ABSTRACTS - Instytut Fizyki JÄ drowej PAN

The Henryk Niewodniczaski
INSTITUTE OF NUCLEAR PHYSICS
Polish Academy of Sciences
152 Radzikowskiego str., 31-342 Kraków, Poland
www.ifj.edu.pl/reports/2009/
Kraków, December 2009
Report No. 2031/AP
XLII Polish Seminar on Nuclear Magnetic Resonance
and Its Applications. Kraków, 1-2 December 2009
ABSTRACTS
Organizing Committee:
Members:
T. Banasik Z.T. Lalowicz /v-chairman/
A. Birczyski K. Majcher
J. Blicharski A. Mynarczyk
S. Heinze-Paluchowska M. Noga /secretary/
J. W. Hennel /chairman/ Z. Olejniczak
M. Jaboska P. Rosicka
A. Jasiski /honorary chairman/ W. Rutkowski
K. Jasiski T. Skórka
A. Krzyak G. Stoch
P. Kulinowski U. Tyrankiewicz
S. Kwieciski W.P. Wglarz /v-chairman/
G. Woniak
Sponsors:
AMX-ARMAR AG,
BRUKER-Polska Sp. z o.o,
PASTWOWA AGENCJA ATOMISTYKI,
VARIAN INTERNATIONAL AG.
KOMITET FIZYKI POLSKIEJ AKADEMII NAUK

Addresses of the sponsors:
AMX-ARMAR AG
Anna Potrzebowska
ul. Bugarska 12a
93-362 ód
tel. (042) 645 00 64
BRUKER POLSKA SP. Z O.O
mgr W. Leszczyski
ul. Budziszyska 69
60-179 Pozna
tel. (061) 868 90 08
fax. (061) 868 90 96
e-mail: sekretariat@bruker.poznan.pl
www.bruker.pl
PASTWOWA AGENCJA ATOMISTYKI
ul. Krucza 36,
00-921 Warszawa
VARIAN NMR
mgr in. W. Komider
ul. Skarbka 21
60-348 Pozna
tel. (061) 867 31 84
tel. kom. 602 287 918
e-mail: woko@eranet.pl
www.varianinc.com
KOMITET FIZYKI POLSKIEJ AKADEMII NAUK
Wydzia III PAN
Instytut Fizyki PAN
Al. Lotników 32/46
02-668 Warszawa

CONTENTS
1. APPLICATION OF 19 F MAGNETIC RESONANCE IMAGING FOR THE STUDY
OF DRUGS EFFICACY EX VIVO.
Dorota Bartusik and Bogusaw Tomanek
2. TOCOTRIENOLS RELATED CHANGES IN HUMAN MCF-7
ADENOCARCINOMA USING 19 F MAGNETIC RESONANCE IMAGING EX
VIVO.
Dorota Bartusik, Boguslaw Tomanek, Danuta Siluk, and Roman Kaliszan
3. NON-CONTRAST BASED MRI OF HUMAN LUNG PERFUSION AND
VENTILATION.
Grzegorz Bauman, Michael Puderbach, Michael Deimling, Julien Dinkel, Christian
Hintze, Lothar R. Schad
4. CARBON-13 NMR RELAXATION STUDY OF THE INTERNAL DYNAMICS
IN CYCLODEXTRINS IN ISOTROPIC SOLUTION.
Piotr Bernatowicz, Katarzyna Ruszczyska-Bartnik, Andrzej Ejchart, Helena
Dodziuk, Ewa Kaczorowska, and Haruhisa Ueda
5. STRUCTURAL CHANGES IN holo-S100A1 AND S100A1(C85M)PROTEIN.
CHEMICAL SHIFT BASED RESULTS.
Monika Budziska, Micha Nowakowski, Igor Zhukov, Agnieszka Belczyk, Andrzej
Bierzyski, and Andrzej Ejchart
6. THE DISPERSION OF WATER PROTON SPIN-LATTICE RELAXATION RATES
IN AQUEOUS HUMAN PROTEIN HC ( 1 -MICROGLOBULIN) SOLUTIONS.
Maria Dobies, Maciej Kozak, Stefan Jurga, and Anders Grubb
7. UNUSUAL STRUCTURE AND NMR SPECTRA OF SOME CYCLOPHANES.
Helena Dodziuk
8. INVESTIGATION OF TAUTOMERISM OF THE SELECTED OXOPURINES
IN NEUTRAL AND BASIC WATER SOLUTIONS BY 13 C NMR AND GIAO-DFT
CALCULATIONS.
Katarzyna Dybiec, Sergey Molchanov, and Adam Gryff-Keller
9. CEREBRAL WHITE MATTER RECOVERY IN ABSTINENT ALCOHOLICS –
A MULTIMODAL MAGNETIC RESONANCE STUDY.
Stefan Gazdzinski, Timothy C. Durazzo, Anderson Mon, Ping-Hong Yeh, and Dieter
J. Meyerhoff
10. INTERACTIVE EFFECTS OF AGE AND EXCESS BODY WEIGHT ON
NEURONAL VIABILITY - A MAGNETIC RESONANCE SPECTROSCOPY
STUDY.
Stefan Gazdzinski, Rachel Millin, Lana G. Kaiser, Susanne G. Mueller, Timothy C.
Durazzo, Michael W. Weiner, and Dieter J. Meyerhoff
1
2
3
4
5
6
7
8
9
11

11. TEMPERATURE INDUCED BOUND WATER IMMOBILIZATION IN
CETRARIA ACULEATA (SCHREB.) FR. THALLI OBSERVED BY PROTON
RELAXATION.
Hubert Haraczyk, P. Nowak, and M.A. Olech
12. MAPPING OF THE B1 FIELD USING ADIABATIC EXCITATION PULSES.
Franciszek Hennel
13. THE MULTIPARAMETRIC CMR-BASED ASSESSMENT OF HEART
FUNCTION IN MURINE MODEL OF HEART FAILURE TGaq*44.
Magdalena Jaboska, UrszulaTyrankiewicz, Tomasz Skórka, Henryk Figiel, Sylwia
Heinze-Paluchowska, Mirosaw Woniak, and Stefan Chopicki
14. MRI APPLICATION OF MICROSTRIP RF COILS.
Krzysztof Jasiski, Peter Latta, Vyacheslav Volotovskyy, Anna Mynarczyk,
Wadysaw P.Wglarz, and Bogusaw Tomanek
15. TAUTOMERISM OF SCHIFF BASES DERIVED FROM (S)-
DIPHENYLVALINOL: THE SOLID STATE NMR AND X-RAY STRUCTURE
STUDIES.
Magdalena Jaworska, Pawe B. Hrynczyszyn, Katarzyna Nowicka, Wodzimierz
Ciesielski, and Marek J. Potrzebowski
16. ADDUCTS FORMATION OF RHODIUM(II) TETRACARBOXYLATES WITH
METHIONINE DERIVATIVES: INVESTIGATIONS BY THE USE OF VIS AND
NMR TECHNIQUES.
Jarosaw Jawiski and Rafa Gaszczka
17. MOLECULAR DYNAMICS AND STRUCTURE IN DIBLOCK COPOLYMERS
STUDIED BY NMR AND BROADBAND DIELECTRIC SPECTROSCOPY.
Jacek Jenczyk, Monika Makrocka-Rydzyk, Aleksandra Wypych, Stanisaw
Gowinkowski, and Stefan Jurga
18. BACKBONE DYNAMICS OF TFE-INDUCED NATIVE-LIKE FOLD OF REGION
4 OF INITIALLY UNFOLDED E. coli RNA POLYMERASE 70 SUBUNIT.
Piotr Kaczka, Agnieszka Polkowska-Nowakowska, Krystyna Bolewska, Igor Zhukov,
Jarosaw Poznaski, and Kazimierz L. Wierzchowski
19. fMRI QUALITY ASSURANCE SYSTEM: PHANTOM AND AUTOMATIC DATA
ANALYSIS.
Karolina Kamiska, Stanisaw Adaszewski, Ewa Pitkowska-Janko, Piotr
Bogorodzki, and Maciej Pisklak
20. NMR RELAXATION MEASUREMENTS OF OXIDATION PROCESSES IN
BLOOD SERUM.
Joanna Kamiska, Jan Kobierski, and Barbara Blicharska
21. THE USEFULNESS OF DIFFUSION-WEIGHTED IMAGING IN
DISTINGUISHING BETWEEN SOME BRAIN PATHOLOGIES.
Al k d li fi d l k
13
14
16
17
18
19
20
21
22
24
25

Aleksandra Klimas, Zofia Drzazga, and Ewa Kluczewska
22. CHEMICAL EXCHANGE PROCESSES IN HYDROGEN PEROXIDE
SOLUTIONS OBSERVED BY NMR RELAXATION.
Jan Kobierski, Hartwig Peemoeller, and Barbara Blicharska
23. DETECTOR FOR PROTON-ELECTRON DOUBLE RESONANCE IMAGING
(PEDRI).
ukasz Koaszewski, Piotr Bogorodzki, Ewa Piatkowska-Janko, Jerzy Piotrowski,
Jerzy Skulski, and Maciej Pisklak
24. RECENT ADVANCES IN MULTIDIMENSIONAL NMR SPECTROSCOPY WITH
SPARSE RANDOM SAMPLING.
Wiktor Komiski, Krzysztof Kazimierczuk, Maria Misiak, Jan Stanek, and Anna
Zawadzka-Kazimierczuk
25. THE ROLE of FULLEREN C 60 , CARBON NANOTUBES AND CARBON
ENCAPSULATED MAGNETIC NANOPARTICLES IN CREATING OF
CREATOL (5-HYDROXYCREATININE) AND UREMIC TOXIN -
METHYLGUANIDINE.
Hanna Krawczyk and Magdalena Popawska
26. DO ESR AND NMR DESCRIBE THE SAME DYNAMICS OF PARAMAGNETIC
SYSTEMS?
Danuta Kruk
27. ANISOTROPIC DIFFUSION PHANTOM FOR B-MATRIX CALCULATION
Artur Tadeusz Krzyak
28. ESR LINESHAPE FOR MULTISPIN SYSTEMS IN THE PRESENCE
OF LAXATION.
Aleksandra Kubica, Artur Mielczarek, and Danuta Kruk
29. ESTIMATION OF PHOSPHOLIPIDS CONCENTRATION IN PLASMA,
MONONUCLEAR CELLS, AND ERYTHROCYTES FROM PATIENTS WITH
HEMATOLOGICAL CANCERS - 31 P MRS IN VITRO STUDY.
Magorzata Kuliszkiewicz-Janus
30. PERFORMANCE OF DFT, SOPPA, SOPPA(CCSD) AND CCSD(T) METHODS IN
PREDICTING NUCLEAR ISOTROPIC SHIELDINGS IN THE COMPLETE BASIS
SET LIMIT.
Teobald Kupka, Micha Stachów, Marzena Nieradka, Jakub Kaminsky, and Tadeusz
Pluta
31. THEORETICAL AND EXPERIMENTAL STUDIES ON CYTOSINE AND 5-
FLUOROCYTOSINE.
Teobald Kupka, Mariana Spulber, Mariana Pinteala, Adrian Fifere, Agnieszka
Raniszewska, and Malgorzata Broda
27
28
29
30
31
32
33
34
36
37

32. THEORETICAL CALCULATION OF NMR SHIELDINGS AND INDIRECT SPIN-
SPIN COUPLING CONSTANTS.
Teobald Kupka
33. CONFORMATIONAL ANALYSIS OF CYCLIC DYNORPHIN ANALOGUES
USING NMR DERIVED DATA.
Maria Kwasiborska, Agnieszka Zieleniak, Micha Nowakowski, Marta Oleszczuk,
Jacek Wójcik, Nga N. Chung, Peter W. Schiller, and Jan Izdebski
34. DYNAMICS OF HYDROXYL DEUTERONS AND BONDED WATER
MOLECULES IN NADY(0.8) ZEOLITE AS STUDIED BY MEANS OF
DEUTERON NMR SPECTROSCOPY.
Zdzisaw T. Lalowicz, Grzegorz Stoch, Artur Birczyski, and M. Punkkinen
35. SPECTRAL DENSITY FUNCTIONS OF COMPLEX MOLECULAR MOTIONS.
Lidia Latanowicz and Zofia Gdaniec
36. SOLID STATE STUDIES OF CYCLODEXTRIN COMPLEXES WITH
ADAMANTANE AND AMANTADINE USING 13 C AND 15 N CP/MAS NMR.
Agnieszka Lis-Cieplak and Wacaw Koodziejski
37. MOLECULAR DYNAMICS AND STRUCTURE OF MIKTOARM STAR BLOCK
COPOLYMERS BASED ON POLY(BUTYL ACRYLATE) AND
POLY(ETHYLENE OXIDE).
Monika Makrocka-Rydzyk, Aleksandra Wypych, Mariusz Jancelewicz, Stefan Jurga,
and Krzysztof Matyjaszewski
38. MOLECULAR DYNAMICS AND NUCLEAR RELAXATION PROCESSES OF
PLANAR CATIONS WITH PSEUDO 5-FOLD SYMMETRY AXES:
IMIDAZOLIUM AND PYRAZOLIUM COMPOUNDS.
W. Medycki, D. Kruk, R. Jakubas, and J. Jadyn
39. THEORY OF SOLID STATE DYNAMIC NUCLEAR POLARIZATION.
Artur Mielczarek, and Danuta Kruk
40. FIELD DEPENDENT RELAXATION PROCESSES IN MULTISPIN SYSTEMS.
Agnieszka Milewska and Danuta Kruk
41. THREE-DIMENSIONAL NMR SPECTROSCOPY OF ORGANIC MOLECULES.
Maria Misiak and Wiktor Komiski
42. APPLICATION OF MR MICROSCOPY FOR ASSESSMENT OF HYDRATION
PROCESSES IN HPMC BASED MATRIX TABLETS.
Anna Mynarczyk, Marco L.H. Gruwel, Piotr Kulinowski, Krzysztof Jasiski,
Przemysaw Doroyski, Bogusaw Tomanek, and Wadysaw P. Wglarz
43. 3D NMR-BASED STRUCTURE OF apo-S100A1 HUMAN PROTEIN.
Micha Nowakowski, ukasz Jaremko, Igor Zhukov, Agnieszka Belczyk, Andrzej
Bierzyski, and Andrzej Ejchart
38
39
40
41
43
44
45
46
47
48
49
51

44. TOWARD CHIRAL CRYSTALS OF BENZODIAZACORONANDS.
Katarzyna Nowicka, Agata Jeziorna, Adam Sobczuk, Janusz Jurczak, Grzegorz D.
Bujacz, Anna Bujacz, Wodzimierz Ciesielski, and Marek J. Potrzebowski
45. OPTIMIZATION METHOD FOR ADIABATIC TAGGING PULSES FOR
ARTERIAL SPIN LABELING.
Wojciech Obrbski, Piotr Bogorodzki, and Ewa Piatkowska-Janko
46. NON-DEBYE RELAXATION AND TEMPERATURE DEPENDENCE
OF THE SECOND MOMENT OF NMR LINE.
Marcin Olszewski and Nikolaj Sergeev
47. ANALYSIS OF BOLD SIGNAL IN THE BRAINSTEM DURING RESTING
STATE AND AFTER CONTINUOUS SOUND ACTIVATION.
Paulina Palowska, Michalina Ry, Uwe Klose, and Zofia Drzazga
48. SOLID STATE NMR STUDIES OF COORDINATION AND
ORGANOMETALLIC COMPLEXES OF NICKEL AND RUTHENIUM.
Piotr Paluch and Marek J. Potrzebowski
49. 1 H, 13 C, 15 N NMR COORDINATION SHIFTS IN CATIONIC Fe(II), Ru(II), Os(II)
WITH 2,2'-BIPYRIDINE AND 1,10-PHENANTHROLINE.
Leszek Pazderski, Tomasz Pawlak, Jerzy Sitkowski, Lech Kozerski, and Edward
Szyk
50. SEEKING A MOLECULAR SWITCH - STUDY ON 7-HYDROXY -4-
METHYLQUINOLINE-8-CARBALDEHYDE IN SOLUTION.
Mariusz Pietrzak, Volha Vetokhina, Jacek Nowacki, Jerzy Herbich, and Andrzej L.
Sobolewski
51. SPIN-LATTICE RELAXATION IN DOPA-MELANIN AND MELANIN
ISOLATED FROM Sepia officinalis - COMPARATIVE EPR STUDIES.
Barbara Pilawa, Magdalena Zdybel, Daria Czyyk, Ewa Chodurek, and Sawomir
Wilczyski
52. SOLID-STATE CHARACTERIZATION OF S(+)CLOPIDOGREL
HYDROGENSULPHATE PHARMACEUTICALS AND THEIR POLYMORPHS.
Edyta Pindelska, Andrzej Mazurek, and Wacaw Koodziejski
53. CONSTRUCTION OF HIGH FREQUENCY COILS FOR MRI AT 0.088 T.
Bartosz Proniewski, Henryk Figiel, and Tadeusz Paasz
54. EPR STUDIES OF PARAMAGNETIC PROPERTIES OF THERMALLY
TREATED SISOMICIN AND VERAPAMIL.
Pawe Ramos, Barbara Pilawa, and Piotr Pepliski
55. THE INFLUENCE OF TRACES OF WATER ON THE CHEMICAL SHIFTS
OF QULAENE IN BENZENE SOLUTION.
Micha Raew, Ewa Kaczorowska, Ewa Kula-wieewska, Ewa Ciepicha, and Jacek
Wójcik
53
54
55
56
58
59
60
61
63
64
65
66

Wójcik
56. APPLICATION OF MRI FOR SPINAL CORD MONITORING IN ANIMAL
MODEL OF PRESSURE IMPACT INJURY.
Paulina Rosicka, Katarzyna Majcher, Wiesaw Marcol, Wojciech lusarczyk, Tomasz
Banasik, Joanna Lewin-Kowalik, and Wadysaw P. Wglarz
57. INTERACTION OF SELECTED BICYCLIC MONOTERPENES WITH -
CYCLODEXTRIN. THERMODYNAMIC CHARACTERISTIC OF
COMPLEXATION.
Katarzyna Ruszczyska-Bartnik, Micha Nowakowski, Helena Dodziuk, and Andrzej
Ejchart
58. SODIUM MR IMAGING.
Lothar R. Schad
59. THE CHANGES OF PHOSPHOLIPIDS IN LIPID RAFTS AT PATIENTS WITH
MYELODYSPLASTIC SYNDROME (MDS). .
Joanna Schiller, Magorzata Kuliszkiewicz-Janus, Izabela Dere-Wagamann, and
Stanisaw Baczyski
60. RELAXATION PROCESSES OF SELECTED BIOMOLECULES.
Aleksandra Skowroska and Danuta Kruk
61. IMPLEMENTATION OF RETROSPECTIVE GATING IN APPLICATION TO MR
ASSESSMENT OF CARDIAC FUNCTION IN MICE.
Tomasz Skórka, Sylwia Heinze-Paluchowska, Urszula Tyrankiewicz, and Magdalena
Jaboska
62. INFLUENCE OF PARAMAGNETIC IONS PRESENCE ON RELAXATION IN
BLOOD SERUM.
Lech Skórski, Mateusz Synowiecki, and Barbara Blicharska
63. THE ITERATIVE APPROACH TO PROCESSING OF RANDOMLY SAMPLED
3D NMR SPECTRA.
Jan Stanek, and Wiktor Komiski
64. STRUCTURE AND STEREOCHEMISTRY OF NTBC AND ITS METABOLITES -
A COMBINED NMR/DFT STUDY.
Przemysaw Szczeciski, Anna Dziadecka and Adam Gryff-Keller
65. MOLECULAR DYNAMIC AND STRUCTURE OF PHOSPHATYDYLOCHOLINE
(DMPC)/CATIONIC GEMINI SURFACTANT (GEM-IK1) SYSTEM.
Kamil Szpotkowski, Aleksandra Wypych, Kosma Szutkowski, ad Maciej Kozak,
Stefan Jurga
66. MOLECULAR DYNAMICS IN PROTON CONDUCTING GEL/H 3 PO 4
ELECTROLYTES STUDIED BY NMR.
L. Szutkowska, M. Dobies, K. Szutkowski, Stefan Jurga, G. ukowska, and
W. Wieczorek
67
68
70
71
73
74
75
76
77
78
80

W. Wieczorek
67. METABOLIC PROFILE OF CEREBROSPINAL FLUID OBTAINED FROM
AMYOTHROPHIC LATERAL SCLEROSIS PATIENTS.
Beata Toczyowska
68. STRUCTURAL STUDIES OF N-TERMINAL SEQUENCE OF DERMORPHIN BY
MEANS OF SOLID STATE NMR SPECTROSCOPY AND XRD.
Katarzyna Trzeciak-Karlikowska, Agata Jeziorna, Grzegorz D. Bujacz, Anna Bujacz,
Wodzimierz Ciesielski, and Marek J. Potrzebowski
69. NEW PERMANENT MAGNETS FOR MRI.
Krzysztof Turek, Piotr Liszkowski, and Boguslaw Tomanek
70. ASSESSMENT OF CARDIAC FUNCTION RESERVE IN MURINE MODELS OF
HEART FAILURE, IN VIVO STUDY.
U. Tyrankiewicz, T. Skórka, S. Heinze-Paluchowska, M. Jaboska, M. Wozniak, L.
Drelicharz, and S. Chlopicki
71. LOCAL AND COOPERATIVE DYNAMICS IN MODIFIED
POLY(DIMETHYLSILOXANE)S STUDIED BY FAST FIELD CYCLING NMR,
BROADBAND DIELECTRIC AND FOURIER TRANSFORM INFRARED
SPECTROSCOPIES.
Wiktor Waszkowiak, Aleksandra Wypych, Maria Dobies, Stefan Jurga, and Hieronim
Maciejewski
72. A NOVEL DETERMINATION OF 13 C NUCLEAR MAGNETIC MOMENT
IN TERMS OF THE 3 He FROM GASEOUS 13 CH 4 NMR SPECTRA
Marcin Wilczek , Anna Szyprowska, and Wodzimierz Makulsk
73. EPR EXAMINATION OF FREE RADICALS IN RADIATIVE STERILIZED
PIPERACILLIN.
Sawomir Wilczyski, Barbara Pilawa, Marta Ptaszkiewicz, Janusz Swako, Pawe
Olko, Robert Koprowski, and Zygmunt Wróbel
74. FREE RADICALS PROPERTIES OF DIETARY SUPPLEMENTS BASED ON
FISH AND PLANT OILS APPLIED IN COSMETOLOGY.
Wilczyski Sawomir, Zdybel Magdalena, Barbara Pilawa, Deda Anna, and
Pierzchaa Ewa
75. RELAXATION PARAMETERS OF GD(III) AND MN(II) IONS IN WATER FROM
THE PERSPECTIVE OF CONTRAST AGENTS.
Milosz Wojciechowski and Danuta Kruk
76. UNIPLANAR GRADIENT COILS - FROM DESIGN TO APPLICATIONS.
Grzegorz Woniak, Tomasz Skórka, Krzysztof Jasiski, Tomasz Banasik,
Mohammad Mohammadzadeh, Dominik von Elverfeldt, Jürgen Hennig, and
Wadysaw P. Wglarz
81
83
84
85
87
88
89
90
91
92

77. DYNAMICS OF POLYSTYRENE OF DIFFERENT MOLECULAR
ARCHITECTURE AS REVEALED BY NUCLEAR MAGNETIC RESONANCE,
BROADBAND DIELECTRIC SPECTROSCOPY AND REOLOGY.
Aleksandra Wypych, Monika Makrocka-Rydzyk, Grzegorz Nowaczyk, Eugeniusz
Szczeniak, Stefan Jurga, Akira Hirao, and Takumi Watanabe
78. SPECTRA OF HIGH DIMENSIONALITY FOR EASY RESONANCE
ASSIGNMENT IN PROTEINS.
Anna Zawadzka-Kazimierczuk, Krzysztof Kazimierczuk, and Wiktor Komiski
79. EFFECT OF TEMPERATURE ON EPR SPECTRA OF DOPA-MELANIN-
NETILMICIN COMPLEXES.
Magdalena Zdybel, Barbara Pilawa, Ewa Buszman, Dorota Wrzeniok, Ryszard
Krzyminiewski, Zdzisaw Kruczyski, Robert Koprowski, and Zygmunt Wróbel
80. SPIN-LATTICE RELAXATION IN THERMALLY STERILIZED
CHLORTALIDONE.
Magdalena Zdybel, Adamczyk, Barbara Pilawa, Magdalena Kocielniak, and Daria
Czyyk
81. DISPERSION MEASUREMENTS OF T 1 IN PROTEIN SOLUTIONS.
ukasz elazny, Dorota Wierzuchowska, and Barbara Blicharska
82. THE ONE NMR TM PROBE FROM VARIAN AND THEIR COMBINATION
TOOLS.
Thomas Zellhofer
93
94
95
97
98
99

APPLICATION OF 19 F MAGNETIC RESONANCE IMAGING FOR THE
STUDY OF DRUGS EFFICACY EX VIVO
Dorota Bartusik 1 and Bogusław Tomanek 1,2,3,4
1 National Research Council Canada, Institute for Biodiagnostics (West), Calgary, Alberta,
Canada; 2 Cross Cancer Institute, Department of Medical Physics, Edmonton, Alberta,
Canada; 3 Institute of Nuclear Physics, Polish Academy of Sciences, Krakow, Poland;
4 University of Alberta, Department of Oncology, Edmonton, Alberta, Canada
Oncology is the major beneficiary of pre-clinical and clinical applications of 19 F
NMR. 19 F nuclei is used in pharmaceutical investigations of anti-cancer drugs because 19 F
stabilizes drugs and is responsible for interactions with intracellular microenvironment, thus
drugs efficacy.
The aim of our study was to apply 19 F Magnetic Resonance Imaging at 9.4 T (Cross
Cancer Institute, Edmonton, Canada) to observe drug efficacy. We labeled Herceptin
(Trastuzumab, Genentech Inc., San Francisco, CA) with fluorine in the form of
perfluorocarbon (PFCE, perfluoro-15-crown-5-ether). For the study we selected human breast
cancer cell line MCF-7 with stable positive over-expression of HER-2 protein. On the cell
surface HER-2 is recognized as Herceptin receptor. As control we used Human mammary
epithelial cells (HMEC). The three dimensional cell cultures were established using Hollow
Fiber Bioreactor (HFB, FiberCell System Inc., Frederick, MD). 19 F MRI was used for
visualization of the cellular uptake of new fluorine labeled Herceptin.
We observed that the oil-water emulsion of Herceptin with PFCE was more efficient
than Herceptin alone in MCF-7 culture. Normal (HMEC) cells did not respond to any
treatment. A significant correlation between duration of treatments and MCF-7 cells viability
was observed. 19 F signal intensity increased due to 19 F uptake, however the cells that were
successfully treated were no longer possible for viability assays with trypan blue. The use of
HFB device allowed high-density 3-D cell cultures in the reproducible experimental setup and
provided controlled conditions during biochemical and MR study.
1

TOCOTRIENOLS RELATED CHANGES IN HUMAN
MCF-7 ADENOCARCINOMA USING 19 F MAGNETIC RESONANCE
IMAGING EX VIVO
Dorota Bartusik 1 , Boguslaw Tomanek 1,2,3,4 , Danuta Siluk 5 , and Roman Kaliszan 5
1 National Research Council Canada, Institute for Biodiagnostics (West), Calgary, Alberta,
Canada; 2 Cross Cancer Institute, Department of Medical Physics, Edmonton, Alberta,
Canada; 3 University of Alberta, Department of Oncology, Edmonton, Alberta, Canada;
4 Polish Academy of Sciences, Institute for Nuclear Physics, Kraków, Poland; 5 Medical
University of Gdańsk, Department of Biopharmaceutics and Pharmacodynamics,
Gdańsk, Poland
Tocotrienols, analogues of vitamin E, are significant sources of antioxidant activity to
all living cells. Cell viability is related to oxygen content and can be used to study the growth of
breast cancer cells. Therefore we study the oxygen content using MR methods to measure
tocotrienols’ efficacy.
In the present study we used 19 F Magnetic Resonance Imaging (MRI) measurements for
assessing oxitometry of MCF-7 cells after α-, γ-, δ- tocotrienols treatments. The cells were
grown in three dimensional (3-D) cultures using MR compatible Hollow Fiber Bioreactor
(HFBR) device. We studied the changes in the intracellular oxygen concentration in 3-D
cultures caused by hexafluorobenzene (HFB) uptake in control and treated cells. The 19 F spin–
lattice relaxation of HFB is highly sensitive to oxygen concentration and minimally to
temperature that is useful to measure intracellular oxygen concentrations. The cells’ viability
was assayed using trypan blue.
The cells were treated with tocotrienols (0-1000 µM) during 72 h. The significant
viability changes were observed for concentrations higher than 250 µM of α-tocotrienol, 200
µM of γ-tocotrienol and 150 µM of δ- tocotrienol, as compared to control. The viability
decreased from 93 ± 4% to 52 ± 4%, 45 ± 9% and 38 ± 6%, respectively. The γ- and δ-
tocotrienols were the most inhibitory compounds concerning effects of tocotrienols on MCF-7
cells’ growth. The IC 50 values for α-, γ-, and δ-tocotrienols was 14 µM, 19 µM and 7 µM,
respectively. The oxygen concentrations in the cultures were 13.0 % O2 , 11.0 % O2 , 9.0 % O2 ,
respectively. Significant correlation between the oxygen and vitamins concentrations, that was
related to MCF-7 cells’ viability, was observed.
2

NON-CONTRAST BASED MRI OF HUMAN LUNG PERFUSION
AND VENTILATION
Grzegorz Bauman 1,4 , Michael Puderbach 2 , Michael Deimling 3 , Julien Dinkel 2 , Christian
Hintze 2 , and Lothar R. Schad 4
1 German Cancer Research Center, Medical Physics in Radiology, Heidelberg, Germany;
2 German Cancer Research Center, Department of Radiology, Heidelberg, Germany;
3 Siemens Healthcare, Erlangen, Germany; 4 Computer Assisted Clinical Medicine, University
of Heidelberg, Mannheim, Germany
Assessment of regional pulmonary perfusion and ventilation has important clinical value as an
indicator of lung function. Current standard MR based methods for evaluation of lung
perfusion and ventilation are dependent on the application of the inhalative or the intravenous
contrast agents. We propose a Fourier Decomposition method for non-contrast-enhanced
functional lung MRI [1]. The method was adapted on a 1.5 T clinical MR scanner using fast
acquisition and submillisecond echo sampling with a Steady-State Free Precession (SSFP)
sequence to produce time-resolved 2D data stacks [2].
All measurements were performed on a 1.5 whole-body MR scanner. Seventeen
healthy volunteers (11 men, 6 women) with mean age of 36.5 ± 14.1 years (age range: 19 - 64
years) were investigated. Set of five coronal slices (S1-S5) was acquired in every subject
using time-resolved scans with a 2D+t SSFP sequence (TR/TE = 1.9 ms/0.8 ms and TA = 116
ms/image) to cover the chest volume. Each time-resolved set consisted of n = 198 images for
every slice location with a total acquisition time T = 59.4 s, spectral resolution ∆f = 1/T =
0.017 Hz and spectral width f B = 1/(2·TA) = 1.667 Hz. The identical measurement protocol
was repeated after 24 hours. Neither breath-holding nor ECG triggering were required.
Images in every data stack containing single slice were corrected for the respiratory motion
using fully automatic non-rigid registration algorithm [3]. Pixel-wise application of Fourier
Transform along the time axis of the data stack converted them into sets of n images
representing spectral frequencies f n ∈{0, ∆f, ... , f B }. After the periodical signal intensity
changes with regard to the cardiac and respiratory cycles were identified on the frequency
spectra, integration of appropriate spectral ranges produced perfusion- and ventilationweighted
images. The images showed a homogenous signal distribution without defects for
every healthy volunteer. Medical and technical reproducibility of the method was examined.
The presented method requires only minimal patient compliance and is not dependent
on triggering techniques. Neither administration of intravenous contrast agents nor inhalative
gaseous media like hyperpolarized helium [4], oxygen [5] is needed. Further studies are
required to determine the clinical impact of the proposed method.
References:
1. Deimling M. et al. Proc. 16th annual meeting ISMRM, 2008, Toronto, Canada, p. 2639;
2. Bauman G. et al. Magn Reson Med, 2009 Sep;62(3):656-64;
3. Chefd’hotel C. et al. VLSM'2001, ICCV Workshop, 2001, Vancouver, Canada;
4. De Lange E.E. et al. Radiology, 1999; 210: 851-857;
5. Edelman R.R. et al. Nat. Med., 1999; 2: 1236-1239.
3

CARBON-13 NMR RELAXATION STUDY OF THE INTERNAL
DYNAMICS IN
CYCLODEXTRINS IN ISOTROPIC SOLUTION
Piotr Bernatowicz a , Katarzyna Ruszczyńska-Bartnik b , Andrzej Ejchart b , Helena
Dodziuk a , Ewa Kaczorowska b , and Haruhisa Ueda c
a Institute of Physical Chemistry, Polish Academy of Sciences, Warsaw, Poland;
b Institute of Biochemistry and Biophysics, Warsaw, Poland; c Department of Physical
Chemistry, Hoshi University, Tokyo, Japan
13 C nuclear spin relaxation processes in 6- to 12-membered cyclodextrins (from
α to η) were investigated in D 2 O solution at multiple magnetic fields. Detailed analysis
of the 13 C longitudinal relaxation, the 13 C relaxation in rotating frame and the 1 H- 13 C
nuclear Overhauser enhancement (NOE) in these molecules yielded their rotational
diffusion tensors and semiquantitative picture of the internal motions these oligosugars
undergo. The dynamics in α- and β- molecules seems to be different than the motional
behaviour of their higher analogues. The results suggest that on the time scale of
molecular tumbling none of the investigated cyclodextrins takes rigid truncated-cone
conformation.
4

STRUCTURAL CHANGES IN holo-S100A1
AND S100A1(C85M)PROTEIN.
CHEMICAL SHIFT BASED RESULTS
Monika Budzińska 1) , Michał Nowakowski 1) , Igor Zhukov 1’2) , Agnieszka Belczyk 1) ,
Andrzej Bierzyński 1) , and Andrzej Ejchart 1)
1) Institute of Biochemistry and Biophysic, Polish Academy of Sciences, Warszawa Poland;
2) Slovenian NMR Centre, National Institute of Chemistry, Ljubljana, Slovenia
S100A1 is a homodimeric calcium binding protein. Each subunit, built up of 93
residues, contains two EF-hand motifs linked with a flexible linker [1]. The EF-hand motif is
composed of a sequence helix –loop – helix. The protein undergoes a conformational change
upon binding calcium in order to interact with protein targets and initiate a biological
response [2].
Possible structural changes among apo-S100A1, holo-S100A1 and S100A1(C85M)
proteins can be detected using ∆δ tot parameter based on the chemical shift data [3]:
∆δ tot ={( ∆δ HN ) 2 + 0.024(∆δ N ) 2 + 0.116(∆δ CO ) 2 + 0.076(∆δ Cα ) 2 } 1/2
On the other hand, secondary structure elements can be predicted via the evaluation of Φ and
Ψ backbone torsion angles using programs: PREDITOR [4] and TALOS [5] which are
essentially based on the backbone chemical shifts as well.
Using double labeled 15 N/ 13 C apo-S100A1 and holo-S100A1 proteins we have
assigned the chemical shifts of almost all backbone nuclei in both proteins. Protein secondary
structure elements have been determined and compared pointing out to distinctive
conformational changes due to the modification of crucial C85 residue or calcium binding.
Acknowledgment:
This work was supported by a grant N301 031234 from the Polish Ministry of Science and
Higher Education
References:
[1] R. Donato, Int. J. Biochem. Cell Biol. (2001)33, 637-668.
[2] N.T. Wright, K.M. Varney, K.C. Ellis, J. Markowitz, R.K. Gitti, D.B. Zimmer,
D.J. Weber, J. Biol. Mol. (2005)353, 410-426.
[3] Ayed, F.A.A. Muller, G.S. Yi, Y. Lu, L.E. Kay, C.H. Arrowsmith, Nature Struct. Biol.
(2001)8, 756-760.
[4] M.V. Berjanskii, S. Neal, D.S. Wishart, Nucleic Acids Res. 2006 Jul 1;34
[5] G. Cornilescu, F. Delaglio, A. Bax, J. Biomol. NMR (1999) 13, 289-302.
5

THE DISPERSION OF WATER PROTON SPIN-LATTICE
RELAXATION RATES IN AQUEOUS HUMAN PROTEIN HC
(α 1 -MICROGLOBULIN) SOLUTIONS
Maria Dobies 1 , Maciej Kozak 1 , Stefan Jurga 1 , and Anders Grubb 2
1 Department of Macromolecular Physics, Faculty of Physics,
Adam Mickiewicz University in Poznań, Poznań, Poland; 2 Department of Clinical Chemistry,
University Hospital, Lund, Sweden
The human protein HC (α 1 – microglobulin) is a low molecular weight heterogeneous
glycoprotein widely distributed in the human body fluids and belonging to the lipocalin
superfamily. This biomolecule is produced by the human liver and used in the clinical routine
as a sensitive indicator of tubular dysfunction. The molecular mass of the glycosylated protein
is about 27 kDa. The monomer of protein HC is characterized by a radius of gyration R G =2.16
nm and the dimer by R G =2.93 nm [1].
The 1 H NMR Fast Field Cycling relaxometry was applied to study the molecular
dynamics of the human protein HC (α 1 -microglobulin), its hydration and aggregation
in solution state. 1 H NMRD data for HC protein solutions at three different concentrations:
0.28 mM, 0.56 mM and 0.83 mM are collected in figure 1.
Figure 1. 1 H NMRD data of human
protein HC in water solution at different
concentrations (0.28 mM, 0.56 mM and
0.83 mM). The best fit model-free
profiles (n=3) and best fit Lorentzian
profiles are assigned by the solid and
dotted lines, respectively.
All 1 H NMRD profiles exhibit strong stretching towards low Larmor frequency and
have a two-step shape. The 1 H NMRD data have revealed the complex nature of the
water/protein HC system resulting from the co-existence of monomer and dimer forms of the
protein in solution as well as the presence of oligosaccharides linked to the polypeptide chain.
A comparison of the average correlation time values obtained from the model-free [2]
fits with the values predicted on the basis of hydrodynamic τ r theory, suggests that the
dynamics in solution state is governed mainly by the dimer form of the protein HC (the
dominant contribution to the water proton-spin lattice relaxation comes from exchanging
protons from the surface of the dimer). The existence of small number of oligomeric forms of
the protein HC in solutions is postulated because of the two-step shape of water proton spinlattice
relaxation rate dispersion profiles [3].
References:
[1] Kozak M., Grubb A., Protein&Peptide Letters, 14 (2007) 425
[2] Halle, B.; Jóhannesson, H.; Venu, K. Journal of Magnetic Resonance. 135 (1998) 1
[3] Dobies M., Kozak M., Jurga S., Grubb A., Protein&Peptide Letters, 16 (2009) 1496
6

UNUSUAL STRUCTURE AND NMR SPECTRA
OF SOME CYCLOPHANES
Helena Dodziuk
Institute of Physical Chemistry, Polish Academy of Sciences,
Warsaw, Poland
Cyclophanes with small bridges, like known 1 – 4, 6 - 9 and some hypothetical ones, like
10 exhibiting a distorted ring structure are exciting objects of stereochemical studies 1,2 . NMR
measurements and the calculations of chemical shifts and coupling constants offer, in addition
to X-ray analysis, a unique opportunity to monitor structural distortions from a standard
geometry. Therefore, combined measurements and calculations of NMR parameters of such
basic systems as para-[2.2]cyclophane 1 and its derivatives, superphanes 4, 5 3 , some
[2.2.2]cyclophanes with three bridges 6 – 9 and para-[3.3]cyclophane 10 seem of importance.
Some calculations have been also carried out for cis- 2 and trans-hexahydro[2.2]paracyclophanes
3 NMR spectra of which has been published 4 . It should be stressed that few
calculations of chemical shifts and coupling constants for strained hydrocarbons have been
published while practically no calculations for NMR parameters of cyclophanes exist 5,6 except
our paper 7 on 6 – 9.
The results obtained indicate that the combination of NMR measurements and the
computation of chemical shifts and coupling constants is a powerful tool in studies of highly
strained hydrocarbons. In particular, the literature data for 3 4 seem to indicate that this
compound has not been sufficiently pure.
H
H
H
H
1 2 3 4 5
6 7 8 9 10a 10b
References:
(1) Modern Cyclophane Chemistry; R. Gleiter, H. Hopf, Eds., Wiley-VCH, Weinheim, 2004;
(2) Hopf, H. In Strained hydrocarbons. Beyond van't Hoff and LeBel hypothesis; H. Dodziuk, Ed. Wiley-VCH,
Weinheim, 2009;
(3) H. Dodziuk, M. Ostrowski, Eur. J. Org. Chem., 2006, 5231;
(4) Lin, S.-T.; Yang, F.-M.; Liang, D. W. J. Chem. Soc. Perkin 1 1999, 1725;
(5) L. Ernst, Ann. Rep. NMR Spectroscopy 2006, 60, 77;
(6) L. Ernst, Progr. Nucl. Magn. Res. 2000, 37, 47;
(7) H. Dodziuk, M. Ostrowski, K. Ruud, J. Jaźwiński, H. Hopf, W. Koźmiński, Magn. Res. Chem., 2009, 47,
407.
7

INVESTIGATION OF TAUTOMERISM OF THE SELECTED OXOPURINES IN NEUTRAL
AND BASIC WATER SOLUTIONS BY 13 C NMR AND GIAO-DFT CALCULATIONS
Katarzyna Dybiec, Sergey Molchanov, and Adam Gryff-Keller
Faculty of Chemistry, Warsaw University of Technology
Warsaw, Poland
2-Oxopurine, 6-oxopurine (hipoxanthine), 8-oxopurine and 2,6-dioxopurine (xanthine)
exist in neutral or basic water solutions as neutral molecules, monoanions and/or dianions
(Fig. 1). Moreover, most of them are equilibrium mixtures of tautomers undergoing fast
mutual transformations.
HN
N
HN
O
H
N
N
H
N
O
HN
O
H
N
O
N
N
H
N
N
N
N
H
O
N
H
N
2OP-CO-N1H-N9H
Hyp-CO-N1H-N7H
8OP-CO-N7H-N9H
Xan-CO-N1H-N3H-N7H
Figure 1. Investigated compounds as represented by the most abundant tautomers of
neutral molecules: 2-oxopurine (2OP), hypoxanthine (Hyp), 8-oxopurine (8OP) and xanthine
(Xan).
In order to determine the structures of the species present in water solutions and to
estimate their populations, the series of 13 C NMR spectra of the investigated compounds in
solutions of various acidities have been recorded. The analysis of the δ(pH) dependences
allowed the spectra of particular forms of a given compound to be retrieved. Then, the 13 C
chemical shifts determined in that way were expressed as the population-weighted sums of the
shielding constants calculated theoretically for the tautomers which were expected to be the
most abundant species in the investigated solutions. Such a two-step analysis allowed the
choice of the important structures to be verified and their populations to be estimated.
The molecular structure optimizations and calculations of shielding constants were
performed for oxopurine species solvated by three hydrogen-bonded water molecules using
the DFT and GIAO-DFT methods with B3LYP functional, 6-311++G(2d,p) basis set and
polarizable continuum model (PCM) for including the impact of the bulk solvent.
8

CEREBRAL WHITE MATTER RECOVERY IN ABSTINENT ALCOHOLICS
– A MULTIMODAL MAGNETIC RESONANCE STUDY
Stefan Gazdzinski, Timothy C. Durazzo, Anderson Mon, Ping-Hong Yeh, and Dieter J. Meyerhoff
Center for Imaging of Neurodegenerative Diseases, DVA Medical Center, and University
of California, San Francisco, California
Introduction: Most previous neuroimaging studies of alcohol-induced brain injury and recovery
thereof during abstinence from alcohol used a single imaging modality. They demonstrated
widespread microstructural, macrostructural, or metabolite abnormalities that were partially
reversible with abstinence [1,2], with cigarette smoking potentially modulating these processes [3].
The goals of this study were to evaluate white matter (WM) injury and recovery thereof
simultaneously with diffusion tensor imaging (DTI), magnetic resonance imaging (MRI) and
spectroscopy in the same cohort and to evaluate relationships between outcome measures of similar
regions.
Methods: Sixteen non-smoking alcohol-dependents (nsALC) and 20 smoking individuals (sALC)
were scanned at 1.5 Tesla, at approximately one week of abstinence from alcohol. Ten nsALC and
11 sALC were rescanned at approximately one month of abstinence. 22 non-smoking light-drinkers
were also scanned only once. Structural and spectroscopy data were acquired with T1-weighted
(TR/TI/TE=10/300/4 ms), and multislice 1 H MRSI (TR/TI/TE=1800/300/25 ms) sequences,
respectively. The latter was obtained in 3 parallel planes through the centrum semiovale, nuclei of
the basal ganglia, and cerebellum. Diffusion weighted images were acquired with a single-shot EPI
sequence (TR/TE/TI=5000/100/3000ms, 2.4x2.4x5mm 3 ) with a double refocusing SE and bipolar
external diffusion gradients [4] to minimize eddy-current artifacts without sacrificing SNR. Six
encoding directions and five b-values (0,160, 360, 640, and 1000 sec/mm 2 ) were used. The T1-
weighted images were segmented into gray matter, white matter (WM), and cerebrospinal fluid
(CSF) of major lobes with automated probabilistic segmentation, aided by an automated atlas-based
region labeling of major lobes, cerebellum, and subcortical structures. Regional atrophy-corrected
metabolite concentrations of N-acetyl-aspartate (NAA, a marker of neuronal viability), cholinecontaining
compounds (Cho), myo-inositol (m-Ino) and creatine containing metabolites (Cr), were
calculated by combining spectroscopic and segmented MRI data. For DTI analyses, median FA and
MD in frontal, parietal, temporal, and occipital WM were calculated using only diffusion voxels
with FA>0.2 and WM>95%.
Results: At one week of abstinence, nsALC had higher MD in frontal, temporal, and parietal WM
(all p

modality provides an incomplete picture of neurobiological processes associated with alcohol
induced brain injury and recovery thereof.
References:
1. Pfefferbaum, A., et al., Neurobiol Aging, 2006. 27(7): p. 994-1009.
2. Sullivan, E.V., NIAAA Research Monograph No. 34:NIAAA, 2000, Bethesda, MD. p. 473-508.
3. Durazzo, T.C., et al., Alcohol Alcohol, 2007. 42(3): p. 174-85.
4. Reese, T.G., et al., Magn Reson Med, 2003. 49(1): p. 177-82.
10

INTERACTIVE EFFECTS OF AGE AND EXCESS BODY WEIGHT
ON NEURONAL VIABILITY – A MAGNETIC RESONANCE
SPECTROSCOPY STUDY
Stefan Gazdzinski, Rachel Millin, Lana G. Kaiser, Susanne G. Mueller, Timothy C. Durazzo,
Michael W. Weiner, and Dieter J. Meyerhoff
DVA Medical Center, University of California San Francisco, San Francisco, USA
Background: Excessive body weight is associated with brain structural alterations, poorer
cognitive function [1], and lower prefrontal glucose metabolism [2], and reports suggest that
excessive body weight accelerates aging processes [3]. We demonstrated widespread
decreases in concentrations of N-acetyl-aspartate (NAA, a marker of neuronal viability,
associated with glucose utilization) in a healthy middle-aged cohort, especially in frontal lobe
[4], and in anterior cingulate cortex (part of frontal lobe) of healthy elderly [5] as a function
of higher body mass index (BMI; a measure of body fat). As NAA was found to depend on
age in cohorts with age range spanning several decades (e.g., [6]), we hypothesized that
elevated BMI is associated with faster NAA decline with advancing age.
Methods: We used data from 46 healthy, highly functioning participants (55.3 ± 19.6 years;
range: 22-84years; BMI = 25.0 ± 3.0 kg/m 2 ; range: 19.0-33.9kg/m 2 ) from studies on brain
aging and Gulf War syndrome. Proton magnetic resonance spectroscopy ( 1 H MRS) at 4 Tesla
measured concentrations of NAA, glutamate (Glu, involved in cellular metabolism), cholinecontaining
compounds (Cho, involved in membrane metabolism), and creatine (Cr, involved
in high energy metabolism) in anterior (ACC) and posterior cingulate cortices (PCC). As
absolute metabolite concentrations were not available, we scaled metabolite concentration to
Cr. Individual ratios were modeled as a function of age, BMI, and interactions between age
and BMI.
NAA/Cr in ACC
1.7
1.6
1.5
1.4
1.3
1.2
1.1
1.0
0.9
0.8
18 20 22 24 26 28 30 32 34
BMI [kg/m 2 ]
Age > 50 years
NAA/Cr in ACC
1.7
1.6
1.5
1.4
1.3
1.2
1.1
1.0
0.9
18 20 22 24 26 28 30 32 34
BMI [kg/m 2 ]
Age < 50 years
Figure: Illustration of the trend for statistical interaction between age and BMI: the inverse
association between BMI and NAA/Cr is stronger among participants older than 50 years
compared to their younger counterparts.
Results: Neither BMI nor age were significant predictors of NAA/Cr in ACC; however, there
was a trend for an interaction between BMI and age (p=0.054), reflecting stronger association
11

etween higher BMI and lower NAA/Cr in older participants than in their younger
counterparts (illustrated in the Figure below); this result can be interpreted as faster NAA/Cr
decline among participant with higher BMI as well. Advancing age (p=0.005), but not BMI or
the interaction between age and BMI, was associated with decreasing NAA/Cr in PCC.
Conclusions: Our results suggest that elevated BMI is associated with faster decline in
neuronal viability with age only in ACC. Conversely, BMI was not associated with NAA/Cr
in PCC; the NAA/Cr decline in PCC depended solely on age. Taken together, the adverse
effects of excess body weight on neuronal viability appear to accumulate over a lifetime.
References:
1. Beydoun MA, et al., Obes Rev 2008;9(3):204-18. 2. Volkow, N.D., et al, Obesity: doi:
10.1038/oby.2008.469. 3. Valdez et al., Lancet 2005; 366: 662–64. 4. Gazdzinski, S., et al., Ann
Neurol, 2008. 63(5): p. 652-7. 5. Gazdzinski S., et al., Obesity, in Press 6. Schuff N, et al.,
Neurobiol Aging 1999;20:279-285.
12

TEMPERATURE INDUCED BOUND WATER IMMOBILIZATION
IN CETRARIA ACULEATA (SCHREB.) FR. THALLI OBSERVED
BY PROTON RELAXATION
Hubert Harańczyk 1 , P. Nowak 1 , and M.A. Olech 2
1 Institute of Physics and 2 Institute of Botany, Jagiellonian University, Cracow, Poland
Numerous Antarctic lichen species may resist low temperatures in their habitat and may
dehydrate to extremely low hydration level and rehydrate from gaseous phase to the
hydration level sufficient to initiate photosynthesis [1-5]. Among them the foliose
species seem to be more effective in low temperature protection [6]. The Antarctic
lichen Umbilicaria aprina reveals the lowest detected photosynthetic activity [1]. The
spontaneous dehydration of thallus is a way to resist very low temperature experienced
by lichens, therefore both drought and cold resistance may have similar molecular
mechanism. The formation of molecular glass may allow cell to survive deep
dehydration [7].
The understanding of the molecular mechanism of the metabolic activity recovery
during rehydration of thallus requires the knowledge on a number and distribution of
water binding sites, sequence and kinetics of their saturation, and the formation of
tightly and loosely bound water fractions at different steps of hydration process. The
rehydration process may be accompanied by the dissolving of the water soluble solid
fraction, and by the swelling of the system [8].
The thalli of fruticose lichen Cetraria aculeata (Schreb.) Fr. were collected in
Arctowski Polar Station, King George Island, Maritime Antarctic.
Proton FID is a superposition of the solid signal and two liquid signal components
∗
coming from tightly bound ( T ≈ 100 µs) and loosely bound water fraction ( T ≈ 1000
∗
2
µs). Gaussian function yields satisfactory approximation of solid signal for hydrated
thalli at higher temperature region, however for the samples at lower hydration levels
much better fits supplies Abragam function (with the line halfwidths equal to 34 kHz).
Such a form of solid signal component is characteristic for carbohydrate systems
forming molecular glass [9]. The liquid component of NMR signal decays smoothly in
intensity with the temperature, suggesting that in the tested range of hydrations (∆m/m 0
between 3.9% to 19.9%) no cooperative water freezing takes place. The amount of free
water is not sufficient to initiate the ice crystallization process.
Address for correspondence: H. Harańczyk, D. Sc., Institute of Physics, Jagiellonian University, ul. Reymonta 4,
30-059 Cracow, e-mail: hubert.haranczyk@uj.edu.pl
References:
[1] H. Harańczyk „On water in etremely dry biological systems”, Wyd. UJ 2003 pp. 276.
[2] H. Harańczyk, J. Grandjean, M. Olech, Colloids & Surfaces, B: Biointerfaces 28/4, 239, (2003).
[3] H. Harańczyk, J. Grandjean, M. Olech, M. Michalik, Colloids & Surfaces, B: Biointerfaces 28/4, 251,
(2003).
[4] H. Harańczyk, A. Pietrzyk, A. Leja, M.A. Olech, Acta Phys. Polon. 109, 411 (2006).
[5] H. Harańczyk, M. Bacior, P. Jastrzębska, M.A. Olech, Acta Phys. Polon. A115, 516 (2009).
[6] H. Harańczyk, M. Bacior, M.A. Olech, Antarctic Science 20, 527 (2008).
[7] T. Kikawada, N. Minawaka, M. Watanabe, T. Okuda, Integr. Comp. Biol. 45, 710 (2003).
[8] H. Harańczyk, J. Czak, P. Nowak, J. Nizioł, “Initial phases of dry DNA rehydration by NMR and
sorption isotherm”, Acta Phys. Polon. in press (2010).
[9] W. Derbyshire, M. van den Bosch, D. van Dusschoten, W. MacNaughtan, I.A. Farhat,
M.A. Hemminga, J.R. Mitchell, J. Magn. Res. 168, 278 (2004).
2
13

MAPPING OF THE B1 FIELD USING ADIABATIC
EXCITATION PULSES
Franciszek Hennel
Bruker-BioSpin MRI, Ettlingen, Germany
Introduction. The knowledge of map of the radio-frequency field (B1) is important for
numerous MRI applications, such as the position-dependent flip-angle calibration, homogenizing
of the B1 field with transmit arrays (“B1-shimming”) or the acceleration of multi-dimensional
selective RF pulses (“Transmit-SENSE”). Existing B1-mapping methods use either the intensity
or the phase of the signal as the source of information. The phase-based approach (1,2) has the
advantage of being independent on the relaxation properties of the object, and, as recently
showed by Morrell, achieves a better sensitivity than in signal amplitude-based methods (3).
Morrell’s method uses two RF pulses: the first one producing a B1-dependent nutation about the
x-axis, the second one rotating this state to the x-y plane and thus linking the nutation angle with
the signal phase. The dynamic range of this method is limited by the ability of the second pulse to
generate transverse magnetization: its effect is best at 90°, and vanishes at 180°. We propose a
way of generating the B1-dependent signal phase with higher dynamic range using an adiabatic
half passage excitation pulse.
Methods. It has been demonstrated (4) that an inverse adiabatic half passage pulse (IAHP), with
the sweep starting on-resonance, also allows deriving B1 maps from the phase of the signal. This
required simulation-based lookup tables and was prone to errors due to resonance offsets. The
improved version presented here uses two IAHP’s, each of them preceded by a short block
section without the frequency sweep. The phase of this block pulse is identical with the starting
phase of the IAHP in one experiment, and the opposite in the other. The phase difference of the
images acquired in the two experiments is affected solely by the block section and allows a
straightforward calculation of the RF field strength: B1 = ∆φ/2γτ, where τ is the block pulse
duration. As a further, optional modification, each of the IAHP pulses can be preceded by a block
rewinder pulse of the phase opposite to that of the IAHP. Its role is to compensate the phase
accrual caused by the adiabatic pulse itself. Although this phase is subtracted upon the phase
difference calculation, it may lead to intra-voxel dephasing (and signal loss) in regions of strong
B1 gradients. The duration of the rewinder pulse was numerically optimized for the expected
range of B1 and the given IAHP shape and sweep. As an alternative way of rephasing, an
adiabatic full-passage (AFP) pulse with double duration, half amplitude and half sweep has been
used to produce a spin echo. All excitation schemes have been combined with a standard 3DFT
imaging gradient sequence. Bloch equations have been numerically solved for all sequences to
verify the B1 vs. ∆φ dependence and its sensitivity to Bo offsets.
Results. The result of the simulations (Fig. 1), for which a 5ms cos-sine IAHP pulse of 5kHz
sweep was taken, show a good agreement of the phase difference with the theoretical value for an
astonishingly high range of B1 values. The expected linear dependence is seen even at B1/2 =
1kHz, where the adiabadicity factor (nutation- to nutation-axis-velocity ratio) is of only 0.6. In
the same range, the sensitivity to Bo offsets becomes marginal. All variants of the method were
used to map the B1 field of a 3cm surface RF coil on a 7T Bruker BioSpec system. The phase
difference was unwrapped and scaled to provide an image of B1 in frequency units. For a
verification, another image was measured with a rectangular 1 ms pulse of the same peak
14

amplitude as the IAHP. This pulse produces black bands where B1/2 = n ×500 Hz, i.e.,
where the effective flip angle is a multiple of 180 degrees. An excellent agreement with measured
B1 contours could be observed. (Fig. 2). A reduction of signal, and artifacts in the B1 map in a
small region close to the RF coil circuit could be observed and disappeared when the rewinder
pulse or the spin echo sequence was used.
Fig.1. Simulated dependence of the phase
difference (vertical, rad) as a function of B1 (kHz) for
a range of B0 offsets (±500Hz).
Fig.2. Measured 500 Hz contours of the B1 field
cross-checked with a Nx180deg signal nulling for a
1ms block pulse.
Discussion/Conclusion. Compared to (2) our method replaces the second RF pulse by an
adiabatic 90 degree plane rotation. This guarantees that the nutation angle caused by the initial
pulse (block section) is perfectly transferred to the signal phase in a high range of B1 values. As a
result, the B1 map can be reconstructed without lookup tables and in a higher dynamic range,
making the method attractive for the evaluation of surface coils. The application of a rewinder
RF pulse compensates the dephasing caused by the IHAP, similarly to the matched-AFP echo.
The inherent 90-degree flip angle of this method has to be admitted as a drawback in the context
of fast 3D acquisition.
References:
1. C.H. Oh et al, Magn. Reson. Imaging, 8, 21-25 (1990)
2. G. R. Morrell, Magn. Reson. Med. 60, 889-894 (2008)
3. G. R. Morrell, ISMRM 2009, 376
4. F. Hennel et al., ISMRM 2009, 2610
15

THE MULTIPARAMETRIC CMR-BASED ASSESSMENT OF HEART
FUNCTION IN MURINE MODEL OF HEART FAILURE TGaq*44
Magdalena Jabłońska 1,2 , UrszulaTyrankiewicz 1 , Tomasz Skórka 1 , Henryk Figiel 2 , Sylwia
Heinze-Paluchowska 1 , Mirosław Woźniak 3 , and Stefan Chłopicki 3
1 Department of Magnetic Resonance Imaging, H. Niewodniczanski Institute of Nuclear
Physics PAN, Krakow, Poland; 2 Department of Medical Physics and Biophysics, AGH
University of Science and Technology, Krakow, Poland; 3 Department of Experimental
Pharmacology, Chair of Pharmacology CMUJ, Krakow, Poland
Introduction: The aims of this work was to assess the feasibility of the selected explorative
data analysis techniques for characterization of the murine model of heart failure (TGaq*44)
and to find homogeneous groups which cluster mice with regard to the mechanisms present
on current stage of pathology progression.
Methods: CMR-based images of the left ventricle were created in the short axis plane at the
papillary muscles level and then segmented semi-automatically for evaluation left ventricle
area and its plot against the time. Dobutamine induced stress was used to unmask potential
alterations in cardiac function at early stage of heart failure, so the cardiac function was
measured twice: at the rest and after dobutamine administration. As the indicators of heart
function some parameters which describe the systolic and diastolic phase of cardiac cycle
were estimated. For the data analysis, methods of unsupervised classification (cluster
analysis) were used. This method allows for using descriptive modeling and exploring
information about character of sample or population.
Results and conclusions: In this work, abilities of multi-parametric heart function
differentiation method facilitate the assessment of induced stress response. This approach
seemed to be adequate with regard to the complicated mechanisms existing in this kind of the
cardiac dysfunction being developed by TGaq*44 transgenic murine strain. Such
dysfunctions can’t be recognize by single variable observation or comparison and need more
sophisticated approaches. The investigation of changes at stress, supported by explorative
techniques, should give better chance for separating homogeneous groups as a preliminary
method of data analysis which provides to simplifying and ordering techniques of results
interpretation and information searching.
Acknowledgements: This work was supported by PMSHE (grant NN 518419733).
16

MRI APPLICATION OF MICROSTRIP RF COILS
Krzysztof Jasiński 1 , Peter Latta 2 , Vyacheslav Volotovskyy 2 , Anna Młynarczyk 1 ,
Władysław P.Węglarz 1 , and Bogusław Tomanek 1,2
1 Institute of Nuclear Physics, Polish Academy of Sciences, Kraków, Poland; 2 Institute for
Biodiagnostics, National Research Council of Canada, Winnipeg, Canada
A microstrip is made of a metallic strip on PCB surface creating a planar RF
transmission line, commonly used in microwave electronics. The RF coils based on microstrip
design, have been already applied to MRI and MRS [1,2]. These coils produce homogenous
RF field only within a very restricted field of view. We present here application of microstrip
microcoil to MR microscopy.
To asses MRI performance of a microstrip, numerical simulations were performed.
Finite Element Method, full 3D quasi static, time harmonic simulations showed a decrease of
the RF field amplitude with the distance from the coil.
The simulation software provided impedance of the coil, spatial distribution of B 1
field and current density which allowed calculation of intrinsic SNR for many configurations
of microcoils, enabling selection of an optimal microstrip coil geometry.
To test the coil performance microstrip coil was constructed, tuned to 500 MHz and
matched to 50 Ohm. Electric parameters measured on-the-bench agreed with numerical
simulations.
For MR imaging experiments, the microstrip coil was positioned inside the 11.7T
vertical bore magnet (Oxford Instr. UK) equipped with a commercial 72mm ID gradient set
(maximum 600 mT/m) and Avance console (Bruker, Germany).
Several MR images were acquired proving the high performance of the microstrip
microcoil for MRI microscopy.
References:
[1] X. Zhang, K. Ugurbil, W. Chen, J. Magn. Reson. 161 (2003) 242–251. [2] P.J.M. van Bentum, J.W.G.
Janssen, A.P.M. Kentgens, J. Bart, J.G.E. Gardeniers, J. Magn. Reson. 189 (2007) 104–113.
17

TAUTOMERISM OF SCHIFF BASES DERIVED
FROM (S)-DIPHENYLVALINOL: THE SOLID STATE NMR
AND X-RAY STRUCTURE STUDIES
Magdalena Jaworska 1 , Paweł B. Hrynczyszyn 1 , Katarzyna Nowicka 2 ,
Włodzimierz Ciesielski 2 , and Marek J. Potrzebowski 2
1 Wydział Chemii, Uniwersytet Mikołaja Kopernika, Toruń, Poland;
2 Centrum Badań Molekularnych i Makromolekularnych, Polska Akademia Nauk,
Łódź, Poland
Schiff bases can be obtained in condensation of aromatic or aliphatic primary amines
with aldehydes or ketones. The tautomerism of o-hydroxy Schiff bases is a very important
phenomena: 1) in biological processes which are concerned in less stable tautomers
responsible for biochemical acivity [1], 2) in catalytic systems often used in asymmetric C-C
bond formation reactions including addition of diorganozincs, cyanosilylation of carbonyl
compounds, nitroaldol reaction, alkynylation etc. Theoretically tridentate o-hydroxy Schiff
bases can exist in two tautomeric forms: phenol-imine and keto-amine.
(S)-Diphenylvalinol (1) was obtained from L-valine according to method previously
reported by Itsuno [3]. A series of eight tridentate Schiff bases was prepared in reaction of (1)
with different salicylaldehydes and 2-hydroxy-1-naphtaldehyde in ethanol at room
temperature. All compounds were yellow or orange crystalline solids.
H
Ph
Ph
H 2 N
OH
(S)-diphenylvalinol
Ph
salicylaldehydes or
2-hydroxy-1-naphtaldehyde
Et OH
R 3
R 2
R 1
OH
N
Ph
2, gdzie:
OH
a: R 1 =R 2 =R 3 = H b: R 1 = t-Bu, R 2 = H, R 3 = t-Bu
c: R 1 = H, R 2 = N,N-dietyloamino, R 3 = H d: R 1 = H, R 2 = H, R 3 = Br
e: R 1 = OH, R 2 = H, R 3 = H f: R 1 = Me, R 2 = H, R 3 = H
g: R 1 = i-Pr, R 2 = H, R 3 = H h: R 1 = H, R 2 and R 3 = phenyl
Scheme 1: Synthesis of Schiff bases from (S)-diphenylvalinol
The crystals of 2a and 2e suitable for the diffraction experiment have been obtained
from EtOH/water solution and X-ray structures were prepared. The significant differences of
some bond lengths have been noticed which was concerned in tautomerism of 2e assisted by
intramolecular hydrogen bond between two hydroxyl groups. This phenomena was confirmed
by CP MAS NMR ( 13 C and 15 N) and PASS-2D experiment. Experimental data were
compared with simulation on WinMas (based on Herzfeld-Berger algorithm). The principal
values of δ 11 , δ 22 and δ 33 of the chemical shift tensor were calculated as well as isotropic
average (δ iso ), Ω and κ for carbons and nitrogen atoms which confirmed the occurrence of
tautomeric keto-amine form for compound 2e.
References:
[1] E. D. Raczyńska, W. Kosińska, B. Ośmiałowski, R. Gawinecki, Chem. Rev. 2005, 105, 3561-3612.
[2] for example.: Y. L. Bennani, S. Hanessian, Chem. Rev. 1997, 97, 3161-3195.
[3] S. Itsuno, K. Ito, A. Hirao, S. Nakahama, J. Org. Chem., 1984, 49, 555-557.
18

ADDUCTS FORMATION OF RHODIUM(II) TETRACARBOXYLATES
WITH METHIONINE DERIVATIVES: INVESTIGATIONS
BY THE USE OF VIS AND NMR TECHNIQUES
Jarosław Jaźwiński and Rafał Głaszczka
Institute of Organic Chemistry, Polish Academy of Sciences
Warszawa, Poland
Dinuclear rhodium(II) salts have unique paddlewheel structure with Rh-Rh single bond
surrounded by four carboxylates 1 . Dirhodium salts are soft bases and can accept one or two
organic ligands with oxygen, sulphur, phosphorus or nitrogen atoms in axial positions
forming 1:1 and 1:2 adducts. Complexes of rhodium(II) salts with organic ligands, obtained in
situ in solution have been widely applied in chemistry, spectroscopy and medicine. 2-5 Our
present work concerned the investigations of rhodium(II) adducts with some derivatives of
R- and S-methioninine in D 2 O and CDCl 3 solutions, by the use of 1 H, 13 C, 15 N NMR and VIS
methods. There were three purposes of the work: (i) whether the complexation occurs in
water solution, i.e in the solvent of biological importance, (ii) what is the complexation site in
the ligand; (iii) what is the adduct stoichiometry.
R 1
R 2 R 3 R 4 R 5
R 1
O O
O O
Rh Rh
O O
R 5
S
O
O
N
R 3 R 4
R 2
H H H ep*
CH 3 H H ep*
CH 3 H CHO ep*
CH 3 CH 3 CH 3 ep*
H R 3 = R 4 = Ft** ep*
O O
R 1
R 1
R 1 = CH 3 , CF 3 , C(CF 3 )(OCH 3 )(C 6 H 5 )
H R 3 = R 4 = Ft** O
ep* – electron pair
Ft** =
O
O
Fig.1. Dirhodium(II) tetracarboxylates and ligands studied.
Our investigations led to the following conclusions: methionine, methionine
hydrochloride and methionine methyl ester hydrochloride formed the adducts with
rhodium(II) tetraacetate in water solution; VIS titration experiments revealed stepwise
formation of two adducts, 1:1 and 1:2. Examination of adduct formation shifts ( 1 H and 13 C
NMR) showed that dirhodium teracarboxylates bonded ligands via sulfur atoms. Amide and
imide derivatives of methionine, insoluble in water, behaved similarly in CDCl 3 solution.
References:
1. Cotton F.A., Hillard E.A., Murillo C.A., J. Am. Chem. Soc. 2002, 124, 5658;
2. Agaskar A., Cotton F.A., Falvello L.R., Han S. J., Am. Chem. Soc. 1986, 108, 1214;
3. Díaz Gómez E., Duddeck H., Magn. Reson. Chem. 2007, 46, 23;
4. Díaz Gómez E., Albert A., Duddeck H., Kozhushkov S. I., Meijere A., Eur. J. Org. Chem. 2006,
2278;
5. Jaźwiński J., J. Mol. Struct., 2005, 750, 7.
19

MOLECULAR DYNAMICS AND STRUCTURE IN DIBLOCK
COPOLYMERS STUDIED BY NMR AND BROADBAND DIELECTRIC
SPECTROSCOPY
Jacek Jenczyk, Monika Makrocka-Rydzyk, Aleksandra Wypych,
Stanisław Głowinkowski, and Stefan Jurga
Department of Macromolecular Physics, Faculty of Physics, Adam Mickiewicz University,
Poznań, Poland
Block copolymers are under great research interest due to their current and potential
applications. Properties of these systems depend on individual blocks characteristics and on
the interaction between the components[1]. We have investigated three nearly symmetric
poly(styrene-b-isporene) diblock copolymers: PS(11500)-b-PI(10500)→SI1, PS(45000)-b-
PI(46000)→SI2 and PS(135000)-b-PI(131000)→SI3. Nuclear Magnetic Resonance and
Broadband Dielectric Spectroscopy studies were applied to characterize molecular motions in
copolymers and their neat components. It appeared that the flexible polyisoprene (PI) and stiff
polystyrene (PS) blocks in copolymer influence one another in terms of molecular dynamics.
As a result the increase of the glass transition temperatures (T g ) for PI chains for all
copolymers was observed [2]. Moreover, there is a substantial broadening in the distribution
of relaxation times connected with the normal mode and glass transition process in
copolymers in comparison with neat PI and it is more pronounced for the copolymer of higher
molecular weight. It is assumed that these changes result from both type chains mutual
interactions, since PS blocks make spatial confinement for PI chains motion, whereas the
moving PI chains act as plasticizer for PS blocks. The size of PS domains were determined by
NMR spin diffusion experiment. The obtained size domain values are in good agreement with
those determined by other experimental techniques. [3,4].
References:
[1] I.W. Hamley Developments in Block Copolymer Science and Technology (2004) John
Wiley&Sons, Ltd.
[2] J. Jenczyk, M. Makrocka-Rydzyk, A. Wypych, S. Głowinkowski, M. Radosz, S. Jurga J.
Non Cryst. Sol. (2009), accepted for publication.
[3] B. Cott Pinheiro, K. I. Winey Macromolecules 31 (1998) 4447-4456
[4] J. Denault, B. Morese-Seguela, J. Prud’homme Macromolecules 23 (1990) 4658-4670
Acknowledgements:
This work is supported by 6th Framework Programme under SoftComp Grant No 502235-2
and research grant No N N202 128536 (Poland).
A. Wypych expresses her appreciation to the Ministry of Science and Higher Education in
Poland for a Postdoctoral Fellowship (POL-POSTDOC III).
20

BACKBONE DYNAMICS OF TFE-INDUCED NATIVE-LIKE FOLD
OF REGION 4 OF INITIALLY UNFOLDED E. coli RNA POLYMERASE
σ 70 SUBUNIT
Piotr Kaczka 1 , Agnieszka Polkowska-Nowakowska 1 , Krystyna Bolewska 1 , Igor Zhukov 1,2 ,
Jarosław Poznański 1 , and Kazimierz L. Wierzchowski 1
1 Institute of Biochemistry and Biophysics PAS; Warszawa, Poland; 2 Slovenian NMR Centre,
National Institute of Chemistry, Ljubljana, Slovenia
Folding of a recombinant protein rECσ 70 4, containing domain 4 of E. coli RNA
polymerase σ 70 subunit, accompanying addition of 2,2,2-trifluoroethanol (TFE) to its aqueous
solution, was monitored by heteronuclear NMR spectroscopy. TFE-induced migration of the
resonance signals in a series of
15 N HSQC spectra displayed sequence-dependent
heterogeneity. A common trend of uniform upfield shift in both 1 H and 15 N dimensions,
indicative of generation of helical structures, breaks down for some residues at 10-15% TFE
(v/v) strongly suggesting the buildup of non-helical regions separating initially induced
helices. Spontaneous organization of the helical regions of the polypeptide into a 3Dstructure,
revealed from structural constraints deduced from 15 N- and 13 C-edited NOESY
spectra, accompany secondary structure formation. The absence of long-range cross-peaks in
15 N-edited NOESY spectra recorded for low pH and at 10% (v/v) TFE protein solutions
clearly indicates that in these conditions the live-time of the low-populated high-order
structures is relatively short. Contrary for 30% TFE protein solution 23 structural cross-peaks
in 15 N- and 13 C-edited NOESY spectra were unequivocally assigned, strongly suggesting that
high TFE solution stabilizes protein tertiary structure. Analysis of CSI descriptors estimated
for NMR spectra recorded at 10% and 30% TFE allowed estimation of the population of the
folded rECσ 70 4. The sequential distribution of TFE-induced secondary chemical shifts clearly
indicates that 2,2,2-trifluoroethanol acts not only as a canonical helix inducer, but also tends
to stabilize a spatial arrangement of the induced secondary structure elements. The postulated
HLHTH folding pattern perfectly agrees with the sequential distribution secondasry structure
elements.
Analysis of 15 N relaxation parameters pointed some regions exhibiting significant
deterioration in fast internal motion in favor of the slower ones. The whole HLHTF fragment,
embracing the H1, H2, and H3 helices and the intervening loop/turn sequences, displays the
increased values of the spectral density function J(0) accompanying the decreased values of
spectral density at high frequence J(0.87ω H ), both indicative for a reduced backbone
flexibility on the pico- to nanosecond time scale and large contributions from slow motional
process on the micro- to millisecond time scale. This supports the hypothesis that the helical
secondary structures become formed cooperatively and exhibit high propensity to fold into a
relatively stable tertiary structure. Summarizing, analysis of the relaxation parameters
demonstrated that at 30% TFE solution the motion of the rECσ 70 4 backbone, with the
exception of the terminal residues, is considerably restricted, as indicated by the values of the
generalized order parameter, showing strong tendency of rECσ 70 4 to fold. Reasonable and
relatively not uniform values of S 2 suggest that rECσ 70 4 exhibit anisotropic behavior. All
together, indicate that TFE induces not only the secondary helical structures, but also a
tertiary one resembling in organization that found in homologous σ 4 domains.
21

fMRI QUALITY ASSURANCE SYSTEM: PHANTOM
AND AUTOMATIC DATA ANALYSIS
Karolina Kamińska 1 , Stanisław Adaszewski 1 , Ewa Piątkowska-Janko 1 ,
Piotr Bogorodzki 1 , and Maciej Pisklak 2
1 Institute of Radioelectronics,Warsaw University of Technology, 2 Faculty of Pharmacy,
Medical University of Warsaw, Warsaw, Poland
Introduction:
In typical functional MRI (fMRI) experiments on 1,5T scanners, blood oxygenation level
dependent (BOLD) signal changes are in the range of a few percent [1]. Hence, system
stability during fMRI acquisition is essential for repeatedly and reliable results. The main aim
of this work was to create a fMRI quality assurance (QA) system for daily scanner
performance monitoring. Proposed system consists of: a) agar head phantom mimicking MRI
RF coil loading caused be human head, b) scanning protocol with typical for fMRI studies
settings and automatic image analysis software.
Subjects and Methods:
The head phantom for QA analysis is a 17cm sphere filled with a doped agar. The purpose of
a doping was to mimic healthy brain relaxation times. We adopted original (Functional
Imaging Research Schizophrenia Testbed - Biomedical Informatics Research Network)
FIRST-BIRN agar phantom recipe [2] in order to reach target relaxation times T 1 = 600 ms,
T 2 = 100 ms (for concentration 0,96% of agar and 1,77mM NiCl 2 [3]). The phantom
relaxation times were evaluated on 1,5 T scanner with the following settings: Multi-Echo TE=
30, 60, 90, 120 ms, TR = 1 s and Inversion Recovery TI = 100, 200, 400, 800, 1600, 2400
ms, TR = 10 s. T 1 and T 2 maps were generated using home written Matlab script.
An automatic analysis software called QAWizzard was developed in C++ using open-source
build system Cmake and libraries, GDCM – a cross-platform library for DICOM medical
files, SQLite - SQL database engine, wxWidgets - a widget toolkit for creating graphical user
interfaces, VTK – an open source graphics toolkit. QAWizzard describes the stability of the
MR scanner giving parameters: a) drift, which defines the signal changes during acquisition,
b) signal to noise ratio (SNR) and signal to fluctuation ratio (SFNR) [2].
Results:
Relaxation times of proposed phantom were evaluated on a 1,5 T Siemens scanner with
setting as described earlier. A relaxation maps show good homogeneity either T 1 or T 2
parameters. A mean relaxation times was calculated for centrally located 10 cm 3 volume
giving T 1 = 560 ms and T 2 = 90 ms. Proposed system was evaluated on three 1,5T MRI
scanners. The results of 14-days long trials are summarized in Tb1.
Medical Mean signal
SNR [a.u.] SFNR [a.u.] Drift [a.u.]
Center value [a.u.]
1 1238,5 ± 23,89 130,62 ± 8,68 129,03 ± 3,09 -0,00067 ± 0,00064
2 1198,49 ± 20,74 216,21 ± 8,04 214,54 ± 4,21 -0,00034 ± 0,00083
3 6553,4 ± 580,39 311,77 ± 21,46 233,07 ± 5,127 -0,03333 ± 0,00193
Table 1. QA data
22

Discussion/Conclusion:
Scanner stability is a key to successful fMRI research. Our example shows that QA system is
a useful tool for scanner stability monitoring. Regular measurements of SNR, SFNR and
signal drift provide important feedback about scanner performance.
References:
[1]Matthews, P.M., 2001, Oxford University Press,p 3-34;
[2] Friedman L., Glover G.H., 2006, J. Magn. Reson. Imaging, 23, p.827-839;
[3] Tofts, P.S., 2003, John Wiley & Sons, Ltd, p. 55-81.
23

NMR RELAXATION MEASUREMENTS OF OXIDATION PROCESSES
IN BLOOD SERUM
Joanna Kamińska, Jan Kobierski, and Barbara Blicharska
Jagiellonian University, Department of Physics, Krakow, Poland
Oxidation reactions are crucial for life. One of reactive oxygen species: hydrogen
peroxide can be produced and consumed in cells. At low concentration aqueous solution of
H 2 O 2 is widely used for bleaching and disinfection (Fenton reaction). NMR relaxation
investigations of Fenton reaction in blood serum may help to understood mechanism of
oxidation of biological systems.
In this communication we present the results of time-changes of relaxation times in
blood serum after addition of H 2 O 2 . It is interesting that behavior of spin-lattice T 1 and spinspin
T 2 relaxation times is different when in blood serum additionally the antioxidants like
Vit.C and Vit.E are present.
24

THE USEFULNESS OF DIFFUSION-WEIGHTED IMAGING
IN DISTINGUISHING BETWEEN SOME BRAIN PATHOLOGIES
Aleksandra Klimas 1 , Zofia Drzazga 1 , and Ewa Kluczewska 2
1 Department of Medical Physics – University of Silesia, Katowice, Poland; 2 Department of Clinical
Radiology and Diagnostics Silesian University of Medicine, Katowice, Zabrze, Poland
Introduction Diffusion-weighted imaging (DWI) has become a powerful, multifaceted tool for
advanced imaging in diagnosis and treatment planning. Diffusion MRI measures the mobility
of the water protons and thus provides a window into tissue microstructure.
DWI is used in imaging the central nervous system (CNS) and quantitatively characterizing
different pathological states of brain (1,2,3)
The aim of this work is to test the ADC values as a function of b-value in distinguishing
between some brain pathologies, such as: schwannoma acoustic neuroma, apoplectic lesion,
sclerosis multiplex, arachnoid cyst.
Materials and methods T1, T2-weighted spin echo sequence and DWI of 4 patients (2 women,
2 man) with different brain diseases and two healthy volunteers were performed on 1,5 T GE
MR unit. DWI was performed using spin echo EPI with diffusion gradient applied in three
orthogonal directions with b=500 s/mm 2 , b=1000 s/mm 2 , b=1200 s/mm 2 and additionally with
b=0 s/mm 2 . The ADC maps were calculated using Functool on the GE console.
Results and Discussion Figure 1 presents ADC map with regions-of-interest (ROIs) marked
manually on slices of patients with: a) schwannoma acoustic neuroma, b) apoplectic lesion, c)
sclerosis multiplex, d) arachnoid cyst. We can see that regions of diseases are characterized
by increase of signal intensity dependent on pathology.
A) C)
B) D)
Figure 1. ADC map with regions-of-interest (ROIs) a) schwannoma acoustic neuroma,
b) apoplectic lesion, c) sclerosis multiplex, d) arachnoid cyst.
25

Figure 2. The ADC values for different pathologies and normal tissue. Insert: Comparison of
ADC values of normal brain tissue between patients and healthy volunteers.
The calculated ADC values as a function of b-value in ROIs are presented in Figure 2. One
can see that ADC values are only slightly dependent on b-values. For healthy brain tissue
ADC parameter is below 1.0 x 10-^3 mm2/s similarly as in other works (4). The highest
values of ADC (about 2,5^x10^-2 mm2/s) was found for arachnoid cyst and apoplectic lesion
what is connected with strong hydration of studied tissue. Smaller changes of ADC
parameters are observed for schwannoma acoustic neuroma and sclerosis multiplex.
Insert in Figure 2 shows comparison between ADC values calculated in areas outside the
pathology of patients and in ROIs of healthy volunteers. It should be noted that calculated
values of ADC are similar in the range of error. It means that it is possible to analyze
pathologic changes respect to suitable healthy part of the brain.
∆ADC = ADC healthy – ADC pathology
b-value sclerosis multiplex schwannoma acoustic neuroma apoplectic lesion arachnoid cyst
500 45,6 86,2 160,7 163,0
100 50,0 90,1 163,7 172,7
1200 44,2 90,0 161,0 175,7
Table 1. Differences between ADC values in the normal and pathological tissue of the brain.
In Table 1 differences between ADC values in the normal and pathological tissue of the brain
(∆ADC = ADC healthy – ADC pathology ) are listed. Quantitative results confirm that the highest
change of the signal intensity was found for arachnoid cyst and the smallest for sclerosis
multiplex and this parameter is nearly independent on b-value.
Conclusions:
∆ADC values obtained from ADC maps for lesion and normal areas of the brain could be
useful for distinguishing between different pathological states.
References:
1. Lutsep HL, Albers GW, de Crespigny A, Kamat GN, Marks MP, Moseley ME. Ann Neurol 1997;41:574–580.
2. Werring DJ, Clark CA, Barker GJ, Thompson AJ, Miller DH. Neurology 1999;52:1626–1632.
3. Mauricio Castillo, J. Keith Smith, Lester Kwock, and Kathy Wilber.AJNR Am J Neuroradiol 2001; 22:60–64.
4. L. Røhl1, J. Sørensen2, A. Obel1, L. Elgaard1, L. Bro1, E. Nielsen1, A. Nehen1. Proc. Intl. Soc. Mag. Reson.
Med. 11 (2004).
26

CHEMICAL EXCHANGE PROCESSES IN HYDROGEN PEROXIDE SOLUTIONS
OBSERVED BY NMR RELAXATION
Jan Kobierski 1 , Hartwig Peemoeller 2 , and Barbara Blicharska 1
1 Jagiellonian University, Department of Physics, Krakow, Poland; 2 University of Waterloo,
Department of Physics, Waterloo, ON, Canada
It is known, that hydrogen in aqueous solution of H 2 O 2 exchanges quickly and
completely between hydrogen peroxide and water. This process is called a chemical
exchange. NMR experiments in high fields enable to observe a chemical exchange. We have
used Selective Inversion Recovery method and 2D experiments to estimate the rate of this
process.
In some cases T 1ρ dispersion measurements can be applied not only for calculation of
molecular dynamics parameters (like correlation time and activation energy} but also for
estimation of time of exchange processes. If exchange time is shorter than correlation time
the relaxation is determined by the time of chemical exchange. In this communication we
present application of NMR methods for observation of chemical exchange for H 2 O 2 solution
samples.
27

DETECTOR FOR PROTON-ELECTRON DOUBLE
RESONANCE IMAGING (PEDRI)
Łukasz Kołaszewski 1 , Piotr Bogorodzki 1 , Ewa Piatkowska-Janko 1 , Jerzy Piotrowski 2 ,
Jerzy Skulski 2 , and Maciej Pisklak 3
1 Institute of Radioelectronics, Faculty of Electronics and Information Technology at the Warsaw
University of Technology, POLAND; 2 Institute of Microelectronics and Optoelectonics, Faculty
of Electronics and Information Technology at the Warsaw University of Technology, POLAND;
3 Department of Physical Chemistry, Faculty of Pharmacy at the Medical University of Warsaw,
POLAND
Purpose/Introduction
PEDRI makes use of a double resonance technique based on the Overhauser-Effect [1], which is
essentially a combination of nuclear magnetic resonance (NMR) and electron paramagnetic
resonance (EPR). Simultaneously irradiation at EPR frequency during NMR signal acquisition can
cause a transfer of magnetic polarization from the electrons to the nuclear spins, resulting in an
enhancement of the measured NMR signal up to 330 times [1].
Subjects and Methods
Detector was designed to Bruker BNT1000 magnetic field of 89 mT. This results in proton Larmor's
frequency 3.728 MHz and 2.454 GHz for electrons. Due to very high electron's gyromagnetic ratio
28 GHz/T [2], compared to proton's 42.6 MHz/T [2], the microwaves rectangular resonator has
been used. The resonator works in mode TE 102 , providing mutual perpendicularity of three magnetic
fields ( B EPR , B NMR , B 0 ). In a geometrical center of the resonator a NMR probe (21 turns of 0.9
mm 2 copper wire solenoid coil) is placed. The probe is connected via impedance transformer to
scanner T/R switch. A 1.4 ml teflon container filled with 4.0 mM water solution of TEMPOL free
radical was placed inside the probe. Thanks to TE 102 mode, there is minimal E field component
within sample volume, which prevents it's heating.
Results
A microwave resonator was measured on AgilentTechnology E5071C vector network analyzer
giving following results: unloaded f 0 =2.45563 GHz, Q=4464 and loaded with sample f 0 =2.45417
GHz, Q=2231. A NMR probe was tuned to 3.7284 MHz, with measured Q-factor about 90.
Thereafter a whole detector was placed in Bruker BNT1000 resistive magnet bore, with mean
magnetic induction of 89 mT and linear sweep of +/-0.82 mT. Signal generator HP33120A was used
to control field sweep and network analyzer via optical isolation. Network analyzer was used as a
receiver in the time domain mode, measuring resonator's S 11 factor during field sweep. Total
spectrum acquisition time was 12 minutes with total 36 averages of 20 seconds lasting acquisitions.
Obtained spectrum is similar to the one obtained on MiniScopeMS200 spectrometer with there are
three absorption lines, which are characteristic for TEMPOL radical [2]. However half height width
1.63 MHz was substantially smaller.
Discussion/Conclusion
Proposed detector was tested on BrukerBNT1000 with good results. Both NMR and EPR blocks are
working as they was indented and it makes potential ability for PEDRI idea.
References:
[1] "Design and evaluation of Overhauser enhanced MRI visible markers"; Raimo Joensuu;
[2] "Dynamic Nuclear Polarization with Nitoxides Dissolved in Biological Fluid"; Daniel Grucker;
28

RECENT ADVANCES IN MULTIDIMENSIONAL NMR
SPECTROSCOPY WITH SPARSE RANDOM SAMPLING
Wiktor Koźmiński, Krzysztof Kazimierczuk, Maria Misiak, Jan Stanek,
and Anna Zawadzka-Kazimierczuk
Faculty of Chemistry, University of Warsaw, Warszawa, Poland
Nuclear Magnetic Resonance is nowadays one of the most efficient spectroscopic
techniques, providing insight into molecular structure and dynamics. The observed
frequencies of oscillatory Free Induction Decay signal are sensitive indicators of electron
surroundings of nuclei and their, often very subtle, changes caused by inter- and
intramolecular interactions. However, especially in studies of biomolecules, complexity of
NMR spectra often causes difficulties in their interpretation. This problem may be solved by
acquisition of multidimensional spectra which increases resolution and enables identification
of nuclei connected by mutual interactions.
The conventional approach to recording multidimensional NMR experiments is
limited by the need for fulfilling of the Nyquist Theorem to avoid aliasing. It determines the
sampling rate to be twice higher than the highest frequency expected in the signal. In
consequence, this is an implicit limit for the maximum evolution time and therefore
obtainable resolution. Because of this, for spectra of higher dimensionality, it is practically
impossible to reach the relaxation limits in indirectly sampled dimensions. The problem of
effective acquisition of multidimensional NMR spectra is getting relatively more important at
high fields, where the required Nyquist rate increases together with spectral width.
Random sampling of evolution time space and Multidimensional Fourier Transform,
enables to obtain, without aliasing, spectra at a very small fraction of number of data points
required conventionally. Moreover, the relative level of artifacts caused by random sampling
does not depend on this fraction. Some new applications of high dimensionality (3-6D) will
be shown where the maximum evolution times are limited only by transverse relaxation rates.
References:
K. Kazimierczuk, W. Koźmiński, I. Zhukov, J. Magn. Reson. 179, 323-328 (2006).
K. Kazimierczuk, A. Zawadzka, W. Koźmiński, I. Zhukov, J. Biomol. NMR, 36, 157-168
(2006)
M. Misiak, W. Koźmiński, Magn. Res Chem., 45, 171-174 (2007)
K. Kazimierczuk, A. Zawadzka, W. Koźmiński, I. Zhukov, J. Magn. Reson., 188, 344-356
(2007)
K. Kazimierczuk, A. Zawadzka, W. Koźmiński, J. Magn. Reson. 192, 123-130 (2008)
K. Kazimierczuk, A. Zawadzka, W. Koźmiński, I. Zhukov, J. Am. Chem. Soc., 130, 5404-
5405 (2008)
M. Misiak, W. Koźmiński, Magn. Res. Chem., 47, 205-209 (2009)
K. Kazimierczuk, A. Zawadzka, W. Koźmiński, J. Magn. Reson., 197, 219-228 (2009)
29

THE ROLE of FULLEREN C 60 , CARBON NANOTUBES AND CRBON
ENCAPSULATED MAGNETIC NANOPARTICLES
IN CREATING OF CREATOL (5-HYDROXYCREATININE)
AND UREMIC TOXIN -METHYLGUANIDINE.
Hanna Krawczyk and Magdalena Popławska
Faculty of Chemistry, Warsaw University of Technology, Warsaw, Poland
Diabetic patients with chronic renal failure accumulate in their sera the creatinine
oxidative metabolites-the creatol (5-hydroxycreatinine), which decomposes into uremic
toxin methylguanidine. The creatol was identified in the urine of patients suffering from
kidney diseases in the beginning of 1990’s [1]. Its significance lies in the fact that in
vivo, it is a direct product of reaction of creatinine with the hydroxyl radical [2].
Hydroxyl radical has a highest reactivity among various species of free radicals. It is
known that in position 5 of creatinine in oxidative conditions, one of the protons at C-5
of the imidazole ring is substituted with a group containing an oxygen atom [3-5] or
amine group [3]. Such transformation proceeds with the presence of activated
carbon[5]. This observation is important as the activated carbon coated with different
types of polymer is widely used as a sorbent in haemoperfusion techniques for the
removal of various toxic products and metabolites [6]. Taking this data into account, we
investigated products of creatinine reaction with water in the presence of other forms of
carbon- fulleren C 60 , carbon nanotubes (CNTs) and carbon encapsulated magnetic
nanoparticels (CEMNPs).
Reference:
1. K. Nakamura and K. Ienaga Experientia 1990, 46, 470-472.
2. K.Nakamura, K.Ienaga ,T. Yokozawa, N. Fujitsuka Nephron. 1994, 68(2):280-1 and Comment on: Nephron.
1991;58(1):42-6.
3. H. Krawczyk, A. Pietras and A. Kraska, Spectrochim. Acta A, 66(1) (2007) 9.
4. K. Nakamura; K. Kawano, (Nippon Zoki Pharmaceutical Co., Ltd., Japan). Eur. Pat. Appl. EP 1195373 A2 10 Apr 2002, 9
pp. DESIGNATED STATES: R: AT, BE, CH, DE, DK, ES, FR, GB, GR, IT, LI, LU, NL, SE, MC, PT, IE, SI, LT, LV,
FI, RO. (English). (European Patent Organization). CODEN: EPXXDW. CLASS: ICM: C07D233-88. ICS: G01N030-
02. APPLICATION: EP 2001-123251 2 Oct 2001. PRIORITY: JP 2000-303172 3 Oct 2000. DOCUMENT TYPE: Patent
CA Section: 9 (Biochemical Methods) Section cross-reference(s): 14.
5. H. Krawczyk, J. Pharm. Biomed. Anal., 49 (2009) 945.
6. J.H. Henderson, C.A.McKenzie, P.J. Hilton, R.M. Leach, Thorax 56 (2001) 242–243.
30

DO ESR AND NMR DESCRIBE THE SAME DYNAMICS OF
PARAMAGNETIC SYSTEMS?
Danuta Kruk
Institute of Physics, Jagiellonian University, Krakow, Poland
In this contribution an attempt is made to describe within one set of parameters multifrequency
ESR spectra for selected Gd(III) complexes and Nuclear Spin Relaxation
Dispersion Profiles (NMRD) for their solutions. So far literature reports say that such a
unified description is very difficult; in fact all attempts turned out to be unsuccessful. The
difficulties are attributed to the fact that electron and nuclear spins are sensitive to frequencies
different by three orders of magnitude and therefore a very accurate modeling of electron spin
spectral densities is needed to describe properly their wings to which nuclear spin is sensitive.
In this work instead of modeling the electron spin spectral densities we go in the direction of
an elaborated description of the relaxation processes beyond validity regimes of perturbation
approaches.
A general treatment of Electron Spin Resonance spectra and nuclear spin relaxation in
paramagnetic systems with S ≥ 1, valid for arbitrary motional conditions and interaction
strength has been applied.. This theory is known in the literature as the ‘slow motion theory’
or the ‘general theory’[1-4]. The name ‘slow motion’ indicates that the motion modulating the
relevant interaction is too slow, compared to the timescale of the spin dynamics, for the
perturbation theory to be valid. This approach is based on a full solution of the Liouville von-
Neumann equation by the multipole representation of tensor operators and the Wigner-Eckart
theorem, and has been developed and modified depending on applied motional models.
As a price for this high computational effort we succeeded for the first time with a
unified description of ESR and NMR features of paramagnetic systems.
References:
1 Westlund P.-O. Dynamics of solutions and fluid mixtures by NMR, J. J. Delpuech (Ed.), Wiley,
Chichester 1995, 173-229
2 Kowalewski J., Kruk D., Parigi G. Advances in Inorganic Chemistry, 2005, 57, 41-104
3 Kruk D., Kowalewski J. J. Chem. Phys. 2002, 116, 4079-4086
4 Kruk D., Kowalewski J., Westlund P.-O. J. Chem. Phys. 2004, 121 (5), 2215-2227
This work has been financed by Polish Ministry of Science and Education,
grant No N N202 105936
31

ANISOTROPIC DIFFUSION PHANTOM FOR B-MATRIX
CALCULATION
Artur Tadeusz KrzyŜak
Department of Magnetic Resonance, H. Niewodniczański Institute of Nuclear Physics PAN
Introduction
Usually the components of the b matrix needed to calculate diffusion tensor (DT) are
determined analytically [1]. The results are estimated only due to complex formula used in the
calculation and unknown the real shapes of MRI’s gradients. We didn’t monitor e.g. the
influence of addition to “cross terms” involved by eddy currents, LC circuits and other
sources of gradient’s distortion. The proposed method eliminates these shortages and enables
the precise and spatial determination of b matrix for any DTI sequence.
Subject and method
The anisotropic diffusion phantom was built in the form of a brick composed as an
array of thin glass plates (100 um) separated with H 2 O layers (20 um). DTI measurements
were performed on 1.5 T GE Signa scanner. The b matrices were calculated for selected ROIs
(2mm 2 x1mm) of phantom’s volume for several directions of diffusion gradient vector. The
calculation carried out using well-known Stejskal-Tanner equation [2],
A ( b)
3 3
ln( ) = − b D
∑ i=
1 ∑ j=
1
A(0)
where: A(b), A(0) - the echo intensities with and without the diffusion gradients, b ij -
components of the b-matrix, D ij - components of the known (as was assumed) diffusion
tensor of phantom. In order to determine the values of b-matrix for any direction of the
diffusion gradient vector, a system of no less six equations was solved for distinct D ij values.
The tensor diffusion D ij values were obtained by rotation of phantom (assumption equipoise
with rotation of tensor D) with various set of Euler’s angels. Data for different ROIs were
compared using two tailed unpaired student’s t-test.
Results
The following diffusion tensor values for phantom in main axes frame D 1 = 1,1 +/-
0,12 [x10 -3 mm 2 /s], D 2 =2,22 +/- 0,21 [x10 -3 mm 2 /s], D 3 =2,12 +/- 0,19 [x10 -3 mm 2 /s] were
registered. Preliminary example results of comparison of b matrices achieved for given ROIs
for several directions of diffusion gradient vector. The statistically significant alterations (p

ESR LINESHAPE FOR MULTISPIN SYSTEMS IN THE PRESENCE OF
RELAXATION
Aleksandra Kubica 1 , Artur Mielczarek 1, 2 , and Danuta Kruk 1
1 Institute of Physics, Jagiellonian University, Krakow, Poland
2
Instytut Fizyki Jądrowej PAN, Kraków, Poland
The main theoretical difficulties in Electron Spin Resonance (ESR) spectroscopy are
caused by the fact that the electron spin system is beyond the validity range of perturbative
treatments. This happens when the electron spin motions are driven by two large, noncommuting
Hamiltonians; for example the Zeeman Hamiltonian, which arises from the
interaction of an electron spin with a laboratory magnetic field, and the zero-field splitting
interaction, which results from spin-orbit couplings for the electron spin quantum numbers S
≥ 1. We present a general treatment of ESR spectra, valid beyond the applicability of
perturbative approaches, for arbitrary motions and interaction strengths. This treatment is
known as the “slow motion theory” or the “general theory” [1-3]. The approach is based on
expressing all relevant interactions and dynamic processes [4,5] (represented first in terms of
Liouville operators) as a (super)matrix in a complete orthonormal basis set [1-3,6], including
all relevant interactions and degrees of freedom of the system. The computationally heavy
step is finding a small number of elements of the inverse of this complex matrix. ESR spectra
are fully determined by only one element of the inverted supermatrix. The method has been
extended to include other interactions, like higher terms of the zero field splitting and the
anisotropy of the g-tensor. This work focuses on the complex influence of the g-tensor
anisotropy on the ESR lineshapes for high electron spin quantum numbers.
References:
[1] Westlund P.-O. Dynamics of solutions and fluid mixtures by NMR, J. J.
Delpuech (Ed.), Wiley, Chichester, 173-229, (1995)
[2] Kowalewski J., Kruk D. and Parigi G. Advances in Inorganic Chemistry, 57, 41-
104, (2005)
[3] Kruk D. Theory of Evolution and Relaxation of Multi-spin Systems.
Application to Nuclear Magnetic Resonance (NMR) and Electron Spin
Resonance (ESR), Abramis Academic, Arima Publishing UK, (2007)
[4] Freed J.H., Bruno G.V. and Polnaszek C. J. Phys. Chem. 55, 5270-5281, (1971)
[5] Freed J.H., Bruno G.V. and Polnaszek C. J. Phys. Chem. 75, 3385-3399, (1971)
[6] Lynden-Bell R.M. Mol. Phys. 22, 837-851, (1971)
This work has been financed by Polish Ministry of Science and Education,
grant No N N202 105936
33

ESTIMATION OF PHOSPHOLIPIDS CONCENTRATION IN PLASMA,
MONONUCLEAR CELLS, AND ERYTHROCYTES FROM PATIENTS
WITH HEMATOLOGICAL CANCERS - 31 P MRS IN VITRO STUDY
Małgorzata Kuliszkiewicz-Janus
Department of Hematology and Transplantology,
Wrocław Medical University, Poland
The investigations of 31 P spectra of sera and cells of patients with hematological malignancies
(acute leukemia, malignant lymphoma, multiple myeloma) and other cancers: renal, thyroid,
esophageal are a clinical trials over the introduction of MRS to monitoring of the therapy. The
changes of concentrations of phospholipids in sera and cancer cells may explain their transport
through cell membranes and significance of this mechanism in apoptosis.
Phospholipids (phosphoglycerides and sphingomyelins) are essential elements of cellular
membranes. Phosphatidylcholine (PC) and sphingomyelin (SM) appear mainly in outer (exterior)
leaflet of membrane. Phosphatidylserine (PS), phosphatidylethanolamine (PE) and
phosphatidylinositol (PI) – are in inner leaflet. In plasma of healthy person PC, SM, LPC and PE
prevail. PI and plasmalogens appear in plasma only in inconspicuous quantity. Cardiolipin (CL) is
essential element of mitochondrial membranes. Sphingomyelin is the most known phospholipid in
cancer disease as an important participant of signal transduction pathway.
In our investigation 31 P NMR spectra were obtained from: sera, sera with added the sodium
salt of cholic acid, methanol-chloroform extracts of phospholipids from: plasma, peripheral blood
mononuclear cells (PBMC), bone marrow mononuclear cells (BMMC). Isopropanol-hexan extracts
of phospholipids we used to extracted: peripheral blood erythrocytes (PBE), bone marrow
erythrocytes (BME). Blood samples were collected by venous puncture after an overnight fast, for
sera – 10 ml, for methanol chloroform extracts of phospholipid from plasma, PBMC, BMMC, PBE,
BME 9 -15 ml. Cellular lipids were isolated from mononuclear cells Ficoll buffy coat
centrifugation, and next underwent methanol-chlorophorm extraction. All spectra came from
phospholipid extracts prepared from 60 x 10 6 cells for PBMC, BMMC and 5 x 10 9 for PBE, BME.
Studies were carried out on AMX 300 Bruker and Avance III Bruker spectrometer 7.05 T.
In our preliminary studies we observed that 31 P NMR spectra of normal serum consist of three
peaks including a downfield peak due to Pi and two additional upfield field peaks from
phospholipids PE+SM and PC. Spectra were performed in healthy volunteers, patients with acute
leukemia, malignant lymphomas and multiple myeloma, at the time of diagnosis and repeated up to
13 times during chemotherapy. The sodium salt of cholic acid added to serum caused separation of
three phospholipid peaks located upfield from inorganic phosphate. Contrary to the earlier studies,
peaks from PE+SM and PC, and also a peak from LPC were observed. The above mentioned
separation method had also been applied in investigations of patients with digestive tract tumors
and with renal cell carcinoma. Changes in phospholipids in the 31 P NMR spectra observed in these
patients were primarily dependent on the advance of the disease. Long-term follow-up studies
showed a good correlation between this 31 P MRS evaluation of sera and the response of the disease
to the therapy. At the time of diagnosis spectra showed strongly reduced peak areas and intensities
from phospholipids (PC, LPC and PE + SM). During chemotherapy important changes in spectra
34

were observed: (1) in responding patients the spectral profile changed to resemble that of normal
serum with increased peak intensities, (2) in non-responding individuals peak intensities were
reduced. Spectra of patients suffering from acute leukemia or HD, who have achieved complete
remission for 4-12 years did not differ from spectra of healthy volunteers. 31 P NMR spectra can
prove the presence of the residual leukemia in patients when the number of leukemic cells equals to
10 9 and is not detectable in laboratory tests of the blood and bone marrow. 31 P NMR spectra can
depict the function of the transplantated. It is possible to estimate the efficiency of cytostatics: e.g.
DHAD & ARA-C as a first course treatment in patients with acute leukemia.
In our study 31 P MRS spectra of normal extract of serum consist of six peaks due to
phospholipids. Beside previously identified, i.e. PC, LPC, PE and SM, some new ones were
observed: PI, CPLAS. 31 P NMR spectra of phospholipid extracts from plasma. In patients with
hematological cancers the values of peaks areas of PC, CPLAS, LPC, SM decreased. In some
patients peaks from PI, PE were not observed. In responding patients the spectral profile changed to
resemble that of normal extract. 31 P MRS spectra of extract of mononuclear cells consist of 9 peaks
due to phospholipids: PC, CPLAS, LPC, SM, PE+PI, PS, CL. The peak of LPC that has the most
prognostic value in sera, in extract of mononuclear cells was observed only in some healthy
volunteers. Important meaning for prognosis of disease course has got a presence of CPLAS (PAF).
In the case of booth the PBMC and BMMC, the PAF concentration was significantly diminished in
patients with ALL relative to the concentration for those with AML and for the healthy
volunteers. No differences were observed in the PAF concentrations for the AML patients and the
healthy volunteers. We focused on the significant difference in the integral intensities and
phospholipid concentrations of SM and PS as well as CPLAS and PI+PE between the ALL and
AML groups.
The experiences already achieved pointed out that 31 P spectra at present didn't allow for
diagnosis of hematological disorders, but were of great importance in monitoring of therapy of the
diseases under consideration. The changes of concentrations of phospholipids in sera and cells in
hematological cancers are probably due to the increased uptake of phospholipid metabolites in
proliferating blast cells, and their disturbed transport through cell membranes. It seems that SM and
CPLAS (PAF) are of great importance in this process. It is very likely that observed reduction of
the level of SM in blast cells’ extraction is due to activation of sphingomyelinase (SMnase) by for
example tumor necrosis factor (TNF) or PAF. SMnase cleaves SM, generating choline phosphate
and ceramide. The latter – among other things – mediates programmed cell death (apoptosis),
induces differentiation and inhibits growth of leukemia cells. 31P MRS it may be useful method for
understanding the meaning of phospholipids in mechanism of cancer’s proliferation. In this review
of our study, it was shown how 31 P MR spectroscopy can be applied for observations of changes of
phospholipid concentrations in sera, plasma, PBMC BMMC, PBE, BME in patients with cancer
diseases. This investigation points out that 31 P MR spectra allow monitoring of phospholipid
concentration in body fluid as well as in cells. Moreover, among acceptable various applications of
31 P MRS, the possibility evaluating an advancement of disease, monitoring the treatment and
forecasting resistance to the therapy seems to be the most promising. Simplicity of presented
methods makes 31 P MR spectroscopy a very useful tool for investigations of phospholipid
metabolism.
35

PERFORMANCE OF DFT, SOPPA, SOPPA(CCSD) AND CCSD(T)
METHODS IN PREDICTING NUCLEAR ISOTROPIC SHIELDINGS
IN THE COMPLETE BASIS SET LIMIT
Teobald Kupka 1 , Michał Stachów 1 , Marzena Nieradka 1 ,
Jakub Kaminsky 2 , and Tadeusz Pluta 3
1 University of Opole, Faculty of Chemistry, Opole, Poland; 2 Department of Molecular
Spectroscopy, Institute of Organic Chemistry and Biochemistry, Prague, Czech Republic;
3 University of Silesia, Institute of Chemistry, Katowice, Poland
Convergence patterns and limiting values of isotropic nuclear magnetic shieldings
were studied for several small molecules (N 2 , CO, CO 2 , NH 3 , CH 4 , C 2 H 2 , C 2 H 4 , C 2 H 6 and
C 6 H 6 ) in the Kohn-Sham limit. Individual results of calculations using dedicated families
of Jensen’s basis sets (pcS-n and pcJ-n) and simple two-parameter formula were fitted
toward the complete basis set limit (CBS) and ZPV correction applied. Several density
functionals were used, and, for comparison purposes, calculations were performed using
RHF, MP2, SOPPA, SOPPA(CCSD) methods and aug-cc-pVTZ-J basis set. Very
demanding CCSD(T) calculations were performed in some cases, too. Finally, the CBS
estimated results were critically compared with earlier reported literature data and
experiment.
Literature
1. A. Auer, J. Gauss, J.F. Stanton, Quantitative prediction of gas-phase 13C nuclear
magnetic shielding constants, J. Chem. Phys. 118, 10407 (2003).
2. T. Kupka, B. Ruscic and R. E. Botto, Toward Hartree-Fock- and Density Functional
Complete Basis-Set Predicted NMR Parameters, J. Phys. Chem. A., 106, 10396 (2002).
3. T. Kupka and C. Lim, Polarization-Consistent vs Correlation-Consistent Basis Sets in
Predicting Molecular and Spectroscopic Properties, J. Phys. Chem. A, 111, 1927 ( 2007).
4. T. Kupka, H 2 O, H 2 , HF, F 2 and F 2 O nuclear magnetic shielding constants and indirect
nuclear spin-spin coupling constants (SSCCs) in the BHandH/pcJ-n and BHandH/XZP
Kohn-Sham limits, Magn. Reson. Chem., 47, 959 (2009).
36

THEORETICAL AND EXPERIMENTAL STUDIES ON CYTOSINE
AND 5-FLUOROCYTOSINE
Teobald Kupka 1 , Mariana Spulber 2 , Mariana Pinteala 2 , Adrian Fifere 2 ,
Agnieszka Raniszewska 1 , and Malgorzata Broda 1
1 University of Opole, Faculty of Chemistry, Opole, Poland;
2 Institute of Macromolecular
Chemistry Petru Poni, Iasi, Romania
Introduction of halogen atom into position 5 of DNA fragments affects their
physico-chemical properties, including conformer stability, protonation states and hydrogen
bonding properties. 5-fluoroderivatives are especially important and are widely used as
drugs in cancer and fungal therapy. Such systems are very promising as controlled delivery
systems in nanomedicine.
This paper discusses possibilities of theoretical methods in predicting structure, IR, Raman
and NMR parameters of cytosine and 5-fluorocytosine in the gas phase and solution.
Literature
1. M. Spulber, M. Pinteala, A. Fifere, V. Harabagiu, B. C. Simionescu, Inclusion
complexes of 5-flucytosine with β-cyclodextrin and hydroxypropyl-β-cyclodextrin:
Characterization in aqueous solution and in solid state, J. Incl. Phenom. Macroc.
Chem., 62 (2008) 117.
2. B. Blicharska and T. Kupka, Theoretical and Experimental NMR Studies on Uracil
and 5-Fluorouracil, J. Mol. Struct., 613, (2000)153.
37

THEORETICAL CALCULATION OF NMR SHIELDINGS
AND INDIRECT SPIN-SPIN COUPLING CONSTANTS
Teobald Kupka
1 University of Opole, Faculty of Chemistry, Opole, Poland
Computational chemistry has been widely used to support assignment of
experimental NMR spectra and for prediction of chemical structure-NMR spectra relation.
The paper will discuss the role of various theoretical methods and basis set selection in
obtaining meaningful NMR parameters without relaying on accidental error cancellation.
The current limits of prediction for small and medium size atomic systems in the gas phase
and solution will be addressed. Some ideas about applying theoretical methods to predict
accurate NMR parameters of selected atoms within larger molecules, for example in active
sites of enzymes, will be outlined, too.
Literature
1. T. H. Dunning, Jr., J. Chem. Phys. 1989, 90, 1007.
2. Auer, J. Gauss, J. F. Stanton, J. Chem. Phys. 2003, 118, 10407.
3. U. Benedikt, A. A. Auer, F. Jensen, J. Chem. Phys. 2008, 129, art. no.
064111.
4. M. J. Frisch, et al., Gaussian 03, Revision E01,. Gaussian, Inc., Wallingford
CT, 2004 (and recent Gaussian 09).
5. T. Helgaker, M. Jaszunski, M. Pecul, Prog. Nucl. Magn. Reson. 2008, 53,
249.
6. K. Dybiec, A. Gryff-Keller, Magn. Reson. Chem. 2009, 47, 63.
7. F. Jensen, J. Chem. Theor. Comput. 2008, 4, 719.
8. T. W. Keal, D. J. Tozer, T. Helgaker, Chem. Phys. Lett. 2004, 391, 374.
9. T. Kupka, B. Ruscic, R. E. Botto, J. Phys. Chem. A. 2002, 106, 10396.
10. T. Kupka, C. Lim, J. Phys. Chem. A. 2007, 111, 1927.
11. T. Kupka, Magn. Reson. Chem. 2008, 46, 851.
12. T. Kupka, Chem. Phys. Lett. 2008, 461, 33.
13. T. Kupka, Magn. Reson. Chem., 2009, 47, 210
14. T. Kupka, Magn. Reson. Chem. 2009, 47, 674.
15. T. Kupka, Magn. Reson. Chem. 2009, 47, 959.
38

CONFORMATIONAL ANALYSIS OF CYCLIC DYNORPHIN
ANALOGUES USING NMR DERIVED DATA
Kwasiborska Maria s , Agnieszka Zieleniak b , Nowakowski Michał a , Oleszczuk Marta c ,
Wójcik Jacek a , Chung Nga N. d , Schiller Peter W. d , and Izdebski Jan b
a Laboratory of Biological NMR, Institute of Biochemistry and Biophysics, PAS, Warszawa,
Poland; b Peptide Laboratory ,Warsaw University, Department of Chemistry, Warszawa,
Poland; c Department of Biochemistry, University of Alberta, Edmonton, Alberta, Canada;
d Laboratory of Chemical Biology and Peptide Research, Clinical Research Institute of
Montreal, Montreal, Canada
Previously we have reported results of our studies of cyclic dermorphin and deltorphin
analogues containing a carbonyl bridge [1-3]. We observed very high dual affinity for both µ
and δ receptors of analogues containing the common (message) 1 - 4 sequence of those two
native peptides. Elongation of the message sequence at the C-terminus to obtain peptides
related to the full sequence of dermorphin [3] resulted in substantial changes of activity and
selectivity for receptors. We found that added sequence effected selectivity of peptides and
changed conformation of their N-terminal portions (message).
In this presentation we report biological activity, NMR and EDMC studies of new opioid
peptide analogues, which contain modified enkephalin sequence (message) [4]: (1) D-
Lys 2 ,Lys 5 ; (2) D-Lys 2 ,Orn 5 ; (3) D-Lys 2 ,Dap 5 ; (4) D-Orn 2 ,Lys 5 ; (5) D-Orn 2 ,Orn 5 ; (6) D-
Orn 2 ,Dab 5 ; (7) D-Orn 2 ,Dap 5 . In addition the C-terminal sequence of dynorphin (address) was
added in each case, namely: -Arg 6 -Arg 7 -Ile 8 -Arg 9 -Pro 10 -Lys 11 -Leu 12 -Lys 13 .
The biological activity of peptides was measured with GPI/MVD tests.
NMR spectra of peptides 1, 3, 4 and 7 were measured at 25 °C on a UNITY500plus (Varian)
spectrometer. TOCSY and gHSQC spectra were measured for assignment of all proton and
carbon signals. ROESY experiments were performed in order to obtain distance restraints.
The conformational space of each peptide was explored using the EDMC method and NMR
restraints were employed to select conformations present in the solution. Comparison of
biological and conformational results allowed drawing SAR conclusions. Possible
biologically active conformation was discussed.
References:
[1] Filip K., Oleszczuk M., Pawlak D., Wójcik J, Chung N.N., Schiller P.W., Izdebski J.
J.Peptide Sci., 9, 649-657 (2003).
[2] Filip K., Oleszczuk M., Wójcik J., Chung N.N., Schiller P.W., Pawlak D., Zieleniak A.,
Parcińska A., Witkowska E., Izdebski J. J. Peptide Sci., 11, 347-352 (2005).
[3] Witkowska E., Nowakowski M., Oleszczuk M., Filip K., Ciszewska M., Chung N.N.,
Schiller P.W., Izdebski J. J. Peptide Sci., 13, 519-528 (2007).
[4] Pawlak D., Oleszczuk M., Wójcik J., Pachulska M., Chung N.N., Schiller P.W., Izdebski
J. J. Peptide Sci., 7, 128-140 (2001).
39

DYNAMICS OF HYDROXYL DEUTERONS AND BONDED WATER
MOLECULES IN NADY(0.8) ZEOLITE AS STUDIED BY MEANS
OF DEUTERON NMR SPECTROSCOPY
Z. T. Lalowicz 1 , G. Stoch 1 , A. Birczyński 1 , and M. Punkkinen 2
1 H. Niewodniczański Institute of Nuclear Physics of PAS, 31–342 Kraków, Poland;
2 Wihuri Physical Laboratory, Department of Physics, University of Turku,
FI–20014 Turku, Finland
Deuteron spin–lattice relaxation and spectra were measured for NaDY(0.8) zeolite
containing some heavy water. Two subsystems of deuterons with different mobility were
disclosed at low temperatures. The corresponding relaxation rates differ by two orders of
magnitude. Spectra exhibit different shapes related directly to a specific motional model.
Hydroxyl deuterons perform incoherent tunneling along the hydrogen bond, then on
increasing temperature jumps to excited states and over the barrier appear. Hydrogen bonded
water molecules perform 180◦ rotational jumps about the twofold symmetry axis. Spectral
amplitudes are consistent with the water content of 13 D 2 O molecules per unit cell. Above
about 240K translational mobility becomes significant and finally water molecules fly across
the free space of cages. It leads to a characteristic change in the slope of the relaxation rate
temperature dependence. Similar features have been observed in the spin-lattice relaxation for
D 2 [1] and CD 4 [2] in cages of various zeolites.
References:
[1] J. S. Blicharski, A. Gutsze, A. M. Korzeniowska, Z. T. Lalowicz, and Z. Olejniczak,
Appl. Magn. Reson. 27, 183 (2004).
[2] A. Birczyński, M. Punkkinen, A. M . Szymocha, and Z. T. Lalowicz, J. Chem. Phys. 127,
204714 (2007).
40

SPECTRAL DENSITY FUNCTIONS
OF COMPLEX MOLECULAR MOTIONS
Lidia Latanowicz a and Zofia Gdaniec b
a
Faculty of Biological Sciences, University of Zielona Góra, Poland; b Institute of Bioorganic
Chemistry, Polish Academy of Sciences, Poznań, Poland
The fundamental claim of Woessner “Although the two types of motion are
independent, their contributions to relaxation are not. If the interaction Hamiltonian is
modulated by series of independent stochastic processes, the total correlation function of
a complex motion should be calculated” is the basis of the calculations of the total spectral
density of complex motion.
The motion of spin pair distanced at R is in liquid state is complex as it consists of the
isotropic (anisotropic) overall motion and the internal motion of molecular groups. The NMR
order parameter is calculated for the case of simultaneous changes in the radial and angular
parts of dipolar Hamiltonian [1]. The radial motion takes place when only one spin in the spin
pair is involved in the motion. Then R is distance undergoes changes simultaneously with the
jump angles Θ is . The effect of the activation parameters of internal motions (known from the
solid state study) on measured T 1 relaxation in the liquid state temperature regime will be
shown. The range of applicability of the model - free approach Lipari and Szabo in the study
of molecular dynamics in solutions is considered. The order parameter, S 2 (model-free),
concerns the angular amplitude of fast internal motion and it is defined as
2
S
m
m
el − free)
= J
is
(ω , complex
motion )/J
is
(ω , isotropic motion )
2τ
iso
2 2
1 + ω τ
iso
(mod in the temperature regime of the
maximum of the function
. The value of S 2 (model-free ) is temperature dependent
because the correlation times of internal τ n and isotropic motions τ iso are temperature
dependent [2]. The influence of the temperature dependencies of spectral densities overlap on
the relation between S 2 (model-free) and the true value of the order parameter S 2 is analyzed.
The temperature regime of the equality S 2 (model-free) = S 2 is estimated.
(a)
J 2 (ω i +ω s
) [s -1 ]
(iso)
(1)
0,8
(4)
0,6
0,4
0,2
0 5 10 15 20
(b)
ORDER PARAMETER S 2 (model - free)
1,1
1,0
0,9
0,8
0,7
0,6
0,5
0,4
(5)
(1)
(2)
(3)
(4)
0,3 S 2 = cos 2 (109.4 0 )
0,2
0,1
0 2 4 6 8
1000/T (K -1 )
1000/T (K -1 )
Fig.1 The illustration of the reductions in the spectral densities in the regime of the maximum of the function
1 +
2τ
iso
2
ω
τ
iso
2
under the influence of internal motion. Exemplary case of the complex motion (C 3 + C iso ); solid
lines – the temperature dependencies of spectral density (a) where (ω s = 2π×75.432 MHz, ω i = 2π×300 MHz),
and order parameter S 2 (model - free) (b). The motional parameters chosen are: R is = 0.109 nm, β is = 70.31 0
(the angle between the R is and C 3 rotation axis),
3
τ
0
− 13
= 3.8 ⋅ 10 s , E 3 = 2.9 kJ/mol (curve (1)), 4.2 kJ/mol
iso − 14
(curve (2)), 6.3 kJ/mol (curve (3)) , 8.4 kJ/mol (4), 12.6 kJ/mol (curve (5)), τ
0
= 1.6 ⋅ 10 s , E iso = 20.5
41
kJ/mol (iso). The spectral densities for the single motion C 3 and isotropic overall motion are shown by pointed
and dashed lines respectively. The dashed arrow indicates room temperature.

The spectral density of a single motion which is C iso goes through a maximum in the
temperature dependence. Such a maximum is split into two maximums for the spectral density
of C aniso motion. This split is due to the fact that the D ⊥ and D ⎢⎢ rotational diffusion
coefficients are dominant in different temperature regimes because of the different rate of
these motions. D ⎢⎢ representing faster motion dominates at lower temperatures. The high
splitting is observed when the activation parameters D ⊥ and D ⎢⎢ differ significantly (curve (1)
in Figure 2). Curve (1) represents the high and curve (3) represents small anisotropy. Curve
(3) is identical with curve (iso) –pointed line, representing the case D ⊥ = D ⎢⎢ . We can
conclude that anisotropic motion is impossible to be recognized in the liquid state temperature
regime, because of the overlapping of the spectral densities of motions represented by D ⊥ and
D ⎢⎢ rotational coefficients. This effect is similar to that of the overlap of spectral densities of
not well-separated isotropic overall and internal motions.
J 0 (ω i -ω s
) [s -1 ]
450
400
350
300
250
200
150
(iso)
(3)
(2)
(1)
T 1s
[s]
0,1
0,01
(iso)
(3)
(2)
(1)
100
50
0 5 10 15 20 25 30
1000/T (K -1 )
0 5 10 15 20 25 30
1000/T (K -1 )
Fig. 2. Exemplary temperature dependencies of the spectral densities J
0
is
( ωi
− ωs
) where (ω s = 2π×75.432
MHz, ω i = 2π×300 MHz) and heteronuclear T 1s temperature dependences for a spin pair (R is = 0.109 nm) which
performs a C aniso motion. The motional parameters chosen are: α is = 30 0 (the angle between main axis of the
⊥ II
13 −1
molecule and R is ), D0 = D0
= 5 × 10 s , E ⊥ = 24.3 kJ/mol, E II = 24.3 kJ/mol (the pointed curve (iso)), E II
= 10.5 kJ/mol (solid curve (1)), E II = 16.8 kJ/mol (solid curve (2)), E II = 21.8 kJ/mol (solid curve (3)). The curve
(iso) is depicted by open circles to distinguish it from curve (3). The dashed arrow indicates room temperature.
The proton spin pairs of the methyl group are known to perform a complex motion
being a resultant of three components involving mass transportation over and through the
3 T
potential barrier [3]. They are characterised by the correlation times τ and τ of the jumps
iso
over the barrier and tunnel jumps in the threefold potential of the methyl group and τ the
correlation time of isotropic rotation. The recently derived equations for the tunnelling rate
constant according to Schrödinger are taken into account in the calculations of the spectral
densities of tunnelling jumps.
PROTON T 1
(s)
1
0,1
#3
90 MHz
#2
#1
Fig. 3 Exemplary case of the proton T 1 for the
complex motion of methyl group (C 3 (classical)
+C 3 (tunneling jumps) + C iso ) - #1, (C 3 (classical) +
C iso ) - #2 and single motion C iso - #3. ω i = 2π×90
MHz, The motional parameters chosen are: R is =
0.178 nm, ω T = 2π×3 MHz (tunnel splitting),
3 − 12 iso − 14 tu − 8
τ
0
= 1 ⋅ 10 s , τ
0
= 7 ⋅ 10 s , τ
0
= 8 ⋅ 10 s ,
E 3 = 3.3 kJ/mol, E iso = 20.9 kJ/mol,
0 10 20 30 40 50 60
References:
1000/T (K -1 )
1. L. Latanowicz, Z. Gdaniec, Mol. Phys. 107 (2009) 1563 – 1576.
2. L. Latanowicz, Z. Gdaniec, J. Phys. Chem. submitted
3. L. Latanowicz, W. Medycki, R. Jakubas, Solid State NMR, accepted
This paper is dedicated to the memory of NMR relaxation specialist,
researcher Donald Edward Woessner 42 (1930 – 2008).

SOLID STATE STUDIES OF CYCLODEXTRIN COMPLEXES WITH
ADAMANTANE AND AMANTADINE USING 13 C AND 15 N
CP/MAS NMR
Agnieszka Lis-Cieplak and Wacław Kołodziejski
Department of Inorganic and Analytical Chemistry, Faculty of Pharmacy, Medical University
of Warsaw, Warsaw, Poland
Cyclodextrins (cyclomaltooligosaccharides) are macrocyclic oligosugars capable of
forming inclusion complexes with small hydrophobic molecules, thus improving their water
solubility [1,2]. Beta-cyclodextrin (β-CD) host is well suited for many guest drug molecules
and this prompts various pharmaceutical applications, especially in drug formulation and
delivery. The complexation with β-CD significantly modifies drug solubility, bioavailability
and stability [3,4]. Therefore, the β-CD/drug complexes seem worthy of careful studies,
including their solid forms used for preparing tablets. Cyclodextrin complexes with various
pharmaceutical compounds were widely investigated using solution and solid-state NMR
[1,3,4]. Amantadine (1-aminoadamantane) is used as an antiviral and antiparkinsonian drug.
We report on the β-CD/amantadine complex, which has been investigated under magic-angle
spinning (MAS) using solid-state 13 C and 15 N NMR spectroscopy with cross-polarization
(CP) from protons. For comparison, we have also studied the β-CD/adamantane complex and
both uncomplexed guest compounds. The NMR spectra supplied us with structural details and
provided interesting information on molecular dynamics. The latter subject has been
successfully studied using kinetics of CP [5]. CP is considerably sensitive to internuclear
distances and to mobility of molecules and/or their functional groups in the solid state. The
CP kinetics has been monitored using variable-contact-time measurements and analyzed in
terms of known CP models [5].
The 1 H → 13 C CP kinetics has indicated that the complexation affects molecular
arrangement and interactions in the solid state, decreases molecular mobility of the guest
molecules, while the host macrocycle becomes less rigid in the macromolecular complex. The
study is to be continued with other drugs containing the adamantyl functional group.
References:
[1] H. Dodziuk, Cyclodextrins and Their Complexes, Wiley-VCH, Weinheim (2006)
[2] W. Saenger, J. Jacob, K. Gessler, T. Steiner, D. Hoffmann, H. Sanbe, K. Koizumi, S.M.
Smith, T. Takaha, Chem. Rev. 98, 1787-1802 (1998)
[3] U. Holzgrabe, I. Wawer, B. Diehl, NMR Spectroscopy in Drug Development and Analysis,
Wiley-VCH, Weinheim (1999)
[4] K. Uekama, F. Hirayama, T. Irie, Chem. Rev. 98, 2045-2076 (1998)
[5] W. Kołodziejski, J. Klinowski, Chem. Rev. 102, 613-628 (2002)
43

MOLECULAR DYNAMICS AND STRUCTURE OF MIKTOARM STAR
BLOCK COPOLYMERS BASED ON POLY(BUTYL ACRYLATE)
AND POLY(ETHYLENE OXIDE)
Monika Makrocka-Rydzyk a , Aleksandra Wypych a , Mariusz Jancelewicz a , Stefan Jurga a* ,
and Krzysztof Matyjaszewski b .
a
Department of Macromolecular Physics, Faculty of Physics, Adam Mickiewicz University,
Poznań, Poland; b Department of Chemistry, Carnegie Mellon University,Pittsburgh,
Pennsylvania, USA
The strategy in modern chemistry is to made large oligomers from relatively simple
building blocks. It follows a way of the nature of performing molecules by joining small units
together through heteroatom links. Use of different types of chains attached to a central core
leads to combinatorial with remarkable modularity and diversity. This class of polymers
embraces miktoarm star copolymers, i.e. star-shaped compound containing arms with
different chemical compositions. They were synthesized by means of the atom transfer radical
polymerization technique [1]. Star-shaped copolymers have applications in drug delivery,
diagnostic assays, nanopatterned structures and photonics [2].
The investigated miktoarm star copolymers are composed of poly(butyl acrylate) -
polyBA and poly(ethylene oxide) - polyEO chains connected to a central core. The molar ratio
of polyBA/polyEO was: 0.80/0.20, 0.50/0.50, 0.80/0.20.
The Broadband Dielectric Spectroscopy (BDS), performed in wide temperature range
allowed to detect in polyBA/polyEO 0.20/0.80 and polyBA/polyEO 0.50/0.50 two relaxation
processes: α relaxation process - assigned to segmental chain motions of polymer backbone
and α c relaxation - coming from chain mobility in amorphous phase, constrained between
crystallites. The BDS studies for polyBA/polyEO 0.80/0.20 do not reveal α c -process probably
due to high amount of amorphous polyBA, which prevents the crystallization process.
Fast Field Cycling NMR studies (FFC) performed for polyBA/polyEO 0.50/0.50 below
melting temperature (T m ) confirm dynamics of polymer chains in confined space, revealing
dispersion of spin-lattice relaxation times obeying power law T 1 ∝ ω 0.75 [3]. However
above T m the frequency dependence of T 1 is described by power laws: T 1 ∝ ω 0.5 and T 1 ∝
ω 0.25 at high and low frequencies, respectively. They reflect chain dynamics, which can be
explained on the basis of the renormalized Rouse model in terms of high-mode and low-mode
number limit [4].
Acknowledgements:
This work is supported by 6th Framework Programme under SoftComp Grant No 502235-2
and research grant No N N202 128536 (Poland). A. Wypych acknowledges the Ministry of
Science and Higher Education in Poland for a Postdoctoral Fellowship (POL-POSTDOC III).
References:
[1] Haifeng Gao and Krzysztof Matyjaszewski; Macromolecules, 2008, 41, 4250-4257.
[2] John E. Moses and Adam D. Moorhouse, Chem. Soc. Rev., 2007, 36, 1249–1262.
[3] Reiner Kimmich and Esteban Anoardo, Prog. NMR Spectrosc. 2004, 44, 257-320.
[4] Nail F. Fatkullin and Reiner Kimmich; J. Chem. Phys. 1994, 101, 822-832.
44

MOLECULAR DYNAMICS AND NUCLEAR RELAXATION
PROCESSES OF PLANAR CATIONS WITH PSEUDO 5-FOLD
SYMMETRY AXES: IMIDAZOLIUM AND PYRAZOLIUM
COMPOUNDS
W. Medycki 1 , D. Kruk 2 , R. Jakubas 3 , and J. JadŜyn 1
1 Institute of Molecular Physics, Polish Academy of Sciences, Poznań, Poland;
2 Institute of Physics, Jagiellonian University, Kraków, Poland; 3 Faculty of Chemistry,
University of Wrocław, Wrocław, Poland
Imidazole molecules are the smallest ones possessing properties favorable for creating
new ferrolectric materials and therefore their dynamics attracts considerable attention.
In this work we study molecular dynamics of selected imidazolium and pyrazolium
compounds: Im 3 Sb 2 Br 9, Im 6 Bi 2 Br 18 , Im 5 Bi 2 Cl 11 , Im 5 Sb 2 Br 11 and Pl 4 [Bi 2 Br 10 ]x2H 2 O, by means
of NMR relaxation experiments.
In some of the previous studies [1] the relaxation features of these systems have been
attributed to two small angle libration modes, at lower temperatures, and to regular C 5 jumps
about the axis perpendicular to the centre of the cation plane, at higher temperatures.
At present we attempt to explain the complex temperature dependencies of the proton
relaxation rates as a result of the quadrupolar interactions leading to efficient relaxation
channels for the nuclei possessing quadrupolar moments [2]. The quadrupolar relaxation
mediates, in turn, the relaxation dynamics of protons via mutual dipole-dipole couplings.
This work has been financed by Polish Ministry of Science and Education, grant No N N202
172135
References:
1. A. Piecha, A. Białońska, R. Jakubas, W. Medycki, Solid State Sciences. 10 (2008) 1469-1479;
2. A. Piecha, R. Jakubas, A. Pietraszko, J. Baran, W. Medycki, D. Kruk, Journal of Solid State Chemistry
182 (2009) 2949–2960.
45

THEORY OF SOLID STATE DYNAMIC NUCLEAR POLARIZATION
Artur Mielczarek 1, 2 , and Danuta Kruk 2
1 Instytut Fizyki Jądrowej PAN, Kraków, Poland; 2 Institute of Physics, Jagiellonian
University, Krakow, Poland
Solid-state dynamic nuclear polarization experiments are commonly performed by
irradiating a sample that contains dilute paramagnetic species with a saturating microwave
field oscillating with, or close to, the electron Larmor frequency. Thereby the polarization of
the nucleai in the sample is enhanced above the equilibrium value, by a factor in the ideal case
given by the electronic divided by the nuclear gyromagnetic ratio. To achieve the
enhancement of the nuclear polarization an effective coupling between the nuclear and the
electronic spins is needed. The interaction leads to eigenfunctions of the electronic – nuclear
spin system being a mixture of ‘ordinary’ Zeeman states. Populations of the individual energy
levels evolve in time following a set of relaxation equations. The relaxation coefficients,
which connect the energy levels, depend on the alternating microwave field and the
absorption spectrum (ESR spectrum) of the electron spin involved. The ESR lineshape
function, is crucial for the dynamic nuclear polarization process. The amplitude of an ESR
spectrum at a given frequency is the fraction of electron spins with transition frequency equal
to that frequency. Therefore the ESR lineshape function gives “weigh factors” for the
transition probabilities (relaxation coefficients). The ESR spectrum is determined by many
parameters and has to be calculated independently starting from an electron spin Hamiltonian
appropriate for the considered case. During the evolution there is a time point at which the
momentary populations of the energy levels are particularly favorable for the nuclear spin
leading to an enhanced nuclear polarization. The enhancement does not last forever; after
some time all populations are equilibrated. This short time period of enhanced nuclear
polarization is very profitable in NMR investigations of biological systems.
We present a perturbative description of Dynamic Nuclear Polarization (DNP) in
solids. It is very common in the literature [eg. 1,2] to distinguish between DNP mechanisms
like ‘solid effect’ or ‘thermal mixing’. This long standing tradition is somewhat confusing
because it suggests a different origin of these processes, while the difference lays, in fact, in
the strength of the electron – nuclear spin dipole – dipole coupling. Here we propose for a
first time (according to our knowledge) an approach based on a perturbation solution of the
Liouville-von Neumann equation, which does not require any ‘a priori’ presumptions
regarding the interaction strengths. The final DNP enhancement factor is obtained as a
solution of population equations including transition probabilities resulting from the
alternating magnetic field and electron- and nuclear spin relaxation. The DNP factor is linked
to a corresponding Electron Spin Resonance (ESR) lineshape, which is simultaneously
evaluated within the same set of parameters.
References:
[1] A. Abragam and M. Goldman Rep. Prog. Phys., 41 395-467 (1978)
[2] T. Maly, G. T. Debelouchina, V. S. Bajaj, K.-N., Hu, C.-G. Joo, M. L. Mak-Jurkauskas, J.
R. Sirigiri, P. C. A. van der Wel, J. Herzfeld, R. J. Temkin and R. G. Griffin J. Chem. Phys.
128(5) (2008)
This work has been financed by Polish Ministry of Science and Education,
grant No N N202 105936
46

FIELD DEPENDENT RELAXATION PROCESSES IN MULTISPIN
SYSTEMS
Agnieszka Milewska and Danuta Kruk
Institute of Physics, Jagiellonian University, Reymonta 4, 30-059 Krakow, Poland
This work is concerned with several aspects of field dependent relaxation processes in
multi-spin systems containing unlike nuclear spins (like poly-alcohols) and nuclear and
electron spins (like radicals and bi-radicals). Multiexponentiality of relaxation processes in
such systems is investigated in detail. The exponentiality problem is a very important issue
for a proper understanding and description of relaxation effects limiting the efficiency of
polarization transfer between electron and nuclear spins. Nuclear spin relaxation resulted from
electron spin interaction for selected radicals is analyzed in detail.
This work has been financed by Polish Ministry of Science and Education,
grant No N N202 105936
47

THREE-DIMENSIONAL NMR SPECTROSCOPY OF ORGANIC
MOLECULES
Maria Misiak and Wiktor Koźmiński
Faculty of Chemistry, University of Warsaw,
Warsaw, Poland
Multidimensional NMR Spectroscopy can be regarded as a powerful technique
used for studies of molecular structure and dynamics of complex organic compounds.
Here we present the application of 3D NMR experiments, which are based on
random sampling of the evolution time space followed by Multidimensional Fourier
Transform (MFT). This approach, applied at first to the strychnine molecule, enables
one to acquire 3D spectra in reasonable experimental time and allows retaining high
resolution in indirectly detected domains.
We also show that by the application of 3D NMR experiments we were able to
do the complete 1 H and 13 C spectral assignment of natural abundance prenol-10, what
was impossible earlier using 1D and 2D techniques because of the complexity of the
spectra.
Furthermore we show here an application of 3D NMR experiments for
measurement of heteronuclear coupling constants of organic compounds yielding
complex spectra. The interpretation of 3D HSQC-TOCSY spectra with E.COSY-type
multiplets allowed us to evaluate heteronuclear coupling constants of strychnine with
high accuracy, what is difficult employing 2D methods owing to signal overlap and
impossible using conventionally recorded 3D NMR spectra in the reasonable time of an
overnight experiment.
References:
[1] Kazimierczuk K, Koźmiński W, Zhukov I, J. Magn. Reson. 179, 323 (2006)
[2] Kazimierczuk K, Zawadzka A, Koźmiński W, Zhukov I. J. Biomol. NMR. 36, 157
(2006)
[3] Kazimierczuk K, Zawadzka A, Koźmiński W, Zhukov I. J. Magn. Reson. 188, 344
(2007)
[4] Misiak M, Koźmiński W, Magn. Reson. Chem, 45, 171 (2007)
[5] Kazimierczuk K, Zawadzka A, Koźmiński W. J. Magn. Reson. 192, 123 (2008)
[6] Kazimierczuk K, Zawadzka A, Koźmiński W, Zhukov I. J. Am. Chem. Soc. 130,
(16) 5404 (2008)
[7] Misiak M, Koźmiński W, Magn. Reson. Chem. 47, 205 (2009)
[8] Misiak M, Koźmiński W, Kwasiborska M, Wójcik J, Ciepichal E, Swiezewska E,
Magn. Reson. Chem., 47, 825 (2009)
48

APPLICATION OF MR MICROSCOPY FOR ASSESSMENT
OF HYDRATION PROCESSES IN HPMC BASED MATRIX TABLETS
Anna Młynarczyk 1 , Marco L.H. Gruwel 2 , Piotr Kulinowski 1 , Krzysztof Jasiński 1 ,
Przemysław DoroŜyński 3 , Bogusław Tomanek 2,1 , and Władysław P. Węglarz 1
1 Department of Magnetic Resonance Imaging, Institute of Nuclear Physics, PAN, Kraków, Poland;
2 National Research Council of Canada, Institute for Biodiagnostics, Winnipeg, Canada;
3 Department of Pharmaceutical Technology and Biopharmaceutics, UJ, Kraków, Poland.
The study of tablets dissolution is needed to ensure efficient and timely drug delivery. In this
work MR microscopy was applied for study of water mobility and concentration in tablets, used for drug
delivery [1,2]. The tablets made of Hydroxypropylmethylcellulose-HPMC with and without addition of
a drug substance of different solubility were tested (Tab.1.).
Tab.1. Tablet formulation with abbreviations used in text
HPMC-1 Metolose60SH10000cP (100%)
HPMC-1 + KT Metolose60SH10000cP + Ketoprofen (50:50%)
HPMC-1 + LD Metolose60SH10000cP + L-dopa (50:50%)
HPMC-2 Metolose65SH400cP (100%)
HPMC-2 + KT Metolose65SH400cP + Ketoprofen (50:50%)
HPMC-2 +LD Metolose65SH400cP + L-dopa (50:50%)
Experiments were performed on a 11.7T vertical bore magnet (Oxford Instruments U.K.)
equipped with a 72mm ID gradient set (Magnex, UK) and an Avance console (Bruker,Germany). MR
images were acquired at 22.0±0.5°C using a MSME sequence with a FOV=15x15mm, matrix size
256x256, TR/TE=4000/6,5ms, NEX=2, total scan time 30min. T 2 and proton density calculations were
carried out using Matlab (The MathWorks, Inc.).
Two-dimensional T 2 and proton density (PD) maps were obtained. T 2 maps for HPMC-1,
HPMC-1+LD and HPMC-1+KT are shown in Fig.1. Histograms of these maps at 120 minutes of
hydration are shown in Fig.2.
A
180 min
B
180 min
C
180 min
120 min
30 min
120 min
30 min
120 min
30 min
60 min
60 min
60 min
Fig.1. Fragments of T 2 maps at 30, 60, 120, 180 minutes: A) HPMC-1, B) HPMC-1+LD, C) HPMC-1+KT .
Fig.2. T 2 map histograms at 120 minutes of hydration
49

Three modes corresponding to different physicochemical properties of polymer-water-drug system are
observed. Spatial distribution of T 2 and proton density along a tablet radius were also analyzed.
Different mechanisms of tablets hydration were observed due to composition of the formulations.
The experiment has shown that the phenomena occurring during drug dissolution are closely related to
the properties of applied substances. It is especially important for the research and the development
studies leading towards more effective drug delivery systems.
REFERENCES
[1] C.Melia, et al, PSTT, 1998, 1(1): 32-39.
[2] M.Vlachou, et al, Polym. Adv. Technol., 2004, 15:683-689.
[3] S.Silva, et al,Journal of Colloid and Interface Science 2008, 327: 333-340.
[4] G.Bajwa, et al, Polymer 2009, 50: 4571-4576.
50

3D NMR-BASED STRUCTURE OF apo-S100A1 HUMAN PROTEIN
Michał Nowakowski 1 , Łukasz Jaremko 1,2 , Igor Zhukov 1,3 , Agnieszka Belczyk 1 ,
Andrzej Bierzyński 1 , and Andrzej Ejchart 1
1 Institute of Biochemistry and Biophysic, Polish Academy of Sciences, Warszawa, Poland;
2 College of Inter-Faculty Individual Studies In Mathematics and Natural Sciences, Warsaw
University, Warszawa, Poland; 3 Slovenian NMR Centre, National Institute of Chemistry,
Ljubljana, Slovenia
S100 is a multigenic family of calcium-modulated proteins of the EF-hand superfamily
implicated in intracellular and extracellular regulatory activities. Within cells, most S100 proteins
exist in the form of antiparallely packed homodimers stabilized by the nonbonding interactions. It
is widely accepted that proteins from S100 family are signalling proteins. S100A1 contains 93
residues per subunit. It is stabilized by noncovalent interactions at its dimer interface; each
subunit contains two calcium binding loops.
Using a number of multinuclear 3D NMR techniques applied to the sample of 13 C, 15 N
double labeled apo-S100A1 human protein measured at 11.7 T, 16.4 T and 18.8 T we have
assigned chemical shifts of almost all backbone and side chain nuclei. Results were deposited in
Biological Magnetic Resonance Bank [1] (entry number 16360).
Secondary structure elements were predicted on the basis of 1 H, 13 C and 15 N chemical
shifts. Program PREDITOR [2] was applied to our chemical shift data confirming, as expected,
existence of four α-helical structures in each subunit.
Preliminary low resolution 3D structure of apo-S100 monomer was predicted using
CS23D [3] program on the basis of chemical shifts and was very close to our expectations.
RMSD calculated in comparison to rat monomer structure aligned by heavy atoms for residues
not interacting with second subunit is 1.52 Å with prediction taking closest homologues from
PDB into account, while it is 2.5 Å when alligned by all heavy atoms respectively.
Figure: Structure of human apo-S100A1 monomer calculated with CS23D (on the left), structure
of rat apo-S100A1 monomer (on the right).
51

This chemical shift based structural information was combined with distance constrains
derived from 13 C and/or 15 N edited NOESY spectra and used to calculate 3D structure of apo-
S100A1 human protein with CNS package [4, 5] and CYANA [6,7]. Calculated structure was
compared with the structures previously determined for rat apo-S100A1 protein [8] and bovine
apo-S100A1 protein modified by disulfide bond formation with β-mercaptoethanol [9].
Acknowledgment
This work was supported by a grant N301 031234 from the Polish Ministry of Science and
Higher Education
References
[1] http://www.bmrb.wisc.edu
[2] M.V. Berjanskii, S. Neal,D.S. Wishart, Nucleic Acids Res. 2006, 34
[3] D.S. Wishart, D.Arndt, M. Berjanskii, P. Tang, J. Zhou, G. Lin, Nucleic Acids Res. 2008,36
[4] A.T. Brunger, P.D. Adams, G.M. Clore, P.Gros, R.W. Grosse-Kunstleve, J.-S. Jiang, J.
Kuszewski, N. Nilges, N.S. Pannu, R.J. Read, L.M. Rice, T. Simonson, G.L. Warren, Acta
Cryst. 1998, D54, 905-921
[5] A.T. Brunger, Nature Protocols 2007, 2, 2728-2733
[6] P. Güntert, C. Mumenthaler, and K Wüthrich J. Mol. Biol. 1997, 273, 283-298
[7] P. Güntert, W. Braun, K Wüthrich J. Mol. Biol. 1991, 217, 517-530
[8] R.R. Rustandi, D.M. Baldisseri, K.G. Inman, P. Nizner, S.M. Hamilto, A. Landar, A.
Landar, D.B. Zimmer and D.J. Webber, Biochemistry 2002, 41(3), 788-796
[9] Zhukov I, Ejchart A, Bierzyński A, Biochemistry 2008 , 47, 640-650
52

TOWARD CHIRAL CRYSTALS OF BENZODIAZACORONANDS
Katarzyna Nowicka 1 , Agata Jeziorna 1 , Adam Sobczuk 2 , Janusz Jurczak 2 , Grzegorz D.
Bujacz 3 , Anna Bujacz 3 , Włodzimierz Ciesielski 1 , and Marek J. Potrzebowski 1
1 Centre of Molecular and Macromolecular Studies, Polish Academy of Sciences, Łódź,
Poland; 2 Institute of Organic Chemistry, Polish Academy of Sciences, Warsaw, Poland;
3 Institute of Technical Biochemistry, Technical University of Łódź, Poland
In this work we present the application of solid state (SS) NMR spectroscopy and
XRD in structural studies of inclusion complexes. On the course of our study we have found
that benzodiazacoronands (host molecules) are able to form molecular complexes with
different guests. The main aim of this project is to develop the methodology of searching of
inclusion complexes in solid state, employing SS NMR and XRD as complementary tools.
13 C CP/MAS spectra of benzodiazacoronand with different guest molecules (acetone, toluene,
methanol) in the crystal lattice are presented below.
53

OPTIMIZATION METHOD FOR ADIABATIC TAGGING PULSES
FOR ARTERIAL SPIN LABELING
Wojciech Obrębski, Piotr Bogorodzki, and Ewa Piatkowska-Janko
Institute of Radioelectronics, Faculty of Electronics and Information Technology at the
Warsaw University of Technology, Warsaw, Poland
Introduction
Magnetically marked water used in MRI imaging allows us to obtain perfusion images as well
as measuring cerebral blood flow (CBF) with comparable spatial resolution and sensitivity to
Blood Oxygenation Level dependent (BOLD) imaging. In this work we deal with a technique
called arterial spin labeling (ASL). ASL is a safe imaging technique working without contrast
agents allowing thus multiple and safe repetitions. A major drawbacks of ASL are limited
time resolution (due to the additional time required to label the blood) and relatively low
signal to noise ratio. In order to obtain acceptable image quality a increased number of
repetitions is required exposing patients on additional dose of SAR.
The aim of this work was to propose a new approach to optimization of adiabatic pulse used
for blood tagging. Optimized with the new method pulses use less power for labeling, so the
tagging can be carried out continuously, regardless of the scanner activity.
Subjects and Methods
The proposed method is based on assumption, that pulse which efficiency is optimized is
non-selective and adiabtic factor should by constant during pulse execution.
Optimized with the new methodology pulses will be compared with the Silver−Joseph−Hoult
(SJH, hyperbolic secant) impulses either by computer simulation or experimental
measurements on a Brucker BNT1000 scanner. Proposed pulses were generated by home
made pulse programmer based on FPGA (Xilinx Spartan3 XC3S200) and direct digital
synthesis (DDS) chips (Analog Devices AD9851).
Results and Conclusions
The optimized pulse showed 60% increase in efficiency in the computer simulations. In
consists of two simple, easy to implement discrete functions. The designed coil is expected to
allow a considerable reduction in the power required for the imaging and realize truly
continuous ASL
54

NON-DEBYE RELAXATION
AND TEMPERATURE DEPENDENCE
OF THE SECOND MOMENT OF NMR LINE
Marcin Olszewski and Nikolaj Sergeev
Institute of Physics, University of Szczecin, Poland
The temperature dependence of the second moment M
2
of NMR line is determined by
expression [1]
δω
1
( T )
π
∫ J
0
−δω
M
2
= ( ω)
dω
, (1)
where the spectral density J
0
( ω)
is the Fourier transform of the dipolar correlation function
h (t) ( t > 0 ) [1]
∞
∫
−∞
J
0
( ω)
= Re h(
t)
⋅ exp( iωt)
dt . (2)
Usually the dipolar correlation function is selected as exponential Debye function
h ( t)
= h(0)exp( − t / τ C
) , where τ
C
is the correlation time described the relaxation of the
dipolar correlation function. The Fourier transform of this function gives the Debye (or
Bloembergen-Purcell-Pound) form of the spectral density J ( ) [1]
( ω ⋅ τ ) 2
C
0
ω
2h(0)
⋅ τC
J ( ω)
= . (3)
0
1+
However the non-exponential relaxation processes are often observed in the different
range of physics (see [2-5] and references there). The non-exponential relaxations give the
spectral densities J
0
( ω)
differed from the Debye form. At present it is well known the
several spectral densities functions described the non-Debye relaxation [2-5]. It have been
used frequently the Cole-Cole function; the Cole-Davidson function; and Havriliak-Negami
function [3-5]. These functions may be wrote by one expression [3]
J
δ
⎡ ( ωτC
) sin( δπ / 2)
ε ⋅ arctan⎢
δ
⎣1+
( ωτC
) cos( δπ / 2)
δ
2δ
( ωτ ) cos( δπ / 2) + ( ωτ )
/
⎡
⎤⎤
2h(0)sin⎢
⎥⎥
⎢⎣
⎦⎥
=
⎦
, 0 < δ ≤ 1, ε ≤ 1/
δ . (4)
ω ⋅
0( ω,
ε,
δ)
ε
[ 1+
2
] 2
C
Spectral density (4) coincides with Debye function (3) if ε = δ = 1 [3].
In the present communication it will be discussed the temperature dependence of the
second moment of NMR line for the case of non-Debye (non-exponential) relaxation. The
temperature dependences of the spin-lattice relaxation time have been considered in [3.]
References:
[1] A.Abragam, The Principles of Nuclear Magnetism (Oxford Univ.Press, Oxford, 1961).
[2] J.C.Phillips, Streched exponential relaxation in molecular and electronic glasses, Report Progress Physics, 59 (1996)
1133-1207.
[3] P.A.Beckmann, Spectral densities and Nuclear Spin Relaxation in solids, Physics Reports, 171 (1988) 86-128.
[4] W.T.Coffey, Dielectric relaxation: an overview, Journal of Molecular Liquids, 114 (2004) 5-25.
[5] W.Dieterich, P.Maass, Non-Debye relaxations in disordered ionic solids, Chemical Physics, 284 (2002) 439-467.
C
55

ANALYSIS OF BOLD SIGNAL IN THE BRAINSTEM DURING
RESTING STATE AND AFTER CONTINUOUS SOUND ACTIVATION
Paulina Palowska 1 , Michalina Ryń 2 , Uwe Klose 2 , and Zofia Drzazga 1
1 Department of Medical Physics – University of Silesia, Katowice, Poland;
2 Department of Diagnostic an Interventional Neuroradiology – University Hospital
Tübingen, Tübingen, Germany
Introduction
Functional magnetic resonance imaging (fMRI) is a powerful method which can detect
discrete areas of slight perfusion changes within the brain resulting from neural activity.
Visualization of functional magnetic resonance imaging activation of subcortical structures
remains challenging because of the cardiac – related pulsatile movement of both the brainstem
and the cerebrospinal fluid and involved special scanning, pre – and postprocessing
techniques.
The purpose of this study is to develop fMRI paradigms. Activations in brainstem
nuclei were investigated during continuous sound (continuous stimulus) and resting state
measurements in correlation analysis. By using special technique like cardiac gatting fMRI it
was possible to obtain activations only in few cases. That why it was used additional regressor
in SPM analysis – signal changes from white matter to obtain better results of activations in
brainstem.
Materials and Methods
Subjects: A total of ten right handed volunteers (aged 26.8 ± 6.7) were examined.
They were all free of neurological disease and did not report a history of hearing problems.
Data acquisition: In this examinations a whole body scanner (3T Trio Tim, Siemens
Erlangen, Germany) was used. Signals were detected by BOLD differences using T 2 * -
weighted EPI technique. A total of 153 data sets were acquired every second heart beat – TR
depend on cardiac rhythm and TE = 30ms or 34ms (conditional on slice thickness). The
volumes consisted of 10 slices with slice thickness 2.5 mm, field of view (FoV) 192mm and
base resolution 64 or with slice thickness 1.7 mm, field of view (FoV) 192mm and base
resolution 96. After functional data T 1 – weighted spin echo anatomical scans were acquired
with parameters: TR = 2300ms, TE = 3.03ms, flip angle = 8 degrees, slice thickness = 1mm,
FoV = 256mm, voxel size 1.0x1.0x1.0mm and base resolution 256.
Data analysis: All data were analyzed and modified by the software package SPM5
and home – written routines in MatLab. Measured variable repetition times were used in
fMRI model specification as a regressor (Fig. 1. a). Since TRs as regressor was insufficient it
was necessity to used signal changes from white matter as additional regressor (Fig. 1. b)
what considerably improved quality of images.
Correlation map
1
a) 0.9 b)
10
0.8
10
Correlation map
1
0.9
0.8
20
0.7
20
0.7
p i x elcoordi n ates
30
40
50
0.6
0.5
0.4
0.3
0.2
p i x elcoordi n ates
30
40
50
0.6
0.5
0.4
0.3
0.2
60
0.1
60
0.1
0
0
10 20 30 40 50 60
10 20 30 40 50 60
pixel coordinates
pixel coordinates
Fig 1. Correlation analysis: a) with TR as regressor b) with TR and white matter.
56

Results and Discussion
The positions of auditory cortex were found and the analysis of correlation was
obtained. The auditory cortex and brainstem nuclei were identified in correlation analysis
with the threshold of correlation coefficient 0.5. Positions of the brainstem were correlated
with the auditory coordinates with the highest value of T (from block design experiments).
Numbers of activated localizations in brainstem nuclei which were found in correlation
analysis during continuous sound and resting state are shown in Table 1. Correlation map for
continuous sound experiment and time course from auditory cortex (left and right side) and
inferior colliculi (right) are demonstrated in Fig. 2.
CONTINUOUS SOUND
RESTING STATE
STRUCTURE
18 measurements 20 measurements
right left right left
IC 12 10 5 6
LLN 10 9 6 2
SOC 10 8 2 4
CN 9 11 5 3
Table 1. Numbers of activated structures of brainstem.
a t e s
o r d i n
o
e lc x
p i
Correlation map
1
a) 10
0.9 b)
20
30
40
50
60
70
80
90
10 20 30 40 50 60 70
pixel coordinates
0.8
0.7
0.6
0.5
0.4
0.3
0.2
0.1
0
S ignalchange
0.08
0.06
0.04
0.02
0
-0.02
-0.04
MR : continous
ACR
ACL
ICR
-0.06
0 50 100 150 200 250 300 350
Time [sec]
Fig. 2. a) Correlation map for continuous sound: slice thickness 1.7mm, TR and white matter
as regressors. b) Time course from auditory cortex and brainstem (IC).
The shape of the curves from auditory cortex demonstrate similar behavior to curve of
brainstem. Although signal changes from brainstem have lower amplitude than those from
auditory cortex.
Presented fMRI acquisition technique allows to visualizing the auditory cortex,
inferior colliculi, lateral lemniscus nuclei, superior olivary complex and cochlear nucleus.
Some of this structures were also obtained during resting state measurements.
Conclusion
After application continuous stimulus (continuous sound) it was possible to detect
cortical and subcortical structures as well as with using conventional block design paradigms.
The difficulty in obtaining fMRI images was concealed in cardiac related pulsatile motion of
brainstem. Others artifacts were partly eliminated by implementation signal changes from
white matter as additional regressor.
57

SOLID STATE NMR STUDIES OF COORDINATION AND
ORGANOMETALLIC COMPLEXES OF NICKEL AND RUTHENIUM
Piotr Paluch and Marek J. Potrzebowski *
Polish Academy of Sciences, Centre of Molecular and Macromolecular Studies,
Sienkiewicza 112, 90-363 Łódź, Poland.
Ni(II) with bis(acetylacetone)ethylenediamine ligand forms complexes which
crystallizes as semi-hydrate (2) with C2/c space group in monoclinic system and anhydrous
form (1) with Pna2(1) space group in orthorhombic system. 13 C and 15 N CP/MAS
experiments were employed for structural characterization of both forms and searching of
process of reversible water exchange in the crystal lattice. The applicability of ONIOM
theoretical calculation in structural analysis of Organometallic Complexes (OMC) is
discussed.
1) 2)
Ruthenium forms many useful organometallic complexes. In our study we concern on
metathesis second generation Hoveyda-Grubbs catalyst (3) and their siloxyl derivatives (4).
13 C CP/MAS , 1 H- 13 C FSLG HETCOR, 1H-1H Back-to-Back (BABA), 2D CRAMPS will be
presented. Problem with detection of carbon atom connected to ruthenium due to extremely
large 13 C CSA will be discussed.
Mes
N
Cl
C (II)
N
Mes
Mes
N
R 3 SiO
C (II)
N
Mes
Cl
Ru
R 3 SiO
Ru
H 3 C
CH 3
O
H 3 C
CH 3
O
3) 4)
58

1 H, 13 C, 15 N NMR COORDINATION SHIFTS
IN CATIONIC Fe(II), Ru(II), Os(II)
WITH 2,2’-BIPYRIDINE AND 1,10-PHENANTHROLINE
Leszek Pazderski, 1 Tomasz Pawlak 1 , Jerzy Sitkowski, 2,3 Lech Kozerski 2,3 , and Edward Szłyk 1
1 Faculty of Chemistry, Nicholas Copernicus University, Toruń, Poland; 2 National Drug
Institute, Warsaw, Poland; 3 Institute of Organic Chemistry, Polish Academy of Sciences,
Warsaw, Poland
1 H, 13 C and 15 N NMR studies of iron(II), ruthenium(II) and osmium(II) tris-chelated
cationic complexes with 2,2’-bipyridine and 1,10-phenanthroline of the general formula
[M(LL) 3 ] 2+ (M = Fe, Ru, Os; LL = bpy, phen) were performed. Inconsistent literature 1 H
signals assignments were corrected. Significant shielding of nitrogen-adjacent protons
(H(6) in bpy, H(2) in phen) and metal-bonded nitrogens was observed, being enhanced in
the series Ru(II) → Os(II) → Fe(II) for 1 H, Fe(II) → Ru(II) → Os(II) for 15 N, and bpy →
phen for both nuclei. The carbons are deshielded, this effect increasing in the order Ru(II)
→ Os(II) → Fe(II).
59

SEEKING A MOLECULAR SWITCH - STUDY ON 7-HYDROXY
-4-METHYLQUINOLINE-8-CARBALDEHYDE IN SOLUTION
Mariusz Pietrzak a , Volha Vetokhina a , Jacek Nowacki b , Jerzy Herbich a ,
and Andrzej L. Sobolewski c
a) Institute of Physical Chemistry, Polish Academy of Sciences, Warsaw, Poland;
b) Department of Chemistry, Warsaw University, Warsaw, Poland;
c) Institute of Physics, Polish Academy of Sciences, Warsaw, Poland
Recently 7-hydroxy-4-methylquinoline-8-carbaldehyde (HMQC) was proposed as a
candidate for an optically driven molecular switch [1]. IR and UV/VIS spectra of the
isolated molecule in a low-temperature matrix as well as theoretical calculations
indicate that this compound exhibits two tautomeric forms separated by a high barrier,
which can be overcome by a photoprocess induced by UV radiation [1].
In this study the behaviour of the HMQC in solution is examined. Our aim is twofold.
Firstly, we would like to confirm (or deny) the existance of the isomers proposed
previously. Secondly, the characteristic of the system in solution is under study.
UV and 1 H NMR spectra in various solvents and at different temperatures were
recorded and analysed. The N-H isomer was identified in the solution. Our results show,
however, that both isomers are in thermal equilibrium depending on the temperature and
the solvent. Mechanism of this phenomenon, which is in apparent contradiction to the
low-temperature matrix results, is discussed.
H
O
O
H
N
O
H
O
H
N
in solution
CH 3
CH 3
[1] L. Lapinski, M. J. Nowak, J. Nowacki, M. F. Rode, A. L. Sobolewski, A Bistable
Molecular Switch Driven by Photoinduced Hydrogen-Atom Transfer, ChemPhysChem
2009, 10, 2290-2295.
Scientific work financed from the science funds for the years 2008-2011 as a research
project.
60

SPIN-LATTICE RELAXATION IN DOPA-MELANIN AND MELANIN
ISOLATED FROM Sepia officinalis – COMPARATIVE EPR STUDIES
Barbara Pilawa 1 , Magdalena Zdybel 1 , Daria Czyżyk 1 , Ewa Chodurek 2 , and Sławomir
Wilczyński 1
1,2 Medical University of Silesia in Katowice, School of Pharmacy and Division of Laboratory
Medicine, 1 Department of Biophysics, Sosnowiec, Poland; 2 Department
of Biopharmacy, Sosnowiec, Poland
Spin-lattice relaxation in model and natural eumelanins was studied by an X-band (9.3
GHz) electron paramagnetic resonance (EPR) spectroscopy. Synthetic DOPA-melanin as the
model eumelanin and eumelanin isolated from Sepia officinalis were tested. The melanin
samples were obtained from Sigma Firm. Chemical structure of eumelanin is presented in
Figure 1 [1]. Melanin biopolymer from Sepia officinalis is used in cosmetics [2] and the
knowledge about its paramagnetic centers system is very important to cosmetology,
dermatology, and esthetical medicine. The aim of this work is to compare magnetic
interactions in DOPA-melanin and melanin isolated from Sepia officinalis.
O
O
HO
HO
N
H
N
H
(COOH)
(COOH)
O
HOOC
N
H
(COOH)
Fig. 1. Chemical structure of eumelanin [1].
EPR measurements were done by the use of electron paramagnetic resonance
spectrometer of RADIOPAN Firm (Poznań). The first-derivative EPR spectra were recorded
with microwave power in the range of 0.7-70 mW. Changes of amplitudes (A) and integral
intensities (I) of EPR spectra with increasing of microwave power were evaluated.
Amplitudes and integral intensities of EPR spectra of the studied melanins increase with
increasing of microwave power, and after reaching the maximum they decrease for the higher
microwave powers. Spin-lattice relaxation time increases with decreasing of microwave
power of saturation of EPR spectra of the sample [3]. It was shown the slow spin-lattice
relaxation processes exist in the both examined eumelanins – DOPA-melanin and melanin
from Sepia officinalis.
References:
[1] Wakamatsu K, Ito S. Advanced chemical methods in melanin determination. Pigment Cell
Res 2002; 15: 174-183.
61

[2] Gibka J. Wykorzystanie melaniny i procesu melanogenezy w kosmetyce. Polish Journal of
Cosmetology 2000; 3: 164-176.
[3] Stankowski J, Hilczer W. Wstęp do spektroskopii rezonansów magnetycznych. Warszawa:
PWN; 2005.
Acknowledgements:
This study was supported by Medical University of Silesia in Katowice (Grant no: KNW 1-
142/09).
62

SOLID-STATE CHARACTERIZATION OF S(+)CLOPIDOGREL
HYDROGENSULPHATE PHARMACEUTICALS AND THEIR
POLYMORPHS
Edyta Pindelska, Andrzej Mazurek, and Wacław Kołodziejski
Department of Inorganic and Analytical Chemistry, Faculty of Pharmacy,
Medical University of Warsaw, Warsaw, Poland
The importance of polymorphism and solvate formation in the crystallization of
organic compounds has an increasing importance in the pharmaceutical industry because
those substances may have chemically identical, but very different physical and biological
properties. Polymorphism is defined as the ability of a substance to exist as two or more
crystalline phases that have different arrangements and/or conformations of the molecules in
the crystal lattice. 1 The “polymorphs of a solvate” means that two solvates of the same
compound with identical solvent molecules and the same stoichiometry can exist in different
crystal structures. 2
S(+)clopidogrel hydrogensulphate - CL (Figure 1) is a pharmaceutical compound with
a novel mechanism of action in the prevention of atherosclerotic events. CL is an example of
polymorph of a solvate. At least six polymorphic forms are known for CL, but only two of
them (form I and II) are used in pharmaceutical formulations. 3
Figure 1. Structural formulae of S(+)clopidogrel hydrogensulphate (CL)
The solid-state studies of the two bioactive polymorphs (I and II) and commercial
available preparations: Plavix®, Areplex® and Zylt® were carried out using 13 C and 15 N
CP/MAS NMR spectroscopy and powder XRD analysis, additionally supported by ab initio
calculations. The assignment of the solid-state NMR spectra was done by comparison of
chemical shifts in solid and liquid phases as well as by analysis of CP kinetics and dipolardephased
spectra. 4
Comparison of the 13 C CP/MAS spectra of pure polymorphic forms I and II to those
from tablets proved that Plavix® contains polymorph II as an active substance, while
Areplex® and Zylt® contains polymorph I. Our study proves that 13 C CP/MAS technique is
sensitive enough for determination of polymorphic content in pharmaceutical tablets.
1 J. Bernstein, In Oxford University Press, Polymorphism in Molecular Crystals, Clarendon Press, Oxford 2002;
2 J. Bernstein, Cryst. Growth. Des. 1661, 5, 2005 ;
3 A. Bousquet, B. Castro, S. Germain, Polymorphic Form of Clopidogrel Hydrogen Sulphate. US Patent no. 6504030, 2003;
4 W. Kołodziejski, J. Klinowski, J. Chem. Rev. 102, 613, 2002;
63

CONSTRUCTION OF HIGH FREQUENCY COILS
FOR MRI AT 0.088 T
Bartosz Proniewski 1 , Henryk Figiel 1 , and Tadeusz Pałasz 2
1 University of Science and Technology, Faculty of Physics and Applied Computer
Science, Kraków, POLAND
2 M. Smoluchowski Institute of Physics, Jagiellonian University
Kraków, POLAND
While most of the research in magnetic resonance imaging is focused on the
development and applications of high field systems, there are certain areas of
diagnostic medicine where imaging at low magnetic field strengths yields comparable
results. Utilization of such systems requires specially designed dedicated rf coils in
order to overcome the technical constrains resulting from working at low magnetic
fields. The main purpose of this work was to determine whether it was possible to
image an elbow at 0.088T using different types of coils not evaluated in this particular
system yet.
For the purpose of this project a total of four different coils have been
designed, constructed and tested, namely two homogeneous resonators: the solenoid
and saddle shape coils, and two heterogeneous resonators: the basic planar surface
coil and half saddle surface coil. The design process consisted of determining the
desired coil dimensions using available anthropometric data, as well as coil
inductance and magnetic field simulations. A series capacitive network has been used
to match the designed coils to the spectrometer at the desired frequency and Q factors
have been evaluated using a spectrum analyzer. Magnetic field induction generated by
the coils has been measured and compared with theoretical expectations and the
theoretical field homogeneity has been assessed using the field histogram method.
Images of six phantoms have been acquired using the constructed coils, which
allowed signal to noise ratio measurements and field profiles to be determined and
compared with field simulations.
After the tests with phantoms, images of an elbow have been successfully
obtained using all of the constructed coils. The solenoid, saddle and planar surface
coils provided good overall image quality. Additionally the ability to focus the
imaging plane at the desired depth with surface coils has been demonstrated. In was
shown that imaging at very low fields can be successfully performed with other than
the solenoid coil configuration.
64

EPR STUDIES OF PARAMAGNETIC PROPERTIES OF THERMALLY
TREATED SISOMICIN AND VERAPAMIL
Paweł Ramos, Barbara Pilawa, and Piotr Pepliński
Medical University of Silesia in Katowice, School of Pharmacy and Laboratory Medicine,
Department of Biophysics, Sosnowiec, Poland,
Properties of paramagnetic centers in thermally treated sisomicin and verapamil were
compared. Sisomicin is an aminoglycoside antibiotic [1-2]. Verapamil is an L-type calcium
channel blocker [1-2]. Chemical structure of the studied drugs is presented in Figure 1 [1].
These two drugs were heated in dry air at the following temperatures: 160 o C, 170 o C, and 180
o C. Times of heating of the samples were: 120, 60, and 30 minutes, respectively. The aim of
this work is to compare effect of thermal sterilization on paramagnetic centers formation in
sisomicin and verapamil. Electron paramagnetic resonance (EPR) spectroscopy was the
experimental method.
NH 2
OH
N
N H 2
H 2
O
O
N
OH
O
NH 2
O
OH
N
H
O
O
O
O
N
Fig. 1. Chemical structure of sisomicin (a) and verapamil (b) [1].
EPR spectra were measured for all the heated samples. Amplitude (A), integral
intensity (I), and linewidth (∆B pp ) of the EPR spectra were analysed. g-Factors were
calculated. Spin-spin and spin-lattice interactions in the heated drugs were characterized.
Concentrations of paramagnetic centers in sisomicin and verapamil were determined. The
obtained results are useful to optymalization of thermal sterilization process of these drugs.
References:
[1] A. Zejc and M. Gorczyca (Red.), Chemia leków, Wydawnictwo Lekarskie PZWL,
Warszawa, 2002
[2] W. Janiec and J. Krupińska (Red.), Farmakodynamika. Podręcznik dla studentów
farmacji. Wydawnictwo Lekarskie PZWL, Warszawa, 2002
Acknowledgements:
This study was supported by Medical University of Silesia in Katowice (Grant no: KNW 1-
142/09).
65

THE INFLUENCE OF TRACES OF WATER ON THE CHEMICAL
SHIFTS OF SQULAENE IN BENZENE SOLUTION
Michał Rażew, Ewa Kaczorowska, Ewa Kula-Świeżewska, Ewa Ciepichał, and Jacek Wójcik
Instytut Biochemii i Biofizyki PAN, Warszawa, Poland
Squalene is a linear polymer composed of six isoprene units (Fig. 1). Squalene
commonly occures in nature. It is produced by all higher organisms and plays an important
biological role as a precursor in the synthesis of all steroids (i.a. cholesterol), as a component
of human sebum with antibacterial and antifungal properties or a compound stimulating the
immune system and increasing the response to a vaccine (immunologic adjuvant). At present,
the Novartis company use squalene as a component of MF59-adjuvanted influenza vaccine
[1], its usefulness in the malaria vaccine is also researched. Squalene has been found effective
in inhibiting the processes of cancer of the skin and lungs of rodents [2]. Squalene is also used
in cosmetics as an antioxidant and a lubricant.
Figure 1. The scheme of squalene.
In the NMR spectra of squalene the 13 C signals are grouped in several ranges: signals
of methyl groups at 10-30 ppm, signals of methylene gropups at 25-50 ppm and signals of
methine groups at 120-130 ppm. Because of the similar chemical character and surroundings
of most of carbon nuclei within a given functional group, the signals arising from different
nuclei overlap impeding interperation of 13 C NMR spectrum. In this work the influence of
traces of water on the chemical shifts of carbon and proton is presented. The results are shown
at the example of 1 H - 13 C correlation spetra. The HSQC spectrum of squalene sample at a
concentration 1µM in C 6 D 6 was recorded an a Varian Inova 400 MHz spectrometer at 25°C.
1 H and 13 C chemical shifts were compared with those obtained under the same conditions, but
with water addition. The calculations of the shielding constants of carbon and hydrogen atoms
in the presence of benzene and mixture of benzene and water were also performed. The
calculations of shielding tensors and geometry optimization were carried out using Gaussian
03 program with HF method.
References:
1. MF59 Adjuvant Fact Sheet, Novartis, June 2009.
2. Desai, K.N.; Wei, H.; Lamartiniere, C.A. The preventive and therapeutic potential of the squalenecontaining
compound, Roidex, on tumor promotion and regression. Cancer Lett. 1996, 19, 93-96,
Molecules 2009, 14 552.
66

APPLICATION OF MRI FOR SPINAL CORD MONITORING IN ANIMAL
MODEL OF PRESSURE IMPACT INJURY
Paulina Rosicka 1 , Katarzyna Majcher 1 , Wiesław Marcol 2 , Wojciech Ślusarczyk 2 , Tomasz Banasik 1 ,
Joanna Lewin-Kowalik 2 , and Władysław P. Węglarz 1
1
H. Niewodniczański Institute of Nuclear Physics PAN, Kraków, Poland
2
Silesian Medical University, Katowice, Poland
After the spinal cord incident, the acute impact causes the primary injury and long lasting
processes of secondary damage which occur weeks later. Time dependent changes of the water
properties in the injured neural tissue may help to understand these processes, as well as monitor
effects of the of neuroprotective drugs treatment.
The experiment was performed on adult male rats. The pressure impactor was used to impair
neural tissue in lower cervical cord. The animals were divide to two groups with different
neuroprotective drug applied to each. As neuprotective substances nicotine and N-acetyl-cysteine
were used. At different time after injury (from 2 weeks to 2 months) MRI measurement were done
on 4.7 T system using SE sequence . The data from 15 axial slices chosen between 10 Th and 12 Th
spine level was aqcuired. The T2- and DWI – weighted images were obtained with diffusion
gradients applied parallel and perpendicular to the spinal cord.
MR images free from any motion artifacts were obtained from spinal cord in vivo. Maps of
diffusion coefficients (longitudinal and transverse diffusion) were calculated for axial slices.
Comprehensive analysis of diffusion and T2- weighted data from ten ROI’s in gray and white
matter gives quantitative description of injury in terms of spatial and temporal changes of the
diffusion parameters.
67

INTERACTION OF SELECTED BICYCLIC MONOTERPENES WITH
α-CYCLODEXTRIN.
THERMODYNAMIC CHARACTERISTIC OF COMPLEXATION
Katarzyna Ruszczyńska-Bartnik 1 , Michał Nowakowski1, Helena Dodziuk2,
and Andrzej Ejchart1
1 Laboratory of Biological NMR, Institute of Biochemistry and Biophysics, Polish Academy of
Sciences, Warszawa, Poland; 2 Institute of Physical Chemistry, Polish Academy of Sciences,
Warszawa, Poland
Chiral recognition by cyclodextrins, CDs, is of considerable importance in the aspect of
enantiospecific action of many drugs. The mechanism of chiral recognition is not yet fully
understood and, therefore, determination and comparison of thermodynamic data for variety of
diastereomeric complexes is necessary. NMR spectroscopy is the method that is well suited for
studying stoichiometry, association constants and geometry of inclusion complexes.
Usually association constants of weak supramolecular complexes are determined under
assumption of chemical exchange between free and complexed states which is fast on the
chemical shift timescale. In such a case the observed NMR parameter value is the mole fraction
weighted average of the parameter values in the free and complexed molecule. However, many of
bicyclic monoterpens and α-CD form diastereomeric complexes with stoichiometry 1 : 2 which
do not fulfill this assumption. Moderately fast chemical exchange in these complexes results in
the broadening of resonances and precludes direct read-out of their frequencies from NMR
spectra. Unjustified assumption of the mole fraction weighted average may, under certain
circumstances, lead to significant underestimation of association constant and, in consequence, to
non-negligible errors in Gibbs free energy under determination. Simultaneous fit of theoretical
exchange broadened lineshapes to all NMR spectra measured in a titration experiment is the only
correct procedure which allows one to determine not only non-biased association constant K a but
also exchange rate k [1, 2]. Despite the more complicated data analysis the exchange broadened
spectra deliver information on the activation parameters of complexation processes which is
unavailable in the fast exchange limit.
Our study of
diastereomeric complexes
formed between bicyclic
monoterpens and α-CD
includes camphor [2], borneol
[3] and isoborneol. Their
structures are shown in Fig. 1.
Figure 1
68

Methodology of the determination of association constants
and exchange rates for the studied complexes from the exchange
broadened 1 H NMR spectra has been described elsewhere [2]. K a
and k parameters can be used to calculate Gibbs free energies ∆G 0
and ∆G # (see Fig. 2) characterizing complex formation according to
formulae:
∆ G = −RT ln( K )
Figure 2
The final results are given in Table 1
∆ G
0 a
#
hk
= −RT
ln( )
k T
Table 1.
Bicyclic 1R4R enantiomer
1S4S enantiomer ∆∆G 0 [kJ/M]
monoterpene ∆G 0 [kJ/M] ∆G # [kJ/M] ∆G 0 [kJ/M] ∆G # [kJ/M] (1S4S-1R4R)
camphor –53.6±0.2 58.82±0.03 –51.4±0.1 59.38±0.02 2.2±0.2
borneol –51.9±0.1 57.19±0.03 –50.9±0.1 57.39±0.03 1.0±0.1
isoborneol –53.4±0.1 56.37±0.06 –53.2±0.1 56.88±0.06 0.2±0.1
Absolute configuration of C1 and C4 chiral centers determines relative stability of studied
diastereomeric complexes. Enantiomers 1R4R form more stable complexes with α-CD than the
1S4S ones. Absolute configuration of an additional chiral carbon C2 does not influence this
regularity. ∆∆G 0 may be regarded as a measure of chiral recognition. It is the largest for camphor
and the smallest for isoborneol. On the other hand, Gibbs free energies of activation, ∆G # , are
larger for the 1S4S containing complexes. In the diastereomeric pairs larger activation energy
corresponds to less stable complex.
References:
[1] J. Feeney, J.G. Batchelor, J.P. Albrand, G.C.K. Roberts (1979) The effects of intermediate
exchange processes on the estimation of equilibrium constants by NMR. J. Magn. Reson.,
33, 519-529.
[2] P. Bernatowicz, M. Nowakowski, H. Dodziuk, A. Ejchart (2006) Determination of
association constants at moderately fast chemical exchange. Complexation of camphor
enantiomers by α-cyclodextrin. J. Magn. Reson., 181, 304-309.
[3] K. Ruszczyńska-Bartnik, M. Nowakowski, H. Dodziuk, A. Ejchart (2008) NMR titration at
intermediate exchange rate conditions: borneol – α-cyclodextrin system. NMR in
Chemistry, Physics and Biological Sciences. 24-26.09.2008, Warszawa. Abstract Book, P-
22.
B
69

SODIUM MR IMAGING
Lothar R. Schad
Lehrstuhl für Computerunterstützte Klinische Medizin, Medizinische Fakultät Mannheim,
Universität Heidelberg, Mannheim, Germany
Sodium ( 23 Na) ions play an important role in cellular homeostasis and cell viability. In
healthy tissue, the extracellular sodium concentration ([Na + ] ex = 145 mM) is about ten times
higher than the intracellular concentration ([Na + ] in = 10-15 mM) [1]. Using sodium MRI,
volume- and relaxation-weighted signal of these compartments can be measured. Thus,
sodium MRI is a promising diagnostic tool, since pathological processes can alter this ion
gradient. A density adapted 3D radial projection reconstruction pulse sequence (DA-3DPR) is
presented [2] that provides a more efficient k-space sampling than conventional 3D projection
reconstruction sequences (3DPR). The gradients of the DA-3DPR sequence are designed such
that the averaged sampling density in each spherical shell of k-space is constant. Due to
hardware restrictions, an inner sphere of k-space is sampled without density adaption.
Benefits for low SNR applications are demonstrated with the example of sodium imaging. In
simulations of the point-spread function (PSF), the SNR is increased by the factor 1.66 for the
DA-3DPR sequence. Using analytical and experimental phantoms, it is shown that the DA-
3DPR sequence allows higher resolutions and is more robust in the presence of field
inhomogeneities. High quality in vivo images of human brain, muscle and kidney are acquired
at 3.0 and 7.0 Tesla. For equivalent scan times, up to a factor of 1.80 higher SNR is observed
and anatomical details are better resolved using DA-3DPR.
References:
1. Hilal SK, Ra JB, Oh CH, Mun IK, Einstein SG, Roschmann P. Sodium Imaging. In: Stark
DD, Bradly WG, editors. Magnetic resonance imaging. St. Louis: C.V. Mosby; 1988.
p. 715-729.
2. Nagel AM, Laun FB, Weber MA, Matthies C, Semmler W, Schad LR. Sodium-MRI using
a density adapted 3D radial acquisition technique. Magn. Reson. Med. 2009, in print.
70

THE CHANGES OF PHOSPHOLIPIDS IN LIPID RAFTS AT PATIENTS
WITH MYELODYSPLASTIC SYNDROME (MDS)
Joanna Schiller 1 , Małgorzata Kuliszkiewicz-Janus 1 , Izabela Dereń-Wagemann 1 , and Stanisław
Baczyński 2
1
Department of Hematology and Transplantology, Wrocław Medical University, Poland,
2
Faculty of Chemistry, University of Wrocław, Poland
INTRODUCTION
The myelodysplastic syndromes (MDS) are a heteregenous group of clonal hematopoietic
stem cell disorder. They are characterized by two main features: an impaired maturation
(dysmyelopoeiesis) and peripheral blood cytopenias resulting from ineffective hematopoiesis.
Recent investigation has shown that impaired lipid rafts occur in neutrophils of patients with MDS.
The lipid rafts are involved in signal transduction, endocytosis, and cholesterol transport in cells.
They are specialized subdomains of the plasma membrane enriched in cholesterol, sphingomyelin,
gangliosides and certain proteins. Phospholipids, which built the lipid rafts, contained saturated
fatty acyl chains which tend to be more tightly packed. Together with high concentration of
cholesterol it results in high order and less fluidity then other part of the membrane. Lipid rafts can
be extracted from cell membranes as detergent-resistant memebranes (DRMs), when we use for
example 1% Triton-X 100.
AIMS
To continue our previous studies on the application of 31 P MRS in vitro to the analysis of
phospholipids changes in erythrocytes from patients with MDS, we decided to evaluate this method
to examine concentration of phospholipids in lipid rafts and disturbances in rafts flotation in a 10%-
40% linear sucrose density gradient.
MATERIALS AND METHODS
The investigation was carried out on erythrocytes from patients suffering from MDS (6
patients) and healthy volunteers (6 persons). The DRMs were isolated from erythrocytes with
modified R. Prohaska procedure [3]. Fractions (500µL) were collected from the top. Phospholipids
were extracted with a modified Folch method. The spectra 31 P were obtained on AMX 500 Bruker
(7.05T) spectrometer. Calculation of phospholipid concentration was based on peak integral
intensities in the 31 P NMR spectra for the individual compounds and methylenediphosphonic acid
(MDPA).
RESULTS
The DRMs from healthy volunteers were always isolated in 2 nd fraction. However in patients
with MDS they were isolated in 3 rd , 4 th , 5 th and 6 th fraction. It may depends on the staging of the
disease (IPSS). The lower fraction in which the DRMs occur may be dependent on advance of
disease. We observed that, the 31 P spectra of phospholipids extracts consisted of 3 resonant peaks.
Two due to phospholipids: PC and SM and another one – due to external reference substance
71

MDPA. Our preliminary data showed that in patients with MDS there is a tendency of decrease of
SM peak and increase of PC peak in comparison with healthy volunteers.
CONCLUSION
Changes observed in phosphholipids concentrations in lipid rafts from eythrocytes in patients
with MDS may result from dyserythropoeisis, which can be observed in this disease. Decrease in
sphingomyelin concentration in this patients may disturb structure of lipid rafts plasma membrane.
It may result that abnormal erythrocytes are being removed or destroyed which can explain anemia
by hypercellular bone marrow.
72

RELAXATION PROCESSES OF SELECTED BIOMOLECULES
Aleksandra Skowronska and Danuta Kruk
Institute of Physics, Jagiellonian University, Reymonta 4, 30-059 Krakow, Poland
In this work relaxation properties of water solutions (H 2 O and D 2 O) of selected
biomolecules are discussed. Due to the nature of the problem the temperature window of the
relaxation studies is rather narrow. Therefore to observe reorientational dynamics of
biomolecules viscosity of the solutions has been adjusted by adding different amounts of
glycerol. Slow motion of large molecules is investigated at low magnetic fields in order to
keep the condition that the Zeeman splitting (in angular frequency units) is comparable to a
correlation time reflecting the time-scale of the molecular dynamics. The issue of anisotropic
motion of large biomolecules in viscous solutions is discussed.
This work has been financed by Polish Ministry of Science and Education,
grant No N N202 105936
73

IMPLEMENTATION OF RETROSPECTIVE GATING
IN APPLICATION TO MR ASSESSMENT
OF CARDIAC FUNCTION IN MICE
Tomasz Skórka 1 , Sylwia Heinze-Paluchowska 1 , Urszula Tyrankiewicz 1 ,
and Magdalena Jabłońska 1,2
1-Zakład Tomografii MR, Instytut Fizyki Jądrowej PAN, Kraków;
2 Katedra Fizyki Medycznej i Biofizyki, Wydz. Fizyki i Informatyki Stosowanej AGH, Kraków
MRI of the heart is sensitive to cardiac and respiratory motions, what may cause
several artifacts in acquired images. In order to achieve images with proper mapping of the
heart ventricle volumes data acquisition is synchronized with the heart motion using ECG or
intrinsic NMR signal (self-gating method).
These measurements must be synchronized with ECG signal to achieve. It’s very important,
especially in cine imaging, where many images per cardiac cycle are achieved. As commonly
used, prospective gating has many advantages, but isn’t possible to cover the entire cardiac
cycle - there is a lapse of time at the end of the diastole. This problem can be eliminated by
the use of the retrospective gating method, where MRI data acquisition is continuous with a
simultaneous ECG recording to reorganize the data during image reconstruction (data are
collected regardless of the cardiac cycle).
In this work we present the implementation of the retrospective triggering method on
the MARAN DRX console. Detection of the R-wave is performed with the use of dedicated
device (SA Instruments Ltd, USA). Written software (IDL, ITT USA) enable for estimation
of the mean heart rate, than the consecutive R-waves are detected and assigned them to the
proper position on the time axis. The information about cardiac motion during imaging is
constantly recorded. After completion of data acquisition, this information is used for
interpolation and sorting of the data according to the corresponding phases of the cardiac
cycle. A number of images depicting different stages of the mouse heart motion were
collected and reconstructed using dedicated software.
Implemented method of reconstruction gives ability for obtaining images from the
entire cardiac cycle for easy changing number of reconstructed images, and potentially can be
more accurate according to the possibility of the R-wave timing post-processing. However,
ECG-based reconstruction is in general less useful in situations, when heart rate isn’t constant
(arrhythmia) or ECG signal is so disturbed that we are unable to detect it in a proper way
(transgenic mice). In these cases the best solution seems to be a method called “selftriggering”.
Acknowledgments: This work was supported by PMSHE, grant NN 518419733 (2007-2010)
74

INFLUENCE OF PARAMAGNETIC IONS PRESENCE ON RELAXATION
IN BLOOD SERUM
Lech Skórski, Mateusz Synowiecki, and Barbara Blicharska
Jagiellonian University, Department of Physics, Krakow, Poland
The presence of paramagnetic ions like Cu, Mn, and Fe influences on the relaxation
times in biological systems. This fact can be used to estimate concentration of free
paramagnetic ions in blood serum. As an exemplary application, we present the results of the
proton spin–lattice relaxation times measurements in blood serum before and after chelation,
which may be used as an alternative method for monitoring the presence of free copper ions
This method may be used in the diagnosis of diseases like leukaemia, liver diseases, and
particularly Wilson’s disease. The advantage is that in contrast to conventional methods, like
spectrophotometry, which records the total number of both bound and free ions, the proton
relaxation technique is sensitive solely to free paramagnetic ions dissolved in blood serum.
75

THE ITERATIVE APPROACH TO PROCESSING OF RANDOMLY
SAMPLED 3D NMR SPECTRA
Jan Stanek, and Wiktor Koźmiński
Faculty of Chemistry, University of Warsaw,
Warsaw, Poland
Multidimensional (nD) NMR has become the routine method in research on the
structure and dynamics of biomolecules. Classical approach to nD-signal sampling and
processing that employs Discrete Fourier Transform (nD DFT), has been gradually replaced
by alternative methods, which use more or less random sampling schemes of evolution time
domain and therefore are not limited by the Nyquist theorem [1]. These new methods are
primarily applicable to sensitive (i.e. featuring high S/N) nD experiments.
Their main advantage is the enhancement of spectral resolution achieved in the reasonable
experimental time. However, irrespective of the particular method, obtained spectra contain
artifacts [2]. They can be substantially suppressed by (a) optimization of the sampling
schedule [3] and/or (b) application of a specialized method of processing. Recently
discovered Multidimensional Fourier Transform seems a very elegant, linear and general
method, however, it is not suitable for spectra of high dynamic range.
In this work the fully automatic iterative cleaning procedure is applied in order to remove
artifacts. It employs a linear and exactly reversible transformation (SVD assisted FT) and
original signal detection algorithm. The benefits of the new approach have been proved
experimentally on 3D 15 N and 13 C-edited NOESY-HSQC spectra of human ubiquitin,
which have been obtained using ca 5 % of samples.
References:
[1] K. Kazimierczuk, A. Zawadzka, W. Koźmiński, I. Zhukov, Random sampling of
evolution time space and Fourier transform processing, J. Biomol. NMR 36 (2006) 157-
168;
[2] K. Kazimierczuk, A. Zawadzka, W. Koźmiński, I. Zhukov, Lineshapes and artifacts in
Multidimensional Fourier Transform of arbitrary sampled NMR data sets, J. Magn. Reson.
188 (2007) 344 – 356;
[3] K. Kazimierczuk, A. Zawadzka, W. Koźmiński, Optimization of random time domain
sampling in multidimensional NMR, J. Magn. Reson. 192 (2008) 123-130.
76

STRUCTURE AND STEREOCHEMISTRY OF NTBC
AND ITS METABOLITES - A COMBINED NMR/DFT STUDY
Przemysław Szczeciński, Anna Dziadecka and Adam Gryff-Keller
Faculty of Chemistry, Warsaw University of Technology, Noakowskiego 3,
00-664 Warszawa, Poland
NTBC, i.e. 2-[2-nitro-4-(trifluoromethyl) benzoyl]cyclohexane-1, 3-dione (Fig. 1), is
a lifesaving drug against an inherited metabolic disease - tyrosinemia type I. It has been found
that this drug is metabolized in organisms of patients and excreted in urine as 5-hydroxy-
NTBC and 6-hydroxy-NTBC (Fig. 1). After separation and chemical identification the
molecular structures of the NTBC metabolites have been investigated in detail using 1 H, 13 C
and 19 F NMR spectroscopy and quantum chemical calculations. The investigated compounds
belong to the class of β,β-triketones that in solutions can undergo various tautomeric and
conformational transformations. The structures of the most abundant forms of the investigated
compounds in CDCl 3 solutions have been established.
O
OH
CF 3
O NO 2
O
O
OH
CF 3
O NO 2
HO
OH
CF 3
O NO 2
HO
Figure 1. NTBC and its metabolites.
The molecular geometry optimizations were done by DFT (PBE1PBE/6-31+G*) method,
whereas the theoretical values of NMR parameters were calculated by GIAO-DFT method
using PBE1PBE/6-311++G(2d,p) level of the theory. The impact of the solvent was taken into
account by PCM method at both calculation stages.
77

MOLECULAR DYNAMIC AND STRUCTURE
OF PHOSPHATYDYLOCHOLINE (DMPC)/CATIONIC GEMINI
SURFACTANT (GEM-IK1) SYSTEM
Kamil Szpotkowski, Aleksandra Wypych, Kosma Szutkowski, ad Maciej Kozak, Stefan Jurga
Department of Macromolecular Physics, Faculty of Physics,
Adam Mickiewicz University, Poznań, Poland
Phospholipids are structural basis of biological membranes, with structures strongly
dependent on temperature and additions, such as cholesterol and surfactants [1]. Gemini
surfactants are a new generation amphiphilic molecules containing two monomer surfactant
molecules linked by a hydrophilic spacer. This group of surfactants have special properties
and potential applications in many areas [2]. Gemini surfactant molecule intercalates between
zwiterionic phospolipid molecules so that the total surface charge is altered which manifests
in change of the structure of the bilayer [3].
Aqueous suspensions of 1,2-dimyrilstoyl-sn-glycero-3-phosphocholine (DMPC) with
different concentration of the surfactant 1,1’-(1,4-butane) bis 3-decyloxymethylimidazolium
chloride (GEM-IK1) have been investigated by Small Angle X-ray Scattering (SAXS), Pulsed
Field Gradient NMR (PGSE NMR), Broadband Dielectric Spectroscopy and Fourier
Transform Infrared Spectroscopy.
The broad distribution of diffusion coefficients was found for pure DMPC, while addition of
gemini surfactant manifested in the significant narrowing of the distribution of diffusion
coefficients as the result of a change in the size distribution of aggregates (1A). This
conclusion is supported by SAXS diffraction patterns (1B).
Dielectric spectroscopy in frequency range from 10 6 Hz to 10 9 Hz was used to study
polar part of DMPC molecule(2A). The observed step-like change in the dependence of
dielectric constant vs. temperature is explained by a transition from gel to liquid crystalline
phase (2B). This is further confirmed by temperature dependence of stretching bands obtained
by FTIR for phosphate and carbonyl groups. Two characteristic bands from IR spectra,
corresponding to symmetric and antisymmetric vibrations of methyl groups, were observed to
detect changes in the non-polar part of phospholipid bilayers. Our results show that the
transition from lamellar to bicellar phase is indeed induced by the addition of the surfactant.
Furthermore the temperatures of observed main- and pre-transitions are decreasing with an
increase of the surfactant concentration.
1A
Amplitude
1
0.1
0.01
1E-3
T=293 K
10% DMPC
10% DMPC + 0.1% Gemini
10% DMPC + 0.5% Gemini
10% DMPC + 1% Gemini
10% DMPC + 5% Gemini
1A
Int (a.u.)
1000000
100000
10000
10% DMPC 5% Gem-IK1
10% DMPC 1% Gem-IK1
10% DMPC 0.5% Gem-IK1
10% DMPC 0.1% Gem-IK1
10% DMPC
1000
1E-4
1E-5
1E-3 0.01 0.1 1 10 100
g 2 δ 2 (∆-1/3δ)
100
0 1 2 3 4 5
s (nm -1 )
78

Figure 1. DMPC and DMPC/GEM-IK1 (0.1%, 0.5%, 1%, 5%) systems at 293K. Log-log plot
of a spin-echo amplitudes as obtained from PGSE NMR (1A). SAXS diffraction patterns
(2A).
2A
2B
10% DMPC/ 5% GEM-IK1/ D 2
O
800
10% DMPC/ 1% GEM-IK1/ D 2
O
700
600
500
400
1.5 MHz
275 K
281 K
287 K
293 K
299 K
305 K
311 K
ε'
10% DMPC/ 0.5% GEM-IK1/ D 2
O
ε'
300
10% DMPC/ 0.1% GEM-IK1/ D 2
O
200
100
0
10 7 10 8 10 9
10% DMPC/ D 2
O
Frequency [Hz]
275 280 285 290 295 300 305 310 315
Temperatura Temperature [K]
Figure 2. Dielectric permittivity vs. frequency for 10% DMPC suspension (2A). Dielectric
spectra of ε’ vs. temperature for DMPC/GEM-IK1/H 2 O systems, with different amounts of
GEM-IK1 measured at 1.5 MHz.
References:
[1] J. Katsaras, T. Gutberlet (Eds.), Springer- Verlag, Berlin – Heidelberg, 2001.
[2] Mingqi Ao, Guiying Xu, Yanyan Zhu, Yan Bai, Journal of Colloid and Interface Science
326 (2008) 490–495.
[3] D. Uhrikowa, G. Rapp, P. Balgavy. Bioelectrochemistry 58 (2002) 87-95.
Acknowledgements:
This work is supported by 6th Framework Programme under SoftComp Grant No 502235-2
and research grant No N202 248935 (Poland). A. Wypych expresses her appreciation to the
Ministry of Science and Higher Education in Poland for a Postdoctoral Fellowship (POL-
POSTDOC III).
79

MOLECULAR DYNAMICS IN PROTON CONDUCTING GEL/H 3 PO 4
ELECTROLYTES STUDIED BY NMR
L. Szutkowska, M. Dobies, K. Szutkowski, Stefan Jurga, G. śukowska*, and W. Wieczorek*,
Department of Macromolecular Physics, Faculty of Physics, Adam Mickiewicz University,
Poznań, Poland; *Faculty of Chemistry, Warsaw University of Technology,
Warszawa, Poland
Migration mechanisms and dynamics of protons are crucial for understanding the
phenomena of the proton conductivity in novel nonaqueous polymer systems. Recently the
new nonaqueous electrolytes have been proposed [1, 2], which are doped with phosphoric
acids. These systems can be used in polymer electrolyte type of fuel cells operating at low
humidity conditions and high temperatures above 100 o C. We have examined nonaqueous gel
consisting of phosphoric acid H 3 PO 4 (PA) dissolved with propylene carbonate (CP),
entrapped within polymer matrix, formed from poly(methylmethacrylate) (PMMA). Two
samples were under investigation with the same amount of PMMA (12% wt.) but with
different content of organic solution of CP/H 3 PO 4 (81/7 and 48/40 % wt.).
Both samples were investigated by NMR. Slow molecular dynamics was studied by
means of T 1 relaxation times as a function of Larmor frequency in the Fast–Field Cycling
NMR experiment. A strong dispersion of relaxation rate 1/T 1 = R 1 (ω), especially at highfrequency
range from 200 to 1 MHz, was observed for gel with higher H 3 PO 4 content. For
the gel with a lower concentration of H 3 PO 4 , the dispersion R 1 (ω) is becoming weaker. The
relaxation behavior may be explained by different interactions (adsorption mechanism) of
organic solution of CP/H 3 PO 4 with a polymer matrix in both samples. Weak adsorption
obviously leads to flat, however, strong adsorption - to steep spin-lattice relaxation
dispersions [3, 4].
The translational diffusion was measured by Pulsed Field Gradient Stimulated Spin Echo
(PGSTE) NMR. Diffusion coefficients turned out to be dependent on diffusion time ∆, giving
an evidence for restricted diffusion. Furthermore diffusion coefficients for all contributing
proton species were analyzed by using pseudo 2D displays of Diffusion Ordered
Spectroscopy (DOSY).
All results are discussed with respect to the ionic conductivity of studied gel electrolyte
systems [5,6]
References
[1] M. Schuster, T. Rager, A. Noda, K. D. Kreuer, J. Maier, Fuel Cells, (2005), 5, 355.
[2] K. D. Kreuer, S. J. Paddison, E. Spohr, M. Schuster, Chem. Rev., (2004), 104, 4637.
[3] S. Stapf, R. Kimmich, and R. O. Seitter, Magn. Reson. Imaging., (1996), 14, 841.
[4] J.-C. Perrin, S. Lyonnard, A. Guillermo, and P. Levitz, J. Phys. Chem., (2006), 110, 5439.
[5] K. R. Jeffrey, W. Wieczorek, D. Raducha, and J.R. Stevens, J. Chem. Phys., (1999), 110
(15), 7474.
[6] K. R. Jeffrey, G. Z. śukowska, J. R. Stevens, J. Chem. Phys., (2003), 119 (4), 2422.
80

METABOLIC PROFILE OF CEREBROSPINAL FLUID OBTAINED
FROM AMYOTHROPHIC LATERAL SCLEROSIS PATIENTS
Beata Toczylowska
Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland;
Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences,
Warsaw, Poland
Introduction
Aim of this study is to determine metabolic profiles of cerebrospinal fluid (CSF) obtained
from the Amyotrophic Lateral Sclerosis (ALS) patients and to compare them against
metabolic profile of samples obtained from control group. CSF surrounds the brain and fills
the ventricles as well as the subarachnoid space. It is the secretion product of various central
nervous system structures and its ultimate composition depends on exchange of metabolites
with the blood and adjacent brain tissue. Since the composition of CSF is directly dependent
upon metabolite production rates in the brain, analysis of the CSF metabolic profile can offer
biochemical insights into central nervous system disorders.
Methods
CSF used for our examination was collected by lumbar puncture from lumbar subarachnoid
space. Two sample categories were collected: control, neurologically healthy, subjects (n=15)
and ALS patients (n=18). Patients without neurological diseases, who underwent varices or
inguinal hernia surgery under spinal anesthesia were qualified to the control group. CSF
samples were centrifuged at room temperature at 15000 rpm for 5 minutes, and then frozen at
-80°C until NMR analysis was performed. In the NMR experiments solvent suppression was
used to minimise strong water signal. 512 scanes and 12 s pulse sequence repetition were used
at 25 C. Small amount of D 2 O was added to each sample in order to provide lock signal. 1mM
trimethylsilyl propionate (TSP) was used as a chemical shift reference.
TSP signal was also used for normalisation of all spectra enabling quantitative statistical and
discrimination analyses. Amounts of metabolites were expressed as relative intensities of their
spectral peaks determined in comparison to the TSP peak, because absolute concentrations
were difficult to determine using the analytical techniques applied here.
Each NMR spectrum was reduced to smaller number of variables, using the target profiling
method. Twenty eight signals from each spectrum were analyzed. Multivariate partial least
square discriminant analysis (PLS-DA) was performed using the software package SIMCA-
P(Version 12, Umetrics AB, Umeå, Sweden)(1).
Results
For the multivariate analysis 28 signal of the following components identified in NMR spectra
were selected: formate, histidine, phenylalanine, tyrosine, myo-inositol, unassigned signals at
δ=4.03 and δ=1.08, glucose, glycine, scyllo-inositol, choline, citruline, creatinine, creatine,
citrate, glutamine, glutamate, pyruvate, acetone, acetoacetate, acetate, GABA, lysine,
arginine, alanine, lactate, isoleucine, 3-OH-butyrate. In order to enhance the identification of
potential differences in biochemical composition caused by neuropathology, PLS-DA
modeling was then employed, using knowledge of the specimen classification. Discriminant
analysis maximizes any separation present between defined memberships classes (Y matrix)
by appropriate weighting of the NMR variables used as the X matrix. In this case, we chose to
analyze for systematic differences between ALS and control samples. The coefficients of the
goodness of fit (R2) and prediction (Q2) were used to assess the model quality i.e. to establish
whether any systematic differences between two groups of patients were present and the
81

variable importance in the projection computed by SIMCA-P were used to define which
metabolites, if any, differentiated both groups. The best model consisted of two components
(R2 = 0.645, Q2 = 0.33). It meant that the model fitted the data well but was not appropriate
for prediction purposes. Validation test indicated that this model was valid for both ALS and
control group. The most important parameters that contributed to class separation were signals
from glutamate, histidine, and lactate as well as signals from acetate and alanine.
Applying PLS-DA, the discrimination of ALS samples from the control samples was obtained
(Fig. 1). In this model all patients from ALS group were classified correctly (100%)as
opposite of only 12 from 15 patients from control group (80%).
PLS-DA
t[Comp. 1]/t[Comp. 2]
ALS
control
6
4
2
t[2]
0
-2
-4
-6
-4 -3 -2 -1 0 1 2 3 4
SIMCA-P+ 12.0.1 - 2009-11-01 17:28:59 (UTC+1)
Fig.1 First two components t[1] and t[2] of the PLS-DA model scores
t[1]
Conclusions
A PLS-DA model was applied splitting the samples in two classes: normal samples formed
class 1 and pathological samples formed class 2. All patients from ALS group were classified
correctly and distinguished from control group on the basis of cerebrospinal fluid component
analysis. Multivariate discriminant analysis performed on the two groups of CSF samples
showed that it was possible to distinguish ALS patients from control patients. Because the
coefficient of goodness of prediction for the model was not good, the further investigation on
larger groups of patients was needed to confirm obtained results.
In conclusion, 1 H NMR data obtained from CSF samples can be treated with multivariate
statistics to provide a useful screening tool. The technique is rapid and eventually could be
used to detect other neuropathologies as well. The potential of the method should be further
validated using greater number of samples and extended to include samples originating from
patients suffering other diseases.
Reference List
1. Ellis,D.I., Dunn,W.B., Griffin,J.L., Allwood,J.W., and Goodacre,R. (2007) Metabolic
fingerprinting as a diagnostic tool, Pharmacogenomics. 8, 1243-1266.
Acknowledgment:
This work was supported by MSHE, Scientific Network 28/E-32/SN-0053/2007 28/E-32/SN-
0053/2008.
82

STRUCTURAL STUDIES OF N-TERMINAL SEQUENCE
OF DERMORPHIN BY MEANS OF SOLID STATE NMR
SPECTROSCOPY AND XRD
Katarzyna Trzeciak-Karlikowska 1 , Agata Jeziorna 1 , Grzegorz D. Bujacz 2 , Anna Bujacz 2 ,
Włodzimierz Ciesielski 1 , and Marek J. Potrzebowski 1
1 Centre of Molecular and Macromolecular Studies, Polish Academy of Sciences, Łódź,
Poland; 2 Institute of Technical Biochemistry, Technical University of Łódź, Poland
Dermorphin is an opioid hepta-peptide (Tyr-D-Ala-Phe-Gly-Tyr-Pro-Ser-NH 2 ) first
isolated from the skin of South American frogs belonging to the genus Phyllomedusa.
Dermorphin is about 30-40 times more potent than morphine but less likely to produce drug
tolerance and addiction. Synthetic analog of dermorphin containing L-alanine is biologically
inactive.
In this paper we present structural studies of two N-terminal sequences of dermorphin: Tyr-D-
Ala-Phe, Tyr-D-Ala-Phe-Gly and their L-alanine analogs Tyr-L-Ala-Phe, Tyr-L-Ala-Phe-Gly.
The application of 1D (CP/MAS) and 2D (PDSD, DAR and DCP/MAS) Solid State NMR
(SS NMR) experiments in spectral assignment of peptides under investigation will be
discussed. The comparative analysis of data obtained from SS NMR and X-ray
crystallography will be shown.
Figure. 2D 13 C- 13 C Dipolar Assisted Recoupling (DAR) correlation of 13 C, 15 N labeled of
tetrapetide Tyr-D-Ala-Phe-Gly
83

NEW PERMANENT MAGNETS FOR MRI
Krzysztof Turek 1 , Piotr Liszkowski 1 , and Boguslaw Tomanek 2
1 AMAG Sp. z o.o., Kraków, Poland; 2 Institute of Nuclear Physics of Polish Academy
of Science, Kraków, Poland
The main application of nuclear magnetic resonance is imaging (MRI) of a human
body. AMAG Sp. o.o. has developed a number of permanent magnets for MRI systems, such
as 0.08 T for MRI of hyperpolarized gases, 0.2 T for animal MRI, whole body 0.2 T 4-post
magnet, 0.2 T “one side” cylindrical magnet and 0.2 T magnet based on Halbach concept. The
latter two structures are dedicated for skin cancer and brain MRI respectively and can be sued
for MRI using gradientless imaging method developed by the Institute for Biodiagnostics of
National Research Council of Canada. The cylindrical magnet consists of 350 Nd-Fe-B
blocks of the total weight 350 kg whereas the Halbach structure comprises about 500 Nd-Fe-
B blocks and 700 non-magnetic elements of the total weight of about 1000 kg. The shape of
the Halbach magnet reminds a typical cylindrical MRI superconducting magnet with the bore
of 500 mm in diameter and 500 mm long. However the direction of the magnetic field is
perpendicular to the magnet geometrical axis. The 0.1% homogeneity required for the
gradientless method is obtained before final shimming in an elliptical cylinder of 185 (axis)
x125 (axis) x 45 mm (length).
84

ASSESSMENT OF CARDIAC FUNCTION RESERVE IN MURINE
MODELS OF HEART FAILURE, IN VIVO STUDY
U. Tyrankiewicz 13 , T. Skorka 1 , S. Heinze-Paluchowska 1 , M. Jablonska 1,2 , M. Wozniak 3 ,
L. Drelicharz 3 , and S. Chlopicki 3
1 Department of Magnetic Resonance Imaging, H. Niewodniczanski Institute of Nuclear
Physics PAN, Krakow, Poland; 2 Departament of Medical Physics and Biophysics, AGH
University of Science and Technology, Krakow, Poland; 3 Department of Experimental
Pharmacology, Chair of Pharmacology CMUJ, Krakow, Poland,
Purpose:
The aim of the present work was to assess the alterations in systolic and diastolic cardiac
function for better understanding of mechanisms leading to cardiomyopathy in two different
murine models of the dilated cardiomyopathy: resulted in genetically induced heart
hypertrophy and being pathology after oncological treatment.
Normalized LV slice area
12
Linear regression
EDA
0
20
8
ER
FR
FAC
40
4
ESA
60
80
0
0 40 80 120 160
Time [ms]
Fig.1. Normalized LV slice area over the time as measured from the R-wave. Ejection (ER)
and Filling (FR) Rates are equal to absolute value of regression line slopes in units of 1/ms.
Fractional Area Change (FAC) is equal to normalized difference between end-diastolic and
end-systolic LV slice areas.
Methods and Materials:
Cardiac MRI was used to assess the area of the short axis left ventricle crossection at the level
of the papillary muscles. Various parameters have been deriven from the resulting area-time
curve to characterize cardiac muscle function (Fig. 1). These were: HR (heart rate), ER
(ejection rate), FR (filling rate) and FAC (fractional area change). Additionally, myocardial
reserve was measured in response to pharmacologically induced (dobutamine) stress.
85

Two murine models of heart failure were used:
(1) Tgαq*44 mice (Tg mice) that develop heart failure (HF) during their lifespan [1,2].
These mice were observed at age of 4, 8, 9,12,13 and 15 months.
(2) Doxorubicin induced HF (DOX mice), C57BL/6 mice, 5 weeks of doxorubicin
treatment 2 mg/kg i.p. injected twice weekly, which were observed 7 weeks after the
beginning of chemotherapeutic treatment;
Experiments were performed on 4.7T/310 magnet (Bruker) system equipped with MARAN
DRX (Resonance Instruments) console. All animal experimental procedures were approved
by Ethic Commission of Jagiellonian University.
Results:
In the control group, dobutamine stimulation improves chronotropic, inotropic and lusitropic
reserve of LV (HR, ER and FR response, respectively). In murine heart pathology models
however, ER and FR response was altered. In Tg mice FR response to dobutamine was not
visible at age of 4 months but was relatively strong in age of 8-9 months, whereas at late stage
of heart pathology the this reserve decreased dramatically. ER response to dobutamine stayed
at approximately the same level despite of decreasing resting value. In DOX-treated mouse
both ER and FR responses to pharmacologically induced stress was impaired.
Acknowledgments: This work was supported by PMSHE (grant NN 518419733)
References:
1. Mende et al, JMCC (2001),
2. Drelicharz et al, BRC (2008)
3. Earls J. P., Ho V. B., et al, JMRI (2002)
4. Wiessman F., Ruff J., et al, Circ. Res. (2001)
86

LOCAL AND COOPERATIVE DYNAMICS IN MODIFIED
POLY(DIMETHYLSILOXANE)S STUDIED BY FAST FIELD CYCLING
NMR, BROADBAND DIELECTRIC AND FOURIER TRANSFORM
INFRARED SPECTROSCOPIES
Wiktor Waszkowiak a , Aleksandra Wypych a , Maria Dobies a , Stefan Jurga a ,
and Hieronim Maciejewski b .
a Department of Macromolecular Physics, Faculty of Physics, Adam Mickiewicz University,
Poznań, Poland; b Poznan Science and Technology Park, Poznań, Poland
Poly(dimethylsiloxane) (PDMS) is an organic polymer made of silicon-oxygen chain
with linked methyl groups. Unique properties of PDMS compared with other organic
polymers make them very interesting class of substances. Although linear PDMS has already
been widely studied, its grafted copolymers are still worth investigating due to their potential
applications. The linear PDMS is partially crystalline, while structure of grafted copolymers
depends on the type of a modifier. An incorporation of following side chains
(a) (-CH 2 ) 3 -O-CH 2 -(CHOCH 2 ), (b) (-CH 2 ) 3 -(OCH 2 CH 2 ) 7 -OCH 3 , (c) (-C 16 H 33 ) to the
backbone influences morphology and molecular dynamics of PDMS. It was found that PDMS
copolymers grafted by side chain of type: (a) has amorphous structure, whereas an
incorporation of the (b) and (c) modifiers leads to semicrystalline morphology formed by an
arrangement of grafted long side chains.
Fast Field Cycling NMR (FFC) studies performed in room temperature suggest the
Rouse dynamics for each investigated copolymers [1]. The Broadband Dielectric
Spectroscopy (BDS), performed in wide temperature range allowed to detect various
relaxation processes like: β relaxation - assigned to local motions, α relaxation - connected to
segmental chain motions of polymer backbone, α c relaxation - coming from restricted chain
mobility by crystalline phase.
In order to determine which part from investigated copolymers of type (b) and (c)
crystallizes, the Fourier Transform Infrared spectroscopy (FTIR) studies were performed in
broad temperature range. It was found that crystallization sensitive bands assigned to long
alkyl or poly(ethylene oxide) chains had appeared in FTIR spectra below melting
temperature [2]. It suggests that crystallization is driven by an arrangement of relatively long
polymer chains grafted to PDMS backbone.
Acknowledgements:
This work is supported by 6th Framework Programme under SoftComp Grant No 502235-2.
A. Wypych acknowledges the Ministry of Science and Higher Education in Poland for
a Postdoctoral Fellowship (POL-POSTDOC III).
References:
[1] R. Kimmich, E. Anoardo, Progress in Nuclear Magnetic Resonance Spectroscopy, 44
(2004) 257–320.
[2] L. Sun, Y. Liu, L, Zhu, B. S. Hsiao, C. A. Avila-Orta, Polymer, 45 (2004) 8181–8193.
87

A NOVEL DETERMINATION OF 13 C NUCLEAR MAGNETIC
MOMENT IN TERMS OF THE 3 He
FROM GASEOUS 13 CH 4 NMR SPECTRA
Marcin Wilczek 1 , Anna Szyprowska 2 , and Włodzimierz Makulski 1
1 Laboratory of NMR Spectroscopy, Department of Chemistry, Warsaw University,
Warsaw, Poland; 2 Pharmaceutical Research Institute, Warsaw, Poland
From the earlier atomic and molecular beam experiments it is known that NMR
spectroscopy is well suited for a precise determination of magnetic properties like nuclear
magnetic dipole moments. Traditional techniques as the static spectroscopy in homogenous
magnetic fields can be applied. Measurements are conducted in bulk materials and always
corrections for susceptibility and shielding effects are needed. These effects can entirely be
omitted when gas phase studies are performed. Extrapolation of appropriate chemical shift
values to the zero-pressure limit is the heart of the matter of this procedure [1]. Slightly
modified version of this method exploits helium-3 atoms, magnetic moment of which is
known with a high accuracy [2]. Very few new results received from gas phase NMR and
theoretical calculations can be found in [3].
In this work we examined the 1 H, 13 C and 3 He chemical shifts in gaseous mixtures of
helium and methane molecules. The gaseous samples containing small amounts of 3 He/ 13 CH 4
mixture in an excess of CO 2 and Xe as buffers were prepared at total densities in the range
0.22 – 1.26 mol/L. The 1 H, 13 C and 3 He NMR resonance frequencies were measured on a
Varian INOVA 500 spectrometer at 300 K.
At a constant external magnetic field B 0 = 11.7578 MHz the measured resonance
frequencies are strictly linearly dependent on CO 2 and Xe gas densities. This is consistent
with the general approach used for the description of gas-phase NMR experiments.
The Larmor precession frequencies observed, which are affected by intermolecular
interactions, are as follows: ν 0 ( 3 He) = 381.3572155(2) MHz, ν 0 ( 13 C) = 125.8762395(7) MHz
and ν 0 ( 1 H) = 500.6068164(5) MHz. Recalculated from those data isotropic experimental
values in methane molecule are: σ 0 ( 1 H) = 30.593(24) ppm, σ 0 ( 13 C) = 195.01(90) ppm in the
absolute shielding scale. Experimental and theoretical shielding constants for helium and 1 H
and 13 C nuclei in methane [4] were also used to receive 13 C nuclear dipole moment for the
free nuclei in its ground state. New experimental value µ( 13 C) is +0.70236943(63)µ N , where
µ N is the nuclear magneton. This more precise quantity is in good agreement with previously
reported results.
REFERENCES:
[1] A.Antušek, K.Jackowski, M.Jaszuński, W.Makulski,, M.Wilczek, Chem.Phys.Lett., 411
(2005) 111.
[2] A.Rudziński, M.Puchalski, K.Pachucki, J.Chem.Phys., 130 (2009) 244102.
[3] http://www.icho.edu.pl/PL/Zespoly/22/
[4] M.Jaszuński, K.Jackowski, Nuclear Magnetic Dipole Moments from NMR Spectra-
Quantum Chemistry and Experiment, Lecture Notes in Physics, Springer: Berlin, 2008;
Vol.745, p.233.
88

EPR EXAMINATION OF FREE RADICALS
IN RADIATIVE STERILIZED PIPERACILLIN
Sławomir Wilczyński 1 , Barbara Pilawa 1 , Marta Ptaszkiewicz 2 , Janusz Swakoń 2 ,
Paweł Olko 2 , Robert Koprowski 3 , and Zygmunt Wróbel 3
1 Medical University of Silesia in Katowice, School of Pharmacy and Laboratory Medicine,
Department of Biophysics, Sosnowiec, Poland; 2 Department of Radiation Physics and
Dosimetry, Institute of Nuclear Physics, Cracov, Poland; 3 University of Silesia, Faculty
of Computer Science and Material Science, Institute of Computer Science,
Sosnowiec, Poland
Free radicals in radiative sterilized piperacillin were studied by electron paramagnetic
resonance (EPR) spectroscopy. Piperacillin sodium salt is the semi-synthetic antibiotics with
broad-spectrum antibacterial activity and have good antibacterial activity against Grampositive
and-negative bacteria. [1]. Chemical structure of piperacillin is presented in Figure 1
[1]. Sterilization of the analyzed antibiotic was performed by gamma irradiation in
THERATRON 780E containing isotope 60 Co. Dose of gamma irradiation of 25 kGy was used.
First-derivative spectra were measured by an X-band (9.3 GHz) EPR spectrometer
with modulation of magnetic field 100 kHz. Microwave power in the range of 0.7-70 mW was
used. Free radical concentration was determined. Ultramarine and a ruby crystal were the
references. Spin-spin and spin-lattice relaxation in the tested antibiotic were analyzed.
Changes in paramagnetic centers system during storage of the drug was obtained. Effect of
oxygen on free radicals in piperacillin during its storage was examined. Changes of integral
intensity of EPR spectra of irradiated piperacillin with increasing time of storage were fitted
by theoretical two-exponential function.
CH
NH
CO
NH
S
CH 3
CH 3
NH
CO
O
N
COONa
N
O
N
O
Fig. 1. Chemical structure of piperacillin [1].
C 2 H 5
Fig. 2. EPR spectrum of gamma irradiated piperacillin. The spectrum was recorded after
irradiation with microwave power of 2 mW.
References:
[1] Chemia leków, A. Zejc, M. Gorczyca, PZWL Warszawa, 2002
Acknowledgements:
This study was financially supported by Medical University of Silesia in Katowice (Grant no.
KNW-2-156/09).
89

FREE RADICALS PROPERTIES OF DIETARY SUPPLEMENTS BASED ON
FISH AND PLANT OILS APPLIED IN COSMETOLOGY
Wilczyński Sławomir 1 , Zdybel Magdalena 1 , Barbara Pilawa 1 , Deda Anna 1,2 ,
and Pierzchała Ewa 2
1 Medical University of Silesia in Katowice, School of Pharmacy and Laboratory Medicine,
Department of Biophysics, Sosnowiec, Poland; 2 Medical University of Silesia in Katowice,
School of Pharmacy and Laboratory Medicine, Department of Esthetical Medicine,
Katowice, Poland.
Free radicals play an important role in the pathogenesis of many diseases [1,2]. Some
of dietary supplement may have protective properties against free radicals. Preventive
efficiency of dietary supplements depend on antioxidants contents [1,2]. To the most popular
dietary supplements preparation containing plant and fish oils belong to. Antioxidative
potential of selected dietary supplements was studied with the aid of electron paramagnetic
resonance spectroscopy at X-band microwave frequencies (9.3 GHz). After adding to
analyzed preparations the standard solution of free radicals (DPPH) the EPR spectra
parameters were estimated. Amplitudes (A), linewidths (∆B pp ) and integral intensities (I) of
the spectra were determined. Oenothera oil (OO), sardine fish oil (SFO) and two different
samples of oli from greenland shark liver (OGS 1 and OGS 2) were analyzed.
All studied dietary supplements are effective in free radicals scavenging. In the
decreasing free radicals scavenging capacity, the order was: OSG 1, SFO, OSG 2 and OO. No
significant changes of linewidths were stated (tab. 1)
Table 1. Amplitudes of EPR lines (A) and linewidths (∆B pp ) for all studied samples.
Sample A [a.u.] ∆B pp (mT) + 0.02
DPPH 1.60 0.13
OO 0.42 0.12
SFO 0.23 0.11
OGS1 0.17 0.12
OGS2 0.39 0.12
It was proven that EPR technique is suitable to antioxidative properties of the studied
dietary supplements determination. Obtain results may indicate that oils of animal origin are
more efficient in free radicals scavenging in comparison with dietary supplements based on
oils of plant origin.
References:
1. Bartosz G. Druga twarz tlenu. Wolne rodniki w przyrodzie. Warszawa: PWN, 2006.
2. Duda-Chodak A, Tarko T. Wolne rodniki – za i przeciw. Laboratorium 2007, 11: 48-50.
Acknowledgements:
This study was financially supported by Medical University of Silesia in Katowice (Grant no.
KNW-2-156/09)
90

RELAXATION PARAMETERS OF GD(III) AND MN(II) IONS IN
WATER FROM THE PERSPECTIVE OF CONTRAST AGENTS
Milosz Wojciechowski and Danuta Kruk
Institute of Physics, Jagiellonian University, Reymonta 4, 30-059 Krakow, Poland
Paramagnetic systems are materials with positive magnetic susceptibility, associated
with unpaired electrons. The paramagnetic solutions of interest for Magnetic Resonance
Imaging contain transition metal complexes. The presence of unpaired electron spins has a
profound influence on Magnetic Resonance Spectra of such solutions. The origin of the
effects is found in the large value of the electronic magnetic moment about 650 times that of
the proton. The paramagnetic species enhance the nuclear spin relaxation rates. The
paramagnetic relaxation enhancement is caused by random variations of the electron spin –
nuclear spin dipole-dipole interactions, which open a pathway for the nuclear spin relaxation.
A very active field of applications of the paramagnetic relaxation enhancement effects is the
development and use of paramagnetic materials as contract agents in Magnetic Resonance
Imaging.
In this work electron spin interactions and relaxation parameters of Gd(III) and Mn(II)
ions are discussed in the contexts of their further effects on proton relaxation of the solvent
molecules. Proton relaxation has been measured at low and high magnetic fields for solutions
of increasing viscosity slowing down reorientational motion of the paramagnetic complexes.
Effects of the molecular tumbling on the paramagnetic relaxation enhancement are
investigated in detail.
This work has been financed by Polish Ministry of Science and Education,
grant No N N202 105936
91

UNIPLANAR GRADIENT COILS – FROM DESIGN TO APPLICATIONS
Grzegorz Woźniak*, Tomasz Skórka*, Krzysztof Jasiński*, Tomasz Banasik*, Mohammad
Mohammadzadeh**, Dominik von Elverfeldt**, Jürgen Hennig**, and Władysław P. Węglarz*
*IFJ PAN Kraków, Poland; **Freiburg University, Freiburg, Germany
Cylindrical gradient coils typically used for MRI imaging have limitations in gradient
strength as well as restrictions in access to sample. In recent years growing interest in coils with
different geometry, allowing for better sample access and/or higher local magnetic field gradient,
is observed [1]. Aim of this study was to design, build and test flat gradient coil set for 3D micro
imaging. Comparison of theoretical results with simulated parameters and physical tests using 4.7
T and 9.4 T MRI scanners are presented.
Flat gradient coil set was designed using stream function approach [2,3]. Coil size was 10 by 10
cm and the assumed field of view (FOV) 1.5x0.15x1.5cm with centre 5.5 mm above coil surface.
The design procedure has been implemented using scripting language of the Comsol Multiphysics
package (Comsol AB, Sweden). Magnetic field generated by current patterns were analysed using
Comsol AC/DC module to verify theoretical predictions. In order to compensate for distortions
introduced by bounding wires and connection paths, an additional current paths were considered.
Temperature simulations were done to assess requirement for cooling method, thermal
conductivity of materials and limits of the current in the coils. Theoretical, simulated and physical
values of coils parameters were compared on a manufactured prototype coil set.
Coils set has been constructed and tested. Overall achieved FOV size and gradient strength was in
agreement compared to original theoretical calculations. Temperature measurements show that
energy deposit can by received by water cooling system at 15 0 C providing temperature below
50 0 C with 50A power supply and 20% filling factor at all three coils on. Results of the
temperature distribution of the probehead surface is presented. Distributions of magnetic field
along work cross-section are presented. MR images of the test sample containing set of parallel
rows filled with water shows regions of gradient linearity.
Results confirm feasibility of manufacturing effective flat gradient coils for MR micro-imaging as
well as usefulness of simulation which allow to determine the best way in every step of design.
Future tests are prepared.
References:
1.see recent ISMRM,e.g.posters #3055-3074,2008,Honolulu,Hawaii,US.
2.R.Lemidasov,R.Ludwig,Conc.Magn.Reson.B.,Vol. 26B(1)(2005)p.67–80.
3.G.Wozniak,T.Skorka,K.Jasinski,W.P.Weglarz,2008,NMR School, Wierzba,Poland
Acknowledgments:
This work was supported by European Community grant FP6-2004-NEST-C-1-028533
92

DYNAMICS OF POLYSTYRENE OF DIFFERENT MOLECULAR
ARCHITECTURE AS REVEALED BY NUCLEAR MAGNETIC
RESONANCE, BROADBAND DIELECTRIC SPECTROSCOPY
AND REOLOGY
Aleksandra Wypych a , Monika Makrocka-Rydzyk a , Grzegorz Nowaczyk a ,
Eugeniusz Szcześniak a , Stefan Jurga* a , Akira Hirao b , and Takumi Watanabe b .
a Department of Macromolecular Physics, Faculty of Physics, Adam Mickiewicz University,
Poznań, Poland; b Graduate School of Science and Engineering, Tokyo Institute of
Technology, Tokyo Japan
Dendrimer-like star-branched polymers have recently
appeared as a new class of hyperbranched polymers
synthesized by means of living anionic polymerization
[1]. Their structure with chain units branching out from
a center cord comprises several layers (so-called
generations).They possess interesting properties
resulting from unusual architectural features obtained
by methods of controlled polymerization [2]. However,
they require a detailed characterization on a molecular
level, what can be realized using modern spectroscopic
methods. The aim of this study is to determine the
influence of the system architecture on molecular
dynamics in linear and dendrimer-like star-branched
polystyrenes.
Dendrimer-like
star-branched polystyrene
The analysis of temperature behavior of:
13 C NMR, Broadband Dielectric
Spectroscopy and Rheological data indicates an existence of three motional processes denoted
as α, β and γ in order of decreasing temperature. The α relaxation is related to the dynamics of
glass transition, the β -relaxation was assigned to a motion of the phenyl rings, while the γ
relaxation was attributed to the local motions of the methylene units (present due to tail-to-tail
or head-to-head sequences). A broadening of the distribution of segmental relaxation times in
the polymer of dendritic structure, with respect to the linear one, was observed.
References:
[1] W.A. Braunecker, K. Matyjaszewski, Prog. Polym. Sci., 32, 93, 2007.
[2] B.Lepoittevin, R. Matmour, R. Francis, D.Taton, Y.Gnanou, Macromolecules, 38, 3120,
2005.
[3] A. Deffieux, M. Schappacher, A. Hirao, T. Watanabe, J. Am. Chem. Soc., 130 (17), 5670,
2008.
Acknowledgements: This work is supported by 6th Framework Programme under SoftComp
Grant No 502235-2 and research grant No N N202 128536 (Poland). A. Wypych
acknowledges the Ministry of Science and Higher Education in Poland for a Postdoctoral
Fellowship (POL-POSTDOC III).
93

SPECTRA OF HIGH DIMENSIONALITY FOR EASY RESONANCE
ASSIGNMENT IN PROTEINS
Anna Zawadzka-Kazimierczuk, Krzysztof Kazimierczuk, and Wiktor Koźmiński
Faculty of Chemistry, University of Warsaw, Warsaw, Poland
Multidimensional experiments, invaluable in case of crowded spectra, may sometimes
not ensure satisfactory results, when performed in a conventional way, where the
evolution time space is sampled at equal intervals. The necessity of fulfilling the
Nyquist Theorem in this case causes that having limited experimental time one has to
compromise between resolution and spectral width. In such situation, when peaks
overlap, it is hard to precisely evaluate chemical shifts. We have proposed the method
which allows overcoming this problem.
Random sampling of evolution time space, followed by Multidimensional Fourier
Transform, provides spectra of very high resolution (it is possible to obtain natural
linewidths) and enables performing experiments of high dimensionality (4D, 5D, 6D).
The processing of such spectra takes advantage of resonance positions form spectra of
lower dimensionality and thus results with a set of 2D spectra, which makes it very
convenient to use.
We propose several novel techniques which allow easy assignment of protein backbone
signals. The experiments were performed on proteins of various sizes: ubiquitin (76
residues), protein interacting with NIMA-kinase from Cenarcheaum symbiosum (96
residues) and maltose binding protein (370 residues).
References:
[1] Zawadzka-Kazimierczuk A, Kazimierczuk K, Koźmiński W
J. Magn. Reson. (in press, doi: 10.1016/j.jmr.2009.10.006)
94

EFFECT OF TEMPERATURE ON EPR SPECTRA
OF DOPA-MELANIN-NETILMICIN COMPLEXES
Magdalena Zdybel 1 , Barbara Pilawa 1 , Ewa Buszman 2 , Dorota Wrześniok 2 ,
Ryszard Krzyminiewski 3 , Zdzisław Kruczyński 3 , Robert Koprowski 4 , and Zygmunt Wróbel 4
1, 2 Medical University of Silesia in Katowice, School of Pharmacy and Division of Laboratory
Medicine, 1 Department of Biophysics, Sosnowiec, Poland; 2 Department of Pharmaceutical
Chemistry, Sosnowiec, Poland; 3 Adam Mickiewicz University, Institute of Physics,
Department of Medical Physics, Poznań, Poland; 4 University of Silesia, Faculty of Computer
Science and Material Science, Institute of Computer Science, Sosnowiec, Poland
Types of paramagnetic centers in DOPA-melanin and its complexes with netilmicin
were analysed by EPR spectroscopy. Free radicals with spin S = 1/2 and biradicals with spin
S =1 were found in the studied melanin samples. Correlations between the integral intensities
(I) of the EPR spectra and the measuring temperature (T) in the range 100-300 K were
searched. Intensities of all the recorded EPR lines do not change with temperature according
the Curie law. Numerical fitting of the experimental data by theoretical functions indicates
that sum of functions for free radicals and biradicals are the best results for the obtained
correlations of I = f(T). Intensities of EPR lines of free radicals fulfill the Curie law (I = C/T).
The other function describes correlation between intensity of EPR lines of biradicals
and temperature.
DOPA-melanin was obtained according to Binn’s method [1]. Concentrations
of complexing agent - netilmicin was 1x10 -3 M. Measurements were done by the use of an X-
band (9.3 GHz) spectrometer of BRUKER Firm. The first derivative EPR spectra were
measured with low microwave power of 0.3 mW to avoid microwave saturation of line.
Changes of amplitude (A), linewidth (∆B pp ), and integral intensity (I) of EPR spectra
with the measuring temperature (T) were determined.
Experimental data IT = f(T) were numerically analysed. The experimental product IT
= f(T) was fitted by sum of the theoretical functions for free radicals and biradicals according
to the formula [2-3]:
IT = C + D/(3 + exp(J/kT))
where: I – integral intensity of EPR line, T – the measuring temperature, and C – Curie
constant, D – constant, J – energy of singlet-triplet excitation, and k – Boltzmann constant.
The parameters C and D, and the energy of singlet-triplet excitation J, were calculated.
The error of the fitting was calculated as follow:
δ = 1/N (Σ T | IT – IT (S) |)
where: IT – experimental product, IT (S) – theoretical product, N – the number of the
experimental points.
The Labview and the program for spectral analysis of Jagmar Firm (Kraków, Poland)
were used to determination of the parameters of the EPR spectra. Integral The Matlab 7.20,
Signal Processing and Statistics were used during the theoretical fitting of the experimental
data.
Free radicals and biradicals in melanin samples were earlier found by Kozdrowska
et al. [4-5]. Free radicals (S = 1/2) and biradicals (S = 1) form complex paramagnetic centers
system in DOPA-melanin and complexes of DOPA-melanin with kanamycin and Cu(II) [4-5].
95

References:
[1] Binns F, Chapman RF, Robson NC, Swan GA, Waggot A. J Chem Soc C 1970: 1128-
1134.
[2] Wertz JE, Bolton JR. New York: Academic Press; 1986.
[3] Hatfield WE. [In:] Boudreaux EA, Mulay LN. New York, London, Sydney, Toronto: John
Wiley & Sons; 1976: pp. 357-359.
[4] Kozdrowska L. Rozprawa doktorska. Uniwersytet Zielonogórski, Zielona Góra, 2005.
[5] Kozdrowska L, Pilawa B, Świątkowska L, Buszman E, Wrzesniok D, Grzegorczyk A,
Więckowski AB, Wojtowicz W, Wilczok T. Abstracts of 21st International Meeting
on Radio- and Microwave Spectroscopy RAMIS 2005. Poznań-Będlewo 2005, p. 33.
[6] Pilawa B, Latocha M, Krzyminiewski R, Kruczyński Z, Buszman E, Wilczok T. Phys
Med 2004; 1: 96-98.
Acknowledgements:
This study was financially supported by Medical University of Silesia in Katowice (Grant no:
KNW- 2-126/09).
96

SPIN-LATTICE RELAXATION IN THERMALLY STERILIZED
CHLORTALIDONE
Magdalena Zdybel, Jakub Adamczyk, Barbara Pilawa, Magdalena Kościelniak,
and Daria CzyŜyk
Medical University of Silesia in Katowice, School of Pharmacy and Laboratory Medicine,
Department of Biophysics, Jedności 8, 41-200 Sosnowiec, Poland,
Spin-lattice relaxation in Chlortalidone was studied by electron paramagnetic
resonance (EPR). Chlorthalidone is a diuretic drug used to treat hypertension. [1]. Chemical
structure of Chlortalidone is shown in Figure 1 [1]. Chlortalidone was sterilized according to
the valid norms [2] in hot air oven at the following conditions: 160 o C by 120 minutes, 170 o C
by 60 minutes, and 180 o C by 30 minutes. The aim of this work was to determine effect of
conditions of thermal sterilization on spin-lattice relaxation in Chlortalidone. Changes of spinlattice
interactions in thermally sterilized drug point out that its chemical structure are
modified. Examination of magnetic interactions in drug is helpful to obtain optimal condition
of its thermal sterilization.
Cl
HO
O
S
NH
O NH 2
O
Fig. 1. Chemical structure of Chlortalidone.
Thermally sterilized Chlortalidone is paramagnetic. EPR spectra were measured for
Chlortalidone heated at 160 o C, 170 o C and 180 o C. The spectra were recorded by the use of
EPR spectrometer produced by RADIOPAN Firm (Poznań). Magnetic modulation was 100
kHz. Microwave frequency from X-band (9.3 GHz) was measured by MCM 101 recorder of
EPRAD Firm (Poznań). EPR spectra were measured in the wide range of microwave power
from 0.7 mW to 70 mW. Continuous microwave saturation of EPR lines was applied to
examination of spin-lattice relaxation processes [3-5].
References:
[1] ZEJC A., GORCZYCA M. (RED.) Chemia leków, PZWL, Warszawa, 2002
[2] Farmakopea VII
[3] STANKOWSKI J., HILCZER W. Wstęp do spektroskopii rezonansów magnetycznych. PWN, Warszawa,
2005
[4] WERTZ J. E., BOLTON J. R. Electron Spin Resonance Theory and Practical Applications. New York,
London 1986
[5] EATON G. R., EATON S. S., SALIKHOV K. M. (Eds.). Foundations of modern EPR, World Scientific,
Singapore, New Jersey, London, Hong Kong 1998
Acknowledgements
This study was supported by Medical University of Silesia in Katowice (Grant no: KNW 1-
142/09).
97

DISPERSION MEASUREMENTS OF T 1ρ IN PROTEIN SOLUTIONS
Łukasz Żelazny 1 , Dorota Wierzuchowska 2 , and Barbara Blicharska 1
1 Jagiellonian University, Department of Physics, Krakow, Poland;
2 Pedagogical University, Institute of Physics, Kraków, Poland
Relaxation time measurements in rotating frame and its dependence on frequency,
called NMR dispersion, may be a source of information about molecular dynamics and
structure changes in aqueous protein solution. Assuming a pure dipole-dipole interaction as a
dominant relaxation mechanism of proton in free and bounded water we are able to calculate
the correlation time from T 1ρ dispersion profile (that is dependence of relaxivity in rotating
frame R 1ρ = 1/T 1ρ on Β 1 ).
These calculations were done for samples of different protein solution ( albumin and
lysozyme) at different protein concentration. The changes of molecular dynamics, which
appear after thermal (irreversible) or alcohol (reversible) denaturation, are discussed.
98

THE ONE NMR PROBE FROM VARIAN AND THEIR
COMBINATION TOOLS
Thomas Zellhofer
Varian Deutschland GmbH, Darmstadt, Germany
The OneNMR Probe represents the most significant advancement in solution-state
probe technology in over a decade. The OneNMR probe uses a hybrid approach to deliver the
performance advantages of both the classic "carbon probe" (direct detect probe) and the highly
sensitive "proton probe" (indirect detect probe). It is simultaneously optimized for both high and
low band frequencies in a single unit. Increased 1H sensitivity is just one component of this new
technology, it also provides excellent RF homogeneity, excellent salt and solvent tolerance,
excellent water suppression, and excellent lock sensitivity compared to traditional probe
technology. This combination offers unprecedented performance and flexibility for all your
routine NMR experiments.
The OneNMR probe provides you with all of the data required for routine structure analysis of
organic molecules, significantly reducing the labor and productivity-loss associated with
switching probes for multiple experiments.
The flexibility and power of the OneNMR probe products is most pronounced when combined
with automation accessories and VnmrJ 3. This latest release of VnmrJ was redesigned with your
workflow in mind. This combination (VnmrJ 3, OneNMR probe, ProTune, and Autosampler) are
the perfect solution for all you routine NMR needs.
99

NUCLEAR RELAXATION IN SELECTED IMIDAZOLIUM
COMPOUNDS: EFFECT OF QUADRUPOLAR INTERACTIONS
Włodzimierz Masierak 1 , Cezary Uniszkiewicz 1 , Alexiei Privalov 2 , Franz Fujara 2 , Wojciech
Medycki 3 , and Danuta Kruk 4
1 Institute of Physics, Bydgoszcz University, Poland; 2 Technische Universität Darmstadt,
Germany; 3 Institute of Molecular Physics, PAS , Poznań, Poland; 4 Institute of Physics,
Jagiellonian University, Krakow, Poland
Imidazolium compounds are very interesting example of systems of mutually coupled
dipolar and quadrupolar spins. Such systems contain several types of dynamically nonequivalent
imidazolium rings. As a consequence of the different dynamics one experimentally
observes proton relaxation as well as polarization transfer effects. Polarization transfer can
take place only if there is an efficient (not averaged out) coupling between the dipolar and
quadrupolar spins. If the magnetic field is set to a value which leads to the Zeeman splitting
of the dipolar spin matching the energy splitting of the quadrupolar spin (determined by the
quadrupolar and Zeeman interactions), the dipole-dipole coupling causes the polarization
transfer. The mutual dipole-dipole coupling links transitions of the dipolar spin to some
transitions of the quadrupolar spin, so they cannot occur independently. The polarization of
the dipolar spins is transferred to the quadrupolar subsystem with an efficiency directly
related to the probability of the joint transitions. The fact that one can clearly see the
polarization transfer effects (as dips of the magnetization curve recorded versus the magnetic
field) allows us to state that some of the imidazolium rings are quite rigid and therefore there
are efficient proton – quadrupolar nuclei dipole – dipole couplings. At the some time proton
relaxation is observed. We attribute this process to a fast movement of protons belonging to
other rings. The relaxation processes are considerably affected by dipole-dipole couplings to
the quadrupolar nuclei.
This work has been financed by Polish Ministry of Science and Education, grant No N N202
172135
100

APPLICATION OF FAST FIELD CYCLING NMR RELAXATION
TO SELECTED IMIDAZOLIUM COMPOUNDS
Cezary Uniszkiewicz 1 , Włodzimierz Masierak 1,2 , Alexiei Privalov 3 , Franz Fujara 3 , Wojciech
Medycki 4 , and Danuta Kruk 5
1 Institute of Physics, Bydgoszcz University, Bydgoszcz, Poland; 2 School of Economy,
Bydgoszcz, Poland; 3 Technische Universität Darmstadt, Germany; 4 Institute of Molecular
Physics, PAS , Poznań, Poland; 5 Institute of Physics, Jagiellonian University,
Krakow, Poland
This work is concerned with several aspects of field dependent relaxation processes
and polarization transfer effects in complex solid state systems containing mutually coupled
quadrupolar and dipolar spins (of the spin quantum number ½). Dipole - dipole interactions
provide the coupling between the two spin subsystems.
These phenomena are illustrated by several experimental examples for various
imidazolium compounds. Such systems are very attractive because of variety of effects caused
by interplay between quadrupolar and dipolar interactions carrying unique information on
molecular dynamics.
This work has been financed by Polish Ministry of Science and Education, grant No N N202
172135
101