Stefan Bluml

Magnetic resonance spectroscopy of the human brain.

Magnetic resonance (MR; synonymous with NMR = nuclear magnetic resonance) is a universal physical technique best known for non‐invasive detection and anatomical mapping of water protons (H). MR‐spectroscopy (MRS) records protons from tissue chemicals other than water, intrinsic phosphorus containing metabolites, sodium, potassium, carbon, nitrogen, and fluorine. MRS is therefore an imaging technique with the potential to record human and animal biochemistry in vivo. As a result of wide availability of MRI equipment in research laboratories and hospitals, MRS is a serious competitor with PET to define normal body composition and its perturbation by pharmacological and pathological events. This article describes practical aspects of in vivo MRS with particular emphasis on the brain, where novel metabolites have been described. A survey of these new aspects of neurochemistry emphasize their practical utility as neuronal and axonal markers, measures of energy status, membrane constituents, and osmolytes, as well as some xenobiotics, such as alcohol. The concept of multinuclear in vivo MRS is illustrated by diagnosis and therapeutic monitoring of several human brain disorders. Although these methods are currently most frequently encountered in human studies, as well as with transgenic and knockout mouse models, MRS adds a new dimension to anatomic and histopathologic descriptions. Anat Rec (New Anat) 265:54–84, 2001.

Expression of Heme Oxygenase-1 in Human Vascular Cells Is Regulated by Peroxisome Proliferator-Activated Receptors

Objective—Activation of peroxisome proliferator-activated receptors (PPARs) by lipid-lowering fibrates and insulin-sensitizing thiazolidinediones inhibits vascular inflammation, atherosclerosis, and restenosis. Here we investigate if the vasculoprotective and anti-inflammatory enzyme heme oxygenase-1 (HO-1) is regulated by PPAR ligands in vascular cells. Methods and Results—We show that treatment of human vascular endothelial and smooth muscle cells with PPAR ligands leads to expression of HO-1. Analysis of the human HO-1 promoter in transient transfection experiments together with mutational analysis and gel shift assays revealed a direct transcriptional regulation of HO-1 by PPAR&agr; and PPAR&ggr; via 2 PPAR responsive elements. We demonstrate that a clinically relevant polymorphism within the HO-1 promoter critically influences its transcriptional activation by both PPAR isoforms. Moreover, inhibition of HO-1 enzymatic activity reversed PPAR ligand-mediated inhibition of cell proliferation and expression of cyclooxygenase-2 in vascular smooth muscle cells. Conclusion—We demonstrate that HO-1 expression is transcriptionally regulated by PPAR&agr; and PPAR&ggr;, indicating a mechanism of anti-inflammatory and antiproliferative action of PPAR ligands via upregulation of HO-1. Identification of HO-1 as a target gene for PPARs provides new strategies for therapy of cardiovascular diseases and a rationale for the use of PPAR ligands in the treatment of other chronic inflammatory diseases.

Three-Point Technique of Fat Quantification of Muscle Tissue as a Marker of Disease Progression in Duchenne Muscular Dystrophy: Preliminary Study

OBJECTIVE Clinical trials involving patients with Duchenne muscular dystrophy are hindered by the lack of suitable objective end points. The purpose of this study was to examine whether muscle lipid infiltration measured with the three-point Dixon MRI technique has value as a marker of disease severity. SUBJECTS AND METHODS Disease severity in nine boys (mean age, 8.6 +/- 2.7 years) with Duchenne muscular dystrophy was determined with the functional ability scale of Brooke and associates. Functional scores were compared with strength measurements obtained by manual testing of muscles of the lower extremities, knee extensor strength measured with an isokinetic dynamometer, and muscle fat percentage in the quadriceps and hamstrings determined with the three-point Dixon MRI technique. RESULTS MRI measurements of fat infiltration had stronger correlation (p < 0.05) with functional grade than did measurements obtained with manual muscle testing (p = 0.07) or quantitative strength measured with the isokinetic dynamometer (p = 0.54). Muscle fat percentage did not correlate with strength measurements from manual or dynamometer muscle testing but increased with age in subjects with Duchenne muscular dystrophy. CONCLUSION Muscle adiposity values obtained with three-point Dixon MRI are accurate in assessment of disease severity in patients with Duchenne muscular dystrophy. Because they are not influenced by patient effort or examiner variability, these measurements are more objective and reproducible than measurements of muscle strength.

Direct determination of the N‐acetyl‐l‐aspartate synthesis rate in the human brain by 13C MRS and [1‐13C]glucose infusion

A non‐invasive 13C magnetic resonance spectroscopy (MRS) technique is described for the determination of the N‐acetyl‐l‐aspartate (NAA) synthesis rate, VNAA, in the human brain in vivo. In controls, the mean VNAA was 9.2 ± 3.9 nmol/min/g. In Canavan disease, where [NAA] is increased (p < 0.001) and [aspartate] is deceased (p < 0.001), VNAA was significantly reduced to 3.6 ± 0.1 nmol/min/g (p < 0.001). These rates are in close agreement with the activity of the biosynthetic enzyme measured in vitro in animals, and with the rate of urinary excretion of NAA in human subjects with Canavan disease. The present result is consistent with the regulation of NAA synthesis by the activity of a single enzyme, l‐aspartate‐N‐acetyltransferase, in vivo, and with its control in Canavan disease by limited substrate supply and/or product inhibition. The 13C MRS technique provides the means for further determination of abnormal rates of neuronal NAA synthesis among neurological disorders in which low cerebral [NAA] has been identified.

Quantitative proton-decoupled 31P MRS and 1H MRS in the evaluation of Huntington's and Parkinson's diseases

Objective: To determine cerebral energy status in patients with Huntingtons disease(HD) and Parkinsons disease (PD). Methods: The study included 15 patients with DNA-proven, symptomatic HD and five patients with medically treated, idiopathic PD, all of whom were candidates for neurotransplant treatment, as well as 20 age-related normal subjects. Quantitative noninvasive, MRI-guided proton MRS was performed of single volumes in putamen of basal ganglia (BG), occipital gray matter, and posterior parietal white matter; in addition, quantitative phosphorus and proton-decoupled phosphorus MRS of superior biparietal white and gray matter was done. Outcome measures were quantitative metabolite ratios and millimolar concentrations of neuronal and glial markers, creatine (Cr) and adenosine triphosphate (ATP), and intracellular pH. Results: In volume-corrected control BG (10.46 ± 0.37 mM), [Cr] was 29%(p < 0.05) higher than in control gray matter (8.10 ± 1.04 mM). In HD and PD, energy metabolism was not abnormal in the four cerebral locations measured by MRS. No increase in cerebral lactate or decrease in phosphocreatine and ATP was detected. Small, systematic abnormalities in N-acetylaspartate (NAA, ddecreased), Cr (decreased), choline-containing compounds (Cho, increased), and myoinositol (mI, increased) were demonstrable in all patients individually and in summed spectra but were insufficient to make diagnosis possible in the individual patient. Conclusion: Previously described failure of global energy metabolism in HD was not confirmed. However, quantitative 1-hydrogen MRS and decoupled 31-phosphorus MRS are sensitive to ±10% alterations in key cerebral metabolites, and may be of value in noninvasive monitoring of appropriate therapies.

Developmental changes in choline‐ and ethanolamine‐containing compounds measured with proton‐decoupled 31P MRS in in vivo human brain

Cerebral phosphorylated metabolites, possibly involved in membrane and myelin sheath metabolism, were measured and quantified using proton‐decoupled 31P ({1H}‐31P) MRS in 32 children and 28 adults. Age‐dependent changes were determined for phosphorylethanolamine (PE), phosphorylcholine (PC), glycerophosphorylethanolamine (GPE), glycerophosphorylcholine (GPC), and phosphocreatine (PCr) concentrations. In the neonate, PE dominates the spectrum and decreases with age along with PC, whereas GPE, GPC, and PCr increase in concentration with postnatal age. PE (1.23 ± 0.13 mM) and GPE (0.57 ± 0.08 mM) co‐resonate with choline in 1H MRS. Together with PC (0.57 ± 0.12 mM) and GPC (0.94 ± 0.13 mM) these four metabolites accounted for all of the visible 1H MRS choline in normal adult brain. Children with diseases that affect myelination were found to have abnormal {1H}‐31P MRS. The new quantitative assay may provide novel insights in determining and monitoring normal and abnormal brain maturation noninvasively. Magn Reson Med 42:643–654, 1999.

Oxidized phospholipids negatively regulate dendritic cell maturation induced by TLRs and CD40.

Maturation of dendritic cells (DCs) induced by pathogen-derived signals via TLRs is a crucial step in the initiation of an adaptive immune response and therefore has to be well controlled. In this study, we demonstrate that oxidized phospholipids (ox-PLs), which are generated during infections, apoptosis, and tissue damage, interfere with DC activation, preventing their maturation. ox-PLs blocked TLR-3- and TLR-4-mediated induction of the costimulatory molecules CD40, CD80, CD83, and CD86, the cytokines IL-12 and TNF, as well as lymphocyte stimulatory capacity. CD40 and TLR-2-mediated cytokine production was also inhibited, whereas up-regulation of costimulatory molecules via these receptors was not affected by ox-PLs. Thus, formation of ox-PLs during the course of an inflammatory response may represent a negative-feedback loop preventing excessive and sustained immune reactions through regulating DC maturation.

Visualization of Cerebrospinal Fluid Movement with Spin Labeling at MR Imaging: Preliminary Results in Normal and Pathophysiologic Conditions

Institutional review board approval and informed consent were obtained for this study. This study was HIPAA compliant. The purpose of this study was to visualize the movement of cerebrospinal fluid (CSF) noninvasively by using an unenhanced magnetic resonance imaging technique. A time-spatial labeling inversion pulse (SLIP) technique was applied to label, or tag, CSF in a region of interest. The tagged CSF was clearly visualized at inversion times of 1500-4500 msec after pulse labeling in both intracranial and intraspinal compartments. Noninvasive visualization of CSF movement, including bulk and turbulent flow, in normal (n = 7) and altered (n = 2) physiologic conditions was possible by using the unenhanced time-SLIP technique.

Human rhinoviruses inhibit the accessory function of dendritic cells by inducing sialoadhesin and B7-H1 expression

Dendritic cells (DC) are professional APCs with an unmatched ability to interact with and activate T cells. There is accumulating evidence that DC not only efficiently stimulate T cell activation but also regulate T cell responses. However, little is known about cell surface structures on DC involved in the regulation of T cell responses. We demonstrate that human rhinoviruses (HRV) can efficiently inhibit the accessory function of DC through induction of inhibitory cell surface receptors. We observed that treatment of DC with HRV14 (R-DC), a member of the major group HRV family, diminished their T cell stimulatory capacity and induced a promiscuous and deep anergic state in cocultured T cells despite high levels of MHC molecules as well as costimulatory molecules, e.g., B7-1 (CD80) and B7-2 (CD86), and independent of inhibitory soluble factors such as IL-10. In contrast, expression of inhibitory B7-H1 molecules was up-regulated and R-DC de novo expressed sialoadhesin (Sn). Most importantly, blocking of B7-H1 and Sn on R-DC with specific mAbs against both receptors reverted the inhibitory phenotype. Thus, inhibitory signals delivered from R-DC to T cells via B7-H1 and Sn were critical for the induction of anergy. These observations suggest that an altered accessory molecule repertoire on DC upon interaction with HRV down-modulates adaptive immune responses during the viral infection.

Magnetic resonance spectroscopy in pediatric neuroradiology: clinical and research applications

Magnetic resonance spectroscopy (MRS) offers a unique, noninvasive approach to assess pediatric neurological abnormalities at microscopic levels by quantifying cellular metabolites. The most widely available MRS method, proton (1H; hydrogen) spectroscopy, is FDA approved for general use and can be ordered by clinicians for pediatric neuroimaging studies if indicated. There are a multitude of both acquisition and post-processing methods that can be used in the implementation of MR spectroscopy. MRS in pediatric neuroimaging is challenging to interpret because of dramatic normal developmental changes that occur in metabolites, particularly in the first year of life. Still, MRS has been proven to provide additional clinically relevant information for several pediatric neurological disease processes such as brain tumors, infectious processes, white matter disorders, and neonatal injury. MRS can also be used as a powerful quantitative research tool. In this article, specific research applications using MRS will be demonstrated in relation to neonatal brain injury and pediatric brain tumor imaging.