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Spatial distribution of persistent organochlorines in ringed seal (Phoca hispida) blubber.

Blubber samples, taken through the entire blubber column, were collected from three different anatomical locations on ringed seals (Phoca hispida). The outer and inner layers of these samples were analysed for concentrations of sigma PCBs (sum of the analysed congeners, polychlorinated biphenyls) and p,p'-DDE (4,4'-dichlorodiphenyldichloroethylene). No significant differences were found in blubber thickness or in per cent extractable lipid when comparing age- and sex-groups, or in extractable lipid content when comparing anatomical locations of the blubber or blubber depth. No significant differences were found between the three different anatomical locations with respect to the concentrations of sigma PCBs or p,p'-DDE. However, the concentrations of both sigma PCB and p,p'-DDE were significantly higher in the outer blubber layer compared to the inner. Higher levels of both sigma PCB and p,p'-DDE were found in males compared to females and juveniles in both the inner and outer parts of the blubber column. The most commonly used blubber sampling devices are biopsy tools that penetrate only the outer layer of the blubber. Analyses of such samples will not reflect the real body burden of organochlorines. Standard methods for sampling blubber of marine mammals should be developed so that comparative studies and longitudinal monitoring programs of pollutants in marine mammals can be conducted in a meaningful way.

Adipose Tissue↗

Simple methods for quantifying the spatial distribution of different categories of motoneuronal nerve endings, using measurements of muscle regionalization.

For skeletal muscles, a well-known match exists between the properties of motoneurones and those of their muscle fibres. Hence, the intramuscular distribution of different kinds of motoneuronal nerve endings (e.g. 'slow' versus 'fast') can be mapped by determining the distribution of the corresponding types of muscle fibre. As a background for further studies of motoneuronal plasticity, we needed precise measures of such distributions. Simple quantitative methods were developed for defining the position and extent of sub-populations of cells within a structure (e.g. the regional distribution of slow versus fast muscle fibres within a muscle cross-section): (a) The 'mass vector method' defined the relative position of the target cell cloud. A line was drawn between the calculated centre of mass for the target cells and that for the whole structure. The direction (a1) and length (a2) of this line gave a measure of the direction and degree of target cell eccentricity within the structure. (b) The 'sector method' delineated the region containing the target fibres. A circle around the centre of mass for the target fibres was subdivided into a number of equal sectors (standard setting: 20). The most remote point was found within each sector and a line joining these points defined the region of the target fibres. When applied to the 'slow' type I fibres of cross-sections from rat hindlimb muscles, the regional area estimates obtained by the sector method were highly correlated with, but approximately 10% lower than those achieved by the well-established 'convex hull' method. Highly significant inter-muscular differences were observed for each one of the three new parameters described in this paper (a1, a2, b).

Animals↗

Spatial distribution, cellular integration and stage development of Parkin protein in Xenopus brain.

Parkin is an ubiquitin-protein ligase molecule abundantly expressed in mammalian brains. Deletional mutations of Parkin protein produce a disease-related parkinsonian phenotype which is inherited with an autosomal recessive mode of transmission. To gain a greater insight into the evolutionary trajectory of the protein among vertebrate species, we describe here the (i) distribution pattern, (ii) sizing of specific fragments and (iii) embryonic development of Parkin in Xenopus laevis utilizing two antibodies to the N- and C-terminal sequence of the human Parkin protein. Parkin immunoreactivity was distributed in a heterogeneous fashion throughout the adult frog brain. The telencephalon, including the olfactory bulb, striatum and nucleus accumbens, harbored high numbers of Parkin-containing cells. High numbers of immunoreactive neurons were also present in discrete regions of the thalamus and hypothalamus. Relatively moderate expression of Parkin protein was noted in the nucleus anterodorsalis tegmenti, nucleus reticularis medius and torus semicircularis. The substantia nigra exhibited a distinctive heterogeneous pattern of Parkin-immunoreactivity, especially within presumptive dopamine neurons. The cerebellum also showed high expression of Parkin-positive material. Characterization of the subcellular distribution of the protein indicated both a cytoplasmic and nuclear integration of Parkin-immunoreactivity. This pattern of subcellular localization was similar to that observed in human brain material, perhaps reflecting distinct structural phosphorylation sites of the Parkin protein. Western blot analysis identified three specific bands with molecular weights varying from 50 to 65 kDa in adult Xenopus brain. However, studies on the temporal expression of Parkin during development showed a complete absence of cellular immunoreactivity which was especially conspicuous during late premetamorphic stages of frog development. These results suggest that the ubiquitination activity of Parkin is limited or non-existent during embryogenesis, but appears to assume a more functional role during adulthood as reflected by the high distribution pattern of the protein within major circuits of the amphibian brain.

Amino Acid Sequence↗

Genetic heterogeneity in human astrocytomas: spatial distribution of P16 and TP53 deletions in biopsies.

Smear preparations of 23 fresh astrocytoma biopsies were analyzed by two-color fluorescence in situ hybridization with cosmids specific for the P16 and the TP53 genes. Additionally, tissue sections of the same tumors were immunostained with the use of a monoclonal antibody that recognizes both wild-type and mutant TP53 protein. In 21 astrocytomas, loss of P16 was observed in a significant proportion of cells. Cells with homozygous P16 loss were present in 13 astrocytomas; 14 astrocytomas showed cells with heterozygous loss of P16. Remarkably, 5 astrocytomas showed a scattered mosaic pattern of cells with homozygous and, respectively, heterozygous p16 loss. Homozygous deletion of TP53 was not observed. Cells with heterozygous TP53 loss were detected in 12 tumors, in 7 of them in association with P16 loss. One tumor showed aberrant cells for neither TP53 nor P16 but strong immunostaining for TP53. Positive TP53 immunostaining was found in 16 astrocytomas. Heterozygous loss of TP53 was significantly correlated with TP53 protein expression. We conclude that, unlike typical tumor suppressor genes, P16 might enhance cellular proliferation after heterozygous loss through a dosage effect and that the distribution of cells with homozygous loss of P16 speaks in favor of a polyclonal loss of the second copy of this gene.

Astrocytoma↗

Analysis of the structure and spatial distribution of ultraviolet-induced DNA repair patches in human cells made in the presence of inhibitors of replicative synthesis.

The repair of ultraviolet-induced damage in the presence of hydroxyurea or hydroxyurea and arabinosylcytosine was investigated in confluent human fibroblasts at the level of DNA loops attached to the nuclear matrix. Estimation of single-strand break frequencies based on the release of DNA from the DNA-nuclear matrix complex after incubation with nuclease S1 revealed the occurrence of multiple incision events per DNA loop in the presence of inhibitors. When both inhibitors were employed, over 90% of the repair-labelled DNA was not ligated within 2 h post-incubation. In the absence of ligation of repair patches, we observed a preferential release of repair-labelled DNA from the nuclear matrix by nuclease S1 compared to prelabelled DNA, regardless of the period of post-UV incubation. The results suggest that repair events are clustered to some extent in a certain area of a DNA loop. However, the position of these clusters relative to the attachment sites of DNA loops at the nuclear matrix is random. The data are discussed in terms of denaturation of a putative repair complex in the presence of hydroxyurea resulting in an excess of incisions over repaired sites.

Cell Line↗

Spatial distribution of photosynthetic response to long-term influence of elevated CO(2) in a mediterranean macchia mini-ecosystem.

During an open-top chamber experiment performed in evergreen 'macchia' ecosystem, which was represented by the clumps of natural vegetation dominated by Quercus ilex trees, the trees were exposed to one of two CO(2) concentrations (ambient CO(2), AC-variant and elevated CO(2), i.e. ambient plus 350 &mgr;mol CO(2) mol(-1), EC-variant) continuously over 5 years. Clumps of natural vegetation were enclosed in open top chambers (OTCs). Within the crowns of investigated Quercus ilex trees in OTCs, two crown layers i.e. sunny (E-leaves) and shaded (S-leaves), were identified as differing in solar radiation environment. To evaluate the effect of elevated CO(2), as well as the functional differentiation in assimilation activity of E- versus S-leaves, gas exchange and chlorophyll ā fluorescence techniques were used. The stimulatory effect of the long-term elevated CO(2) on the A(N)-PPFD relation was evident in E- and S-leaves of investigated Quercus trees. The A(Nmax) sensitivity of AC-variant leaves to the sudden application of elevated CO(2) was higher for S-leaves (42%) than for E-leaves (24%). The PPFD saturated rate of regulated thermal energy dissipation (ERD) confirmed the foliage differentiation caused by the long-term influence of elevated CO(2). The ERD of E- and S-leaves in the AC-variant were 1.11 times of that in the EC-variant. However, the estimated rates of photochemistry (ERP) of E- and S-leaves in the EC-variant were 1.35 and 1.22 times of E- and S-leaves in the AC-variant. The achievement of the critical value of q(P)=0.4 in E- and S-leaves from the EC-variant under lower values of PPFD compared to the AC-variant indicates a higher degree of PSII over-reduction. Thus, elevated CO(2) can be responsible for an increased susceptibility of photosynthetic apparatus to high irradiance. The obtained results support the hypothesis on the foliage vertical distribution effect on the whole canopy response to elevated CO(2). The S-layers of the canopy can play an important role in providing storage space for photosynthesis under elevated CO(2).

Journal Article↗

Progression of natural attenuation processes at a crude oil spill site: II. Controls on spatial distribution of microbial populations.

A multidisciplinary study of a crude-oil contaminated aquifer shows that the distribution of microbial physiologic types is strongly controlled by the aquifer properties and crude oil location. The microbial populations of four physiologic types were analyzed together with permeability, pore-water chemistry, nonaqueous oil content, and extractable sediment iron. Microbial data from three vertical profiles through the anaerobic portion of the contaminated aquifer clearly show areas that have progressed from iron-reduction to methanogenesis. These locations contain lower numbers of iron reducers, and increased numbers of fermenters with detectable methanogens. Methanogenic conditions exist both in the area contaminated by nonaqueous oil and also below the oil where high hydrocarbon concentrations correspond to local increases in aquifer permeability. The results indicate that high contaminant flux either from local dissolution or by advective transport plays a key role in determining which areas first become methanogenic. Other factors besides flux that are important include the sediment Fe(II) content and proximity to the water table. In locations near a seasonally oscillating water table, methanogenic conditions exist only below the lowest typical water table elevation. During 20 years since the oil spill occurred, a laterally continuous methanogenic zone has developed along a narrow horizon extending from the source area to 50-60 m downgradient. A companion paper [J. Contam. Hydrol. 53, 369-386] documents how the growth of the methanogenic zone results in expansion of the aquifer volume contaminated with the highest concentrations of benzene, toluene, ethylbenzene, and xylenes.

Bacteria, Aerobic↗

Temporal trends for 99Tc in Norwegian coastal environments and spatial distribution in the Barents Sea.

The objective of this study was to reassess 99Tc transit times and transfer factors, from Sellafield to northern Norway, and to determine the extent of 99Tc migration to the Barents Sea. Filtered seawater samples were collected on a monthly basis from Hillesøy, northern Norway, and in February 1999 from the Barents Sea. Results showed an increase in levels of 99Tc at Hillesøy where activity concentrations have increased from a baseline of 0.2-0.4Bq m(-3) to a maximum of 1.6 Bq m(-3). A transit time of 42 months and a transfer factor of 6Bq m(-3) per PBq a(-1) have been derived, using cross-correlation analysis. The current study predicts that future levels are unlikely to increase dramatically over the levels observed in 1998. Levels of 99Tc in the Barents Sea ranged from 0.2 Bq m(-3) to 1.1 Bq m(-3) showing the influence of new 99Tc inputs by early 1999.

Environmental Pollutants↗

Selective migration from deprived areas in Northern Ireland and the spatial distribution of inequalities: implications for monitoring health and inequalities in health.

Much of the evidence suggesting that inequalities in health have been increasing over the last two decades has come from studies that compared the changes in relative health status of areas over time. Such studies ignore the movement of people between areas. This paper examines the population movement between small areas in Northern Ireland in the year prior to the 1991 census as well as the geographical distribution of migrants to Northern Ireland over the same period. It shows that deprived areas tended to become depopulated and that those who left these areas were the more affluent residents. While immigrants differed a little from the indigenous population, the overall effect of their distribution would be to maintain the geographical socio-economic status quo. The selective movement of people between areas would result in the distribution of health and ill-health becoming more polarized, i.e. produce a picture of widening inequalities between areas even though the distribution between individuals is unchanged. These processes suggest potential significant problems with the area-based approaches to monitoring health and inequalities in health.

Censuses↗

Spatial distribution of NA+ and K+ channels in spinal dorsal horn neurones: role of the soma, axon and dendrites in spike generation.

Spinal dorsal horn neurones play an important role in processing sensory information received from primary afferent fibers. The application of the patch-clamp technique to thin slices of rat spinal cord has enabled the study of ionic channels in visually identified dorsal horn neurones. The small soma of these neurones isolated from the slice by means of a novel method of 'entire soma isolation' has become a convenient model for investigating the properties and distributions of ionic channels. The present review summarizes results of recent experiments studying different types of voltage-gated Na+ and K+ channels expressed in dorsal horn neurones. Uneven distribution of the channels between the soma. axon and dendrites appears to play a major role in determining the neuronal excitability. The contribution of the soma, axon and dendrites to generation and propagation of the action potentials in central neurones is discussed.

Action Potentials↗

Temporal and spatial distribution of corticosteroidogenic enzymes immunoreactivity in developing human adrenal.

Steroid hormones secreted by fetal adrenocortical cells are considered to be a requirement for a fetus to maintain intrauterine life, but, to date, the regulation of steroid hormone secretion has not been studied in detail in the human fetal adrenal gland. In this study, we examined the immunolocalization of steroidogenic enzymes and their local regulation, including adrenal 4-binding protein (Ad4BP or NR5A1), steroidogenic acute regulatory protein (StAR), P450 cholesterol side-chain cleavage (P450scc or CYP11A1), P450 17alpha-hydroxylase/17,20-lyase (P450c17 or CYP17), 3beta-hydroxysteroid dehydrogenase/isomerase (3beta-HSD), P450 21 hydroxylase (P450c21 or CYP21), dehydroepiandrosterone sulfotransferase (DHEA-ST), P450 oxidoreductase and cytochrome b5, in the human fetal adrenal gland (n=31) obtained from fetuses ranging in ages from 14 to 40 weeks of gestation. Ad4BP immunoreactivity was detected in all adrenocortical zones throughout gestation, suggesting that this nuclear protein is likely to be essential in the development of the human adrenal. Immunoreactivity for StAR, P450scc, P450c21, P450 oxidoreductase and cytochrome b5 was detected only in fetal and transitional zone between 14 and 22 weeks of gestation, but was detected in all three zones after 23 gestational weeks. 3beta-HSD immunoreactivity was not detected in any of the three cortical zones prior to 22 weeks of gestation, but became discernible in the transitional zone and definitive zone after 23 weeks. Immunoreactivity for P450c17 and DHEA-ST was detected in the transitional and fetal zones throughout gestation, but not in the definitive zone. These results suggest that the human adrenal cortex may produce dehydroepiandrosterone (DHEA) in the transitional and fetal zones throughout gestation, and cortisol in the transitional zone after the 23rd week of gestation.

Adrenal Cortex Hormones↗

A comparative analysis of structure and spatial distribution of decorin in human leiomyoma and normal myometrium.

Leiomyoma is a benign smooth muscle tumor of the uterus that affects many women in active reproductive life. It is composed by bundles of smooth muscle cells surrounded by extracellular matrix. We have recently shown that the glycosylation of extracellular matrix proteoglycans is modified in leiomyoma: increased amounts of galactosaminoglycans with structural modifications are present. The data here presented show that decorin is present in both normal myometrium and leiomyoma but tumoral decorin is glycosylated with longer galactosaminoglycan side chains. Furthermore, these chains contain a higher ratio D-glucuronate/L-iduronate, as compared to normal tissue. To determine if these changes in proteoglycan glycosylation correlates with modifications in the extracellular matrix organization, we compared the general structural architecture of leiomyoma to normal myometrium. By histochemical and immunofluorescence methods, we found a reorganization of muscle fibers and extracellular matrix, with changes in the distribution of glycoproteins, proteoglycans, and collagen. Thin reticular fibers, possibly composed by types I and III collagen, were replaced by thick fibers, possibly richer in type I collagen. Type I collagen colocalized with decorin both in leiomyoma and normal myometrium, in contrast to type IV collagen that did not. The relative amount of decorin was increased and the distribution of decorin and collagen was totally modified in the tumor, as compared to the normal myometrium. These findings reveal that not only decorin structure is modified in leiomyoma but also the tissue architecture changed, especially concerning extracellular matrix.

Adult↗

Spatial distribution of myosin heavy chain isoforms and lactate dehydrogenase M4 in the limb musculature of two crossbred lambs.

Sixteen different skeletal muscle samples were distributed in the cross-section of eight hip and thigh muscles. Contractile characteristics were assessed by measuring myosin heavy chain (MHCI, MHCIIa, MHCIIb) composition by electrophoresis. Glycolytic capacity was estimated by immunochemical quantitation of the LDH-M4. Histochemistry was used as a reference. The MHC isoform composition of most of the muscles in this study was heterogeneous. When an intramuscular transversal regionality was observed (semitendinosus, vastus lateralis, vastus medialis and rectus femoris muscles), MHCI percentage increased toward deeper layers, while MHCIIb and LDH-M4 decreased. The pattern of MHCIIa isoform distribution was less evident. Within semimembranosus and gluteus medius muscles, proportions of MHC isoforms were constant. Gradients of variation of MHCI and MHCIIb isoforms across rectus femoris and vastus medialis muscles were sharper than those of semitendinosus and vastus lateralis muscles. For the vastus lateralis muscle, these gradients may also be modified according to the breed. Breed effect was mainly shown by MHCIIb and MHCI isoforms and was not observed at all the sampling points of the muscles. These observations show that breed effect on muscle contractile and metabolic characteristics is not uniformly expressed throughout the muscle. Results of a comparison may differ according to the muscle and sampling location.

Animals↗

Non-random spatial distribution of intermembraneous particles in red blood cell membrane.

A combined analysis using digital image processing and quantitative evaluation of computed radial distribution of freeze fracture P face particles irradiated and nonirradiated has been carried out. Important quantitative features of the arrangement of particles can be described by computing a statistical measure, the radial distribution function g(r), commonly used to study the structure of liquids. The coordinates for calculating g(r) for each sample were measured automatically by a digital image processing system. The results for the radial distribution function were found to closely resemble g(r) for fluids, indicating the presence of shortrange order. This measure distinguishes three different patterns: i) random distribution, ii) dilute gas like and iii) a liquid like model. The mean number of first nearest neighbours surrounding each particle (coordination number) was found to be of the order of three. The RBC intramembraneous particles are heterogeneous in their behavioural pattern: two thirds random; one fifth, dilute gas; and one tenth, liquid.

Animals↗

The spatial distribution of pulmonary lesions in severe ARDS. An autopsy study of 35 cases.

The present study was undertaken in order to describe the local distribution and temporal course of pulmonary lesions in severe ARDS. We investigated a total of 35 patients (22 females), ranging in age from 2 to 51 years, who suffered from ARDS III and IV and were treated by extracorporeal CO2 removal and low frequency positive pressure ventilation (ECCO2-R). The extent of acute and chronic diffuse alveolar damage was assessed on histologic gross sections in the ventral, central and dorsal zone of the upper and lower lobes. The lesions showed a characteristic uniform distribution. Areas with chronic DAD were predominantly situated in the ventral portions of the upper lobes. Acute DAD predominated in the dorsal and basal areas of the lung. The extent of acute and chronic DAD was virtually independent of the duration of disease. Hemorrhage occurred at the interface zone between chronic and acute DAD and made up a significant volume portion of the lung tissue, ranging between 8% (lower lobes) and 42% (upper lobes). We conclude that the progression of acute DAD to chronic DAD is mainly determined by local factors (hydrodynamic and hydrostatic forces, intraalveolar pressure) that differ within the lung, whereas the duration of disease plays a minor role. Parenchymal hemorrhage occurs at the interface between areas of acute and chronic DAD and may therefore primarily be due to an increased susceptibility of the pulmonary parenchyma to mechanical stress.

Adolescent↗

[Influence of a priori parameters on bayesian relative risks estimations. Spatial distribution of bladder cancer in the urban area of Grenoble].

Bayesian estimates of disease relative risks is currently the gold standard in disease mapping when the disease is rare and/or when the geographical area is small. Its use has become quite easy with adhoc software. However, the implicit mechanisms of the choices made by the user must be clearly identified. We were interested here in the consequences of the choice of the hyper a priori parameters. We have compared results obtained using various hyper a priori parameters. The consequences of these choices are illustrated through the example of the incidence of bladder cancer among men in the urban area of Grenoble. We show that the risks can appear weak from a statistical point of view but important from an epidemiologic point of view in the presentation of the results.

Aged↗

MRI and x-ray CT study of spatial distribution of core breakdown in 'Conference' pears.

Two non-destructive tomographic techniques, X-ray CT imaging and magnetic resonance imaging (MRI), were applied to study the development of core breakdown disorder in 'Conference' pears (Pyrus communis cv. Conference). This disorder, which is characterized by brown discoloration of the tissue and development of cavities, is induced by elevated CO(2) and decreased O(2) levels during controlled atmosphere storage. Tomographic images of pears stored for 10 months under disorder inducing conditions, were acquired with both techniques and compared to the actual slices. Both X-ray and MRI were able to differentiate between unaffected tissue, brown tissue and cavities. A simple image-processing program, based on threshold values, was developed to determine the area percentage of affected and unaffected tissue as well as the cavity and core area per slice. For all three imaging techniques the area percentage brown tissue per slice increased with the diameter of the pear, but was systematically underestimated by 12% and 6% for, respectively, X-ray and MRI, compared to the actual slices. The area percentage cavity corresponded very well for all techniques. It was also found that the contours of the brown tissue were parallel to the fruit boundaries, suggesting a relation between the disorder symptoms and gas diffusion properties of the fruit. It was concluded that MRI is the most appropriate technique to study the development of core breakdown disorder during postharvest storage in future experiments.

Magnetic Resonance Imaging↗

Spatial distribution of perylenequinones in lichens and extended quinones in quincyte using confocal fluorescence microscopy.

The application of confocal fluorescence microscopy and microspectrofluorimetry to the characterization of the distribution of organic compounds in bulk lichens and mineral structures is demonstrated. Perylenequinones and extended quinones were chosen as both model compounds and as the naturally occurring fluorophores. These molecules occur, respectively, in corticolous microlichens and in a pink-colored mineral called quincyte. The structures of quincyte and of the lichens Cryptothelium rhodotitton and Graphis hematites are described, and the possibilities of energy dissipation and photoprotection mechanisms in these lichens are discussed. This study also illustrates how, for a wide range of specimens, naturally occurring quinone fluorophores in the specimen can be exploited directly to yield chemical and structural information without using fluorescent labelling. These intrinsic quinonoid compounds have molecular fluorescence yields and laser damage thresholds comparable or superior to common microscopy dyes, and can therefore be used to obtain high-contrast 3D fluorescence imaging without the complications introduced by dye labelling.

Lichens↗