Susceptibility to radiation-induced leukaemia/lymphoma is genetically separable from sensitivity to radiation-induced genomic instability

PURPOSE: To determine whether there is a relationship between the genetics underlying the susceptibility to radiation-induced leukaemia in CBA/H (acute myeloid leukaemia, AML) and C57BL/6 (thymic lymphoma, TL) mice, and the genetics underlying the sensitivity of CBA/H (sensitive) and C57BL/6 (resistant) mice to radiation-induced chromosomal instability. MATERIALS AND METHODS: CBA/H, (CBA/H x C57BL/6)F1, F1 x CBA/H, F1 x C57BL/6 and F1 x F1 mice were exposed to a single acute dose of 3.0 Gy X-rays. AML and TL were diagnosed over the subsequent 30 months. RESULTS: There was no statistically significant difference in the incidence of AML in F1, F1 x F1, F1 x CBA/H and F1 x C57BL/6 mice, which was approximately 50% that in CBA/H mice. AML susceptibility is therefore a dominant polygenic trait, and both susceptibility and resistance (variable penetrance) CBA/H and C57BL/6 loci are involved. The incidence of TL in the FM and F1 x CBA/H mice was negligible, indicating that TL susceptibility is a recessive trait. As the TL incidence in the F1 x C57BL/6 mice was about half that in C57BL/6 mice, one recessive locus is probably involved. CONCLUSIONS: AML susceptibility in CBA/H mice is a dominant trait in contrast to the recessive inheritance of CBA/H sensitivity to radiation-induced chromosomal instability. TL-susceptibility in C57BL/6 is a recessive trait in contrast to the dominant inheritance of C57BL/6 resistance to radiation-induced chromosomal instability.     

Susceptibility to radiation-induced leukaemia/lymphoma is genetically separable from sensitivity to radiation-induced genomic instability

Purpose : To determine whether there is a relationship between the genetics underlying the susceptibility to radiation-induced leukaemia in CBA/H (acute myeloid leukaemia, AML) and C57BL/6 (thymic lymphoma, TL) mice, and the genetics underlying the sensitivity of CBA/H (sensitive) and C57BL/6 (resistant) mice to radiation-induced chromosomal instability. Materials and methods : CBA/H, (CBA/H ×C57BL/6)F 1, F 1 ×CBA/H, F 1 ×C57BL/6 and F 1 ×F 1 mice were exposed to a single acute dose of 3.0 Gy X-rays. AML and TL were diagnosed over the subsequent 30 months. Results : There was no statistically significant difference in the incidence of AML in F 1, F 1 ×F 1, F 1 ×CBA/H and F 1 ×C57BL/6 mice, which was ~50% that in CBA/H mice. AML susceptibility is therefore a dominant polygenic trait, and both susceptibility and resistance (variable penetrance) CBA/H and C57BL/6 loci are involved. The incidence of TL in the F 1 and F 1 ×CBA/H mice was negligible, indicating that TL susceptibility is a recessive trait. As the TL incidence in the F 1 ×C57BL/6 mice was about half that in C57BL/6 mice, one recessive locus is probably involved. Conclusions : AML susceptibility in CBA/H mice is a dominant trait in contrast to the recessive inheritance of CBA/H sensitivity to radiation-induced chromosomal instability. TL-susceptibility in C57BL/6 is a recessive trait in contrast to the dominant inheritance of C57BL/6 resistance to radiation-induced chromosomal instability.     

Spectral simplification for resolved glutamate and glutamine measurement using a standard STEAM sequence with optimized timing parameters at 3, 4, 4.7, 7, and 9.4T.

The C4 multiplet proton resonances of glutamate (Glu) around 2.35 ppm and glutamine (Gln) around 2.45 ppm usually overlap in MR spectra, particularly at low- and mid-field strengths (1.5-4.7T). A spectral simplification approach is introduced that provides unobstructed Glu and Gln measurement using a standard STEAM localization sequence with optimized interpulse timings. The underlying idea is to exploit the dependence of response of a coupled spin system on the echo time (TE) and mixing time (TM) to find an optimum timing set (TE, TM), at which the outer-wings of C4 "pseudo-triplet" proton resonances of Glu and Gln are significantly suppressed while the central peaks are maintained. The spectral overlap is thus resolved as the overlap exists exclusively at the outer-wings and the central peaks are readily separated due to the approximate 0.1-ppm difference in chemical shift. Density matrix simulation for Glu, Gln, and other overlapping metabolites at 2.3-2.5 ppm was conducted to predict the optimum timing sets. The simulated, phantom, and in vivo results demonstrated that the C4 multiplet proton resonances of Glu and Gln can be resolved for unobstructed detection at 3T, 4T, and 4.7T. For simplicity, only simulated data are illustrated at 7T and 9.4T.     

Developmental and regional changes in the neurochemical profile of the rat brain determined by in vivo 1H NMR spectroscopy.

Sixteen metabolites were quantified from 11-24 micro l volumes in three different brain regions (hippocampus, striatum, and cerebral cortex) during postnatal development. Rat pups from the same litter were repeatedly measured on postnatal days 7, 10, 14, 21, and 28 using a completely noninvasive and longitudinal study design. Metabolite quantification was based on ultra-short echo-time (1)H NMR spectroscopy at 9.4 T and LCModel processing. Most of the brain metabolites were quantified with Cramer-Rao lower bounds (CRLB) less than 20%, which corresponded to an estimated concentration error <0.2 micro mol/g. Taurine and total creatine were quantified with CRLB < or = 5% from all 114 processed spectra. The resulting high reliability and reproducibility revealed significant regional and age-related changes in metabolite concentrations. The most sensitive markers for developmental and regional variations between hippocampus, striatum, and cerebral cortex were N-acetylaspartate, myo-inositol, taurine, glutamate, and choline compounds. Absolute values of metabolite concentrations were in very good agreement with previously published in vitro results based on chromatographic measurements of brain extracts. The current data may serve as a reference for studies focused on developmental defects and pathologies using neonatal rat models.     

High-resolution in vivo CBV mapping with MRI in wild-type mice.

NMR microimaging has the potential to elucidate cerebrovascular abnormalities in mouse models. In this study, the relative regional cerebral blood volume (CBV) map is presented for C57BL6/J wild-type mice. The CBV mapping was based on changes in the steady-state NMR transverse relaxation rate (DeltaR(2)) associated with the presence of a superparamagnetic intravascular contrast agent (MION) with a long blood halflife. The experiments were performed at 9.4 T at a voxel size of 100 microm x 100 microm x 600 microm. Fine details, such as the hippocampal and olfactory bulb area, were visualized in the CBV map. The relative regional CBV values of various brain regions were measured. The DeltaR(2) dosage dependency and MION tissue clearance in mouse are also reported.     

Localized proton MRS of cerebral metabolite profiles in different mouse strains.

Localized proton MR spectroscopy (MRS) was used to quantify cerebral metabolite concentrations in NMRI (n = 8), BALB/c (n = 7), and C57BL/6 (n = 8) mice in vivo and 1 hr after global irreversible ischemia (2.35 T, STEAM, TR/TE/TM = 6000/20/10 ms, 4 x 3 x 4 mm(3) volume, corrections for cerebrospinal fluid). Anatomical MRI and proton MRS revealed significant differences of the C57BL/6 strain in comparison with both BALB/c and NMRI mice. While MRI volumetry yielded larger ventricular spaces of the C57BL/6 strain, proton MRS resulted in elevated concentrations of N-acetylaspartate (tNAA), creatine and phosphocreatine (tCr), choline-containing compounds (Cho), glucose (Glc), and lactate (Lac) relative to BALB/c mice and elevated Glc relative to NMRI mice. Apart from the expected decrease of Glc and increase of Lac 1 hr post mortem, C57BL/6 mice presented with significant reductions of tNAA, tCr, and Cho, whereas these metabolites remained unchanged in BALB/c and NMRI mice. The results support the hypothesis that the more pronounced vulnerability of C57BL/6 mice to brain ischemia is linked to strain-dependent differences of the cerebral energy metabolism.     

Clinical 1H magnetic resonance spectroscopy of brain metastases at 1.5T and 3T

Purpose: To investigate whether improvements in signal-to-noise ratio (SNR) and spectral resolution are found in spectra from patients with brain metastases obtained at higher magnetic field strengths using standard clinical instrumentation.

Material and Methods: Six patients with brain metastases, 13 healthy volunteers, and a phantom containing brain metabolites were examined using two clinical MR instruments operating at 1.5T (Siemens) and 3T (Philips) with standard clinical head coils. Spectra were obtained using a point resolved spectroscopy pulse sequence, echo times (TE) 32 ms and 144 ms, and repetition time 2000 ms from a volume-of-interest (VOI) of size 15×15×15 mm³. SNR and spectral resolution of the metabolites N-acetylaspartate, choline, and creatine compounds in spectra from 3T were compared to the 1.5T spectra.

Results: In general, spectral resolution was improved by 25-30% at higher magnetic field strength. Only minor improvements in SNR were obtained at 3T using short echo time and 20-50% at long echo time.

Conclusion: SNR and spectral resolution were improved at higher magnetic field strength, especially with TE 144 ms, including spectra from patients with heterogeneous brain tumors. However, differences in the defined effective VOI, particularly at short echo time, reduced the expected effect of increased magnetic field strength on the measured SNR.     

Modification of an Incompatible Graft-versus-host Relationship by Admixture of Grafts with F1 Haemopoietic Cells

Summary

When lethally-irradiated CBA.T6T6 mice were grafted with mixtures of C57BL and (C57BL × CBA.T6T6)F₁ cells of bone-marrow or foetal liver the incidence of secondary disease was considerably reduced as compared with mice receiving the incompatible C57BL cells only. Cytogenetic examination of bone-marrow, spleen, thymus and lymph node and identification of CFU in blood showed that F₁ cells were not eliminated as anticipated and that C57BL cells persisted in similar numbers, whether or not the host had secondary disease. The mechanism of avoidance of secondary disease in these experiments appears to be adaptation of the incompatible allogeneic graft, the host being carried through the periods of haemopoietic and immunological insufficiency by the F₁ cells. The findings constitute not only a therapeutic advance in a hitherto incompatible graft-versus-host relationship but also require reappraisal of the dominant role accorded to GVHR in secondary disease.     

ACE: a single-shot method for water-suppressed localization and editing of spectra, images, and spectroscopic images

A versatile method for localized (1H) NMR spectroscopy is presented. The method intrinsically combines B0-based spatial localization with the possibility of water suppression and spectral editing. With this sequence it is feasible to localize not only single spectra but also phase-encoded images and spectroscopic images. The technique essentially integrates the "Hahn spin-echo" with the "stimulated echo" sequence and is therefore called ACE (acquiring combined echoes). It realizes water-suppressed three-dimensional localization in a single shot and can be used for localized shimming. Studies in which the new method is applied to phantoms with metabolites diluted at low concentrations are presented. Discrimination between lactate and alanine, employing an adapted spectral editing method with complete inversion, combined with simultaneous water suppression and localization of a 0.06-cc volume is shown. The suppression of signals from outside the selected volume is greater than or equal to 24,000. Also, the method is demonstrated by in vivo experiments at 6.3 T. Localized water-suppressed 1H spectra are obtained completely noninvasively, leaving scalp and fur intact, from well-defined volumes of 0.15 cc in the brain of a living rat. Water-suppressed spectroscopic imaging over a localized volume with "body" coil excitation and noninvasive surface coil detection yielded spectra from voxels as small as 25 microliters in the in vivo rat brain.     

Radiosynthesis and biodistribution of a histamine H3 receptor antagonist 4-[3-(4-piperidin-1-yl-but-1-ynyl)-[11C]benzyl]-morpholine: evaluation of a potential PET ligand

The potent histamine H(3) receptor antagonist JNJ-10181457 (1) was successfully labeled with (11)C in a novel one-pot reaction sequence, with high chemical yield (decay-corrected yield, 28+/-8%) and high specific radioactivity (56+/-26 GBq/mumol). The binding of [(11)C]1 to H(3) receptors was studied in vitro in rat brain and in vivo in rats and mice. The in vitro binding of [(11)C]1 in rat coronal brain slices showed high binding in the striatum, and this binding was blocked by histamine and by two known H(3) antagonists, JNJ-5207852 (2) and unlabeled Compound (1), in a concentration-dependent manner. The biodistribution of [(11)C]1 in rats was measured at 5, 10, 30 and 60 min. The uptake of [(11)C]1 in regions rich in H(3) receptors was highest at 30 min, giving 0.98%, 1.41%, 1.28% and 1.72% dose/g for the olfactory bulb, hippocampus, striatum and cerebral cortex, respectively. However, the binding of [(11)C]1 in the rat brain could not be blocked by pretreatment with either Compound (2) (30 min or 24 h pretreatment) or cold Compound (1) (30-min pretreatment). The biodistribution of [(11)C]1 in a second species (Balb/c mice) showed a higher overall uptake of the radioligand with an average brain uptake of 8.9% dose/g. In C57BL/6-H(3)(-/-) knockout mice, a higher brain uptake was also observed. Analyses of metabolites and plasma protein binding were also undertaken. It appeared that [(11)C]1 could not specifically label H(3) receptors in rodent brain in vivo. Possible causes are discussed.     

In vivo high-resolution volume-selected proton spectroscopy and T1 measurements in the dog brain

Successful in vivo NMR spectroscopy requires a combination of techniques to address the problems of volume selection, water suppression, and resolution. All this needs to be done in the very heterogeneous environment found in living organisms. Previously published techniques are used to obtain 1H spectra from a dog brain, observing metabolites with concentrations below 1 mM. Measurements of spin-lattice relaxation times (T1) are also presented. The 1H relaxation times are long (T1 greater than 1.0 s) yielding information about the fluidity of the molecular environment. Comments are made concerning the achievable linewidth in vivo and the deficiencies that phase-encoding spectroscopic methods may have in obtaining high-resolution 1H spectra.     

High magnetic field water and metabolite proton T1 and T2 relaxation in rat brain in vivo.

Comprehensive and quantitative measurements of T1 and T2 relaxation times of water, metabolites, and macromolecules in rat brain under similar experimental conditions at three high magnetic field strengths (4.0 T, 9.4 T, and 11.7 T) are presented. Water relaxation showed a highly significant increase (T1) and decrease (T2) with increasing field strength for all nine analyzed brain structures. Similar but less pronounced effects were observed for all metabolites. Macromolecules displayed field-independent T2 relaxation and a strong increase of T1 with field strength. Among other features, these data show that while spectral resolution continues to increase with field strength, the absolute signal-to-noise ratio (SNR) in T1/T2-based anatomical MRI quickly levels off beyond approximately 7 T and may actually decrease at higher magnetic fields.     

1H-localized broadband 13C NMR spectroscopy of the rat brain in vivo at 9.4 T.

Localized (13)C NMR spectra were obtained from the rat brain in vivo over a broad spectral range (15-100 ppm) with minimal chemical-shift displacement error (<10%) using semi-adiabatic distortionless enhancement by polarization transfer (DEPT) combined with (1)H localization. A new gradient dephasing scheme was employed to eliminate unwanted coherences generated by DEPT when using surface coils with highly inhomogeneous B(1) fields. Excellent sensitivity was evident from the simultaneous detection of natural abundance signals for N-acetylaspartate, myo-inositol, and glutamate in the rat brain in vivo at 9.4 T. After infusion of (13)C-labeled glucose, up to 18 (13)C resonances were simultaneously measured in the rat brain, including glutamate C2, C3, C4, glutamine C2, C3, C4, aspartate C2, C3, glucose C1, C6, N-acetyl-aspartate C2, C3, C6, as well as GABA C2, lactate C3, and alanine C3. (13)C-(13)C multiplets corresponding to multiply labeled compounds were clearly observed, suggesting that extensive isotopomer analysis is possible in vivo. This unprecedented amount of information will be useful for metabolic modeling studies aimed at understanding brain energy metabolism and neurotransmission in the rodent brain.     

Concentrations of human cardiac phosphorus metabolites determined by SLOOP 31P NMR spectroscopy.

Human cardiac 31P nuclear magnetic resonance (NMR) spectra are usually quantified in relative terms, i.e., the ratio of metabolite signals is calculated. If 31P NMR spectroscopy of the heart is to emerge as a clinically relevant diagnostic modality, reliable quantification of absolute concentrations of 31P metabolites is required. We applied spectral localization with optimal point spread function (SLOOP) 31P NMR spectroscopy to measure absolute concentrations of phosphocreatine (PCr) and adenosine triphosphate (ATP) in human myocardium. The accuracy of the quantification was first validated in a phantom study. Seven healthy volunteers (aged 19-29 years) were then examined at 1.5 T using a nominal spatial resolution of 25 mL. SLOOP allowed us to obtain localized spectra from compartments anatomically matched to the left ventricular wall. The a priori knowledge of the anatomical structure was obtained from 1H images. The spatially varying effects of saturation, off-resonance, and sensitivity were considered during the reconstruction process. Metabolites were quantified with reference to an external 31P standard. Concentrations of 9.0 +/- 1.2 and 5.3 +/- 1.2 mmol/kg wet wt (mean +/- SD, n = 9) were determined for PCr and ATP in normal heart, respectively. The influence of nuclear Overhauser enhancement on metabolite quantification is discussed.     

Experimental approaches to image localized human 31P NMR spectroscopy

Experimental procedures for obtaining localized 31P NMR spectra of humans by means of the ISIS sequence are discussed in detail. The technique is optimized for use with volume coils and with surface coils in order to measure localized 31P NMR spectra of different tissues and organs. Selective frequency-modulated (FM) inversion and excitation pulses are applied for optimal inversion or excitation despite B1 inhomogeneity. Pulse imperfection may lead to spurious signal contributions from outside the selected volume; this contamination is reduced by using long pulse intervals, by properly ordering the ISIS acquisitions, and by using FM excitation pulses. Simultaneous measurement of multiple volumes was implemented by including an additional selective inversion pulse, and an extension of the ISIS addition/subtraction scheme. Localized T1 measurements with surface coils are implemented by using a B1-insensitive inversion pulse in the inversion recovery sequence. The quantitative reproducibility of localized 31P NMR spectra was verified. Absolute metabolite concentration can be determined after a suitable calibration of the 31P NMR spectrum. Localized shimming is required to obtain localized 31P NMR spectra of excellent spectral resolution. This is done by monitoring the 1H NMR signal from water by a single-shot localization technique. The techniques discussed can be applied to obtain spectra of brain, liver, heart, and other organs. 31P NMR spectra of intracranial tumors demonstrate its applicability in the examination of patients.     

Fast (13)C-glucose metabolite mapping in rat brain using (1)H echo planar spectroscopic imaging technique at 2T

For fast (13)C metabolite mapping in rat brains, (1)H-detected (13)C NMR spectroscopy using gradient-enhanced heteronuclear multiple-quantum coherence and (1)H echo planar spectroscopic imaging were combined. (13)C glucose and 3-/4-(13)C-Glu/Gln images of rat brain were successfully constructed with 35-minute temporal resolution under a 2T magnetic field. In the ischemic region of the suture middle cerebral artery occlusion model, glucose and Glu/Gln signals decreased and lactate signals appeared. J. Magn. Reson. Imaging 2001;13:787-791.     

Investigating brain metabolism at high fields using localized 13C NMR spectroscopy without 1H decoupling.

Most in vivo 13C NMR spectroscopy studies in the brain have been performed using 1H decoupling during acquisition. Decoupling imposes significant constraints on the experimental setup (particularly for human studies at high magnetic field) in order to stay within safety limits for power deposition. We show here that incorporation of the 13C label from 13C-labeled glucose into brain amino acids can be monitored accurately using localized 13C NMR spectroscopy without the application of 1H decoupling. Using LCModel quantification with prior knowledge of one-bond and multiple-bond J(CH) coupling constants, the uncertainty on metabolites concentrations was only 35% to 91% higher (depending on the carbon resonance of interest) in undecoupled spectra compared to decoupled spectra in the rat brain at 9.4 Tesla. Although less sensitive, 13C NMR without decoupling dramatically reduces experimental constraints on coil setup and pulse sequence design required to keep power deposition within safety guidelines. This opens the prospect of safely measuring 13C NMR spectra in humans at varied brain locations (not only the occipital lobe) and at very high magnetic fields above 4 Tesla.     

31P NMR spectroscopy of the in vivo metabolism of an intracerebral glioma in the rat

The in vivo high-energy phosphorus metabolic profile and pH of an experimental intracerebral C6 glioma in rats was examined using surface coil 31P NMR spectroscopy. Initially, phosphorus-containing metabolites of the glioma were characterized by in vivo 31P surface coil spectroscopy of subcutaneously implanted tumors and by high-resolution NMR studies of perchloric acid (PCA) extracts of both freeze-clamped subcutaneous tumor tissue and cultured cells. These studies demonstrated that the C6 glioma has reduced levels of phosphocreatine (PCr) compared to the levels found in normal rat brain. Thus, reduced spectral PCr levels were useful as a metabolic indicator for monitoring the spatial selectivity of tumor metabolism distinct from that of adjacent normal brain tissue. To maximize 31P NMR signals from intracerebral tumors, tumor cells were stereotaxically placed superficially in the brain. Proton magnetic resonance imaging (1H MRI) was used to determine the size and location of the resultant brain tumors in order to preselect rats with large superficial tumors for spectroscopic study. 31P NMR spectra of the glioma tumors revealed a consistent reduction in the PCr/ATP ratio, an increase in the Pi/ATP ratio, and a slightly increased tissue pH. No correlation was found between levels of Pi/ATP and tumor pH in subcutaneous or intracerebral gliomas and the amount of necrosis as determined histologically. This study demonstrates that phosphorus metabolites of an experimental brain tumor in the rat can be monitored in vivo with minimal contributions from adjacent normal brain tissue metabolites using surface coil 31P NMR spectroscopy.