Absolute quantification of phospholipid metabolites in brain-tissue extracts by 1H NMR spectroscopy.
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Animals↗
Biomedical subjects
S R Williams
Publications and source records attributed to S R Williams.
Regional and developmental variations in metabolite concentration in the rat brain and eye: a study using 1H NMR spectroscopy and high performance liquid chromatography.
Regional and developmental changes in metabolite concentrations were measured by 1H NMR spectroscopy and HPLC of perchloric acid extracts from rat brain and eye. The highest concentrations of N-acetylaspartate were found in grey matter as opposed to white matter with concentration increasing with age from neonate to adult, while the related compound N-acetylaspartylglutamate was highest in adult optic nerve. Creatine and choline-containing compounds were present in all regions throughout development, with higher levels of creatine found in grey matter compared to other regions. Choline-containing compounds were present at the highest concentrations in the eye at all ages examined, and tended to decrease in concentration to minimum values in adulthood in all regions. The presence of hypotaurine in corpus callosum and optic nerve was consistent with the metabolic profiles of O-2A progenitor cells and oligodendrocytes, which are cells composing these tissues. The neurotransmitters glutamate and GABA reached their highest concentrations in the olfactory bulb (higher than in adult cortex).
Aging↗
A genome-wide search for human non-insulin-dependent (type 2) diabetes genes reveals a major susceptibility locus on chromosome 2.
Non-insulin-dependent (type 2) diabetes mellitus (NIDDM) is a common disorder of middle-aged individuals characterized by high blood glucose levels which, if untreated, can cause serious medical complications and lead to early death. Genetic factors play an important role in determining susceptibility to this disorder. However, the number of genes involved, their chromosomal location and the magnitude of their effect on NIDDM susceptibility are unknown. We have screened the human genome for susceptibility genes for NIDDM using non-and quasi-parametric linkage analysis methods in a group of Mexican American affected sib pairs. One marker, D2S125, showed significant evidence of linkage to NIDDM and appears to be a major factor affecting the development of diabetes mellitus in Mexican Americans. We propose that this locus be designated NIDDM1.
Chromosomes, Human, Pair 2↗
Proton nuclear magnetic resonance spectroscopy of primary cells derived from nervous tissue.
Cell culture techniques, high-resolution in vitro 1H nuclear magnetic resonance (NMR) spectroscopy, and chromatographic analyses were used to compare the properties of purified cell populations derived from the PNS and cortical neurones. Cell cultures were immunocytochemically characterised with specific antibodies to ensure purity of the individual cultures. Spectra of perchoric acid extracts of cultured Schwann cells, perineural fibroblasts, dorsal root ganglion neurones, and cortical neurones displayed several common features. However, statistically significant differences were found by 1H NMR spectroscopy in most metabolites among the cell types studied. In addition, cells could be distinguished by the presence or absence of certain amino acids. For example, N-acetylaspartate was present in dorsal root ganglion neurones and cortical neurones, gamma-aminobutyric acid was present in large amounts in cortical neurones, and Schwann cell spectra displayed a large signal from glycine. These results extend our earlier findings that different cell types of the CNS exhibit highly characteristic metabolite profiles to now include the major cell types of the PNS. These latter cell types also exhibit characteristic metabolite compositions, such that even Schwann cells and oligodendrocyte type 2 astrocyte (O-2A) progenitor cells-precursors of the myelinating cells of the CNS and PNS, respectively-can be readily distinguished from each other.
Animals↗
Bioenergetic recovery following ischemia in brain slices studied by 31P-NMR spectroscopy: differential age effect of depolarization mediated by endogenous nitric oxide.
Proximate neurotoxic mechanisms during postischemic recovery may be influenced by stage of development and complicating factors such as cortical spreading depression or secondary brain insult. Using 31P nuclear magnetic resonance spectroscopy, we have monitored pH and cellular energy metabolites phosphocreatine (PCr) and ATP in the ex vivo rat cerebral cortex before, during, and after substrate and oxygen deprivation, which represents "in vitro ischemia." There were important developmental differences in resistance and response to an ischemic insult. Twenty-one-day-old (P21) rat cortical slices had no detectable beta-ATP or PCr at the end of a 20-min insult, while 7-day-old (P7) slices had 50 +/- 13.7% (mean +/- SD, n = 12) and 17 +/- 14.8% relative to preischemia levels, respectively. Postischemic depolarization resulted in age-dependent effects on PCr (p < 0.05): In the older tissue, depolarization significantly worsened the recovery of PCr, whereas in young tissue it ameliorated recovery. This amelioration could be prevented by inhibiting nitric oxide production with methylene blue (depolarization-methylene blue interaction, p < 0.05) and enhanced by administration of the nitric oxide donor glyceryl trinitrate (GTN; p < 0.01). However, in P21 tissue, GTN further exacerbated injury (age-GTN interaction, p < 0.01). Therefore, in this vascular-independent preparation, a neuronal or glial nitric oxide-dependent mechanism appears to confer improved postischemic bioenergetic recovery in the developing brain compared with the mature brain.
Adenosine Triphosphate↗
The use of ondansetron in the treatment of nausea and vomiting associated with acetaminophen poisoning.
BACKGROUND: Nausea and vomiting associated with poisoning can complicate treatment and in some cases delay potential antidote administration. Side effect such as lowering the seizure threshold may at times discourage the use of traditional phenothiazine and butyrophenone antiemetics. METHODS: We performed a prospective, single arm, observational study examining the effectiveness of the 5HT3 receptor antagonist ondansetron in the management if nausea and vomiting associated with acetaminophen poisoning. Patients with a history or laboratory evidence of acetaminophen poisoning were eligible for inclusion in the study. Exclusion criteria included age less than 18 or greater than 65, use of other antiemetic therapy within the previous 12 hours, history of preexisting hepatic or hematologic disease, pregnancy, or significant ingestion of other substances. Upon meeting entry criteria, patients were administered 8 mg of intravenous ondansetron. Nausea was graded on a 100 mm scale with number of emetic episodes recorded before and after treatment. RESULTS: Six patients were entered in the study. All patients had nausea and at least one emetic episode prior to ondansetron and prior to administration of N-acetylcysteine. All patients reported relief of nausea after ondansetron. The degree of nausea decreased by an average of 52% at 30 min and 88% at 60 min following ondansetron administration. No significant vital sign changes were recorded in any patient, and there were no complications related to therapy. Three patients were administered N-acetylcysteine, and all tolerated this therapy without vomiting after ondansetron. CONCLUSIONS: Ondansetron appears to be a potentially useful adjunct in the management of nausea and vomiting associated with acetaminophen poisoning.
Acetaminophen↗
Hypermagnesemia following an acute ingestion of Epsom salt in a patient with normal renal function.
CASE REPORT: Hypermagnesemia is a rare condition that commonly follows excessive therapeutic administration of magnesium sulfate to treat eclampsia of pregnancy. Signs and symptoms of this condition include extreme muscle weakness, loss of deep tendon reflexes, mental status depression, and cardiac dysrhythmias. Clinically significant hypermagnesemia following oral or rectal administration of magnesium containing products in patients with normal renal function is rare. We report a case of hypermagnesemia following massive Epsom salt ingestion that resulted in extreme musculoskeletal weakness and altered mentation.
Central Nervous System Diseases↗
Cell type-specific fingerprinting of meningioma and meningeal cells by proton nuclear magnetic resonance spectroscopy.
We compared the properties of six human meningiomas with normal rat meningeal cells using cell culture techniques, high resolution in vitro 1H-NMR (nuclear magnetic resonance) spectroscopy, and chromatographic analysis. Cell cultures were immunocytochemically characterized at all stages with specific antibodies. Quantitative and qualitative metabolite assessments in cell extracts were obtained from 1H-NMR spectra and chromatographic analysis. Human meningioma cells expressed a characteristic spectrum of metabolites including free amino acids, compounds related to membrane phospholipid metabolism, energy metabolites, and other intermediary products. These spectral characteristics, although different in some respects, were strikingly similar to the ones of rat meningeal cells. Particularly, several metabolites that allow discrimination between meningeal cells and other cell types of the central nervous system were preserved in meningiomas. These similarities suggest that the regulation of intracellular levels of such metabolites is so intrinsic to the identity of cell type as to be conserved across species and through transformation. Additionally, human meningioma cultures expressed some spectroscopic characteristics that enabled them to be clearly distinguished from primary rat meningeal cultures. Thus, human meningiomas may be both specifically recognizable by 1H-NMR spectroscopy and also distinguishable from normal rat meningeal tissue. Our results raise the eventual possibility of using NMR in the noninvasive diagnosis of brain tumors in vivo.
Amino Acids↗
Characteristic metabolic profiles revealed by 1H NMR spectroscopy for three types of human brain and nervous system tumours.
Cell culture techniques, high-resolution in vitro 1H NMR spectroscopy, and chromatographic analyses were used to compare the properties of three types of human brain and nervous system tumours. Cell lines were immunocytochemically characterized at all stages in culture with specific antibodies. Intracellular metabolites present in cell extracts were analysed by 1H NMR spectroscopy and by high performance liquid chromatography (HPLC). The spectra from meningiomas, neuroblastomas, and glioblastomas displayed, in addition to similarities-including the presence of signals from leucine, isoleucine, valine, threonine, lactate, acetate, glutamate, choline-containing compounds and glycine-certain distinguishing metabolic features. Spectra from meningiomas featured relatively high signals from alanine. Intense signals from creatine were present in neuroblastoma spectra, while in spectra from glioblastoma they were not detectable. We found statistically significant differences by 1H NMR spectroscopy in the amounts of alanine, glutamate, creatine, phosphorylcholine and threonine among the types of tumours examined. HPLC determinations confirmed that there were also other metabolites specific to a type of tumour, such as taurine, gamma-aminobutyric acid, and serine. We suggest that these findings have potential relevance for the development of non-invasive diagnosis of tumour lineage by 1H NMR spectroscopy in vivo.
Alanine↗
Gamma-hydroxybutyrate promotes oscillatory activity of rat and cat thalamocortical neurons by a tonic GABAB, receptor-mediated hyperpolarization.
The actions of gamma-hydroxybutyrate, a drug known to lead to an increase in nocturnal slow wave sleep and induce epileptic states following systemic application, on the membrane properties of thalamocortical neurons from brain slices of the rat and cat dorsal lateral geniculate nucleus were studied using sharp electrode intracellular recordings. Gamma-hydroxybutyrate applied by addition to the perfusion medium led to a concentration-dependent and reversible hyperpolarization of the membrane potential accompanied by a decrease in apparent input resistance (0.1 mM: 2.3 +/- 0.3 mV, 9.5 +/- 1.0%; 10 mM: 11.3 +/- 1.3 mV, 37.5 +/- 10.8%, respectively). In six of seven neurons the iontophoretic or bath (0.1-0.2 mM) application of low concentrations of gamma-hydroxybutyrate led to a hyperpolarization accompanied by the appearance of low-frequency (< 4 Hz) membrane potential oscillations crowned by bursts of action potentials, when the membrane potential of these neurons was initially set depolarized to the range where ongoing oscillatory activity occurred. The gamma-hydroxybutyrate-elicited hyperpolarization was reversibly antagonized by the co-application of the GABAB receptor antagonist CGP 35348 (0.4-1 mM), but was not affected by the putative gamma-hydroxybutyrate receptor antagonist NCS 382 (0.1-5 mM) or tetrodotoxin (1 microM), suggesting that gamma-hydroxybutyrate tonically activates postsynaptic GABAB receptors. The gamma-hydroxybutyrate-induced promotion of oscillatory activity and action potential burst firing of thalamocortical neurons may be one mechanism by which gamma-hydroxybutyrate leads to an increase in the deep stages of sleep and the generation of electroencephalogram and behavioural patterns typical of absence epilepsy.
Action Potentials↗
Quality assessment in in vivo NMR spectroscopy: VI. Multicentre quantification of MRS test signals.
In this paper the results are presented of a joint European quantitative data-analysis study on a series of in vivo NMR time-domain test signals. The purpose of this study was to investigate, whether the quantitative MRS results from the various European biomedical research-centres can be made more comparable and reproducible. From the results of the study it can be concluded that in case of heavily overlapping NMR lines the quantifications should be done by some form of model function fitting and that, whenever possible, prior knowledge on the quantitative parameters should be incorporated into the quantification algorithms.
Magnetic Resonance Spectroscopy↗
Formic acid skin burns resulting in systemic toxicity.
Systemic toxicity developed in a 3-year-old girl burned by formic acid over 35% of her total body surface area. The patient presented with profound metabolic acidosis and a serum formate level of 400 micrograms/mL, the highest reported in the literature for poisoning by any route. The patient was successfully treated with hemodialysis, IV bicarbonate, and supportive measures. Reports of formic acid skin burns and systemic toxicity are limited. We discuss the pathophysiology and treatment of these cases.
Acidosis↗
Contact dermatitis associated with capsaicin: Hunan hand syndrome.
"Hunan hand" is a contact dermatitis resulting from the direct handling of chili peppers containing capsaicin. Capsaicin also is found in an over-the-counter topical agent for treatment of postherpetic neuralgia, diabetic neuropathy, and arthritis. We present the case of a patient with capsaicin-induced dermatitis and discuss the pathophysiology, therapy, and current uses of capsaicin.
Adult↗
High resolution proton NMR spectroscopy of multiple sclerosis lesions.
Tissue from postmortem multiple sclerosis and normal control brains was extracted with perchloric acid and analysed using proton NMR spectroscopy. The content of N-acetyl-derived groups (the sum of N-acetyl-aspartate, acetate, and N-acetylaspartylglutamate) was decreased in multiple sclerosis plaques compared with normal control white matter (mean, 4.36 vs. 6.64 mumol/g wet weight). In normal appearing white matter adjacent to plaques a corresponding decrease was seen, with no change in white matter distant from plaques. A decrease in the content of total creatine was observed in multiple sclerosis plaques in comparison with normal control white matter (mean, 4.64 vs. 6.56 mumol/g wet weight), which correlated strongly with the decrease in N-acetyl-derived groups. No changes in other metabolites such as total choline or myo-inositol were seen. The decreases in content of N-acetyl-derived groups are in agreement with observations from in vivo proton NMR spectroscopy in multiple sclerosis patients. The decrease in total creatine is in contrast to most of the observations made in vivo where total creatine is assumed to be unchanged and metabolite levels are often expressed as a total creatine ratio. The use of a total creatine ratio in vivo could lead to an underestimation of reductions in N-acetylaspartate and an apparent increase in other metabolites in the multiple sclerosis lesion.
Adult↗
Rapid detection of homologous recombinants in nontransformed human cells.
Gene targeting is a technique by which a preselected site in the genome of a living cell can be modified by inserting, deleting, or exchanging DNA sequences. The application of this technology to cells with a limited life-span, such as nontransformed human somatic cells, requires the development of simplified and efficient procedures to allow the isolation of correctly modified cells from the much larger pool of random integrants. The current study describes the development of a widely applicable strategy for detecting homologous recombinants in human cells by using an ELISA-based screen. When this system is used accurately targeted clones can be detected with high efficiency as soon as 14 days following transfection. Data are presented demonstrating the utility of this detection system in isolating targeted recombinants at the beta 2-microglobulin locus in both human retinal pigmented epithelial cells and human keratinocytes.
Base Sequence↗
Compartmentation of cerebral glutamate in situ as detected by 1H/13C n.m.r.
Incorporation of 13C label from either [1-13C]glucose to glutamate C-4 and lactate C-3 or from [2-13C]acetate to glutamate C-4 was monitored in situ in a superfused brain slice preparation by using 1H-detected/13C-edited (1H/13C) n.m.r. spectroscopy. The fractional enrichments of both metabolites were determined by this means in both brain slices and acid extracts of the preparations in order to assess their 1H-n.m.r. detectabilities. The 1H/13C satellite resonances from glutamate C-4 and lactate C-3 in brain tissue were followed from 4 min onwards in the presence of 5 mM [1-13C]glucose. Fractional enrichment of glutamate C-4 in the slice preparations was higher than in their acid extracts throughout the incubation of 100 min; at 30 min the enrichment was 15.9 +/- 0.6% in the slice preparations and 10.6 +/- 0.9% in extracts and at 100 min 24.5 +/- 1.7% compared with 19.7 +/- 0.4%, respectively. In contrast, lactate C-3 reached a steady-state fractional enrichment of approx. 43% by 15 min and there was no difference between the values determined in the slice preparations and the acid extracts. There was a significant difference between the glutamate C-4 fractional enrichments in the brain slices (7.4 +/- 0.6%) and extracts (5.1 +/- 0.3%) after 60 min of incubation with [2-13C]acetate. Thus 13C label from both glucose and exogenous acetate enters a pool of glutamate that is more amenable to 1H n.m.r. detection than total acid-extracted brain biochemical glutamate, whereas lactate is labelled with full 1H n.m.r. visibility. The results are discussed in the light of the biochemical factors that affect glutamate 1H-n.m.r. susceptibility and thus its n.m.r. visibility.
Animals↗
q-Space imaging of the brain.
q-Space imaging (Callaghan, J. Magn. Reson. 88, 493 (1990)) has been used to obtain mouse brain water displacement profiles. These profiles take the form of a unidirectional incoherent-displacement probability density distribution. Two groups of mice were studied, a normal group and one in which surgery had been performed to reduce the supply of blood to the forebrain. In the normal group the incoherent displacement of water was reduced postmortem. Four of the surgically treated mice yielded displacement profiles that resembled those obtained postmortem; the remaining two were near normal. This study demonstrates the feasibility of in vivo q-space imaging. The displacement profile changes that occur subsequent to an interruption of the forebrain blood supply are consistent with the hyperintensity changes seen in diffusion-weighted imaging.
Animals↗
Monitoring the initial expansion of focal ischaemic changes by diffusion-weighted MRI using a remote controlled method of occlusion.
The purpose of the present study was to quantify the early changes which occur on diffusion-weighted imaging following focal cerebral ischaemia. We have developed a method for the remote occlusion of the middle cerebral artery in the rat, allowing early changes to be monitored and images to be acquired before and after ischaemia under identical conditions. Sequential diffusion-weighted images were acquired before and for up to 6 h following the occlusion. The data show that a hyperintense area appears soon after occlusion which expands during the first 2 h. It has previously been shown that the hyperintensity observed after ischaemia is linked to events associated with energy failure. Our findings are in good agreement with previous pathophysiological studies of focal ischaemia, which suggest a growing of the energy depleted area. In conclusion, our data extend the usefulness of diffusion-weighted imaging to the study of the early spatial evolution of the most compromised area after focal ischaemia. Diffusion-weighted imaging may constitute a very powerful tool to investigate pharmacological therapies which may interact with this evolution, both in experimental and clinical studies.
Animals↗