Post-mortem assessment of rat spinal cord injury and white matter sparing using inversion recovery-supported proton density magnetic resonance imaging.

STUDY DESIGN: This was an experimental study. OBJECTIVES: White matter sparing influences locomotor recovery after traumatic spinal cord injury (SCI). The objective of the present post-mortem magnetic resonance imaging (MRI) investigation was to assess the potential of a simple inversion recovery (IR) sequence in combination with high-resolution proton density (PD) images to selectively depict spared white matter after experimental SCI in the rat. SETTING: This study was conducted at University of Liège and Centre Hospitalier Universitaire, Liège, Belgium and Hasselt University, Diepenbeek, Belgium. METHODS: Post-mortem 9.4 tesla (T) MRI was obtained from five excised rat spines 2 months after compressive SCI. The locomotor recovery had been followed weekly using the standardized Basso-Beattie-Bresnahan scale. IR MRI was used to depict normal white matter as very hypo-intense. Preserved white matter, cord atrophy and lesion volume were assessed, and histology was used to confirm MRI data. RESULTS: MRI showed lesion severity and white matter sparing in accordance with the degree of locomotor recovery. IR MRI enhanced detection of spared and injured white matter by selectively altering the signal of spared white matter. Even subtle white matter changes could be detected, increasing diagnostic accuracy as compared to PD alone. MRI accuracy was confirmed by histology. CONCLUSION: High-resolution IR-supported PD MRI provides useful micro-anatomical information about white matter damage and sparing in the post-mortem assessment of chronic rat SCI.

Detailed visualization of the functional regions of the rat pituitary gland by high-resolution T2-weighted MRI.

This high-resolution MRI study focuses on the visualization of the detailed morphology of the rat's pituitary gland by means of post-mortem as well as in vivo MRI at 9.4 T. Determination of the local T1- and T2-relaxation decay times allows to explain the regional image intensities which reflects the degree of tissue organization at the molecular level. Detailed characterization of the molecular level of the pituitary gland, as provided by the relaxation decay times, can offer a rigid platform with respect to functional or pathological explorations. It is demonstrated that T1-weighted imaging, as is routinely used in the clinic, can differentiate between the posterior and anterior lobe but not between the posterior and intermediate lobe. T2-weighted images, however, clearly show the three distinct lobes of the rat pituitary gland without the use of contrast agents, i.e. the posterior, the intermediate and the anterior lobe. Histological analysis of the rat's pituitary gland confirms the morphological structures seen on the MR images. Although the intermediate lobe is less defined in humans and can neither be differentiated by T1-weighted MRI, its clinical visualization might be possible in T2-weighted images.

Microanatomical and histological assessment of the content of superior genial spinal foramen and its bony canal.

OBJECTIVES: To assess histologically the true content of the superior genial spinal foramen and to match these findings to the microanatomical canal content observed using high-resolution magnetic resonance images of the same region. METHODS: Ten human mandibular specimens were imaged using a 9.4 Tesla MRI unit. Afterwards, eight specimens were decalcified, embedded in paraffin and serially sectioned (7 microm) for histological examination; the remaining two were embedded in methylmethacrylate and sectioned (50-60 microm). All sections were examined using routine light microscopy to inspect the superior genial spinal foramen region and its content. RESULTS: Histological observations of the superior genial spinal foramen confirmed the presence of a well-defined neurovascular bundle, with branches of the lingual nerve and lingual artery. The canal had an average diameter exceeding 1 mm and could thus be considered significant. CONCLUSIONS: The present study confirms the existence of a true superior genial spinal foramen's bony canal with neural and blood vessel content. These findings imply that surgical procedures should consider a proper preoperative assessment of the neurovascular trajectory of the superior genial spinal foramen.

The Arnold-Chiari type II malformation at midgestation.

The Arnold-Chiari malformation type II (ACMII) is reported to be reversible after closure of a myelomeningocele at midgestation. To elucidate the developmental state of the ACMII malformation at the approximate time fetal surgery is performed, the ACMII of a 20-week human fetus was investigated in vitro using high-field magnetic resonance microscopy at 9.4 T and compared with the hindbrain of a neurologically intact fetus of the same gestational age. Up to 20 weeks of gestation, the developmental failures caused by the early embryonic herniation of the posterior fossa contents are the dominant feature of fetal ACMII, but after 20 weeks, the accelerated and disproportionate growth of the cerebellum dominates. As midgestational surgery stops the leakage of cerebrospinal fluid, the posterior fossa will expand in time to allow further normal growth of both the cerebellum and brain stem. Some early developmental anomalies already present in the primitive rhombencephalon due to early embryonic hindbrain herniation as well as some intra-axial anomalies are probably not reversible.

Scope and limitations of a new highly selective synthesis of unsymmetrical monomers for the synthesis of precursors toward poly(arylenevinylene)s

In our laboratory a precursor route to poly(p-phenylenevinylene) derivatives is developed in which unsymmetrically substituted p-xylene derivatives, possessing a benzylic sulfinylalkyl group, are used as monomers. Because of this unsymmetry, we were forced to investigate thoroughly the synthesis of these sulfoxides, as we start from symmetric and readily accessible molecules, namely, bis(halomethyl)-p-xylene derivatives. In a former publication, a new extremely effective route for the production of these unsymmetrically substituted sulfinyl monomers was presented. This paper expands upon these previously reported results. To examine the scope and limitations of this elegant route, this new method was applied to the synthesis of various derivatives not included in the initial work.

Metabolites in feces can be important markers for the abuse of anabolic steroids in cattle.

In Belgium, to control the abuse of anabolic steroids in cattle, urine samples have been gradually replaced by feces samples, because the latter can be obtained more easily from living animals. Urine and feces samples were collected from heifers after administration of boldenone, norethandrolone or ethylestrenol. Metabolites present in feces or urine were determined by GC-MS. Large qualitative and quantitative differences in the metabolic profiles were observed. In feces, in contrast to urine, the parent compounds or their major metabolites were detectable only shortly after administration. On the other hand, metabolites resulting from the reduction of the 3-oxo group and the unsaturated carbon-carbon bonds, present on the A-ring, allow for long-term detection in feces. A-ring reduced metabolites have been identified in samples found positive for norgestrel, boldenone, methylboldenone and methyltestosterone, respectively. These results are in agreement with concomitant in vivo experiments.

In vitro liver models are important tools to monitor the abuse of anabolic steroids in cattle.

Current veterinary residue analysis mainly focuses on the monitoring of residues of the administered parent compound. However, it is possible that larger amounts of metabolites are excreted and that they can have a prolonged excretion period. In order to unravel specific metabolic steps and to identify possible biological markers, two in vitro liver models were used, i.e. monolayer cultures of isolated hepatocytes and liver microsomes, both prepared from liver tissue of cattle. Chostebol, boldenone, norethandrolone (NE) and ethylestrenol (EES) were used as model substrates. Results show that the metabolic profiles derived from in vitro experiments are predictive for the in vivo metabolic pathways of the steroids evaluated in this study. By means of this strategy, it is possible to identify 17 alpha-ethyl-5 beta-estrane-3 alpha,17 beta-diol (EED) as a common biological marker for NE and EES. By in vivo experiments it was shown that EED is particularly important for the detection of the abuse of NE or EES because of its high excretion levels and its prolonged presence as compared with the parent compounds or any other metabolite.

Identification of some important metabolites of boldenone in urine and feces of cattle by gas chromatography-mass spectrometry.

17 alpha-Boldenone (17 alpha-BOL) and/or 17 beta-boldenone (17 beta-BOL) appear occasionally in fecal matter of cattle. In addition to 17 alpha-BOL, a whole array of boldenone related substances can be found in the same samples. In vitro experiments with microsomal liver preparations and isolated hepatocytes combined with the excretion profiles found in urine and feces samples of in vivo experiments made it possible to identify several metabolites of 17 beta-BOL in 17 beta-BOL positive feces samples. In one animal treated with 17 beta-BOL, no 17 beta-BOL or its metabolites were present before treatment and most of these compounds disappeared gradually in time after the treatment was stopped. It is not clear what the origin is of 17 alpha-BOL and boldenone metabolites in samples screened routinely for the abuse of anabolic steroids and considered to be 'negative' because of the absence of 17 beta-BOL since other workers showed some evidence that 17 alpha-BOL can be of endogenous origin. However, in our hands, most of these 17 alpha-BOL positive samples, obtained during routinely performed screenings of cattle, contained large amounts of delta 4-androstene-3,17-dione (AED), which normally is absent from routinely screened negative samples. Furthermore, AED was absent in all samples obtained from the animals treated with 17 beta-BOL. We have no direct evidence that 17 alpha-BOL or 17 beta-BOL is of endogenous origin.

Deformation of the cervicomedullary junction and spinal cord in a surgically treated adult Chiari I hindbrain hernia associated with syringomyelia: a magnetic resonance microscopic and neuropathological study. Case report.

The lower brainstem and cervical spinal cord from an ordinarily treated case of Chiari Type I hindbrain hernia associated with syringomyelia was examined using high-resolution magnetic resonance microscopy and standard neuropathological techniques. Magnetic resonance microscopy allows total screening and visualizes the disturbed internal and external microanatomy in the three orthogonal planes with the resolution of low-power optical microscopy. An additional advantage is the in situ visualization of the shunts. Afterwards the intact specimen is still available for microscopic examination. Part of the deformation of the medulla is caused by chronic tonsillar compression and molding inside the foramen magnum. Other anomalies, such as atrophy caused by demyelination, elongation, and unusual disturbances at the level of the trigeminal and solitary nuclear complexes contribute to the deformation. At the level of the syrinx-free upper part of the cervical cord, anomalies of the dorsal root and the dorsal horn are demonstrated.

High field magnetic resonance imaging of normal and pathologic human medulla oblongata.

High field proton magnetic resonance (MR) imaging has been applied to depict the MR appearance of the normal excised human cervicomedullary junction, based on which neuropathologic specimens can be described. More specifically, two normal cases and one case of Chiari deformity were imaged in the transverse, sagittal, and coronal dimensions using a 9.4 Tesla vertical bore magnet. The MR images of the normal specimens reveal most of the neuroanatomical microstructures described in literature. An accurate description of the Chiari deformity could be made by comparing the MR reference images with those of the pathologic specimen. All MR detected abnormalities were confirmed by histopathology, by which no additional lesions could be found.

Microanatomy of the excised human spinal cord and the cervicomedullary junction examined with high-resolution MR imaging at 9.4 Tesla.

PURPOSE: To study in detail the MR anatomy of the spinal cord and the cervicomedullary junction that could serve as a reference for clinical MR studies. METHODS: Specimens of fresh human spinal cord and formalin-fixed cervicomedullary transition zones were imaged with a 9.4-T vertical bore magnet. Using a multisection spin-warp pulse sequence the parameters were selected to produce essentially proton density images. RESULTS: The images obtained depict the microanatomical organization of the spinal cord and cervicomedullary junction. In the spinal cord, the central gray has the expected higher signal intensity compared with the white matter, which is, apart from its darker general appearance, characterized by the presence of a dense radially structured neuroglial framework of high signal intensity. Anatomically more complex regions such as the dorsal root entry zone, the adjacent posterior horn complex, and the crossing fibers of the cervicomedullary junction are seen as well as parts of the microvascular system. CONCLUSION: Although cellular details are still beyond the limits of this investigation, the images at 9.4 T show the spinal cord and cervicomedullary junction with detail comparable to low-power microscopic images of fixed sections, especially with respect to distinguishing gray and white matter, nuclei, tracts, and angioarchitecture.

Investigation of protein structure by means of 19F-NMR. A study of hen egg-white lysozyme.

A 19F-labeled derivative of hen egg-white lysozyme, in which the six epsilon-amino groups are trifluoroacetylated (LF6), was prepared by reaction of lysozyme with S-ethyltrifluorothioacetate. The reaction mixture was fractionated by cation-exchange chromatography at pH 7.3. A comparison of the circular dichroic spectra and the activity towards Micrococcus lysodeikticus of both LF6 and native lysozyme reveals that the labeling causes no major conformational changes of the polypeptide backbone. Assignment of the six resonances present in the 19F-NMR spectrum of LF6 was accomplished by using a variety of techniques: specific chemical modifications, the effect of the inhibitor (GlcNAc)3, 19F-shift/pH information and relaxation parameters.