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Magnetic cell separation using antibody binding with protein a expressed on bacterial magnetic particles.

Bacterial magnetic particles (BacMPs) are efficient platforms of proteins for surface display systems. In this study, mononuclear cells from peripheral blood were separated using BacMPs expressing protein A on the BacMP membrane surface (protein A-BacMPs), which were complexed with the Fc fragment of anti-mouse IgG antibody. The procedure of positive selection involves incubation of mononuclear cells and mouse monoclonal antibodies against different cell surface antigens (CD8, CD14, CD19, CD20) prior to treatment with protein A-BacMP binding with rabbit anti-mouse IgG secondary antibodies. Flow cytometric analysis showed that approximately 97.5 +/- 1.7% of CD19(+) and CD20(+) cells were involved in the positive fraction after magnetic separation. The ratio of the negative cells in the negative fraction was approximately 97.6 +/-1.4%. This indicates that CD19(+) and CD20(+) cells can be efficiently separated from mononuclear cells. Stem cell marker (CD34) positive cells were also separated using protein A-BacMP binding with antibody. May-Grunwald Giemsa stain showed a high nuclear/cytoplasm ratio, which indicates a typical staining pattern of stem cells. The separated cells had the capability of colony formation as hematopoietic stem cells. Furthermore, the inhibitory effect of magnetic cell separation on CD14(+) cells was evaluated by measurement of cytokine in the culture supernatant by ELISA when the cells were cultured with or without lipopolysaccharide (LPS). The induction of IL1-beta, TNFalpha, and IL6 was observed in the presence of 1 ng/mL LPS in all fractions. On the other hand, in the absence of LPS, BacMPs had little immunopotentiation to CD14(+) cells as well as that of artificial magnetic particles, although TNFalpha and IL6 were slightly induced in the absence of LPS in the positive fraction.

Binding Sites, Antibody↗

Chiral magnetic metal-organic frameworks of dimetal subunits: magnetism tuning by mixed-metal compositions of the solid solutions.

The isostructural, chiral molecular magnetic materials with the formula [MxM'(2-x)(ca)2(1,4-dimb)]n [H2ca = D-(+)-camphoric acid, 1,4-dimb = 1,4-di-(1-imidazolyl-methyl)-benzene, M = Ni(II), M' = CoII, 0 < or = x < or = 2] consist of ca-bridged (4,4) layers with [M2(O2CR)4] as secondary building units that are pillared by the 1,4-dimb ligands into a unique 3D framework. The high-spin octahedral symmetry and the proportions of the mixed-metal ions were characterized by UV-vis spectroscopy. The compounds exhibit the onset of antiferromagnetic ordering at 7.5 approximately 23 K, as well as weak ferromagnetism, spin-flop, and glassy behavior that result from the randomness of the mixed-metal pairs, magnetic anisotropy of the metallic cations, and antisymmetric exchange. The composites should be regarded as molecular alloys of the pure Ni(II) and Co(II) compounds. The magnetic behavior of the solid solutions shows unambiguously that the organic bridges, bond angles, and bond distances greatly influence the effective interactions and bring about cooperative magnetic behavior in the chiral 3D frameworks.

Cobalt↗

DNA-based magnetic nanoparticle assembly acts as a magnetic relaxation nanoswitch allowing screening of DNA-cleaving agents.

Monodisperse magnetic nanoparticles conjugated with complementary oligonucleotide sequences self-assemble into stable magnetic nanoassemblies resulting in a decrease of the spin-spin relaxation times (T2) of neighboring water protons. When these nanoassemblies are treated with a DNA cleaving agent, the nanoparticles become dispersed, switching the T2 of the solution back to original values. These qualities render the developed nanoparticles and their nanoassemblies as magnetic relaxation switches capable of screening for DNA-cleaving compounds by magnetic resonance methods such as MRI and NMR.

Biosensing Techniques↗

Selective loss of progenitor subsets following clinical CD34+ cell enrichment by magnetic field, magnetic beads or chromatography separation.

In this preclinical evaluation we have compared the efficacy of three clinical CD34+enrichment procedures with respect to purity, yield and recovery, as well as risk of selective loss of CD34+ lineage-specific subsets. The three devices work by different principles and have several different manipulation steps: The magnetic field separator uses paramagnetic iron-dextran particles; the magnetic microbead selection is based on the advantage of a large surface area for immobilisation of the monoclonal antibody within a very small volume; the original immunoabsorption technique is based on the use of biotinylated antibody applied to a column of avidin-coated sephadex beads. The results of this evaluation gave a median purity 96% (88-98%), 86% (62-97%), and 49% (18-85%), and median yield of 65% (54-100%), 40% (21-74%), and 30% (8-55%), respectively. Subset analysis recognised a selective loss of CD34+/61+ after enrichment, most likely due to class I-II antibodies used for the enrichment step or, alternatively, nonspecific binding of megakaryocytic progenitors. Tumour cell spiking experiments on a clinical scale documented an expected 2-4 log reduction resulting in a number of potentially malignant cells in the CD34 enriched product. Our data support four major conclusions: First, that magnetic field separation is superior to magnetic beads and chromatography selection, mainly due to the risk of cell loss and insufficient recovery with the two latter methods. Second, that late differentiated progenitors with CD34 class III epitopes present are lost during the enrichment procedures. The third major conclusion is that chromatography selection results in a selective loss of CD34bright cells, which are most likely uncommitted early progenitors. This was an unexpected finding which may be a consequence of an imbalance between the strong forces between biotin-avidin and insufficient physical manipulation for CD34+ cell release. Finally, the data document that CD34 selection alone is an inappropriate way to eliminate tumour cells due to the uncontrolled variables and the inconsistent outcome. The only products which can be expected to be purged free of tumour cells are the ones with very minimal (<10-5) contamination in the starting products, ie products documented tumour free with the most sensitive techniques for quantitation. If this is not the case, the optimal purging strategy may be a two-step procedure including CD34 selection and subsequent depletion of the tumour cells in question.

Antigens, CD34↗

Interventional catheter magnetic resonance angiography with a conventional 1.5-T magnet: work in progress.

Magnetic resonance contrast enhancement depends on the relative timing of image acquisition. Limited human trials have demonstrated efficacy of intra-arterial gadolinium diethylenetriaminepentaacetic acid (Gd-DTPA) in delineating vascular anatomy with X-rays. The present study assessed the ability of dynamic MR during intra-arterial Gd-DTPA administration to demonstrate vascular anatomy compared to conventional angiography as the gold standard. As interventional MR techniques using dedicated magnets proliferate, the ability to perform invasive MR angiography with a conventional magnet would be of great utility at established sites. Four subjects referred for different types of angiography underwent dynamic MR studies, including one with iliac artery stenting (Palmaz P204, Johnson and Johnson). All were examined with conventional angiography, and again after dynamic intra-arterial (IA) Gd-DTPA infusion. Coronal MRI images of the body were acquired using a 1.5-T superconducting magnet (three with a GE Signa, one with Philips NT), fast spoiled gradient echo (FSPGR); echo time (TE) = 4.2 msec, repetition time (TR) = 68-150 msec, flip = 75 degrees, 0-600 s after dilute Gd-DTPA IA bolus injection during sequential breath-hold acquisitions of 13-32 s each. All arteries were detected with dynamic MR. The FSPGR MRI with IA Gd-DTPA administration can provide adequate time and spatial resolution to demonstrate arterial anatomy and arterial stent patency.

Carotid Artery Diseases↗

Magnetic resonance imaging and diseases of the liver and biliary tract. Part 2. Magnetic resonance cholangiography and angiography and conclusions.

Magnetic resonance cholangiography (MRC) relies on the strong T2 signal from stationary liquids, in this case bile, to generate images. No contrast agents are required, and the failure rate and risk of serious complications is lower than with endoscopic retrograde cholangiopancreatography (ERCP). Data from MRC can be summated to produce an image much like the cholangiogram obtained by using ERCP. In addition, MRC and conventional MRI can provide information about the biliary and other anatomy above and below a biliary obstruction. This provides information for therapeutic intervention that is probably most useful for hilar and intrahepatic biliary obstruction. Magnetic resonance cholangiography appears to be similar to ERCP with respect to sensitivity and specificity in detecting lesions causing biliary obstruction, and in the diagnosis of choledocholithiasis. It is also suited to the assessment of biliary anatomy (including the assessment of surgical bile-duct injuries) and intrahepatic biliary pathology. However, ERCP can be therapeutic as well as diagnostic, and MRC should be limited to situations where intervention is unlikely, where intrahepatic or hilar pathology is suspected, to delineate the biliary anatomy prior to other interventions, or after failed or inadequate ERCP. Magnetic resonance angiography (MRA) relies on the properties of flowing liquids to generate images. It is particularly suited to assessment of the hepatic vasculature and appears as good as conventional angiography. It has been shown to be useful in delineating vascular anatomy prior to liver transplantation or insertion of a transjugular intrahepatic portasystemic shunt. Magnetic resonance angiography may also be useful in predicting subsequent variceal haemorrhage in patients with oesophageal varices.

Biliary Tract Diseases↗

Proton magnetic resonance and magnetic susceptibility characterization of ferredoxin I from Bacillus polymyxa.

Magnetic susceptibility and proton magnetic resonance spectra are reported for the oxidized and reduced forms of the iron-sulfur protein Bacillus polymyxa ferredoxin I. The magnetic susceptibility of the oxidized form indicates antiferromagnetic exchange coupling between component iron atoms that is quantitatively similar to that observed for the clostridial ferredoxins and for the [(C(2)H(5))(4)N](2) [Fe(4)S(4)(SCH(2)C(6)H(5))(4)] analog. Contact-shifted resonances observed in the proton-magnetic-resonance spectra of oxidized and reduced forms of the B. polymyxa protein can be correlated with the contact-shifted resonances of corresponding redox forms of the clostridial ferredoxins. Characteristics of the contact-shifted resonances observed in partially reduced B. polymyxa ferredoxin I are compatible with a "slow" rate of electron exchange between redox forms, which suggests that the "fast" electron exchange earlier observed in the eight-iron clostridial ferredoxins may derive from an intramolecular component.

Bacillus↗

Effective magnetic labeling of transplanted cells with HVJ-E for magnetic resonance imaging.

Magnetic labeling of transplanted cells permits us to monitor their localization non-invasively using MRI. Since most transfection agents for magnetic labeling have the same cationic charge as Fe(3+), the efficiency may be reduced. The hemagglutinating virus-envelope has no charge and utilizes membrane fusion activity to deliver internalized materials. In this study, we investigated the feasibility of using the envelope to incorporate paramagnetic Fe(3+) particles into PC12 cells and astrocytes. The envelope effectively labeled both cells with Fe(3+), which showed significant decreases of signal intensity in T2-weighted MRI. Labeled cells transplanted into the rat striatum were clearly visualized by T2*-weighted MRI at a magnetic field of 2 T. The results indicate that the hemagglutinating virus-envelope is a powerful tool for magnetic labeling.

Animals↗

Experimental hepatic tumor necrosis. Comparison of spin-echo and pulsed magnetization transfer contrast magnetic resonance imaging.

RATIONALE AND OBJECTIVES: We compared the effectiveness of pulsed magnetization transfer contrast (MTC) magnetic resonance imaging (MRI) and spin-echo MRI in detecting tumor necrosis. METHODS: Adenocarcinoma cells were transplanted in the livers of 12 syngenic BDIX rats. To induce various degrees of tumor necrosis, the rats were randomly assigned to the following groups: 1) control; 2) localized hyperthermia; 3) intralesional cisplatin; and 4) hyperthermia plus intralesional cisplatin. At day 7 after treatment, the rats were imaged using a 1.5-T imager with 1) multiplanar gradient-recalled echo sequence (MPGR) 500/8/20 degrees with and without magnetization transfer contrast (MTC); 2) spin-echo 2500/20,80, and 3) spin-echo 300/20 pulse sequences. The rats were then sacrificed and pathologic specimens were prepared using MR images as guidance. T2 and ratios of signal intensity after saturation to signal intensity before saturation (Ms/Mo ratios) of the necrotic and granulation tissues and viable tumors were determined in 10 rats. RESULTS: Compared with standard MPGR images, MPGR images with MTC provided better contrast between the pathologic tissues and normal liver. However, T2 values were more useful than Ms/Mo ratios in distinguishing necrotic areas from viable tumor. The T2 values of coagulative necrosis and granulation tissue were significantly different from that of viable tumor. No significant difference between the Ms/Mo ratios of the different pathologic tissues and normal liver was found. CONCLUSION: Pulsed magnetization transfer contrast MRI was inferior to spin-echo MRI in distinguishing necrotic from viable tumors in rat livers using the pulse sequences described, and none of the sequences studied was thought to be reliable enough for this purpose.

Adenocarcinoma↗

Measurement of lumbar spine flexion-extension using a low-field open-magnet magnetic resonance scanner.

RATIONALE AND OBJECTIVES: The authors investigated the feasibility of using a low-field open-magnet magnetic resonance (MR) scanner to acquire functional flexion-extension images for range of motion (ROM) measurements on the lumbar spine. METHODS: Seventeen healthy subjects with no symptoms of back pain (age range, 22-59 years) were scanned in a low-field open-magnet MR scanner in the flexed, neutral, and extended positions. Each image was downloaded to a computer workstation for subsequent flexion-extension, lordosis, and ROM measurement. RESULTS: Data from two subjects were not analyzed because their images did not show all the lumbar vertebrae. For the remaining 15, there was a large variation in the magnitude of the ROM values (range, 9 degrees-70 degrees; mean 36.4 degrees, SD 16.5 degrees). However, there was a significant correlation between age and ROM (r = -0.63; P < 0.05). CONCLUSIONS: The low-field open-magnet MR scanner provides a method for noninvasive imaging of the lumbar spine, allowing the subject freedom of movement in the horizontal plane. This enables functional flexion-extension images of the lumbar spine to be acquired.

Adult↗

Magnetic properties of middle ear and stapes implants in a 9.4-T magnetic resonance field.

HYPOTHESIS: A 9.4-T magnetic resonance (MR) field may cause motion displacement of the middle ear and stapes implants not previously observed with 1.5- and 3.0-T magnets. BACKGROUND: Publications have described the safety limitations of some otologic implants in 4.7-T field and resulted in several companywide patient safety-related recalls. To date, no studies have been reported for otologic implants in a 9.4-T MR field nor have comparisons been made with 4.7-T field strengths. METHODS: Twenty-three commonly used middle ear and stapes prostheses were selected and exposed to 9.4-T MR fields in vitro within petri dishes, and eight of the 23 implants were further studied ex corpus in human temporal bones (TBs) in a 9.4-T MR field. This study has been approved by the institutional review board. RESULTS: Eight prostheses in petri dishes grossly displaced at 9.4 T, three of which had not previously moved in either the 1.5- or 3.0-T magnets. The eight TB preparations showed no avulsions or motion indicators after exposure at 9.4 T. CONCLUSION: Middle ear and stapes implants can move dramatically in petri dishes at 9.4-T MR field, more so than at 1.5 and 3.0 T. The absence of avulsions in the TB group strongly suggests that the surgical means used to fixate the middle ear implants to the middle ear structures successfully overcomes the magnetic moment produced at MR field strengths up to 9.4 T. The use of MR imaging is not contraindicated by this study's findings.

Cadaver↗

Automatic magnetic resonance tissue characterization for three-dimensional magnetic resonance imaging of the brain.

Computer-assisted diagnostic systems enhance the information available from magnetic resonance imaging. Segmentations are the basis on which three-dimensional volume renderings are made. The application of a raw data-based, operator-independent (automatic), magnetic resonance segmentation technique for tissue differentiation is demonstrated. Segmentation images of vasogenic edema with gross and histopathological correlation are presented for demonstration of the technique. A pixel was classified into a tissue class based on a feature vector using unsupervised fuzzy clustering techniques as the pattern recognition method. Correlation of fuzzy segmentations and gross and histopathology were successfully performed. Based on the results of neuropathological correlation, the application of fuzzy magnetic resonance image segmentation to a patient with a brain tumor and extensive edema represents a viable technique for automatically displaying clinically important tissue differentiation. With this pattern recognition technique, it is possible to generate automatic segmentation images that display diagnostically relevant neuroanatomical and neuropathological tissue contrast information from raw magnetic resonance data for use in three-dimensional volume reconstructions.

Adult↗

In vivo heating of magnetic nanoparticles in alternating magnetic field.

We have evaluated heating capabilities of new magnetic nanoparticles. In in vitro experiments they were exposed to an alternating magnetic field with frequency 3.5 MHz and induction 1.5 mT produced in three turn pancake coil. In in vivo experiments rats with injected magnetic nanoparticles were also exposed to an ac field. An optimal increase of temperature of the tumor to 44 degrees C was achieved after 10 minutes of exposure. Obtained results showed that magnetic nanoparticles may be easily heated in vitro as well as in vivo, and may be therefore useful for hyperthermic therapy of cancer.

Electromagnetic Fields↗

The magnetic field dependence of water proton T1 in aqueous solutions: implications for magnetic imaging contrast media.

The magnetic field dependence of the water proton nuclear magnetic relaxation induced by a nitroxide radical both free and covalently bound to serum albumin has been measured from 0.23 mT to 0.7 T. The field dependence of the water proton 1/T1 shows only a minor dependence on the degree of radical immobilization, though the electron spin resonance (ESR) spectrum is very sensitive to the correlation time of the radical. The results and theory are described in more detail elsewhere (Journal of Chemical Physics, in press). The observed relaxation behavior arises because the long electron relaxation time of the nitroxide forces the correlation time for the magnetic interaction driving the water proton relaxation to become the correlation time for the relative translational motion of the water and the nitroxide. The relative insensitivity of the water proton relaxation to nitroxide immobilization should not change if the nitroxide is used as a contrast reagent for magnetic imaging in vivo.

Cyclic N-Oxides↗

Dynamic MR imaging of the pelvic floor performed with patient sitting in an open-magnet unit versus with patient supine in a closed-magnet unit.

PURPOSE: To compare open-magnet magnetic resonance (MR) imaging performed with the patient sitting with dynamic closed-magnet MR imaging of the pelvic floor performed with the patient supine. MATERIALS AND METHODS: Thirty-eight patients underwent dynamic 1.5-T closed-magnet pelvic floor MR imaging while in the supine position. Midsagittal T2-weighted single-shot fast spin-echo and T1-weighted multiphase spoiled gradient-recalled-echo (SPGR) MR images were obtained before and after rectal contrast agent administration, respectively, with the patient at rest, straining, and maximally contracting the sphincter. Subsequently, the patient was transferred to an open 0.5-T system. Midsagittal multiphase T1-weighted SPGR MR images were then obtained every 2 seconds with the patient sitting while at rest, maximally contracting the sphincter, straining, and defecating. Images were analyzed with regard to presence of enteroceles, anterior rectoceles, intussusceptions, rectal descents, bladder descents, and vaginal vault descents. RESULTS: All intussusceptions were missed at supine MR imaging. With sitting MR imaging as the reference standard, the sensitivity of supine MR imaging was 79% for depiction of bladder descents. When MR findings were graded and clinically irrelevant MR findings were excluded, sensitivity increased to 100% for depiction of bladder descents and anterior rectoceles and to 96% for depiction of rectal descents. CONCLUSION: Dynamic supine MR imaging performed with a closed-configuration unit before and after rectal contrast agent administration appears to be an alternative to sitting MR defecography performed with an open-configuration unit for diagnosis of clinically relevant pelvic floor abnormalities.

Adult↗

Circumferential myocardial shortening in the normal human left ventricle. Assessment by magnetic resonance imaging using spatial modulation of magnetization.

BACKGROUND: Conventional cardiac imaging methods do not depict true segmental myocardial shortening, since they cannot determine segment length between fixed points in the myocardium. METHODS AND RESULTS: We used electrocardiographically gated magnetic resonance imaging with spatial modulation of magnetization to noninvasively "tag" the myocardium with dark stripes at uniform 7-mm intervals center to center at end diastole. We then determined end-systolic stripe separation and thereby calculated circumferential shortening. When end systole was not reached in the first image series, a second temporally overlapped series starting in late systole was used to determine late-systolic shortening. Septal, anterior, lateral, and inferior segments were assessed at endocardium, midwall, and epicardium on five midventricular short-axis sections each in 10 normal volunteers. A transmural gradient in circumferential shortening was observed, with the percentage of endocardial segment shortening consistently greater than epicardial segment shortening (epicardial, 22 +/- 5%; midwall, 30 +/- 6%; and endocardial, 44 +/- 6%; p less than 0.0001 by analysis of variance). Circumferential shortening varied from apex to base with slices closer to the base of the left ventricle showing less shortening at the midwall (28 +/- 9%) and endocardium (39 +/- 6%) than more apical slices at the midwall (34 +/- 13%) and endocardium (49 +/- 9%) (p less than 0.05 and p less than 0.01, respectively, by analysis of variance). CONCLUSIONS: Transmural and longitudinal heterogeneity of circumferential shortening is present in the normal human left ventricle. Magnetic resonance imaging with spatial modulation of magnetization is a powerful new tool for assessment of circumferential shortening and provides information unobtainable with conventional imaging methods.

Adult↗

Effects of surface topography on magnetization reversal of magnetic thin films.

The influence of the created surface roughness on the coercivity of magnetic thin films has been investigated. The magnetic thin films (CoFe and alternatively NiFe) are sputtered on top of smooth substrates that were previously covered with an array of considerably rougher lines with one of these materials Pt, Cu, CoFe, and NiFe. The lines have been patterned using optical lithography into arrays that are deposited with different thicknesses varying between 5 nm-15 nm. The lines have been designed to have a very rough edge and seated in two different angles relative to the wafer edge (zero and 45 degrees). Magneto-optic Kerr effect (MOKE) measurements showed two distinct switching fields in the hysteresis loops that are due to magnetic domain wall trapping created by the surface roughness. The magnetization reversal showed a strong dependence on the height, the orientation angle, and the material's type of the created surface roughness (the lines).

Cobalt↗

The prevalence of abnormal magnetic resonance imaging findings in asymptomatic knees. With correlation of magnetic resonance imaging to arthroscopic findings in symptomatic knees.

The purpose of this study was to prospectively evaluate the prevalence of abnormal magnetic resonance imaging scans of the knees of asymptomatic subjects. A prospective analysis of magnetic resonance imaging to arthroscopic findings in symptomatic knees was also performed. The prevalence of meniscal tears found in asymptomatic knees was 5.6% (medial meniscus, 1.9%; lateral meniscus, 3.7%). Other abnormal findings included a prevalence of 1.9% for degenerative changes of the medial femoral condyle and 3.7% both for ganglion cysts and patellofemoral joint articular cartilage degenerative changes. There was also a prevalence of 24.1% of Grade II signal changes of the posterior horn of the medial meniscus. Statistical comparison of our results to previous studies revealed that the magnetic resonance imaging scan readings on the asymptomatic knees in this study were accurate and lesions were correctly identified. We recommend that clinicians match clinical signs and symptoms with magnetic resonance imaging findings before instituting surgical treatment because of a 5.6% prevalence of meniscal tears in the asymptomatic population. The significance of the high percentage of posterior horn medial meniscal Grade II signal changes is unknown.

Adult↗