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Cell cycle-dependent changes in Golgi stacks, vacuoles, clathrin-coated vesicles and multivesicular bodies in meristematic cells of Arabidopsis thaliana: a quantitative and spatial analysis.

Cytokinesis in plants involves both the formation of a new wall and the partitioning of organelles between the daughter cells. To characterize the cellular changes that accompany the latter process, we have quantitatively analyzed the cell cycle-dependent changes in cell architecture of shoot apical meristem cells of Arabidopsis thaliana. For this analysis, the cells were preserved by high-pressure freezing and freeze-substitution techniques, and their Golgi stacks, multivesicular bodies, vacuoles and clathrin-coated vesicles (CCVs) characterized by means of serial thin section reconstructions, stereology and electron tomography techniques. Interphase cells possess approximately 35 Golgi stacks, and this number doubles during G2 immediately prior to mitosis. At the onset of cytokinesis, the stacks concentrate around the periphery of the growing cell plate, but do not orient towards the cell plate. Interphase cells contain approximately 18 multivesicular bodies, most of which are located close to a Golgi stack. During late cytokinesis, the appearance of a second group of cell plate-associated multivesicular bodies coincides with the onset of CCV formation at the cell plate. During this period a 4x increase in CCVs is paralleled by a doubling in number and a 4x increase in multivesicular bodies volume. The vacuole system also undergoes major changes in organization, size, and volume, with the most notable change seen during early telophase cytokinesis. In particular, the vacuoles form sausage-like tubular compartments with a 50% reduced surface area and an 80% reduced volume compared to prometaphase cells. We postulate that this transient reduction in vacuole volume during early telophase provides a means for increasing the volume of the cytosol to accommodate the forming phragmoplast microtubule array and associated cell plate-forming structures.

Arabidopsis↗

Spatial analysis of limb bud myogenesis: elaboration of the proximodistal gradient of myoblasts requires the continuing presence of apical ectodermal ridge.

Myogenic tissue from embryonic chick wing and leg buds is composed of several subpopulations of myoblasts. These clonally distinct subpopulations first appear at different developmental stages, and are distributed differently along the proximo-distal axis of the buds, giving the appearance of a gradient of myoblast cell types. This myoblast distribution pattern has been utilized to investigate the dependence of muscle tissue outgrowth and development on the presence of the apical ectodermal ridge (AER). Wing buds which have had the AER removed at stages 17-18 (2 days) subsequently develop normal proximal regions, but fail to elaborate skeletal structures distal to the humerus. The myoblast pattern of operated buds is also normal proximally, but distal portions of the pattern are not observed. Removal of the AER at stage 20 (3 days) results in buds which develop slightly more distal skeletal structures and the coinciding portions of the myoblast pattern, but in which the more distal portions of the normal myoblast gradient are truncated. These data suggest that elaboration of the myogenic pattern in early limb buds is dependent on the continuing presence of the AER, and that early removal of the AER leads to the subsequent cessation of myoblast pattern specification.

Animals↗

Temporal and spatial analysis of cartilage proteoglycan core protein gene expression during limb development by in situ hybridization.

As limb mesenchymal cells differentiate into chondrocytes they initiate the synthesis of a cartilage-specific sulfated proteoglycan, cartilage-characteristic type II collagen, and other cartilage-specific proteins. In the present study, in situ hybridization with a 32P-labeled cloned cDNA probe complementary to mRNA encoding the core protein of cartilage proteoglycan has been used to visualize and localize the accumulation of cartilage proteoglycan core protein mRNA sequences during development of the chick limb bud in vivo. When the probe was hybridized to sections through 7-day (stage 32) limbs, an intense hybridization signal was observed over the well-differentiated cartilage rudiments of the limb, while no signal above background was observed over nonchondrogenic tissues including muscle, loose connective tissue, and epidermis. At early stages of limb development, an accumulation of silver grains representing hybridizable core protein mRNA first became detectable in the proximal central core of the limb where the prechondrogenic condensation of mesenchymal cells that characterizes the onset of cartilage differentiation was occurring. In fact, the pattern of silver grain accumulation closely followed the pattern of mesenchymal cell condensation, and no hybridizable core protein mRNA sequences were detectable in the limb bud prior to condensation. Cartilage-characteristic type II collagen mRNA was colocalized with core protein mRNA in the condensing central core of the limb suggesting that the genes for these two major constituents of cartilage matrix are coordinately regulated at the onset of chondrogenesis. Furthermore, the appearance of hybridizable core protein mRNA was closely followed by the appearance of the protein for which it codes as detected by immunohistochemical staining with monospecific antibody. These observations support the hypothesis that at the initial stages of limb chondrogenesis core protein gene expression is controlled primarily at the transcriptional level.

Aggrecans↗

A spatial analysis of on- and off-ganglion cells in the cat retina.

Using reduced silver staining methods it was possible to stain all alpha-ganglion cells of the cat retina. The dendritic trees of alpha-cells are unistratified in either of two laminae within the inner plexiform layer. This stratification difference was shown physiologically to correspond to the on-off dichotomy. For all alpha-cells recorded, the dendritic field was covered by the corresponding receptive field centre. In addition the general shape of the receptive field centre corresponded to the shape of the dendritic field. The size of the dendritic tree was always smaller than the receptive field centre. The topographical distribution of on- and off-alpha cells could be studied. They were found to occur in about equal numbers. Both on- and off-alpha-cell perikarya form a regular lattice and both lattices are superimposed independently. The dendritic branches of neighbouring alpha-cells overlap and each retinal point is covered by the dendritic field of at least one on- and one off-alpha-cell. After horseradish peroxidase (HRP) injection into the lateral geniculate nucleus all beta-cells were labelled. In this way it is shown that about 55% of all ganglion cells are beta-cells. The mosaic of on- and off-beta-cells was studied from the HRP-labelled material. It is commonly assumed that beta-cells are associated with the resolution of fine detail in the cat visual system. The mosaic of beta-cells imposes some constraints and permits some predictions to be made with respect to the cat's visual discrimination.

Action Potentials↗

Spatial analysis of surgery locations in general practice.

Consideration is given to some of the theoretical issues involved in applying location allocation analysis to the problem of location of general practitioner's surgeries. Alternatives criteria for minimization can be used in the algorithm to represent different planning objectives. The technique is applied to data for two Medical Practice Areas in East Kent. The travel cost for the population, as measured by the procedure, is compared for different types of surgery distributions in the two areas. The technique is proposed as an additional method to that already used to assess medical manpower distribution for planning purposes.

England↗

Journey patterns of the elderly and disabled in the Cotswolds: a spatial analysis.

The decline of services and transport provision in the North Cotswolds has led to a progressive deterioration in the welfare of the elderly and disabled. This study, based on interviews with 498 individuals in eight typical parishes, relates the travel behaviour of these groups to health status, personal circumstances, the grade and location of services, and mode of transport. The repercussions of the 1985 Transport Act for the car-less and severely disabled are then examined. Given the increased loadings on domiciliary support systems, the case is argued for coordinated initiatives in alternative transport provision which relate, sensitively, to the needs of the different groups among the elderly and disabled.

Activities of Daily Living↗

Quantitative spatial analysis of the distribution of NADPH-diaphorase-positive neurons in the developing and mature rat retina.

NADPH-diaphorase (NADPH-d) histochemistry labels a subpopulation of nitric oxide-synthesizing amacrine cells in the inner nuclear layer of the rat retina. We have studied their morphology and distribution in postnatal and adult rats in whole-mounted retinae. NAPDH-d-positive neurons are detected as early as postnatal day (P)5, especially in the peripheral retina; intense labeling of somata and long lengths of dendrites is obtained between P10 and P18, after which only the somata exhibit NADPH-d activity. The density and number of these cells increase progressively from P7 to P14, with a significantly higher density in the central retina as compared to the periphery. The sociology of these cells was analyzed quantitatively studying the Voronoi domains: a polygon area can be drawn that delineates the territory of the map that is closer to the cell than to any other cell of the map. In addition, we calculated the conformity ratio of Cook, i.e., the mean nearest neighbor distance/standard deviation of all the nearest neighbor distances, in order to reveal whether or not these cells are regularly distributed through the retina. We find that the distribution of the NADPH-d-positive cells tends to be regular throughout the retina: the local coefficient of variation (obtained by comparing the size of each Voronoi polygon area to those of its neighbors) tends to regularity at P14 and remains unaltered through maturity. Therefore, as other cell types, NADPH-d-positive amacrine cells are almost regularly distributed from the time of eye opening and nitric oxide may play a role in the development of retinal circuitry and in the regulation of retinal blood flow.

Age Factors↗

Neural activity associated with metaphor comprehension: spatial analysis.

Though neuropsychological data indicate that the right hemisphere (RH) plays a major role in metaphor processing, other studies suggest that, at least during some phases of this processing, a RH advantage may not exist. The present study explores, through a temporally agile neural signal--the event-related potentials (ERPs)--, and through source-localization algorithms applied to ERP recordings, whether the crucial phase of metaphor comprehension presents or not a RH advantage. Participants (n=24) were submitted to a S1-S2 experimental paradigm. S1 consisted of visually presented metaphoric sentences (e.g., "Green lung of the city"), followed by S2, which consisted of words that could (i.e., "Park") or could not (i.e., "Semaphore") be defined by S1. ERPs elicited by S2 were analyzed using temporal principal component analysis (tPCA) and source-localization algorithms. These analyses revealed that metaphorically related S2 words showed significantly higher N400 amplitudes than non-related S2 words. Source-localization algorithms showed differential activity between the two S2 conditions in the right middle/superior temporal areas. These results support the existence of an important RH contribution to (at least) one phase of metaphor processing and, furthermore, implicate the temporal cortex with respect to that contribution.

Adult↗

Spatial analysis of bovine spongiform encephalopathy in Galicia, Spain (2000-2005).

In Spain, the first bovine spongiform encephalopathy (BSE) case was detected in 2000 in a cow born in the Galicia region (Northwestern Spain). From then and until October 2005, 590 cases were detected, 223 of them in Galicia. In 1994, meat and bone meal (MBM) was banned on ruminant feed and, in 1996, an EU decision mandating an overall change in MBM processing was implemented. This decision was gradually applied in the territory and not enforced before July 1998. The objective of this study was to explore clustering of BSE cases and estimate the standard incidence ratio (SIR) of BSE in Galicia. Our study was based on the BSE cases detected during the surveillance period 2000-2005 in the Galicia region. These cases were divided, based on birth date, into two periods: animals born from 1994 to July 1998, and those born after July 1998. We tested the role of cross-contamination on the geographical SIR distribution for both periods. Hierarchical Bayesian models were used to model the overdispersion and lack of independence of the SIR estimates. The geographical distribution of the standard incidence ratio of BSE between both periods was different. In the second period, the SIR was reduced in some areas. The reduction in these areas could be attributable to the changes in the processing of MBM. We did not find any statistical link between the poultry population and the standard incidence ratio, but pig population had a positive effect.

Animal Feed↗

Spatial analysis of annual air pollution exposure and mortality.

The aim of this study was to relate ambient air pollution levels to mortality in Auckland, New Zealand. We used urban airshed modelling and GIS-based techniques to quantify long-term exposure to ambient air pollution levels and associated mortality. After adjusting for age, sex, ethnicity, socio-economic status, and urban/rural domicile there was a 1.3% (95%CI: 1-1.5%) increase in non-external cause mortality, and 1.8% (95%CI: 1.5-2.1%) increase in circulatory and respiratory causes per 1 microg/m(3) increase in annual average NO(2). Based on these exposure-response relationships and applying an annual average threshold of 13 microg/m(3), the average annual (for 1996-1999) number of people estimated to die from non-external causes and circulatory and respiratory causes attributable to air pollution in Auckland is 268 (95% range: 227-310) (3.9% of total all cause deaths) and 203 (95% range: 169-237) (5.9% of total circulatory and respiratory deaths) per year, respectively. The number of attributable deaths found in this study are consistent with a previous New Zealand risk assessment using a different methodology, and is approximately twice the number of people dying from motor vehicle accidents in the region, which is on average (1996-1999) 103 per year. The GIS-based exposure maps identify high exposure areas for policy developers and planners in a simple and realistic manner. Taken together with overseas studies the study provides additional evidence that long-term exposure to poor air quality, even at levels below current standards, is a hazard to the public health.

Air Pollutants↗

Spatial analysis of grain size in Santa Monica Bay.

Maps are useful scientific tools for presenting environmental information, but the statistical techniques necessary to prepare scientifically rigorous maps have primarily focused on terrestrial habitats. This study compares three popular techniques (triangulation, kriging, and co-kriging) to map sediment grain size in Santa Monica Bay, California. Two grain size data sets, one collected in 1994 (79 sites) and one collected in 1997 and 1998 (149 sites) were used for model development. A bathymetric data set collected in 1997 was used as a model covariate. A third grain size data set (40 sites) collected in 1996 from independent sites was used for model evaluation. Predictions were compared to validation data by average difference, prediction mean square error (PMSE), and a goodness-of-prediction measure, G. The average difference between prediction and truth was similar for all methods, but the PMSE for triangulation was more than twice that for kriging or co-kriging, which were similar. The G measure also shows triangulation to be a far worse predictor than kriging and co-kriging. Small-scale differences were observed between kriging and co-kriging at steep depth contours, where co-kriging predicted values commensurate with the expected depth-defined grain size.

California↗

High-resolution spatial analysis of stomatal ozone uptake in arable crops and pastures.

Ozone effects on plants depend on atmospheric transport and stomatal uptake. Thus, ozone-risk assessments should use measured ozone concentrations and account for the influence of atmospheric conditions and soil moisture on stomatal and nonstomatal ozone deposition. This requires disaggregated data for the physical input parameters and species-specific data for specific stomatal conductance (g(s)). In this study, an approach was developed based on a resistance analogue transport model. This model requires interpolated routine-measuring data for ozone concentration at 3-5 m height, wind speed, precipitation, and soil moisture content as inputs to estimate the amount of ozone taken up by wheat (Triticum aestivum) and grass/clover pastures with a 1x1-km resolution. The model was applied to the area under agricultural production in Switzerland. Using data for June 1994, the calculations revealed that the median of the distribution of stomatal ozone uptake was 88% higher in wheat compared to grassland. This was mainly due to the higher maximum stomatal conductance in wheat. Because ozone flux to soil and to external plant surfaces was comparable in both vegetation types, the difference in the stomatal fluxes was mainly responsible for distinct differences in flux partitioning. In both cases, only about 11% of the total cumulative flux was absorbed by external plant surfaces, whereas the soil was a strong sink responsible for as much as 50% of the total flux into grasslands. The higher-ozone flux to wheat resulted in clearly lower-ozone concentrations at canopy height, but no significant correlation between cumulative canopy-level ozone exposure, expressed as accumulated exposure above 40 ppb (AOT40), and stomatal uptake was found. Thus, to estimate the ozone risk for crops using a flux-based approach may lead to results that differ substantially from those obtained with a concentration-based approach.

Ecosystem↗

Subject loss in spatial analysis of breast cancer.

Possible selection bias from assignment of latitude-longitude coordinates to the place of residence of all Connecticut women diagnosed with breast cancer from 1992 to 1995 (N = 11,470) was evaluated. Exact address-matching was accomplished for 8,121 records (70.8%) and an additional 1,722 records (15.0%) were matched using relaxed criteria. We did not address-match 1,627 records (14.1%) due to missing address information or limitations of the Geographic Information Systems street file. The age-adjusted likelihood of address-matching records was significantly greater for women of color, those born within Connecticut, residents of urban locales or census tracts with low median family incomes and those cases diagnosed nearer to 1992. Few differences in address-matching were attributable to tumor characteristics or therapeutic modality.

Adult↗

A spatial analysis of the blood-brain barrier damage in experimental allergic encephalomyelitis.

Experimental allergic encephalomyelitis was induced in young male Lewis rats. Following the development of neurological signs, the local distribution of perivascular inflammatory cellular infiltrates and the local blood-to-tissue transfer constants (K1) of alpha-aminoisobutyric acid (AIB) were determined, and these results were compared. Perivascular infiltrative lesions were generally found near areas of the CNS that normally lack an effective blood-brain barrier (BBB) such as the choroid plexus and the entry zones of the cranial and spinal nerve roots. This distribution pattern indicates that the entry of the causative agent into CNS tissue may be by way of the permeable microvessels of these structures. In tissue around inflamed veins, the mean transfer constant was slightly but significantly increased (2.8 +/- 1.5 microliter g-1 min-1) compared with uninvolved regions (0.9 +/- 0.2 microliter g-1 min-1) and similar areas in control animals (0.9 +/- 0.3 microliter g-1 min-1). Analysis of the autoradiographic method of determining transfer constants suggested that the AIB influx rate in the lesion areas may actually be manyfold larger than measured, that BBB permeability may be greatly increased at such sites, and that the areas of lymphocytic infiltration and increased K values may be virtually identical.

Aminoisobutyric Acids↗

The use of flash photolysis for a high-resolution temporal and spatial analysis of bacterial chemotactic behaviour: CheZ is not always necessary for chemotaxis.

The purpose of this work was to develop a high-resolution analysis of behaviour as an assay of the physiological consequences of mutations in the che genes and also to examine the role of CheZ in chemotaxis. Recent advances in flash photolysis have made it possible to expose cells to an unstable chemical gradient created by a square-wave increase in attractant concentration. The response of individual cells can be tracked in the order of milliseconds using real-time motion analysis. The tumble frequency of wild-type Escherichia coli exposed to photoreleased aspartate falls very quickly to smooth-swimming levels, and the swimming speed of these cells rises. As a consequence of these behavioural changes, there is an increase in the number of bacteria present in the centre of the flashed area, that is the bacteria's response to the transient gradient generated by flash photolysis was to swim into the centre of the flash area. This allowed the rapid quantitative measurement of chemotaxis. Deletion of various che genes resulted in predictable changes in chemotactic behaviour. cheZ null mutants are non-chemotactic when measured by classical techniques but demonstrate a definite chemotactic response to photoreleased attractant.

Bacterial Proteins↗