structure of the target molecules.
Acknowledgements ‘Fundação para a Ciência e Tecnologia’ is acknowledged for funding (IF/01488/2013,
IF/00069/2014 and CQB strategic project UID/MULTI/00612/2013).
References
1. S. S. A. Shah, G. Rivera, M. Ashfaq, Mini Rev. Med. Chem. 13 (2013) 70-86.
2. M. Lopez, B. Paul, A.Hofmann et al., J. Med. Chem. 52 (2009) 6421-6432.
3. J.-Y. Winuma, P. A. Colinas, C. T. Supuran, Bioorg. Med. Chem. 21 (2013) 1419-1426.
74
CQB-Day 2015
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SYNTHESIS AND BIOACTIVITY EVALUATION OF PURINE AND PYRIMIDINE 6´-ISONUCLEOSIDES
D. Batista,a R. Jorda,b S. Schwarz,c A. Loesche,c R. Csuk,c N. M. Xaviera
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, Ed. C8, 2º Piso, Campo Grande, 1749-016
Lisboa,;Laboratory of Growth Regulators, Palacký University & Institute of Experimental Botany ASCR, 78371 Olomouc, Czech
Republic:cBereich Organische Chemie, Martin-Luther-Universität Halle-Wittenberg, Kurt-Mothes-Str. 2, D-06120 Halle (Saale),
Germany
([email protected]; [email protected])
Isonucleosides, regioisomers of nucleosides in which the nucleobase is linked to the carbohydrate moiety
at a non-anomeric position, are among the nucleoside analogs that have attracted much interest, due to
their propensity to display antiviral and antitumor activities.1,2 This bioactivity profile arise from their
ability to interfere in biological processes in which natural nucleosides are involved, such as nucleic acid
synthesis and cell division, which are deregulated in diseases such as cancer or viral infections.[3,4] In
adition, such molecules present better stability towards enzymatic hydrolysis that their natural
counterparts.
The reported isonucleosides mostly encompass furanosyl derivatives comprising the nucleobase at C-2 or
C-3. Thus, we were motivated to explore the access to new types of isonucleosides, based on
hexopyranosyl units linked through C-6 to the nucleobase.
In this communication, the synthesis of 6´-isonucleosides (Figure 1), embodying purine or pyrimidine
motifs is presented. The synthetic approach was based on the Mitsunobu coupling of partially protected
glycosides containing a free OH-6 with a nucleobase. Variations on the substitution and configuration of
the sugar moiety were made, extending the panel of compounds for further bioactivity screening.
Pyrimidine nucleobases, such as uracil, allowed the access to disaccharide isonucleosides.
The compounds were subjected to biological evaluation, focusing firstly their cytotoxicity to tumor and to
healthy cells. To have a broader knowledge on the molecules’ bioactivity potential, their ability to inhibit
cholinesterases was further studied. The synthetic work and the results of the biological assessment will
be disclosed.
Figure 1. General structure of the synthesized 6´-isonucleosides.
Acknowledgements The authors thank Fundação para a Ciência e a Tecnologia for funding through the projects
IF/01488/2013 and UID/MULTI/00612/2013 and for the research grant to Daniela Batista.
References
1. V. Nair, Antiviral isonucleosides: discovery, chemistry and chemical biology. In Recent Advances in
Nucleosides: Chemistry and Chemotherapy (Ed. C.K. Chu), Elsevier, 2002, 149-166.
2. H.-W. Yu, H.-Y. Zhang, Z.-J. Yang et al., Pure Appl. Chem. 70 (1998) 435-438.
3. C. M. Galmarini, J. R. Mackey, C. Dumontet, Lancet Oncol., 3 (2002) 415–424;
4. L. P. Jordheim, D. Durantel, F. Zoulim, C. Dumontet, Nat. Rev. Drug. Discov. 12 (2013) 447-464.
75
P62
CQB-Day 2015
BIOMASS-DERIVED ACTIVATED CARBONS: PROMISING GREEN MATERIALS FOR
ENVIRONMENTAL REMEDIATION
M. A. Andradea,b, A. S. Mestrea, C. O. Aniab, A. P. Carvalhoa
a Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal b ADPOR
Group, Instituto Nacional del Carbón, INCAR – CSIC, 33080, Oviedo, Spain ([email protected])
Lignocellulosic biomass is a renewable resource that can be used to produce energy by several
technologies, such as pyrolysis, liquefaction, and gasification. Among all biomass conversion processes,
hydrothermal carbonization (HTC) is a technology that enables the preparation of carbon materials, being
widely used due to its simplicity, low-cost, greenness and high efficiency.1,2
In this context, the goals of this work were to evaluate the effect of an initial hydrothermal carbonization
(HTC) in the properties of the activated carbon obtained by further chemical activation with K2CO3 and to
investigate the ability of the prepared carbons for the adsorption of caffeine and paracetamol.
The K2CO3 activation of the sisal-derived hydrochar allows to obtain activated carbons with better
developed porous structures than those obtained by direct activation of sisal. Although the use of a twostep procedure increases the time needed to obtain these materials, the advantages can be reflected in a
higher adsorption capacity and/or affinity towards caffeine and paracetamol, (performance compares
favourably with that of the commercial carbon). Based on these promising preliminary results, this work
points out that sisal-derived hydrochars are excellent precursors for the preparation of low-cost
nanoporous carbons with superior performance in the adsorption of pharmaceutical compounds from
aqueous media.
(a)
(b)
Figure 1. a) SEM micrograph of a sisal-based hydrochar; (b) Caffeine adsorption isotherms at 30 ºC on the activated
carbons selected: NS-commercial carbon; SHC-carbon obtained by chemical activation of a sisal-derived hydrochar;
SC- carbon obtained by direct activation of sisal. Symbols correspond to the experimental data, dotted lines to the
fitting to the Langmuir equation. Error bars are included.
Acknowledgements The authors acknowledge the financial support of FCT and MINECO (projects
UID/MULTI/00612/2013 and grant CTM2011/23378, respectively). MAA and ASM thank FCT for a PhD and a
Post-doc grant, SFRH/BD/71673/2010, SFRH/BPD/86693/2012.
References
1. Titirici, M.M., White, R.J., Falco, C. Sevilla, M. (2012) “Black perspectives for a green future: hydrothermal
carbons for environment protection and energy storage”, Energy Environ. Sci., 5, 6796–6822.
2. Hoekman, S.K., Broch, A., Robbins, C. (2011) Hydrothermal Carbonization (HTC) of Lignocellulosic Biomass”,
Energy Fuels, 25, 1802–1810.
76
CQB-Day 2015
P63
C-GLUCOSYLATION REACTIONS IN THE SYNTHESIS OF NEW SUGAR-LINKED MOLECULES WITH
POTENTIAL ANTI-AMYLOID ACTIVITY
A. M. Matos, A. P. Rauter
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected]).
Type 2 diabetic patients have a 2-fold higher risk of developing Alzheimer’s disease, thus being urgent to
search for a new antidiabetic therapy that is simultaneously able to prevent neurodegeneration. 8-β-Dglucosylgenistein, the major component of Genista tenera ethyl acetate extract, was found to possess a
notable antidiabetic activity in STZ-induced diabetic Wistar rats and to interact in vitro with both hIAPP
and Aβ1-42 toxic oligomers, pointing towards a multitarget mechanism of action. Our latest STD-NMR
studies highlighted the importance of the sugar moiety for its anti-amyloid activity, and reinforced the
already expected importance of both aromatic rings [1].
A C-glucosylphenolic 8-β-D-glucosylgenistein analogue scaffold was rationally designed with the support
of molecular modelling analyses, in the light of the structural requirements found for the lead compound,
aiming to achieve new bioactive molecular entities in fewer synthetic steps and using simpler
experimental methodologies. The TMSOTf-promoted reactions that gave rise to the C-glucosylphenols
used as precursors in this approach will be disclosed and discussed, together with the importance of the
sugar protecting groups for the success of the C-glycosylation step. Ultimately, structure-activity
relationships carried out with new analogues will provide more detail on key structural features for
activity optimization and will constitute a step forward in the investigation on innovative amyloiddisrupting therapeutic strategies against diabetes and Alzheimer’s disease.
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OR1
L
OH
O
O
HO
R1O
HO
OR1
OH
OH
OR2
Figure 1. TMSOTf-promoted synthesis of protected C-glucosylphenols for subsequent derivatization.
R1 = protecting group; L = leaving group; R2 = acyl group.
Acknowledgements The authors thank Fundação para a Ciência e a Tecnologia (FCT) for financial support of the
PhD grant SFRH/BD/93170/2013, and of the projects UID/MULTI/00612/2013 and PEST
UID/MULTI/00612/2013.
References
1. Jesus A.R., Dias C., Matos A.M., Almeida R.F.M., Viana A.S., Marcelo F., Ribeiro R.T., Macedo M.P., Airoldi
C., Nicotra F., Martins A., Cabrita E.J., Jimenez-Barbero J., Rauter A.P. J. Med. Chem. 2014, 57(22), 9463-9472.
77
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CQB-Day 2015
ALKYL DEOXY GLYCOSIDES AS ANTIMICROBIAL AGENTS: SYNTHESIS, SURFACE PROPERTIES
AND MECHANISM OF ACTION
C. Dias,a J. P. Pais,a P. Serra,a A. Almeida,a R. F. M. Almeida,a A. Martins,a M.S.C.S. Santos,a
A.S. Viana,a A. Pelerito,b A. P Rautera
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected]). bInstituto Nacional de Saúde Doutor Ricardo Jorge, Avenida Padre Cruz, 1749-016 Lisboa, Portugal
The search for new antimicrobial drugs is currently one of the major ongoing research areas due to the
spread of multidrug-resistance, making research on new antibacterial agents with new mechanisms of
action highly relevant. Another matter of concern is the fact that infections in elderly populations are
known to be not only more frequent but also more severe, being this susceptibility often related to
neurodegenerative diseases such as dementia and Alzheimer’s [1]. Alkyl 2-deoxy/2,6-dideoxy-arabinohexopyranosides with a potent antimicrobial activity in several Bacillus species have been previously
described by our research group [2, 3]. Moreover, promising preliminary results arising from interaction
studies of some of these 2,6-dideoxyglycosides with cystatin B amyloid fibrils, assessed by NMR
spectroscopy, show their potential for neurodegenerative diseases as well.
We present now the synthesis and the antimicrobial activity of related glycosides differing in the glycon
structure (D- and L-series, hexo- and pentopyranosides, arabino and threo configuration) and in the
lipophilic chain (chain size, fluorinated chain, S-linked chain, amide containing chain), for the recognition
of the structural features that determine the selectivity for Bacillus species. The synthetic approach is
based on the reaction of glycals with alcohols catalysed by triphenylphosphane hydrobromide. Moreover,
aiming at a better understanding of the importance of the deoxygenation pattern, synthetic methodologies
towards new alkyl 3-deoxy, 4-deoxy and 6-deoxy glycosides were also investigated.
The surface properties of the most active compounds, in terms of adsorption and aggregation parameters,
were assessed and suggested that surface activity is required for the bioactivity. However, one of the most
promising surface active glycosides was not active. In order to have insights into the relationship between
surface activity and bioactivity of this family of compounds expected to target bacterial membranes,
collaborative work based on biophysical methods for the study of the interaction of these molecules with
lipossomes was encouraged.
This work clearly demonstrates the uniqueness of carbohydrates which stereochemistry and chemical
structure can tune the bioactivity exhibited by stereoisomers.
Acknowledgements This work was supported by FEDER-QREN-SI I&DT, Co-Promotion Project nr. 21457. The
authors thank the FCT for financial support (PEST UID/MULTI/00612/2013), and FCT and Cipan are also
gratefully acknowledged for the Ph.D. grants of C. Dias (SFRH/BDE/51998/2012) and J. P. Pais
(SFRH/BDE/51957/2012).
References
3. G. Gavazzia, K.-H. Krause. Lancet Infec. Dis. 2 (11) (2002) 659.
4. A. P. Rauter, S. Lucas, T. Almeida, D. Sacoto, V. Ribeiro, J. Justino, A. Neves, F. V. M. Silva, M. C.
Oliveira, M. J. Ferreira, M. S. Santos, E. Barbosa. Carbohydr. Res. 340 (2005) 191.
5. F. Silva, M. Goulart, J. Justino, A. Neves, F. Santos, J. Caio, S. Lucas, A. Newton, D. Sacoto, E. Barbosa, M. S.
Santos, A. P. Rauter. Bioorg. Med. Chem. 16 (2008) 4083.
78
CQB-Day 2015
P65
GLYCOLIPIDS: ANTIBACTERIAL ACTIVITY AND INSIGHTS INTO MECHANISM OF ACTION VIA
GENETIC AND METABOLIC APPROACHES
J. Pais,a,b A. S. Viana,a A. P. Rauter,a L. Sobral,b R. Dias,b R. Tenreirob
aCentro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal;
Instituto de Biosistemas & Ciências Integrativas, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa,
Portugal;([email protected])
b
In the present work, the study of a new family of 2,6-dideoxy alkyl glycosides is presented, namely their
antibacterial properties against Bacillus species and the mechanism of action associated.
The compounds were synthesized using a fairly simple methodology, involving the synthesis of glycals,
in particular that derived from L-rhamnal, and further conjugation with an aliphatic chain. The lead
compound (dodecyl 2,6-dideoxy-α-L-arabino-hexopyranoside) (Fig. 1), was submitted to several
biological assays, in order to understand the mechanism of action (MoA). Multiple approaches were used,
dived mainly in a genetic approach and a metabolic one. Genetic dissection was carried out by testing
several mutant libraries by random transposition and knock-out of specific membrane related targets,
using as a model strain B. cereus ATCC 14579. Also, the compounds impact on the bacterial vitality,
viability and bacterial metabolism (reconstruction using phenotypic microarrays Biolog®) as well as the
effects of the compound in the bacterial sporulation cycle and in different cellular ultra-structures,
assessed using protoplasts and spheroplasts, provided essential insights into its mechanism of action.
Finally, imaging techniques, namely atomic force microscopy and fluorescence microscopy allowed to
observe and verify the compounds biological activity.
1'
O
5
H3C
HO 6
4
3
OH
1
O
3'
2'
5'
4'
7'
6'
9'
8'
11'
10'
2
Figure 1. Dodecyl 2,6-dideoxy-α-L-arabino-hexopyranoside
79
CH3
12'
P66
CQB-Day 2015
SYNTHESIS AND IN VITRO STUDY OF C-GLUCOSYL DIHYDROCHALCONES AS POTENTIAL
SODIUM-GLUCOSE CO-TRANSPORTER (SGLT) INHIBITORS FOR THE TREATMENT OF DIABETES
A. R. Jesus,a,b T. Doreb, A. P. Rautera
a
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, Campo Grande, 1749-016 Lisbon,
Portugal,; b Saadiyat Campus, Experimental Research Building, New York University Abu Dhabi, Saadiyat Island, Abu Dhabi,
United Arab Emirates ([email protected], [email protected])
In our body, glucose is transported into the cells by a family of proteins called glucose transporters. There
are two sub-families, GLUT and sodium-glucose co-transporters (SGLTs). SGLT1 and SGLT2, two
isoforms of SGLTs, are responsible for > 99% of the reabsorption of glucose in the kidneys and
intestines.1 For the past couple of years scientists have been studying the inhibition of SGLTs as a
treatment for diabetes as the inhibition of this reabsorption process would be advantageous for diabetic
patients. However, SGLT1 has also affinity to galactose and its inhibition leads to gastrointestinal side
effects. To avoid this problem, scientists have been searching for selective SGLT2 inhibitors in the last
couple of years.2
Phlorizin, a dihydrochalcone glucoside, was the first SGLT1/SGLT2 inhibitor, discovered in 1835. 3 This
compound did not go to clinical trials due to its SGLT1 inhibition and also because it is hydrolyzed by
gastrointestinal enzymes. However, C-glycosyl derivatives are stable against them.
In this work we present the synthesis of a small library of C-glucosyl analogs of phlorizin as new
potential SGLT2 inhibitors (Scheme 1). Their synthesis comprises the preparation of the glycosyl donor
(2,3,4,6-tetra-O-benzyl-D-glucopyranose) and the glycosyl acceptors (dihydrochalcones) followed by the
C-glycosylation of dihydrochalcones in the presence of catalytic amount of TMSOTf, benzyl groups’
removal with Pd/C and Et3SiH by in situ hydrogenation.
Cell viability revealed that none of these C-glucosyl compounds are cytotoxic in HEK293 cell line (cell
viability > 95%). The aglycones (chalcones and dihydrochalcones) showed, in some cases, a slightly
higher cytotoxicity with 60-70% cell viability after 24 h incubation.
Scheme 1. Synthesis of C-glucosyl dihydrochalcones. i) TMSOTf (0.5 equiv.), DCM/ACN, Drierite (40-60% yield);
ii) Pd/C, Et3SiH, EtOAc/MeOH (90-99% yield).
Glucose uptake assays are being currently carried out but preliminary results showed that the C-glucosyl
dihydrochalcones are active against both SGLT1 and SGLT2 proteins but with a slightly higher
selectivity for SGLT2 (> 500-fold). The aglycones chalcones and dihydrochalcones inhibit both proteins
as well, but with no selectivity (< 10-fold). These also seemed to inhibit GLUT, while the C-glucosyl
compounds do not affect GLUT proteins.
References
(1) Wright, E. M.; Loo, D. D.; Hirayama, B. A. Physiological reviews 2011, 91, 733.
(2) Wright, E. M.; Turk, E. Pflugers Archiv : European journal of physiology 2004, 447, 510.
(3) Ehrenkranz, J. R.; Lewis, N. G.; Kahn, C. R.; Roth, J. Diabetes/metabolism research and reviews 2005, 21, 31
80
CQB-Day 2015
P67
PREPARATION OF SPHERICAL NANOPOROUS CARBONS BY K2CO3 ACTIVATION OF BIOMASS ACID
LIQUORS DERIVED CHARS
F. Hesse,a,b A. S. Mestre,a,c C. Freire,c C. O. Ania,d A. P. Carvalhoa
a Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal b Institut für
Anorganische Chemie, Christian-Albrechts-Universität zu Kiel, Max-Eyth-Strasse 2, D-24118 Kiel, Germany c REQUIMTE,
Departamento de Química e Bioquímica, Faculdade de Ciências, Universidade do Porto, 4169-007 Porto, Portugal d ADPOR
Group, Instituto Nacional del Carbón, INCAR – CSIC, 33080, Oviedo, Spain ([email protected])
The search for new methodologies to obtain valuable nanoporous carbons from biomass residues is a hot
research topic in material science. In this sense, several approaches have been assayed as it is the case of
hydrothermal treatment of carbohydrates which allows the preparation of microspherical carbons with an
incipient pore structure. To develop the porosity and retain the spherical morphology, the choice of the
activating agent is crucial. In a recent study we proved the potentialities of K 2CO3 to obtain spherical
activated carbons with tailored microporosity from a sucrose-derived biochar.1,2 The use of
lignocellulosic biomass as source for carbohydrate units is however a more sustainable synthetic route
and literature studies prove that activated carbon materials can been prepared from acid digestion of
biomass followed by KOH-activation.3
In this study, a new more sustainable method was applied using sisal residues (rejects of rope industry)
and corn stalks as precursors. The first step was inspired in ref. 3 and consisted in the extraction of the
saccharic units present in the biomass precursors by digestion with H2SO4 followed by a
polycondensation reaction of the saccharic acid solution at 90 C. The obtained biochars were then
activated with K2CO3 (biochar:K2CO3 weight ratio of 1:3, heating at 800C for 1h under N2 flow of 5
cm3s-1).1 While the biochars have an incipient porous network (S72 in Fig. 1(a)) the carbons obtained are
mainly microporous solids with apparent surface areas reaching 1750 m2g-1, and total pore volumes
between 0.65 and 0.84 cm3g-1. The results of CO2 adsorption proved that in all the cases monomodal
micropore size distributions centered in widths between 0.5 and 0.7 nm are obtained (Fig.1(b)). SEM
micrographs (Fig. 1(c)) reveal that our activation procedure allowed the preparation of nanoporous
carbons with spherical morphology, with particle diameters between 50 and 100 nm. DRIFT data show
that the surface chemistry of the biochar is rich is oxygen surface groups that disappear after activation.
Preliminary studies reveal that these porous solids are promising adsorbents for pharmaceutical
compounds and their spherical morphology and high thermal stability allows also to envisage their use for
the synthesis of functional carbon-based materials.
(a)
(b)
Differential volume / cm3 g-1 nm-1
20
nads / mmol g-1
(c)
0.14
25
15
CS48/800
S48/800
CS72/800
S72/800
S72
10
5
CS48/800
S48/800
CS72/800
S72/800
S72
0.12
0.10
0.08
0.06
0.04
0.02
0.00
0
0.30
0
0.2
0.4
p/p0
0.6
0.8
1
0.80
1.30
Pore width / nm
1.80
500 nm
Figure 1. (a) N2 isotherms at -196 ºC; (b) Micropore size distributions, obtained by fitting CO2 adsorption data at 0 ºC to the
method described in [4] (c) SEM image of the activated carbon obtained from sisal treated with H 2SO4 at 48% (sample S48/800).
Acknowledgements The authors thank the Fundação para a Ciência e Tecnologia, Portugal, for financial support (Project PEstOE/QUI/UI0100/ 2014) and a fellowship to ASM (SFRH/BPD/86693/2012). Cordex, is also acknowledged for providing the
sisal wastes.
References
1. A. S. Mestre, C. Freire, J. Pires, A. P. Carvalho, M. L. Pinto, J. Mat. Chem .A, 2 (2014) 15337
2. A.S. Mestre, E. Tyszko, M A. Andrade, M. Galhetas, C. Freire, A. P. Carvalho, RSC Adv. 5 (2015) 19696
3. L. Wang, Y. Guo, Y. Zhu, Y. Li, Y. Qu, C. Rong, X. Ma, Z. Wang, Bioresour. Technol. 101 (2010) 9807
4. M. L. Pinto, A. S. Mestre, A. P. Carvalho, J. Pires, Ind. Eng. Chem. Res., 49 (2010) 4726
81
P68
CQB-Day 2015
HIERARCHICAL ZEOLITES: A GREEN ALTERNATIVE TO CONVENTIONAL FRIEDEL-CRAFTS
ACYLATION OF HETEROAROMATICS
R. Aleixo,a N. Nunes,a,b R. Leitão,a,b F. Martins,b A. P. Carvalho,b A. Brigas,c A. Martinsa,b
a Área
Departamental de Engenharia Química, ISEL, IPL, R. Conselheiro Emídio Navarro, 1959-007 Lisboa, Portugal. b Centro
de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016, Lisboa, Portugal ([email protected],
[email protected]). c Departamento de Química Farmacêutica, UAlg, Campus da Penha, Estrada da Penha, 8005-139,
Faro, Portugal.
Friedel-Crafts acylation is an important industrial reaction to produce reaction intermediates of
pharmaceuticals, fragrances, dyes, flavours and agrochemical products. Traditionally, this reaction is
performed at high temperatures with large amounts of homogenous catalysts, such as AlCl3 and FeCl3 that
are harmful to the environment. Zeolites are proposed as alternative catalysts 1. However, the purely
microporous nature of these materials limits its application due to mass transfer limitations and
accessibility of active sites. The use of hierarchical zeolites, possessing two levels of porosity, the native
micropores and an additional mesopore system, is a promising solution.
In this study, three commercial zeolite structures, BEA (Si/Al=12.5), MOR (Si/Al=10) and MFI
(Si/Al=15), were submitted to desilication treatments with NaOH using previously optimized protocols. 2
In some cases, a subsequent acid treatment with HCl was made. The XRD diffraction patterns showed
that the modified materials preserve the crystallinity whereas the low temperature N2 adsorption
isotherms revealed the development of mesoporosity. The catalytic behaviour was investigated in the
acylation of furan by acetic anhydride (molar ratio 1:5), using 150 mg of zeolite sample, at 60 ºC. The
reaction mixture was analysed by GC and the results are expressed as yields of 2-acetylfuran vs. reaction
time. When comparing the three zeolite structures, higher yields were obtained for BEA. Fig. 1 shows the
catalytic results obtained for commercial (BEA), desilicated (BEA_D) and desilicated + acid treated
(BEA_D/AT) samples. The results obtained show that the desilication treatment is not enough to improve
the catalytic behaviour (BEA_D) due to possible deposition of extra-framework species deposited at the
pore mouths of the zeolite. These species are removed upon acid treatment (BEA_D/AT) improving the
mass transfer and the access to the active sites, with consequent higher yield of 2-acetylfuran.
100
Yield (%)
80
60
40
BEA
BEA_D
BEA_D /AT
20
0
0
10
20
30
time (min)
40
50
Figure 1 – Yield of 2-acetylfuran vs. reaction time.
Acknowledgements: We thank FCT for financial support under PEst UID/MULTI/00612/2013.
References
1. V.F.D. Álvaro, A. Brigas, E.G. Derouane, J.P. Lourenço, B.S. Santos, J. Mol. Catal. A: Chem. 305
(2009) 100.
2. J.C. Groen, L.A.A. . Peffer, J.A. Moulijn, J. Pérez-Ramírez, Microporous and Mesoporous Mater. 69 (2004) 29.
82
CQB-Day 2015
P69
REMOVAL OF MICROCYSTIN-LR BY CORK-BASED AND COMMERCIAL ACTIVATED CARBONS
C. Menezes,a S. José,a P. Pereira,a S. P. Silva,b A. P. Carvalhoc, A. S. Mestrec
a Laboratório de Biologia e Ecotoxicologia, Departamento de Saúde Ambiental, Instituto Nacional de Saúde Doutor Ricardo
Jorge, Avenida Padre Cruz, 1649-016 Lisboa, Portugal b Corticeira Amorim, S.G.P.S., S.A., Rua do Ribeirinho, 202, 4536-907
S. Paio de Oleiros, Portugal; c Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016
Lisboa, Portugal ([email protected])
Cyanotoxins and mainly microcystin-LR (MCLR) is a potent and ubiquitous freshwater toxin that poses a
risk for human and animal health. According to the Guidelines for Drinking Water Quality published by
WHO in 2011, the maximum concentration of MCLR allowed in water for human consumption is 1 g L1
. The methods currently used to achieve effective MCLR removal are highly costly and so, the search for
new, cost effective adsorbents to be incorporated in the water treatment technologies is an important issue
that has been addressed by the scientific community.
The present study is focused in this thematic being the first description of the application of industrial pretreated cork derived activated carbons for the removal of MCLR. The material selected for the study was
steam activated at 800 C, presenting a mesopore (0.22 cm3g-1) and micropore (0.28 cm3g-1) network.1
Commercial activated carbons with different textural characteristics were also tested.
Batch kinetic adsorption assays were performed in controlled conditions of temperature, light and
agitation, and MCLR was quantified by HPLC-DAD. For MCLR concentrations between 0.5 and 8 g
cm-3, after 1h of contact time the cork derived material presents removal efficiencies > 90 % (Fig. 1(a)).
These results are identical to those of the best commercial samples (Fig. 1(b)), highlighting the influence
of the mesopore network in the adsorption process of MCLR.
The successful removal of MCLR from aqueous solutions with cork derived activated carbons may
constitute a cost efficient approach to water treatment and an alternative to commercially available
activated carbons.
(a)
(b)
Figure 1. (a) Removal efficiencies obtained using 0.5mg/mL cork activated carbon at 2, 4 and 8µg/mL MCLR; (b)
Removal efficiencies obtained using 1mg/mL commercial activated carbons at 1µg/mL MCLR.
Acknowledgements Agência de Inovação (AdI) – QREN is acknowledge for research funding grants attributed to
CM, SJ and ASM in the scope of the WaterCork project– Valorization of subproducts from the cork industry in the
removal of organic contaminants in waters. This work was also supported by FCT pluriannual program of CQB
through strategic project UID/MULTI/00612/2013. ASMestre acknowledge FCT for the Post-Doc grant
SFRH/BPD/86693/2012.
References
1. A.S. Mestre, R.A. Pires, I. Aroso, E.M. Fernandes, M.L. Pinto, R.L. Reis, M.A. Andrade, J. Pires, S.P. Silva, A.P.
Carvalho, Chem Eng J, 253 (2014) 408
83
CYSTOSEIRA TAMARISCIFOLIA AS A POTENTIAL BIOMEDICAL SOURCE FOR ANTIOXIDANT AND
ANTI-HEPATOCARCINOMA COMPOUNDS
C. Vizetto-Duartea, L. Custódioa, J. H. G. Lagob, T. R. Moraisb, C. B. de Sousaa, K. N. Gangadhara,c,
L. Barreiraa, F. Albericiod, A. P. Rautere, J. Varelaa
aCentre of Marine Sciences, University of Algarve, Faculty of Sciences and Technology, Ed. 7, Campus of Gambelas, 8005-139
Faro, Portugal ([email protected]) bInstitute of Environmental, Chemical and Pharmaceutical Sciences, Federal University of
Sao Paulo, 09972-270, Sao Paulo, Brazil cInstituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Avenida
da Republica, 2780-157 Oeiras, Portugal dInstitute for Research in Biomedicine, Barcelona Science Park, Baldiri Reixac 10,
08028, Barcelona, Spain eCenter of Chemistry and Biochemistry, Department of Chemistry and Biochemistry, Faculty of
Sciences University of Lisbon, Campo Grande, Ed. C8, Piso 5, 1749-016 Lisbon, Portugal
Organic extracts (hexane, diethyl ether, ethyl acetate and methanol) from three Cystoseira species (C.
humilis, C. tamariscifolia and C. usneoides) were evaluated for their radical scavenging activity (RSA)
against 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid
(ABTS) radicals, and cytotoxic activity against a human hepatocarcinoma cell line (HepG2 cells). C.
tamariscifolia extracts had the highest RSA and cytotoxic activity (IC50 = 2.31 µg/mL for C.
tamariscifolia hexane extract). C. tamariscifolia hexane extract was further evaluated in four human
tumoural cell lines (cervical adenocarcinoma: HeLa; gastric adenocarcinoma: AGS; colorectal
adenocarcinoma: HCT-15; neuroblastoma: SH-SY5Y), and two non-tumoural cell lines (murine stromal
cells: S17 and human umbilical vein endothelial cells: HUVEC). This extract strongly and selectively
reduced the viability of all cell lines, especially when compared with HUVEC cells [IC 50 = 29.04 µg/mL
and selectivity index (SI) = 31.2]. In addition, it was capable to inhibit cell proliferation (decrease of
BrdU incorporation from 25.8% to 13.6%) and had pro-apoptotic effects in HepG2 cells. C.
tamariscifolia hexane extract was submitted to a bio-guided fractionation leading to the chemical
characterization of a cytotoxic fraction whose major component was identified as demethoxy cystoketal
chromane [1], a derivative of the meroditerpenoid cystoketal.
16
15
14
17
18
13
3'
1
12
2'
10
11
2
4'
9
3
5'
1'
5
6'
4
20
7
8
6
19
Figure 1 - Demethoxy cystoketal chromane.
Acknowledgements The authors would like to acknowledge SEABIOMED (PTDC/MAR/103957/2008) and
XtremeBio projects (PTDC/MAR-EST/4346/2012) funded by the Foundation for Science and Technology (FCT).
Vizetto-Duarte C. is a FCT doctoral research fellow (SFRH/BD/81425/2011) as well as Bruno de Sousa C.
(SFRH/BD/78062/2011) and Katkam N. Gangadhar is a post-doctoral research fellow (SFRH/BPD/81882/2011).
Custódio L. was supported by FCT Investigator Programme (IF/00049/2012). Morais T. R. and Lago J. H. G.
acknowledge CAPES and CNPq, respectively, for scientific research support.
References
1. Valls R., Mesguiche V., Piovetti L., Prost M., Peiffer G. Meroditerpenes from the brown alga Cystoseira
amentacea var. stricta collected off the French mediterranean coast. Phytochemistry, 1996; 41: 1367-1371.
84
CQB-Day 2015
P71
BIOMASS CONVERSION INTO CARBON BIOSORBENTS
P. S. de Velasco,a A. S. Mestre,b V. Hernández,a A. P. Carvalhob
a Departamento de Ingeniería Química y Bioquímica, Instituto Tecnológico de Aguascalientes, 20256 Aguascalientes, México b
Centro de Química e Bioquímica, Faculdade de Ciências, Universidade de Lisboa, 1749-016 Lisboa, Portugal
([email protected])
The search of low-cost and environmentally friendly technologies to produce carbon materials from
renewable sources has received great attention from scientific community. In this context hydrothermal
carbonization is a relatively new way to convert biomass into higher value carbonaceous materials.1
The biomass used in this study was sisal which is an autoctone plant from Mexico, being the strongest
vegetal fiber known. Its main use is for rope manufacture that produces a large amount of residues that
can be re-used to produce ropes of low quality or as fuel. In our group we have been interested in
transform this residue into more valuable material, i.e. carbons materials that can be successfully applied
as adsorbents and catalysts supports.
In this study, we explored the possibility of performing hydrothermal carbonization of the acid liquor
obtained after digestion of discarded sisal ropes with H2SO4. This protocol is a modification of the
methodology followed by Wang et al., 2010 aiming a faster and more energetically efficient process.2
The results obtained (Fig. 1(a)) proved that if hydrothermal carbonization is used, the 6 h treatment of the
acid liquor at 90 C and atmospheric pressure, mentioned in the literature protocol, can be shortened to 3
h, with no loss of yield. No increased yield was observed when the liquor was autoclaved at 120 C for 3
and 6h. The FTIR spectra of the hydrochars presents bands at approximately 1700 and 1600 cm-1,
attributed to C=O and C=C vibrations, whereas the bands in the range 1000-1450 cm-1 include the C-O
stretching and O-H bending vibrations (see Fig. 1(b)).
The materials were characterized by N2 and CO2 adsorption isotherms, X-ray diffraction, elemental and
thermogravimetric analysis, scanning electron microscopy and determination of ash and pH of the point
of zero charge (pHPZC).
(a)
(b)
Figure 1. (a) Effect of temperature and time on the yields of hydrochars synthesis. (b) FTIR spectra of hydrochars
obtained from sisal treated with H2SO4 at 48% by hydrothermal treatment (SH/120/3) and methodology according to
ref. 2 (S/90/6).
Acknowledgements The authors acknowledge the financial support of FCT (UID/MULTI/00612/2013). ASM
thanks FCT for a Post-doc grant SFRH/BPD/86693/2012.
References
1. M.M. Titirici, M. Antonietti. Chem Soc Rev, 39 (2010) 103
2. L. Wang, Y. Guo, Y. Zhu, Y. Li, Y. Qu, C. Rong, X. Ma, Z. Wang. Bioresour Technol, 101 (2010) 9807
85
86
Science at CQB
Index of Authors
CQB-Day 2015
A
Agapito, F.
Ahmad, S. M.
Albericio, F.
Aldeias, J. B.
Aleixo, R.
Almeida, A.
Almeida, C.
Almeida, I.
Almeida, L. C.
Almeida, R. F. M. de
Alves, A. F.
Andrade, M.
Ania, C. O.
Antunes, C. A. C.
Antunes, F.
Araújo, M. E. M.
Assis, S.
P17; P55
O4; P22
P70
P13
O6; P23; P68
P64
O4
P9
P29
O6; P3; P8; P9; P10; P11; P24; P25; P34; P64
P12
P61
P21; P62; P67
P10
P7
P40; P41
P31
B
Baby, A. R.
Bagulho, A.
Barata, S.
Barreira, L.
Barrocas, B.
Barroso, S.
Bastos, A. E. P.
Batista, D.
Batista, M.
Bellili, S.
Bento, A.
Bento, R. N.
Bernardes, C. E. S.
Bettaieb Ben-Kaab, L.
Bordado, J. C. M
Borges dos Santos, R. M.
Bou-Chacra, N. A.
Branchadell, V.
Bravo, C.
Brigas, A.
Brito, E.
Bronze, M. R.
P33
P11
P5
P70
P19; P47
P6
P25
P61
P43
P41
P27
P7
P7; P26; P30
P41
P1
P17; P55
P33
P53
O1; P28
P23
P5
P49
C
Cachatra, V.
Caio, J. M.
Calhorda, M. J.
Câmara, A.I.
Camelo, C.
Capelli, S. C.
Carmona. B.
P54
P36
O2; O3; P1; P6; P13; P18; P45; P51; P53
O7; P52
O7; P52
P4
O7; P52
89
CQB-Day 2015
Carreira, A.C.
Carvalho, A. P.
Carvalho, M. D.
Carvalho S.
Ciríaco, S.
Coelho, J.
Cordeiro, C.
Cordeiro, M.
Correia, J. P.
Corvo, M. L.
Costa, M. C.
Costa, P. J.
Custódio, L.
Cruz, M. M.
Cruz e Silva, O.
Csuk, R.
P8: P29
O2; P16; P21; P23; P43; P49; P62; P67; P68; P69; P71
O3; P12
P38
O8; P42
P44
P38
P34
P29; P57
P3
P20
P35; P60
P70
P12
P36
P59; P61
D
Dario, M. F.
Delgado, I.
Dias, C.
Dias, P.
Dias, R.
Diogo, H. P.
Dore, T.
Duarte, F. J. S.
Duarte, T. A. G.
P33
P44
O6; P64
P56
P21; P65
P30
P66
P18
P16
E
Esteves, T.
P36
F
Falé, P.L.
Félix, V.
Fernandes, A. S.
Fernandes, C. I.
Fernandes, J.
Ferreira, A. E. N.
Ferreira, C.
Ferreira, F.
Ferreira, L. P.
Ferreira, O.
Figueiredo, A.
Fitzpatrick, A.
Fitzpatrick, A. A.
Flor, A. G.
Florêncio, M. H.
Fonseca, I.
Fortuna, A.
Frade, T.
Francisco, S.
90
O5; P5; P56
P36
P10
O1; P4
P37
P38
O5
P5
O3; P6; P12
P1
P38
P6
O3
P24
P5; P39; P49
O2; P43
P60
P2
P44
CQB-Day 2015
Freire, C.
Freire, R.
P67
O2
G
Gangadhar, K.N.
Gano, L.
Garcia, M. H.
Gaspar, H.
Gil, A.
Gonçalves, C.
Gonçalves, E. M.
Godinho, M.
Gomes, A.
Graça, Angelica
P70
P54
P3; P31
O8; P42
P53
O8; P42
P17
P12
P2
P5; P14
H
Hernández, V.
Herrmann, A.
Hesse, F.
P71
P34
P67
I
Ide, A. H.
P22
J
Jerónimo, A.
Jesus, A.R,
Jin, G.
Jorda, R.
José, S.
Joseph, A.
P11
P66
P9
P59; P61
P69
O3; P26
L
Lago, J. H. G.
Leal, C.
Leitão, A.
Leitão, R.
Li, W.
Lima, P. A.
Lobato, K.
Loesche, A.
Lopes, P.
Lorion, M. M.
Lourenço, J. P.
Lucas, S. D.
P70
O8; P42
P44
P23; P68
P9
P25
P2; P14
P59 ; P61
P5
P18
P20
P59
M
Machuqueiro, M.
Maia, A.
Maia, M.
Marcuzzo, J.
O2; P15; P35; P48; P58
P5
P38
P21
91
CQB-Day 2015
Marinho, H. S.
Marques, A. P.
Marques, A. P.
Marques, F.
Marques, N.
Marques, S.
Marquês, J,
Marquês, J. M
Marquês, J. T.
Matias, A.
Matos, A. M.
Matos, I.
Martinho, P. N.
Martinho Simões, J. A.
Martins, A.
Martins, A
Martins, F.
Martins, L. M. D. R. S.
Martins, M. J.
Melato, A. L.
Melo Jorge, M. E.
Mendo, S. G.
Mendonça, M. H.
Menezes, C,
Mestre, A. S.
Minas da Piedade, M. E.
Mnif, W.
Moiteiro, C.
Monteiro, F.
Monteiro, O. C.
Morais, T. R.
Morais, T. S.
Morgan, G. G.
O7; P10; P34; P52
P38
P39
P31
P5
P21
O5
P25
P9; P10; P34; P46
O8; P42
P63
P43
O3; P6; P13; P51
P17; P55
P23; P68
O6; P56; P64
P14; P23; P68
P16
P36
P51
P14
P12
P12
P69
O2; P21; P43; P62; P67; P69; P71
O3; P7; P26; P30; P33
P41
P36
P38
O1; P19; P20; P28; P47
P70
P31
O3
N
Neng, N. R.
Niu, Y.
Nogueira, C.
Nogueira, J. M. F.
Nolasco, S.
Nunes, C. D.
Nunes, N.
Nunes, R.
O4; P22
P9
P50
O4; P22
P44
O1; P4; P19; P27; P28
P23; P68
P58
O
Oliveira, B.
Oliveira, M. C.
Ortet, O.
Osawa, R. A.
92
P40
P54
P50
P49
CQB-Day 2015
P
Pacheco, R.
Pais, J.
Pais, J. P.
Paiva, A. P.
Paiva, T. M.O.
Paulo, A.
Pelerito, A.
Pena, A.
Peneda, C.
Pereira, I.
Pereira, L. C. J.
Pereira, P.
Peres, J.
Piedade, M. F. M.
Pinto, M.
Pires, J.
Poeta, A. C.
Poli, G.
Pombeiro, A. J. L.
Ponces Freire, A.
O5; P5; P46
P58; P65
O6; P64
P50
P9; P24, P25
P54
O6; P64
P11
O7; P11; P52
P57
P6
P69
O5; P5
P26; P30
P37
P37
P31
P18
P16
P38
R
Rauter, A. P.
Real, C.
Realista, S.
Redmond, G.
Reis, J. C.R.
Reis, P. B. P. S.
Ressaissi, A.
Rezgui, M.
Robalo, M. P.
Rodrigues, J.
Rodrigues, V.
O6; P54; P56; P58; P63; P64; P65; P66;P70
P11
P6; P13; P51
O3
P32
P48
P5
P41
P31
O8; P42
O3
S
Sabino, C. P.
Salas, M.
Sanches, A.
Sánchez, C.
Santos, F. C.
Santos, F. C.
Santos, G.
Santos, M. S. C. S.
Santos, S.
Santos, T. C.
Saraiva, M. S.
Sebastiana, M.
Sério, S.
P24
P5
P27
P39
P3; P11
P10
P6
O2; O6; P3;P14; P32; P33;P64
O8; P31; P42
P3
P45
P38
P14
93
CQB-Day 2015
Serra, P.
Serralheiro, M. L.
Shannon, S. P.
Silva, A.
Silva, E. R.
Silva, L. C.
Silva, M.
Siva, S. P.
Silvestre, A. J.
Simões, R. G.
Siopa, D.
Soares, H.
Sobral, L.
Sousa, A. D. de
Sousa, C. B. De
Sousa Silva, M.
Schwarz, S.
O6; P64
O5; P5; P39; P46
O3
O5; P5; P46
P1
P8
O2
P69
P19
P30
P14
O7; P44; P52
P65
P3
P70
P38
P54; P59; P61
T
Tavares, A.
Teixeira, V. H.
Tenreiro, R.
Tomás, A. M.
Tomaz, A. I.
P44
P15; P48
P65
P38
P3; P31
V
Valente, A.
Varela, J.
Vaz,, P. D.
Velasco, M. V. R.
Velasco, P.
Viana, A. S.
Vicente, A. I.
Videira, A.
Vieira, B.
Vilas-Boas, F.
Vila-Viçosa, D.
Vital, J.
Vitor, G.
Vizetto-Duarte, C.
P31
P70
P4; P27
P33
P71
O5; O6; P9; P10; P25; P26; P29; P34; P46; P64; P65
O3
P10
P20
P11
P15; P35; P48; P58
P43
P20
P70
X
Xavier, N. M.
94
P59; P60; P61