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| LabRMN-CNC – Nuclear Magnetic Resonance Laboratory Center of Neurosciences and Cell Biology, University of Coimbra |
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The LabRMN-CNC is part of the Area E “Biophysics and Biomedical NMR” of the CNC. It results from the merge in January 2008 of the researchers of the former NMR Spectroscopy Unit of the Faculty of Science and Technology of the University of Coimbra (FCTUC) to the CNC.
It is part of the Portuguese National Network of Nuclear Magnetic Resonance Spectroscopy, under the supervision of the Foundation for Science and Technology (FCT), Portugal.
Coordinator:
Carlos F. G. C. Geraldes, PhD
Contact: geraldes@bioq.uc.pt
Spectrometer Administrator:
Emeric Wasielewski
Contact: emeric@cnc.uc.pt
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| Research Interests: |
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A. Inorganic Complexes for Medical Diagnostics: MRI and Molecular Imaging:
Studies focus on inorganic compounds for medical diagnostic imaging, in particular contrast agents for magnetic resonance imaging (MRI CAs). They include design, development and physico-chemical evaluation of metal based nanoparticles and small metal chelate agents for multimodal targeted medical imaging agents, followed by in vitro and animal model evaluation using MRI and nuclear imaging techniques. |
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Contact person:
Carlos F. G. C. Geraldes
geraldes@bioq.uc.pt
Representative Publications:
Pereira GA, Norek M, Peters JA, Ananias D, Rocha J, Geraldes CFGC. NMR Transversal Relaxivity of Aqueous Suspensions of Particles of Ln3+ -Based Zeolite Type Materials. Dalton Trans., 2241-2247 (2008).
Sousa PL, Livramento JB, Helm L, Merbach AE, Même W, Doan BT, Beloeil JC, Prata MIM, Santos AC, Geraldes CFGC, Tóth E. In vivo MRI assessment of a novel GdIII-based contrast agent designed for high magnetic field applications Contrast Media Mol. Imaging 3, 78-85 (2008).
Torres S, Prata MIM, Santos AC, André JP, Martins JA, Helm L, Tóth E, García-Martín ML, Rodrigues TB, López-Larrubia P, Cerdán S, Geraldes CFGC. Gd(III)-EPTPAC-16, a new self-assembling potential liver MRI contrast agent: in vitro characterization and /in vivo/ animal imaging studies. NMR Biomed. 21, 322–336 (2008).
Peter Caravan, Carlos F.G.C. Geraldes, Matthew T. Greenfield, Garry E. Kiefer, Mai Lin, Kenneth McMillan, M. Isabel M. Prata, Ana C. Santos, A. Dean Sherry, Xiankai Sun, Jufeng Wang, Mark Woods, Shanrong Zhang and Piyu Zhao, The effect of charge on the biodistribution and CEST properties of lanthanide(III) DOTA-tetraamide derivatives, Investig. Radiol., 43, 861-870 (2008).
Zuzana Kotková, Giovannia A. Pereira, Kristina Djanashvili, Jan Kotek, Jakub Rudovský, Petr Hermann, Luce Vander Elst, Robert N. Muller, Carlos F.G.C. Geraldes, Ivan Lukeš and Joop A. Peters, Lanthanide(III) complexes of phosphonic acid monoester and phosphinic acid analogues of H4DOTA as model compounds for evaluation of the second-sphere hydration, Eur. J. Inorg. Chem., 119-136 (2009).
Carlos F.G.C. Geraldes, Sophie Laurent, Classification and Basic Properties of Contrast Agents for Magnetic Resonance Imaging, Contrast Media Mol. Imaging, 4, 1-23 (2009).
Sara Silvério, Susana Torres, André F. Martins, José A. Martins, João P. André, Lothar Helm, M. Isabel M. Prata, Ana C. Santos and Carlos F.G.C. Geraldes, (bis)-Hydroxymethyl-substituted DTTA skeleton: a new lead for the synthesis of high relaxivity MRI contrast agents? Dalton Trans., 4656-4670 (2009).
NMR expertise:
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Design, development and physico-chemical characterization of paramagnetic chelates through relaxometric (T1, T2, T2*) and chemical shift studies using multinuclear NMR (17O, 1H, 2H), NMRD techniques, EPR and simultaneous fitting of paramagnetic data. |
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Relaxometric (T1, T2, T2*) characterization of paramagnetic nanoparticulate systems. |
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In vitro (cells) and in vivo (small animals) MRI characterization of paramagnetic nanoparticulate systems. |
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Paramagnetic shifts (1H, 13C, 31P...) study to obtain solution structure and dynamics of lanthanide chelates and, together with residual dipolar couplings (RDCs), of covalently tagged oriented proteins. |
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Interaction study of ligands and complexes with biomacromolecules using STD technique. |
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Structural characterization of small molecules using 1D and 2D NMR techniques. |
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B: Inorganic Biochemistry: Inorganic Complexes and Species for Therapy and Environment:
We are interested on the design and study of inorganic compounds as drugs for medical therapy (lithium Li+ and bipolar disorder; vanadium V(IV/V) complexes as oral insulin-mimetic agents). We focus on their mechanisms of action using cell and animal models. We also study the environmental impact of chromium Cr(V) and reduced species.
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Contact person:
M. Margarida Castro (gcastro@bioq.uc.pt)
M. Carmen Alpoim (mcalpoim@iav.uc.pt)
Representative Publications:
Fonseca CP, Sierra A, Geraldes CFGC, Cerdán S, Castro MMCA. Mechanisms underlying Li+ effects in glutamatergic and GABAergic neurotransmissions in the adult rat brain and in primary cultures of neural cells as revealed by 13C NMR. J. Neurosc. Res., 87,1046–1055 (2009).
H. Faneca, V.A. Figueiredo, Isabel Tomaz, Gisela Gonçalves, Fernando Avecilla, M.C. Pedroso de Lima, Carlos F.G.C. Geraldes, João Costa Pessoa and M. Margarida C.A. Castro, Vanadium compounds as therapeutic agents: some chemical and biochemical studies, J. Inorg. Biochem., 103, 601-608 (2009)
Crystal structure of the V(V) dimer [V2O2(µ-O)(dmpp)2(OCH3)2] and its equilibrium with the V(V) trimer [V3O3(µ-O)3(dmpp)3(H2O)](H2O)2 in methanol/water solutions, Fernando Avecilla, Carlos F. G. C. Geraldes, Anjos, L. Macedo and M. Margarida C. A. Castro, Eur. J. Inorg. Chem., 3586-3594 (2006).
Vanadium (IV and V) complexes of reduced Schiff bases derived from the reaction of aromatic o-hydroxyaldehydes and diamines containing carboxylic groups, J. Costa Pessoa, S. Marcão, I. Correia, G. Gonçalves, A. Dörnyei , T. Kiss, T. Jackush, I. Tomaz, M.M.C.A. Castro, C.F.G.C. Geraldes Eur. J. Inorg. Chem., 3595-3606 (2006).
Tricarboxylic acid cycle inhibition by Li+ in the human neuroblastoma SH-SY5Y cell line: a 13C NMR isotopomer analysis, Carla P. Fonseca, John G. Jones, Rui A. Carvalho, F. Mark H. Jeffrey, Liliana P. Montezinho, Carlos F.G.C. Geraldes, M. Margarida C.A. Castro, Neurochem. International, 47, 385-393 (2005).
Uptake and metabolic effects of insulin mimetic oxovanadium compounds in human erythrocytes, Teresa C. Delgado, A. Isabel Tomaz, Isabel Correia, João Costa Pessoa, John G. Jones, Carlos F.G.C. Geraldes and M. Margarida C.A. Castro, J. Inorg. Biochem., 99, 2328-2339 (2005).
NMR expertise:
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Characterization of the structure, speciation and cell interactions of complexes in solution and cell suspensions using multinuclear NMR and EPR. |
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Study of the effects of inorganic compounds on cell transport metabolism using metal (eg. 7Li, 23Na) through 31P and 13C NMR (with isotopomer analysis of 13C-enriched metabolites). |
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C. Polymers and their complexes:
We focus on the development and characterization of conjugated polymers and biopolymers for applications like biosensors.
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Contact person:
Luisa Ramos (mlramos@ci.uc.pt)
Licínia Justino (liciniaj@ci.uc.pt)
Representative Publications:
Burrows HD, Knaapila M, Monkman AP, Tapia MJ, Fonseca SM, Ramos ML, Pyckhout-Hintzen W, Pradhan S, Scherf U. Structural studies on cationic poly{9,9-bis [6-(N,N,N - trimethylammonium) alkyl] fluorene-co-1,4-phenylene} iodides in aqueous solutions in the presence of the nonionic surfactant pentaethyleneglycol monododecyl ether (C12E5). J. Phys. Condensed Matter. 20, 104210-104216 (2008).
Knaapila M, Almásy L, Garamus VM, Ramos ML, Justino LLG, Torkkeli M, Dias FB, Scherf U, Burrows HD, Monkman AP. An effect of side chain length on the solution structure of poly(9,9-dialkylfluorene)s in toluene. Polymer 49, 2033-2038 (2008).
Sobral AJFN, Justino LLG, Santos ACC, Silva JA, Arranja CT, Silva MR, Beja AM. Synthesis and Structural Characterization of a New Self-Assembled Disulphide Linked meso-Tetrakis-Porphyrin Macromolecular Array. J. Porphyr. Phthalocya. 12, 845 – 848 (2008).
Costa D, Ramos ML, Burrows HD, Tapia MJ, Miguel MG. Using lanthanides as probes for polyelectrolyte-metal ion interactions and hydration changes on binding of trivalent cations to nucleotides and nucleic acids. Chem. Phys. 352, 241-248 (2008).
NMR expertise:
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NMR characterization of polymers in solution, combined with DFT calculations. |
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D. NMR studies of metabolic fluxes:
We study intermediary metabolism using 2H, 13C and 15N stable isotope tracers. Glucose, protein and lipid homeostasis are studied in human subjects, in animal models of human diseases, and in isolated perfused animal organs.
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Contact person:
John G. Jones (jones@cnc.uc.pt)
Madalena Caldeira (mcaldeira@ci.uc.pt)
Rui Carvalho (carvalho@bioq.uc.pt)
Representative Publications:
Jones, J.G., Garcia, P., Barosa, C., Delgado,T.C. and Diogo,L. 2009. Hepatic anaplerotic outflow fluxes are redirected from gluconeogenesis to lactate synthesis in patients with Type 1a Glycogen Storage Disease. Metabolic Engineering 11, 155-162.
Soares A.F., Viega, F.J., Carvalho, R.A. and Jones, J.G. 2009. Quantifying hepatic glycogen synthesis by direct and indirect pathways in rats under normal ad-libitum feeding conditions. Magn. Res. Med. 61, 1-5.
Jones, J.G., Garcia. P., Barosa, C., Delgado, T.C., Caldeira, M.M. and Diogo, L. 2008. Quantification of hepatic transaldolase exchange activity and its effects on tracer measurements of indirect pathway flux in humans. Magn. Res. Med. 59, 423-429.
Alves, T., Nunes, P, Palmeira, C., Jones, J.G. and Carvalho, R. 2008. Estimating gluconeogenesis by NMR isotopomer distribution analysis (NMR-IDA) of [13C]bicarbonate and [1-13C]lactate. NMR in Biomedicine, 21, 337-344.
Jones, J.G., Fagulha,A., Barosa, C., Bastos, M., Barros, L., Baptista, C., Caldeira M., and Carvalheiro M. 2006. Noninvasive analysis of hepatic glycogen kinetics before and after breakfast with deuterated water and acetaminophen. Diabetes, 55, 2294-2300.
Ribeiro, A., Caldeira, M.M., Carvalheiro, M., Bastos, M., Baptista, C., Fagulha, A., Barros, L., Barosa, C. And Jones, J.G. 2005. A simple measurement of gluconeogenesis by direct 2H NMR analysis of menthol glucuronide enrichment from 2H2O. Magn. Res. Med. 54, 429-434.
NMR expertise:
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Analysis of metabolic flux. |
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13C NMR isotopomer analysis of 13C-enriched metabolites from perfused organs, plasma glucose and urinary metabolites by direct and indirect detection methods. |
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2H NMR analysis of metabolite positional, 2H-enrichments from 2H2O in perfused organs, plasma glucose and urinary metabolites. |
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| Areas of NMR expertise in metabolic flux analysis: |
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13C-NMR isotopomer analysis of 13C-enriched metabolites from perfused organs, plasma glucose and urinary metabolites by direct and indirect detection methods.
2H-NMR analysis of metabolite positional 2H-enrichments from 2H2O in perfused organs, plasma glucose and urinary metabolites.
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| Equipment: |
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 Varian VNMR 600 System with 3 observe channels (Installed in 2007)
Probes:
5-mm PFG triple resonance I.D. (optimal sample volume = 600-700 μl)
5-mm PFG Broadband (optimal sample volume = 600-700 μl)
3-mm PFG triple resonance I.D. (optimal sample volume = 180-200 μl)
3-mm PFG Broadband (optimal sample volume = 180-200 μl)
4-mm PFG nanoprobe (HR-MAS)
Note on the use of 3-mm microprobes: The 3-mm probes offer a ~40% increase in sensitivity over their 5-mm counterparts provided that the molecule of interest is soluble in the smaller volume and that the signal linewidths are preserved. For many biological samples that contain salt, e.g. protein solutions or tissue extracts, the smaller volumes result in higher salt concentrations leading to longer pulse widths, less efficient 1H-broadband decoupling, and increased sample heating. This latter effect may result in a drifting lock signal particularly if the lock solvent is D2O.
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 Varian UNITY 500 System (Installed in 1992)
Probes:
5-mm I.D. (optimal sample volume = 600-700 μl)
10-mm Broadband (optimal sample volume = 1ml)
5-mm Low frequency (optimal sample volume = 600-700 μl)
5-mm PFG Broadband (optimal sample volume = 600-700 μl)
 
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Varian NMR probes
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Guide for NMR access and use:
1) Location: The NMR Center is located in the ground floor of the Medicine Faculty at the University of Coimbra. It opens directly into the courtyard of the Medicine Building aside the principal personnel entrance into the courtyard which is opposite the entrance for vehicles. Parking inside the courtyard is reserved for Faculty cardholders and there is no visitor car park. The street parking in and around the University fills up quickly after 8:30 am. Please be aware that cars that are either parked improperly or are in designated sites for any length of time will almost certainly be clamped. There are regular bus services connecting the University with main bus and train stations (SMUTC).
2) Scheduling: NMR time is scheduled for use every 2 weeks from a Monday (week 1) to Sunday (week 2) cycle. Unless there are exceptional circumstances, external users will be scheduled during non-holiday weekdays only. Requests for NMR time must be received by 12:00 pm on the previous Friday and the online forms are to be used. The distribution of time will be posted on the website or sent by e-mail to the users by Sunday. Cancellations of NMR time should be immediately notified to the NMR administrator.
3) Materials and Sample Preparation: The NMR user must supply his/her own tubes and NMR solvents. For nanoprobe users, rotors from the NMR Center are available for use. NMR samples may be prepared in the laboratory next to the NMR room. This is equipped with pipettes, a balance and microcentrifuge. There is also a fridge/freezer for perishable samples. This lab does not have a fume hood or solvent evaporator.
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| Pricing informations for the use of the LabNMR spectrometers (starting November 1st 2009) |
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| I – Costs of the service for Academics (Non-Profit)/NMR Network |
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| Spectrometer |
Spectral Acquisition (€ / hour) |
| With operator* |
Without operator** |
| VNMRS 600 |
9,00 |
5,00 |
| Unity 500 |
7,50 |
3,20 |
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* Experiments preparation time with operator charged.
** Cost for acquisition extra time after preparation.
Costs include the legal taxe IVA of 20%.
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| II – Costs of the services for companies (For-Profit)a |
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| Spectrometer |
VNMRS 600 |
Unity 500 |
| Cost (€) |
Cost (€) |
| 1H |
60,30 |
38,00 |
| 13C |
173,00 |
108,00 |
| 31P |
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54,00 |
| 19F |
- |
108,00 |
| 13C-DEPT |
144,00 |
90,00 |
| COSY |
173,00 |
108,00 |
| HSQC |
288,00 |
180,00 |
| HMBC |
518,00 |
324,00 |
| NOESY |
615,00 |
378,00 |
| TOCSY |
173,00 |
108,00 |
| Package 1D* |
356,00 |
222,00 |
| Package 1D+2D** |
1067,00 |
667,00 |
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*includes 1H, 13C, 13C-DEPT
**includes 1H, 13C, 13C-DEPT, COSY, HSQC, HMBC
Costs include the legal taxe IVA of 20%.
a Costs are based on an average time of preparation and acquisition of the experiments. They include the use of the equipments as the preparation of sample and standard deutered solvants (CDCl3, D2O, DMSO, aceton).
Related with the sample quantity (MM), costs are valid for:
1H and 2D > 10 mg; 13C and 13C-DEPT > 200 mg; 31P > 10 mg; 19F > 10 mg
In case more acquisition time is needed than established for some experiments additionnal hours are charged (600 MHz 80€/h and 500 MHz 65€/h). Analysis of sample in relatively small quantity (concentration between 2 and 5 mg) is possible under conditions and a budget will be given after consultation of the service.
Processed spectra are send in electronic format (PDF) and acquisition files are available.
Elaboration of reports of spectra interpretations is possible. But due to projects specificities reports are charged in accordance with the time of their execution (100 € / hour).
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| III - Bio-NMR |
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| Spectrum |
VNMRS 600 |
| Cost (€)a |
| 15N-HSQC |
38,00 |
| 13C-HSQC |
536,00 |
| 3D-HNCO |
1358,00 |
| 3D-HN(CA)CO |
1358,00 |
| 3D-HN(CO)CACB |
1358,00 |
| 3D-HNCACB |
1358,00 |
| 3D-15N-NOESY |
2637,00 |
| 3D-15N-TOCSY |
1358,00 |
| 3D-hCCH-TOCSY |
2637,00 |
| 3D-HNHA |
1358,00 |
| 3D 13C-NOESY |
2637,00 |
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| a Costs are based on an average time of preparation and acquisition of the experiments for labelled samples (13C and 15N) with a standard concentration of 0.5 mM and an approximative molecular mass of 20 kDa. |
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