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Ultrasound-assisted extraction and on-line LC-GC-MS for determination of polycyclic aromatic hydrocarbons (PAH) in urban dust and diesel particulate matter.

A method has been developed for determination of polycyclic aromatic hydrocarbons (PAH) in particulate matter from ambient air and diesel exhaust emissions. It is reproducible and accurate and, compared with similar methods for analysis of individual PAH components in complex matrices, it is relatively fast and simple. Single PAH components can be determined in samples of particulate matter from ambient air and diesel exhaust emissions with LOD of approximately 1 pg/sample. Further, sample throughput is high, because more than 20 samples can be extracted and prepared for analysis in one working (8-h) day. The particulate matter is subjected to ultrasound-assisted extraction, a technique that is shown to extract PAH from particulate material with efficiencies fully comparable with those of Soxhlet extraction. An aliphatic/PAH-enriched fraction is obtained by solid-phase extraction before isolation, separation, and identification/quantification of PAH by on-line liquid chromatography-gas chromatography-mass spectrometry. The method was validated by analysis of US National Institute of Standards and Technology (NIST) Standard Reference Materials (SRM) 1649a, Urban Dust, and 2975, Diesel Particulate Matter. Results from the method are in good agreement with the NIST-certified PAH concentrations and with NIST reference PAH concentrations.

Air Pollutants↗

Fast quantitative diffusion-tensor imaging of cerebral white matter from the neonatal period to adolescence.

We investigated the isotropic diffusion coefficient (D') and fractional anisotropy (FA) in white matter (WM) during brain development, using an optimised diffusion-tensor imaging (DTI) method with whole brain coverage in a clinically acceptable time. We images 52 children with no evident neurological abnormality (30 boys, 22 girls aged 1 day-16 years) using high-angle DTI with optimised temporal gradient performance. D' and FA were calculated in 10 regions of interest in white matter. We saw that the age-related reduction in D' and increase in FA follow a mono- or biexponential model in white matter, probably depending on the compactness and myelination rate of the fibre tracts. In contrast to other areas, in which adult values were reached during the third year, there is a trend to continuous increase in FA in all deep white-matter areas, suggesting continuing maturation and organisation of deep tracts not detected on conventional MRI.

Adolescent↗

Incidental white-matter foci on MRI in "healthy" subjects: evidence of subtle cognitive dysfunction.

The clinical significance of incidental white-matter foci seen on MRI is controversial. Mainly using a computer-assisted neuropsychological test battery, we tested the hypothesis that there is a clinical correlate of these foci. We studied 41 individuals aged 45-65 years with no history of neurological or psychiatric disorder, in whom no indication of central nervous system abnormalities was found on standardised neurological examination. A computer-assisted neuropsychological test battery, with the advantage of precise measuring of both time and deviation (e.g. in position memory tests), and rating scales for emotional dysfunction were administered; selected soft neurological signs were assessed. In 16 subjects (39%) MRI showed high-signal foci in the white matter on spin-echo sequences. White-matter foci not adjacent to the lateral ventricles were found to be related to performance on immediate visual memory/visuoperceptual skills, visuomotor tracking/psychomotor speed and, to a lesser degree, learning capacity and abstract and conceptual reasoning skills. Subtle cognitive dysfunction would appear to be a clinical correlate of punctate white-matter foci on MRI of otherwise "healty" individuals.

Aged↗

Unusual MRI findings in grey matter heterotopia.

We report unusual MRI patterns in patients with grey matter heterotopia. Standard T1- and T2-weighted spin-echo and inversion-recovery sequences were used in 22 patients presenting with seizures or developmental delay. The images were reviewed for signal change surrounding white matter and for atypical size, morphology or topography. We found 10 cases of subependymal heterotopias 11 of focal subcortical heterotopia and of diffuse subcortical heterotopia. On clinical or MRI grounds, 8 cases were considered unusual: 2 of the subependymal type, 2 of focal subcortical heterotopia with white matter abnormalities, 2 of focal subcortical heterotopia with no clinicoradiological correlation 1 of extensive hemispheric subcortical heterotopia and 1 of diffuse subcortical heterotopia confined to the frontal lobe. The classical classification of heterotopia enables easy radiological diagnosis even in cases with unusual patterns. In some cases, heterogeneity and high signal in surrounding white matter can be found. Cortical dysplasia is the most frequent associated malformation.

Adolescent↗

Influence of catchment-scale military land use on stream physical and organic matter variables in small southeastern plains catchments (USA).

We conducted a 3-year study designed to examine the relationship between disturbance from military land use and stream physical and organic matter variables within 12 small (<5.5 km2) Southeastern Plains catchments at the Fort Benning Military Installation, Georgia, USA. Primary land-use categories were based on percentages of bare ground and road cover and nonforested land (grasslands, sparse vegetation, shrublands, fields) in catchments and natural catchments features, including soils (% sandy soils) and catchment size (area). We quantified stream flashiness (determined by slope of recession limbs of storm hydrographs), streambed instability (measured by relative changes in bed height over time), organic matter storage [coarse wood debris (CWD) relative abundance, benthic particulate organic matter (BPOM)] and stream-water dissolved organic carbon concentration (DOC). Stream flashiness was positively correlated with average storm magnitude and percent of the catchment with sandy soil, whereas streambed instability was related to percent of the catchment containing nonforested (disturbed) land. The proportions of in-stream CWD and sediment BPOM, and stream-water DOC were negatively related to the percent of bare ground and road cover in catchments. Collectively, our results suggest that the amount of catchment disturbance causing denuded vegetation and exposed, mobile soil is (1) a key terrestrial influence on stream geomorphology and hydrology and (2) a greater determinant of in-stream organic matter conditions than is natural geomorphic or topographic variation (catchment size, soil type) in these systems.

Carbon↗

1H chemical shift imaging of the brain in guanidino methyltransferase deficiency, a creatine deficiency syndrome; guanidinoacetate accumulation in the gray matter.

MR spectroscopy results in a mild case of guanidinoacetate methyltransferase (GAMT) deficiency are presented. The approach differs from previous MRS studies in the acquisition of a chemical shift imaging spectral map showing gray and white matter with the corresponding spectra in one overview. MR spectroscopy revealed guanidinoacetate (GAA) in the absence of creatine. New is that GAA signals are more prominent in gray matter than in white. In the prevailing view, that enzyme deficiency is localized in liver and pancreas and that all GAA is transported into the brain from the blood and the cerebrospinal fluid, this would be compatible with a more limited uptake and/or better clearance of GAA from the white matter compared to the grey matter.

Aspartic Acid↗

(1)H MR spectroscopy of the brain in multiple sclerosis subtypes with analysis of the metabolite concentrations in gray and white matter: initial findings.

Many MR spectroscopy (MRS) studies of multiple sclerosis (MS) have focussed on metabolism in normal-appearing white matter (NAWM) and in white matter lesions (WML). In this study, eight patients suffering from primary or secondary progressive MS (PPMS/SPMS) and seven patients with relapsing/remitting MS (RRMS) were examined by (1)H-MRS to assess metabolite levels in gray matter (GM) as well. (1)H-MRS chemical-shift imaging of a cerebral volume of interest of 8x8x2 cm(3) above the lateral ventricles revealed differences between the metabolite concentrations in the three groups varying from almost significant [NAWM, choline (cho); P=0.0547] to highly significant [GM, N-acetylaspartate (NAA); P=0.0003]. In PPMS/SPMS patients, the decreases in choline, creatine (Cr) and N-acetylaspartate compared with six healthy controls were significant in GM and to a lesser extent, in NAWM. No significant differences in metabolite concentrations were found between RRMS and controls. In WML, all metabolites were reduced compared with white matter in controls (Cho: P=0.0020; Cr and NAA: P<0.0001, both). In conclusion, the concentrations of Cho, Cr and NAA are reduced in PPMS/SPMS patients, especially in GM and in WML. Despite contrary observations in previous studies, increases in the concentrations of Cr and/or Cho were not observed.

Adult↗

A novel leukoencephalopathy associated with tau deposits primarily in white matter glia.

A 79-year-old woman had a 10-year history of dementia, initially presenting as non-fluent aphasia. Magnetic resonance imaging showed frontal atrophy (left greater than right) and hyperintense foci within white matter. Neuropathologically, there was severe frontal atrophy due to cortical neuronal loss with spongy change and to an even greater loss of white matter that contained prominent eosinophilic deposits. The deposits were immunoreactive for phosphorylated tau, non-reactive for Abeta and alpha-synuclein and equivocally or weakly reactive for ubiquitin. They stained with the Gallyas, Bielschowsky, and Bodian techniques. Ultrastructural examination revealed the deposits to be composed of straight filaments with a diameter of approximately 10 nm, primarily in white matter glia. Moderate loss of neurons in substantia nigra and numerous argyrophilic threads in gray and particularly white matter were noted. The precise relationship between this disorder and other frontotemporal degenerations/tauopathies, as well as the pathogenetic basis of the leukoencephalopathy, remains to be determined.

Aged↗

Experimental cerebral hypoperfusion induces white matter injury and microglial activation in the rat brain.

Though cerebral white matter injury is a frequently described phenomenon in aging and dementia, the cause of white matter lesions has not been conclusively determined. Since the lesions are often associated with cerebrovascular risk factors, ischemia emerges as a potential condition for the development of white matter injury. In the present study, we induced experimental cerebral hypoperfusion by permanent, bilateral occlusion of the common carotid arteries of rats (n=6). A sham-operated group served as control (n=6). Thirteen weeks after the onset of occlusion, markers for astrocytes, microglia, and myelin were found to be labeled by means of immunocytochemistry in the corpus callosum, the internal capsule, and the optic tract. The ultrastructural integrity and oligodendrocyte density in the optic tract were investigated by electron microscopy. Quantitative analysis revealed that chronic cerebral hypoperfusion caused mild astrogliosis in the corpus callosum and the internal capsule, while astrocytic disintegration in the optic tract increased by 50%. Further, a ten-fold increase in microglial activation and a nearly doubled oligodendrocyte density were measured in the optic tract of the hypoperfused rats as compared with the controls. Finally, vacuolization and irregular myelin sheaths were observed at the ultrastructural level in the optic tract. In summary, the rat optic tract appears to be particularly vulnerable to ischemia, probably because of the rat brain's angioarchitecture. Since the detected glial changes correspond with those reported in vascular and Alzheimer dementia, this model of cerebral hypoperfusion may serve to characterize the causal relationship between ischemia and white matter damage.

Analysis of Variance↗

Cerebral white matter damage in HIV infection demonstrated using beta-amyloid precursor protein immunoreactivity.

We have examined brain sections from 55 autopsy cases of AIDS for the prevalence and severity of axonal damage, assessed using beta-amyloid precursor protein (beta APP) immunoreactivity as a marker of such damage. The cases were subdivided into cases with HIV encephalitis with multinucleated giant cells (MGC), cases with other specific pathology, such as cerebral toxoplasmosis or lymphoma, cases with non-specific pathology and cases with no pathology. Significantly more foci containing beta APP+ axons were found in cases with HIV encephalitis with MGC (80%) and in cases with other specific pathology (58%) than in those with non-specific (30%) or no pathology (30%). The prevalence and abundance of beta APP+ axons generally paralleled the severity of pallor of myelin staining of cerebral white matter in cases without other specific pathology but in 4 cases without any pallor of myelin staining beta APP+ axons were present, suggesting that it may be a more sensitive marker of some forms of white matter damage in HIV infection than myelin pallor. Foci of beta APP+ axons were found in subcortical and deep white matter but did not convincingly co-localise with foci of demonstrable HIV infection as indicated by the presence of MGC and HIV p24 immunoreactivity. In contrast, they showed an approximately perivascular distribution at some sites in all of the disease categories studied. We consider this localisation to be more suggestive of a vascular pathogenetic mechanism of deep white matter damage in HIV infection than a mechanism dependent on diffusion of local myelinotoxic products from foci of cerebral HIV infection.

Acquired Immunodeficiency Syndrome↗

Assessment of white matter injury following prolonged focal cerebral ischaemia in the rat.

The ability of putative neuroprotective compounds to protect against white matter injury remains poorly investigated due to the lack of suitable methods for assessing white matter injury. This study was therefore designed to investigate the utility of Tau 1 (oligodendrocytes/axons), myelin basic protein (MBP; myelin) and amyloid precursor protein (APP; axons) immunohistochemistry in assessing white matter injury at various times following middle cerebral artery occlusion (MCAO) in the rat. Focal cerebral, ischaemia was induced in halothane-anaesthetised rats using an intraluminal thread model. At 24 h, 1 and 2 weeks following MCAO, white matter injury was assessed using Tau 1, APP, MBP and Luxol-fast blue staining and neuronal injury with cresyl fast violet (CFV). In histologically normal tissue MBP immunoreactivity was detected in myelinated fibre tracts, while Tau 1 and APP were axonally located. At 24 h following permanent MCAO, MBP, and Tau 1 staining remained relatively unchanged within the myelin and axonal compartments of the ischaemic region. In contrast, increased Tau 1 staining was apparent in oligodendrocytes within ischaemic tissue, while APP accumulated in axons surrounding the lesion. At 1 and 2 weeks following transient MCAO, Tau 1 and APP staining was markedly decreased within ischaemic tissue. Marked reduction in MBP levels within ischaemic tissue were not detected until 2 weeks following MCAO. The area of axonal injury as assessed by reduced Tau 1 or APP staining correlated with the area of neuronal damage as assessed by CFV staining. This study shows that MBP, Tau 1 and APP immunohistochemistry can be utilised to assess myelin and axonal integrity following sustained ischaemia using standard image analysis techniques.

Amyloid beta-Protein Precursor↗

Breakdown of the blood-brain barrier to proteins in white matter of the developing brain following systemic inflammation.

Compromised blood-brain barrier permeability resulting from systemic inflammation has been implicated as a possible cause of brain damage in fetuses and newborns and may underlie white matter damage later in life. Rats at postnatal day (P) 0, P8 and P20 and opossums (Monodelphis domestica) at P15, P20, P35, P50 and P60 and adults of both species were injected intraperitoneally with 0.2-10 mg/kg body weight of 055:B5 lipopolysaccharide. An acute-phase response occurred in all animals. A change in the permeability of the blood-brain barrier to plasma proteins during a restricted period of postnatal development in both species was determined immunocytochemically by the presence of proteins surrounding cerebral blood vessels and in brain parenchyma. Blood vessels in white matter, but not grey matter, became transiently permeable to proteins between 10 and 24 h after lipopolysaccharide injection in P0 and P8 rats and P35-P60 opossums. Brains of Monodelphis younger than P35, rats older than P20 and adults of both species were not affected. Permeability of the blood-cerebrospinal fluid (CSF) barrier to proteins was not affected by systemic inflammation for at least 48 h after intraperitoneal injection of lipopolysaccharide. These results show that there is a restricted period in brain development when the blood-brain barrier, but not the blood-CSF barrier, to proteins is susceptible to systemic inflammation; this does not appear to be attributable to barrier "immaturity" but to its stage of development and only occurs in white matter.

Animals↗

Fluoro-Jade B staining following zymosan microinjection into the spinal cord white matter.

1. The fluorescein derivate Fluoro-Jade B (FJB), which primarily labels dead or dying neurons, was used to study the acute focal inflammation in the spinal cord white matter. Inflammation was induced by microinjection of the yeast particulate zymosan to evaluate the biological effects of intraspinal macrophages activation without the confounding effects of physical trauma. 2. A single bolus of zymosan (Sigma, 75 nL) was stereotaxically injected at the thoracic level into the lateral white matter of rat spinal cord. A standard Fluoro-Jade B staining protocol was applied to spinal cord sections at 6, 12, 24 h and 2, 4 days postinjection. Neutral Red, NADPH-diaphorase, Iba1-IR, and DAPI staining protocols accomplished examination of the cells participating in the acute inflammatory response. 3. Zymosan caused formation of clearly delineated inflammation lesions localized in the lateral white matter of the spinal cord. Fluoro-Jade B stained cells in the area of inflammation were not observed at 12 h postinjection while mild FJB staining appeared at 24 h and intense staining was observed at 2 and 4 days postinjection. 4. This study shows that the acute response to zymosan-induced inflammation in the rat spinal cord white matter causes a gradual appearance of phagocytic microglia/macrophages and delayed FJB staining of the inflammatory cells. 5. FJB, a reliable marker of dying neurons, is a more universal agent than formerly believed. One possible explanation for the gradual appearance of FJB-stained cells in the area of inflammation is that specific time is required for sufficient levels of proteins and/or myelin debris of axonal origin to appear in the cytoplasm of phagocytic microglia/macrophages.

Animals↗

A case study of spatial variation and enrichment of selected elements in ambient particulate matter around a large coal-fired power station in central India.

The dominant use of coal in power sectors has been associated with adverse environmental impacts. Ambient air monitoring for the two size fractions of particulate matter, respirable suspended particulate matter (RSPM) and non-respirable suspended particulate matter (NRSPM) in the downwind and upwind directions of a large coal-fired power station in central India, was carried out. Collected samples of ambient particulate matter were analysed atomic absorption spectrophotometrically for 15 elements. Spatial variability in elemental composition of RSPM and NRSPM and the degree of enrichment of these toxic metals in RSPM were investigated. A significant spatial variability for the elements in RSPM and NRSPM and higher degrees of enrichment of the elements were observed.

Air Pollutants↗

Migraine and white matter hyperintensities.

Patients with migraine are at increased risk for white matter hyperintensities detected on magnetic resonance imaging. The presence of nonspecific white matter hyperintensities may cause uncertainty for physicians and anxiety for patients. The pathophysiology and long-term consequences of these lesions are unknown. Occasionally, white matter lesions in a migraineur may indicate an underlying disease such as cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), mitochondrial encephalopathy with lactic acidosis and stroke-like episodes (MELAS), or central nervous system vasculitis. The ability to distinguish between nonspecific and disease-specific patterns of white matter hyperintensities in migraine sufferers is important for the practicing clinician.

Brain↗

Depth profile of local oxygen tension and blood flow in rat cerebral cortex, white matter and hippocampus.

Microregional oxygenation and blood flow were measured in rat cerebral cortical lamina, subcortical white matter and hippocampus. This was done to examine relationships between oxygenation and blood flow at a local level, to determine effects of craniotomy, and to consider whether flow/oxygenation relationships might be predictive of selective vulnerability known to accompany anoxia or ischemia. Blood flow and oxygen tension were measured with closely apposed polarographic microelectrodes. Oxygen tension was highest in white matter, lower in the region of cortical laminae IV-V and lowest in the hippocampus. Blood flow in the hippocampus was higher than that in white matter or laminae IV-V of cerebral cortex. Ratios of blood flow to oxygenation were similar throughout the cortex, higher in white matter but oxygenation in hippocampus was significantly less than expected from measurement of hippocampal blood flow reflecting increased oxygen consumption or relative hypoxia due to increased diffusion distances for oxygen in hippocampus. Comparison of data from closed vs opened skull animals indicated that diffusion of oxygen and hydrogen influenced data to approximately 1000 micron below the cerebral surface.

Animals↗

Changes in permissivity for neuronal attachment and neurite outgrowth of spinal cord grey and white matters during development: a study with the 'cryoculture' bioassay.

We have used the recently developed cryoculture bioassay (Carbonetto et al., J. Neurosci., 7 (1987) 610-620) to document changes during development of CNS tissular ability to support nerve fiber growth. Neuronal attachment and neurite outgrowth of purified neurons cultured on tissue sections of rat spinal cord at various stages of development were quantified. Nerve fiber growth permissivity increased during embryonic stages, reaching as postnatal days 2-4 (P2-P4) a maximum value, higher than that found on adult PNS tissue sections. This permissivity diminished rapidly thereafter, indicating that early postnatally, the nerve fiber growth supporting ability of the CNS environment shifts abruptly from an increasingly permissive mode to an increasingly non-permissive status. Furthermore, after P4, neurite outgrowth permissivity diminished in parallel on white and grey matters, whereas neuronal attachment declined much more drastically on white matter than on grey matter. This indicates that the progressive loss of spinal cord ability to support nerve fiber growth is attributable to both grey and white matters. In several instances it also appeared that neuronal adhesion was not necessarily followed by a comparable level of nerve fiber growth, suggesting that these two processes could be regulated by different factors.

Aging↗

Microglial cell reaction in the gray and white matter in spinal cords from jimpy mice. An enzyme histochemical study at the light and electron microscope level.

Jimpy is a genetic disorder which results in a severe hypomyelination in the central nervous system associated with a variety of astroglial and oligodendroglial abnormalities. In this study, we examined the morphology and distribution of microglial cells in spinal cord sections from jimpy and normal mice at 10-12 and 20-22 days postnatal using a specific microglial marker, the nucleoside diphosphatase staining. Compared to those of normal littermates, the spinal cords of jimpy mice showed an intense microglial cell reaction in white and gray matter, as revealed by quantitative analysis and light and electron microscope study. Microglial reactivity was apparent in all spinal cord areas, although it was more pronounced in white than in gray matter. The mean microglial densities in the jimpy white matter were about threefold (10-12 days) and fivefold (20-22 days) higher than in the normal, whereas in the gray matter, microglial density in jimpy was about 60% higher than in normal at both ages. Morphologically, microglial cells in the normal spinal cord showed a ramified appearance, similar in size and ramification pattern to those reported in other normal CNS areas. In contrast, microglial cells in the jimpy spinal cord showed a reactive morphology, characterized by a shortening and coarsening of their cell processes, swelling of their cell body and accumulation of lipid inclusions. Reactive microglial cells were found in close association with axons and oligodendroglial cells. The possible role of microglial cells in hypomyelination is discussed.

Acid Anhydride Hydrolases↗