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Biomedical subjects

P Bendel

Publications and source records attributed to P Bendel.

At least 19 recordsLinked to original sources

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.

Biophysical Phenomena↗

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.

Animals↗

Developmental changes and water status in tulip bulbs during storage: visualization by NMR imaging.

Magnetic Resonance Imaging (MRI) and light and scanning electron microscopy (SEM) were used to follow time-dependent morphological changes and changes in water status of tulip bulbs (Tulipa gesneriana L., cv. 'Apeldoorn') during bulb storage for 12 weeks at 20 degrees C (non-chilled) or 4 degrees C (chilled) and after planting. MR images reflecting the water content, the relaxation times T1 and T2 (or their reciprocal values, the relaxation rates R1 and R2), and the apparent self-diffusion coefficient of water molecules (ADC), were obtained for intact bulbs. After planting, scape elongation and flowering occurred only in chilled bulbs, while elongation in non-chilled bulbs was retarded. Microscopic observations showed different structural components and high heterogeneity of the bulb tissues. MRI revealed the elongation of the flower bud during storage, which was significantly faster in the chilled bulbs. In addition, MRI demonstrated a redistribution of water between different bulb organs, as well as significant differences in the pattern of this redistribution between the chilled and non-chilled bulbs. Generally, R2 relaxation rates became faster in all bulb organs during storage. At the same time, ADC values remained constant in the chilled bulbs, while exhibiting a significant increase in the non-chilled bulbs.

Magnetic Resonance Imaging↗

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.

Animals↗

Line narrowing of I = 12 spins coupled to quadrupolar nuclei in liquids: effects of weak decoupling fields

In this paper an exact description of the observed transverse magnetization of spin 12 nuclei, coupled to quadrupolar spins which are subjected to RF irradiation, is presented. It is shown that for on-resonance CW decoupling at weak to intermediate irradiation levels, the transverse decay of the spin 12 magnetization is modulated with a period of 1/nu(2), where nu(2) is the amplitude of the decoupling irradiation. When the spin 12 signal is created as a spin echo, and the quadrupolar resonance continuously irradiated during the echo evolution, the echo amplitudes experience much stronger modulation with period 2/nu(2). In previous treatments of such spin systems, the regime of weak decoupling power was usually neglected, and approximate analytical expressions seeking to define the "adequate" or "minimal" decoupling power, necessary to achieve the collapse of the spin 12 multiplet into a single narrow line, were derived. It is demonstrated here, both by experiments and by simulations using a full Redfield formalism, that simple analytical predictions for the T(2) decay of the spin 12 magnetization are still possible, even when the scalar relaxation is not in the "fast exchange" limit and the transverse decay is considerably modulated due to insufficient decoupling power. In this case, the expected single exponential decay rate is obtained for the nonmodulated component of the signal. The theoretical solution for spin I = 12 coupled to S = 3 is derived, and results for the proton decay in (10)B-enriched sodium borocaptate in aqueous solution are presented. The effects of irradiation by several composite pulse decoupling sequences are also considered. Copyright 1999 Academic Press.

Journal Article↗

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.

Animals↗

Boron-11 NMR of borocaptate: relaxation and in vivo detection in melanoma-bearing mice.

Longitudinal and transverse relaxation rates for the 11B resonances in sodium borocaptate (BSH) at varying concentrations were measured in undiluted horse serum in a 4.7 Tesla field. The results could be fit by a model that assumes fast exchange of the BSH molecule between a free and a bound state, using values of 0.77+/-0.7 MHz for the 11B quadrupole coupling constant and (6.3+/-0.9) x 10(-9) s for the rotational correlation time in the bound state. These results were used as a basis for assessing the requirements and limitations of quantitative determination of BSH concentrations in vivo, using 11B NMR. Surface coil 11B NMR spectroscopy was performed on a total of 14 mice injected with BSH. Some of the animals (n=9) had implanted M2R melanoma tumors grown to various sizes in the rear thigh, in which case the surface coil was placed against the tumor, whereas for the other animals (without tumor), the coil was placed against the rear thigh muscle. NMR spectra were acquired under fully relaxed conditions. The spectra were quantitated by peak integration; apparent absolute BSH concentrations were derived by comparison with spectra from a phantom with known BSH concentration, using extrapolation of the time-domain data to zero preacquisition delay. The results indicate significantly higher 11B BSH signal intensities in tumors, compared with muscle tissue, whereas the uptake and clearance kinetics were similar.

Algorithms↗

Choline metabolism in breast cancer; 2H-, 13C- and 31P-NMR studies of cells and tumors.

Choline metabolism in breast cancer cells and tumors has been investigated by multinuclear NMR in order to provide the biochemical basis for the presence of high phosphocholine in breast carcinoma relative to benign breast tumors and normal breast tissue. Choline was found to be transported into MCF7 human breast cancer cells and rapidly phosphorylated to phosphocholine which was then accumulated in the cells to high concentrations. The increased level of phosphocholine did not affect the rate of synthesis of phosphatidylcholine, indicating tight regulation of this pathway. The incorporation of [1,2-13C]choline (100 microM) into phosphocholine and phosphatidylcholine after 24 h was 69.5 and 36% of the total respective pools. Incorporation of 2H9-choline to tumors implanted in nude mice was achieved by infusing the deuterated choline to the blood circulation. The metabolism of deuterated choline was then monitored by 2H localized MRS. The blood level of choline before the infusion was 58.6 +/- 10.3 microM (measured by 1H-NMR of plasma samples) and increased approximately 5-fold during the infusion (measured by 2H-NMR). This increase in the blood level resulted in a gradual increase of a signal at 3.2 ppm due to deuterated choline metabolites. It appears that the increased availability of choline in the blood circulation leads to accumulation of phosphocholine in the tumors by the same mechanism as in the cells.

Animals↗

Quantitative diffusion imaging in implanted human breast tumors.

Quantitative diffusion measurements were performed in tumors arising from inoculation of nude mice with two human breast cancer cell lines (MCF7 and T47D) to evaluate the specificity of this technique for characterizing solid tumors. ADC maps were compared to histology and correlated well with gross tumor morphology. Measured ADCs were highly specific for viable and necrotic tumor in the five T47D tumors included in this study (P < 0.02), while only two of the five MCF7 tumors studied developed distinguishable areas of necrosis. No statistically significant difference was observed in ADCs from viable tumor between the different cell lines (P > 0.05).

Animals↗

Quadrupolar relaxation of spin 3 in the intermediate omega0tauc regime

The equations for the quadrupolar relaxation of spin 3 were derived for the Redfield limit where the molecular reorientation rate is much faster than the size of the quadrupolar interaction. In the extreme narrowing regime (omega0tauc << 1), the results converge to the analytical expressions for the relaxation rates available in the literature. For slower motions, both longitudinal (spin-lattice) and transverse (spin-spin) relaxations are described by a superposition of three exponentials, where both the rates themselves and their relative weights are functions of omega0tauc. Numerical calculations of the relevant relaxation parameters in the intermediate omega0tauc regime are presented. Spin-lattice relaxation is described to very good approximation by a single exponential for all values of omega0tauc, with the weight of the dominant decay mode exceeding 0. 97 for the entire range. The predictions of these simulations were found to be in good agreement with experimentally measured relaxation rates of the 10B resonances in the sodium salt of Na2B12H12S, mercaptoundecahydro-closo-dodecaborane (sodium borocaptate or BSH) dissolved in glycerol, determined at omega0 = 53. 73 MHz, between temperatures of 268 and 323 K. The fit to the experimental results yielded a value of 1.25 MHz for the average 10B quadrupolar coupling constant in this molecule. Copyright 1997 Academic Press. Copyright 1997Academic Press

Journal Article↗

The role of boron MRI in boron neutron capture therapy.

Boron magnetic resonance imaging (MRI) and spectroscopy (MRS) are being developed for use in boron neutron capture therapy (BNCT). With continued development, boron MRI may be used to locate BNCT agents in vivo, evaluate the pharmacokinetics of BNCT agents, and aid in treatment planning.

Animals↗

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.

Animals↗

In vivo detection of a boron-neutron-capture agent in melanoma by proton observed 1H-10B double resonance.

The Cesium salt of BSSB (Cs4B24H22S2), a common boron-neutron-capture-therapy (BNCT) agent, was injected into M2R mouse melanoma xenografts, and detected in vivo by 1H-observed, 10B-edited NMR spectroscopy. The technique of spin-echo difference spectroscopy, in which a proton spin-echo is detected following the alternating presence and absence of a 10B 180 degrees pulse was used. This method provides much higher sensitivity than direct 10B NMR detection, and should thus be suitable for in vivo detection in patients about to undergo BNCT treatment, where the infused agents are 95% isotopically enriched in 10B.

Animals↗

Snapshot MRI with T2*-weighted magnetization preparation.

A method for encoding T2*-dependent contrasts into the preparation period of magnetization-prepared ultrafast MRI sequences is demonstrated and validated. The preparation block consists of a 90 degrees---tau---90 degrees p sequence, where p is shifted by 90 degrees for successive images. By adding two such successive images in quadrature, a combined image is obtained, in which the pixel intensities depend on the incoherent intravoxel dephasing that occurred during tau in a way that is similar to the dependence of TE in a gradient-echo sequence. The method could therefore be useful for applications that require very short repetition times for good temporal resolution, together with detection of T2*-weighted contrasts, such as functional MRI in the brain.

Brain↗

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.

Animals↗

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.

Animals↗

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.

Animals↗

In vivo studies by magnetic resonance imaging and spectroscopy of the response to tamoxifen of MCF7 human breast cancer implanted in nude mice.

Magnetic resonance 1H-imaging and 31P-localized spectroscopy were utilized to monitor, noninvasively, MCF7 human breast cancer tumors implanted in immunodeficient mice. The tumors were followed during estrogen dependent growth and tamoxifen induced remission. Early after tamoxifen administration enhanced necrosis developed, extending to most of the tumor volume. This was followed by growth of repair tissue along with tumor regression. The short-term tamoxifen treatment also modified the content of the phosphate metabolites, increasing the nucleoside triphosphate to inorganic phosphate ratio from 0.41 +/- 0.15 (n = 14) before treatment to 1.10 +/- 0.70 (n = 8, P less than 0.025) and to 1.75 +/- 0.66 (n = 9, P less than 0.0002) 4-7 days and 9-19 days, respectively, after treatment. This change was attributed to the growth of reparative tissue. The results provide new information regarding the response and the mechanism of action of tamoxifen.

Animals↗