Deuterium NMR in the solid-state and in solution of the molecular motion of the bases in poly(I) and poly(I) . poly(C).

To provide information regarding the conformational flexibility of nucleic acids, in particular the rate and amplitude of base motions, we have observed the deuterium NMR from single-stranded and double-stranded polynucleotides. Poly(I) was deuterated at the 8-position of the base, and the deuterium NMR was examined in solution (at 23.0 and 55.4 MHz) and for hydrated and dry fibers (at 23.0 MHz). In the solid state, the deuterium signal of dry poly(I) exhibits a powder pattern with the maximal expected quadrupolar splitting, while the relatively short spin-lattice relaxation time indicates the presence of a rapid internal reorientation of the C-D bond with an amplitude of that motion of at least +/- 2.4 degrees. Hydrating the poly(I) fibers to the extent of eight molecules of water per nucleotide results in the disappearance of the deuterium signal, apparently due to a decreased spin-spin relaxation time shorter than the instrumental dead-time (even using the quadrupolar echo technique); this could occur if conformational fluctuations are occurring at a rate comparable to the deuterium quadrupole interaction strength, i.e., 175 kHz. In solution, a theoretical fit to the measured Lorentzian linewidths and spin-lattice relaxation times necessitates the inclusion of at least two motional correlation times, with a subnanosecond internal motion. Double-stranded poly(I) . poly(C) yielded a solid state spectrum similar to poly(I), albeit with a longer T1, which reduced the lower limit for the amplitude of an internal motion to +/- 1.9 degrees. The 2H signal from the poly(I) . poly(C), hydrated to a degree of approx. eight molecules of water per base pair, retained its solid-state lineshape (with a reduced T1 value, indicating increased internal mobility of the bases with a lower limit on amplitude of +/- 4.7 degrees). In solution, however, the 2H-NMR signal from poly(I) . poly(C) became virtually undetectable, even in solid-echo experiments, when the echo was observed after 52 microseconds. This indicates that the spin-spin relaxation time of the deuterium nucleus must be close to its theoretical minimum of about 9 microseconds, and the correlation time for an isotropic reorientation of the C-D vector can be estimated to be between 0.2 and 200 microseconds.

[Study of optical absorption of sensitizers in vivo].

The subject of the paper is study of optical absorption of sensitizers in biological tissue. The study shows that absorbance can be used as a tool that allows studying biodistribution of sensitizers and their interaction with tissue in vivo. The article presents a simple technique of determining biological tissue absorption in vivo, and discusses the results of experimental animal studies of some sensitizers.

One- and two-dimensional 90.5-MHz 13C-NMR spectroscopy of the N-linked triantennary oligosaccharide units of calf fetuin.

Complete assignments of all anomeric resonances in the proton and carbon spectra of the N-linked oligosaccharide units of fetuin were made using one- and two-dimensional NMR spectroscopy. We are able to confirm the presence of microheterogeneity in the N-acetylneuraminic acid linkages to the galactose residues and the presence of a unique triantennary structure which carries a side chain: NeuAc alpha(1-3)Gal beta(1-3)GlcNac beta(1-4)-. Anomeric carbon chemical shifts changes resulting from long-range conformational effects were observed.

Significant differences in the retention of the borocaptate monomer (BSH) and dimer (BSSB) in malignant cells.

The (10)B enriched form of Na(2)B(12)H(11)SH (BSH) is used in a binary cancer radiation treatment, known as boron neutron capture therapy (BNCT). The BSH anion can also form a dimer, [B(24)H(22)S(2)](4-) (BSSB), previously used in animal model studies. It is demonstrated here that the retention of BSSB in mouse M2R melanoma and rat C6 glioma cells is significantly longer than that of the monomer BSH. The washout of the BNCT agents from cultured cells was followed using (11)B nuclear magnetic resonance spectroscopy. One hour after switching to boron-free culture medium, the relative amount of BSSB retained in M2R cells was at least about six times higher than that of BSH. The initial washout rate (over the first approximately 4 h) of BSSB was 4x10(-3) min(-1) (t(1/2), approximately 3 h) for M2R cells, and of the same order of magnitude for glioma cells.

Uptake and washout of borocaptate sodium and borono-phenylalanine in cultured melanoma cells: a multi-nuclear NMR study.

The cellular uptake and washout of the two principal boron neutron capture therapy (BNCT) agents, borocaptate sodium (BSH) and borono-phenylalanine (BPA), were monitored on-line, noninvasively, using nuclear magnetic resonance (NMR) spectroscopy. The uptake and washout of inorganic borate (B(i)) was also followed for comparison. M2R mouse melanoma cells grown on polystyrene microspheres were perfused inside the NMR sample tube. (11)B NMR was used to detect the presence of B(i), BSH and BPA, and (19)F NMR was applied to detect fluorinated BPA ((19)F-BPA). The results revealed chemical modifications of BSH due to spontaneous formation of the borocaptate dimer, BSSB, in the culture medium. BPA readily formed a complex with glucose contained in the culture medium but was also converted in the cells to a yet unidentified compound in a reaction that probably involves the hydrolysis of BPA and the release of B(i). The cellular accumulation ratio for BPA was significantly higher than 1 and was also significantly higher than that for BSH. On the other hand, the cellular retention time observed for BSH was much longer than for BPA, indicating a strong trapping of BSH in cells.

Tamoxifen induced changes in MCF7 human breast cancer: in vitro and in vivo studies using nuclear magnetic resonance spectroscopy and imaging.

The effects of 17 beta-estradiol versus tamoxifen on the growth and metabolism of MCF7 human breast cancer cells, in culture and in tumors implanted in nude mice, were studied by 31P and 13C nuclear magnetic resonance spectroscopy and by proton magnetic resonance imaging. In culture, the content of the phosphate metabolites including nucleoside triphosphates (NTP), phosphomonoesters, phosphodiesters and inorganic phosphate (Pi) were not affected by tamoxifen treatment. However, in the presence of estrogen the rate of glucose consumption and lactate production via glycolysis (270 and 280 fmol/cell.h, respectively) were twice that of tamoxifen treated cells. Estrogen rescue of tamoxifen treated cells indicated that glycolysis induction occurs at the early stages of the hormonal response. The in vivo studies included recording of proton images that provided an accurate measure of tumor size and distribution of tumor cells, necrotic regions and stromal tissue. Tamoxifen caused enhanced necrosis extending from the center of the tumor during the first two days of treatment (12 h to 6 days). This was followed by growth of reparative tissue along with tumor regression. Tamoxifen also modified the content of the phosphate metabolites, increasing markedly (P less than 0.0002) the ratio of NTP to Pi from 0.41 before treatment to 1.75 9-19 days after treatment. This change was attributed to the enhanced growth of repair tissue. The results provide new information regarding the response of human breast cancer to hormonal treatment and suggest a mechanism for the induction of tumor regression by tamoxifen.

Quantitation of ventricular size in normal and spontaneously hypertensive rats by magnetic resonance imaging.

Magnetic resonance imaging (MRI) performed at high field (4.7 Tesla), and high spatial resolution (0.6 mm slice thickness, 0.18 mm inplane) enabled noninvasive quantitative measurement of the ventricular vol. in live rats. Comparing the results for 15 male Wistar-Kyoto (WKY) rats, aged 2.5-10 months, with those from 17 spontaneously hypertensive rats (SHR), clearly confirmed the previously reported elevated ventricular vols. in the SHR strain. A significant difference in ventricular vol. between the two strains was detected above the age of 3 months. For mature animals above the age of 6 months the mean vol. in the SHR strain was elevated by about a factor of two compared to the WKY control animals.

Altered brain glucose metabolism in transgenic-PFKL mice with elevated L-phosphofructokinase: in vivo NMR studies.

The gene for the liver-type subunit of phosphofructokinase (PFKL) resides on chromosome 21 and is overexpressed in Down syndrome (DS) patients. Transgenic PFKL (Tg-PFKL) mice with elevated levels of PFKL were used to determine whether, as in DS, overexpression of PFKL was also associated with altered sugar metabolism. We found that Tg-PFKL mice had an abnormal glucose metabolism with reduced clearance rate from blood and enhanced metabolic rate in brain. Transgenic-PFKL mice exhibited elevated activity of phosphofructokinase in both blood and brain, as compared to control non-transgenic (ntg) mice. Following glucose infusion, the rate of glucose clearance from the blood of Tg-PFKL mice was significantly slower than that of control ntg mice, although the basal blood glucose levels were similar. However, unlike the slower rate of glucose metabolism in blood, the initial rate of glucose utilization in the brain of the transgenic mice, was 58% faster than in control ntg mice. This was determined by infusion of [1-13C]-glucose followed by in vivo nuclear magnetic resonance (NMR) measurements of brain glucose metabolism. The faster utilization of glucose in Tg-PFKL brain is similar to the increased rate of cerebral glucose metabolism found in the brain of young adult DS patients, which may play a role in the etiology of their cognitive disabilities.

The ontogeny of a neurotoxic lesion in rat brain revealed by combined MRI and histology.

We have used magnetic resonance imaging (MRI) of the living rat brain to longitudinally analyze the ontogenesis of an ibotenic acid lesion targeted at the piriform cortex. The MRI data were systematically compared with data obtained from a battery of histopathological techniques, including Nissl stain, hematoxylin stain, and a stain for cytochrome oxidase activity. Two days after the lesioning, widespread and heterogeneous damage was detected in, around and distant from the toxin-targeted area. Some damage apparently diminished within approximately 10 days, whereas other damage remained throughout the length of this study (60 days). We found that the small-animal MRI technology used by us is useful in determining the initial, transient impact of surgery and neurotoxic lesioning, and in delineating the gross effects of the lesion over time. This is particularly useful for early elimination of animals from the protocol of physiological and behavioral experiments in which the lesion exceeds the target area. Our data also indicate that, in order to avoid confounding effects of transient post-lesioning phenomena, behavioral and physiological tests should be carried out in neurotoxically lesioned animals > 2 weeks after infliction of the lesion.

The effects of simultaneous pulsing in different gradient coils on the nuclear magnetic resonance imaging of oblique slices.

The definition and mapping of oblique planes by magnetic resonance imaging (MRI) requires the simultaneous application of two or three orthogonal gradients to define the desired intermediate direction of the frequency encoding or "readout" gradient. Each of the three main gradient coils produces different patterns of eddy currents. Consequently, the application of dephasing and rephasing lobes of these gradients will produce echoes at slightly different times for each gradient. If two or three gradients are applied simultaneously to create an arbitrary view direction, the resulting echo will therefore be shifted in time and considerably reduced in intensity. In this article, we present an analysis of the behavior of the magnetization in a typical two-dimensional Fourier transform pulse sequence for the imaging of oblique slices. The theoretical displacements in time and reduction in intensity of the echo amplitudes are calculated and compared to the experimental behavior. We show that, in spite of this phenomenon, the final image suffers only marginally in signal-to-noise ratio, provided the slice width is small compared to the field of view. This is due to the fact that there always exists a cycle in the sequence in which the phase-encoding gradient almost completely compensates for the above described effect.

Optimal detection of the neutron capture therapy agent borocaptate sodium (BSH): a comparison between 1H and 10B NMR.

Boron Neutron Capture Therapy (BNCT), an experimental binary cancer treatment modality, requires selective targeting of 10B containing compounds to tumors. One of the compounds under evaluation in an EORTC phase I trial, and used in Japan for patient treatments for many years, is borocaptate sodium (BSH, also known as sulfhydril boron hydride). To optimize the clinical applications, a noninvasive method is needed to monitor the distribution of the boron compound, and NMR may offer such a possibility. A comparison between the relative sensitivities for detecting BSH by 10B or 1H NMR was conducted at two magnetic field strengths: 2 and 4.7 T. At each field strength, similar-sized radio frequency (rf) coils were used for both nuclei. Theoretical predictions for the intrinsic signal to noise (S/N) advantage of 1H over 10B detection vary between a factor of 5.4 and a factor of 28.9, depending on whether the effective resistance is dominated by coil losses or sample losses. Our tests, conducted on relatively small aqueous samples, which loaded the coils less than expected for animal or human subjects, resulted nevertheless in advantage factors close to the lower limit of this range. The measured S/N detection advantage factors for 1H were about 5.2 at 4.7 T, using a dedicated 1H coil, and 7.7 at 2 T, where the measurements were conducted with a double-tuned coil. However, when predicting the expected performance for in vivo MRS or MRI, one should bear in mind that proton detection has to be conducted by spectral-editing pulse sequences with an inherent S/N loss by at least a factor of 2, and that the T1 relaxation time for 10B in BSH is about 30 times shorter than the 1HT1 value. In view of these considerations, direct 10B detection could well be the preferred strategy for MRI/MRS of BSH in vivo.

Antivascular treatment of solid melanoma tumors with bacteriochlorophyll-serine-based photodynamic therapy.

We describe here a strategy for photodynamic eradication of solid melanoma tumors that is based on photo-induced vascular destruction. The suggested protocol relies on synchronizing illumination with maximal circulating drug concentration in the tumor vasculature attained within the first minute after administrating the sensitizer. This differs from conventional photodynamic therapy (PDT) of tumors where illumination coincides with a maximal concentration differential of sensitizer in favor of the tumor, relative to the normal surrounding tissue. This time window is often achieved after a delay (3-48 h) following sensitizer administration. We used a novel photosensitizer, bacteriochlorophyll-serine (Bchl-Ser), which is water soluble, highly toxic upon illumination in the near-infrared (lambda max 765-780 nm) and clears from the circulation in less than 24 h. Nude CD1 mice bearing malignant M2R melanotic melanoma xenografts (76-212 mm3) received a single complete treatment session. Massive vascular damage was already apparent 1 h after treatment. Changes in vascular permeability were observed in vivo using contrast-enhanced magnetic resonance imaging (MRI), with the contrast reagent Gd-DTPA, by shortening spin-spin relaxation time because of hemorrhage formation and by determination of vascular macromolecular leakage. Twenty-four hours after treatment a complete arrest of vascular perfusion was observed by Gd-DTPA-enhanced MRI. Histopathology performed at the same time confirmed primary vascular damage with occlusive thrombi, hemorrhage and tumor necrosis. The success rate of cure of over 80% with Bchl-Ser indicates the benefits of the short and effective treatment protocol. Combining the sensitizer administration and illumination steps into one treatment session (30 min) suggests a clear advantage for future PDT of solid tumors.

Echo projection imaging-a method to obtain NMR images undistorted by magnetic field inhomogeneities.

It is proposed to use a modified form of Mansfield's echo planar imaging to obtain NMR images which are entirely undistorted by background inhomogeneities of the magnetic field. In the proposed method, a train of 180 degrees pulses is applied in the presence of a periodically switched or sinusoidally modulated linear field gradient. The time-domain signal is sampled at half the distance between the 180 degrees pulses. At these points the magnetization will be modulated by the gradient, but will be independent of any mechanisms of inhomogeneous broadening, such as static field inhomogeneities, local susceptibility effects, or chemical shifts. A Fourier transform of the function comprising these points will therefore yield a faithful projection of the spin density, although the magnitude of the superimposed gradient need not be large compared to the inhomogeneous broadenings. This paper demonstrates the application of the proposed pulse sequence to a small-scale one-dimensional phantom. The major problem in upscaling this technique to human-scale dimensions lies in the limited available and allowed RF power, which in turn limits the maximal tolerable field inhomogeneities as well as the maximal practical field strength. An analysis of the tolerance of the proposed technique to these factors is presented, based on numerical simulation of its performance, using the Bloch equations. It is concluded that its use may be feasible on low-field systems, providing the advantages of increased signal-to-noise, lower required gradient strength, and drastically reduced sensitivity to the homogeneity and stability of the magnetic field, at the expense of larger RF power.

Magnetization transfer and double-quantum filtered imaging as probes for motional restricted water in tulip bulbs.

Parameter sensitive MRI experiments were performed on tulip bulbs before and after storage at two different temperatures, 4 degrees C (chilled), and 20 degrees C (non-chilled). Quantitative measurements of the amount of magnetization transfer (MT) in the storage scales of the bulbs, were compared to the average values of the relaxation rates R(1) and R(2), and the apparent normalized spin density (NSD). At the end of the storage period, bulbs were also scanned using 1H double quantum (DQ) filtered imaging. Both MT and DQ filtered imaging revealed significant differences between chilled and non-chilled bulbs, which were consistent with the differences observed in the average values of NSD, R(1,) and R(2.) The results indicated a smaller fraction of solid protons (e.g., starch, sugars, and possibly bound water), or less contact between these solid protons and (free) water in the storage scales of the chilled bulbs, after 8 weeks of storage at low temperature.

In vivo boron-11 MRI and MRS using (B24H22S2)4- in the rat.

In vivo boron-11 magnetic resonance imaging (MRI) and magnetic resonance spectroscopy (MRS) were performed on a rat that had been infused with a potential boron neutron capture therapy agent, Na4B24H22S2, using methods for detecting nuclei with a short T2 relaxation time. MRI and MRS were also performed on a euthanized rat that had been similarly infused in vivo. Boron-11 spectral intensities decreased in the living rat over a 25-h period. The results demonstrate the capability of MRI and MRS to noninvasively monitor the distribution and excretion of boron agents in vivo.

Observation of electro-osmotic flow echoes in porous media by nuclear magnetic resonance.

A method for assessing the time reversibility of molecular displacements in fluids is presented. The method utilizes pulsed field gradient NMR experiments, in which the flow driving force is inverted during the magnetization lifetime in each measurement cycle. The method is suitable for opaque three-dimensional systems and short displacements, and provides inherent separation between thermal diffusion and displacements driven by externally controlled forces. This approach was applied to study the time reversibility of an electric-field-driven flow of water in natural sand samples, over time scales of up to 0.4 s and displacement scales of the order of one particle diameter. It is demonstrated that the intensity loss of the NMR signal, caused by flow-induced phase dispersion, is fully refocused upon inversion of the polarity of the applied electric field, resulting in flow echoes.

Atrophic enlargement of CSF volume after subarachnoid hemorrhage: correlation with neuropsychological outcome.

BACKGROUND AND PURPOSE: Ventricular dilation and sulcal enlargement are common sequelae after aSAH. Our aim was to quantify the late ventricular dilation and volumes of the CSF spaces after aSAH and to determine if they correlate with neurologic and cognitive impairments frequently detected in these patients. MATERIALS AND METHODS: 3D T1-weighted images needed for volumetry were available in 76 patients 1 year after aSAH, along with 75 neuropsychological assessments. Volumes of CSF segments and ICV were quantified by SPM in 76 patients and 30 control subjects to determine CSF/ICV ratios. The mCMI was calculated to roughly evaluate the ventricular dilation. The contributing factors for enlarged ventricles and CSF volumes were reviewed from radiologic, clinical, and neuropsychological perspectives. RESULTS: The mCMI was higher in patients with aSAH (0.23 +/- 0.06) compared with control subjects (0.20 +/- 0.04; P = .020). In line with these planimetric measurements, the SPM-based CSF/ICV ratios were higher in patients with aSAH (35.58 +/- 7.0) than in control subjects (30.36 +/- 6.25; P = .001). Preoperative hydrocephalus, higher HH and Fisher grades, and focal parenchymal lesions on brain MR imaging, but not the treatment technique, were associated with ventricular enlargement. The clinical outcome and presence of neuropsychological deficits correlated significantly with CSF enlargement. CONCLUSIONS: Ventricular and sulcal enlargement, together with reduced GM volumes, after aSAH may indicate general atrophy rather than hydrocephalus. Enlarged CSF spaces correlate with cognitive deficits after aSAH. A simple measure, mCMI proved to be a feasible tool to assess the diffuse atrophic brain damage after aSAH.

Motor cortex and thalamic atrophy in Unverricht-Lundborg disease: voxel-based morphometric study.

OBJECTIVE: To evaluate possible changes in the gray matter volume of patients with Unverricht-Lundborg disease (EPM1) compared with healthy controls. METHODS: Thirty-four genetically verified patients with EPM1 and 30 healthy controls matched for age and sex underwent MRI (T1-, T2-, fluid-attenuated inversion recovery-, and T1-weighted 3-dimensional images). T1-weighted 3-dimensional images were analyzed with voxel-based morphometry (VBM) to compare the regional differences in gray matter volumes between patients and controls. The patients with EPM1 were also clinically evaluated for myoclonus severity using the Unified Myoclonus Rating Scale. RESULTS: VBM analysis revealed atrophy in the bilateral primary, premotor, and supplementary motor cortex. The thalamus and precuneus were also bilaterally affected. No infratentorial changes were detected in the group analysis. CONCLUSION: The cortical motor areas of the brain are particularly affected in EPM1, correlating with the motor symptoms of this disease. The combination of detailed imaging with neurophysiologic evaluation may help to reveal the pathogenesis of Unverricht-Lundborg disease.

Study of manganese bacteriopheophorbide as a potential contrast agent for magnetic resonance tomography.

The use of manganese bacteriopheophorbide characterized by a high relaxation capacity and selectively accumulating in the tumor as a contrast agent for magnetic resonance tomography significantly improves tumor contrasting against the background of normal tissues. The pharmacokinetics and selectivity of accumulation were studied by diffuse reflection spectroscopy.

Subarachnoid hemorrhage is followed by temporomesial volume loss: MRI volumetric study.

OBJECTIVE: To assess whether subarachnoid hemorrhage (SAH) and its treatment is followed by volume loss in temporomesial structures. METHODS: One hundred fifty-five consecutive patients with aneurysmal SAH were randomly assigned to surgical or endovascular treatment. Volumetric MRI was performed in 77 SAH patients with good or moderate clinical outcome 1 year after hemorrhage. A comprehensive neuropsychological test battery was used to evaluate the cognitive performance of the subjects. Thirty healthy individuals were imaged as MRI controls. RESULTS: The normalized hippocampal (HC) volumes were 24.7/23.7 (right/left), and the amygdaloid (AM) volumes were 21.0/20.5 in the matched control population. In SAH patients, the corresponding volumes were smaller, HC 23.2/21.3 (p = 0.072/0.002) and AM 18.4/18.7 (p = 0.012/0.045). In addition, the AM ipsilateral to the ruptured aneurysm was smaller in patients who had undergone surgical treatment (15.7) vs endovascular treatment (20.3; p < 0.001). Treatment modality did not significantly affect the measured HC volumes. The hippocampal but not amygdaloid volumes correlated with the scores of several neuropsychological tests. CONCLUSION: Subarachnoid hemorrhage and its treatment may be followed by atrophy in temporomesial structures. A clear correlation was demonstrated between neuropsychological performance and reduced temporomesial volumes.