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PubMed1994

Gene targeting and development of the nervous system.

Gene targeting provides a means of directly assaying the function of specific genes during mouse nervous system development. Generation of targeted mutant mice has provided the first evidence of developmental roles for genes whose function was suggested based on their expression, but for which appropriate assay systems were lacking. In other cases, where gene function was known, targeted mutations have revealed in which cell population, and at what developmental stage, particular genes are first indispensable. The existing targeted mutants suggest that an early mechanism of pattern formation in mammals involves regional control of proliferation or survival of neural precursors, and that later general functions, such as the control of differentiation of precursors, may be performed by different genes in distinct neural lineages. As many genes display complex temporal and spatial patterns of expression, analysis of the full range of functions of such genes will require the generation of a series of alleles, including stage- and tissue-specific mutations.

Animals↗
PubMed2003

Receptor-regulated dynamic interaction between endothelial nitric oxide synthase and calmodulin revealed by fluorescence resonance energy transfer in living cells.

The endothelial isoform of nitric oxide synthase (eNOS), a key regulator of vascular tone, is activated in endothelial cells by diverse Ca(2+)-mobilizing agonists, including vascular endothelial growth factor (VEGF). Although the activation state of eNOS and the subcellular localization of the enzyme are both highly regulated, the relationship between enzyme activity and subcellular targeting remains obscure. We aim here to elucidate this relationship by direct dynamic imaging analysis of Ca(2+)/CaM-dependent eNOS activation in living endothelial cells, using high-resolution confocal microscopy and donor dequenching fluorescence resonance energy transfer (FRET) techniques. Confocal images show a complex pattern of eNOS subcellular distribution; the enzyme is concentrated in both the plasma membrane and internal membranes, with robust expression in the perinuclear region. We construct a fusion protein between eNOS and the FRET-based calcium sensor cameleon, and analyze the temporal and spatial pattern of VEGF-mediated calcium mobilization using donor dequenching FRET methods. We find that VEGF promotes rapid mobilization of intracellular calcium throughout the regions of the cell in which eNOS is distributed. We further create a series of fusion proteins and use FRET imaging methods to study the interactions between eNOS and its obligate allosteric activator protein calmodulin. We clone the FRET acceptor EYFP (enhanced yellow fluorescent protein) at the C-terminus of calmodulin, and the FRET donor ECFP (enhanced cyan fluorescent protein) into eNOS at a site adjacent to its calmodulin-binding domain. FRET imaging analysis of individual endothelial cells cotransfected with eNOS-ECFP and calmodulin-EYFP shows that VEGF induces interactions between eNOS and calmodulin wherever both are present in the cell. Our studies provide evidence that the pool of rapidly responsive receptor-activated eNOS is distributed throughout endothelial cells in both plasma membrane and internal membrane structures, and that this distribution parallels the localization of agonist-induced intracellular Ca(2+) changes in the vicinity of eNOS.

Animals↗
PubMed1996

The large-scale vortex structures in plasma-like media and the electric explosion of conductors.

Formation of large-scale hydrodynamic convective patterns in plasma-like current-carrying media is considered. This process is shown to be described by the same equations, as Benard rolls, except that a temperature field must be replaced by a magnetic field. A simple low-mode model of spatial pattern formation for a case of cylindrical liquid-metal conductor with current is proposed and investigated. Nonlinear interaction of perturbations of the magnetic field and the velocity field results in an increase of effective conductor resistance even when transport coefficients are constant. In our opinion, it is this instability, that is of first importance at the initial stages of the electric explosion of conductors. In particular, it leads to conductor stratification and electric current interruption. (c) 1996 American Institute of Physics.

Journal Article↗
PubMed2004

Spatial correlations of mapped malaria rates with environmental factors in Belize, Central America.

BACKGROUND: The purposes of this study were to map overall malaria incidence rates from 1989 through 1999 for villages in Belize; to assess the seasonal distribution of malaria incidence by region; and to correlate malaria incidence rates with vegetation cover and rivers in villages, using geographic information system technology.Malaria information on 156 villages was obtained from an electronic database maintained by the Belize National Malaria Control Program. Average annual malaria incidence rates per 1000 population over 10 years were calculated for villages using the 1991 population census as a denominator. Malaria incidence rates were integrated with vegetation cover from a 1995 vegetation map, and with river data from a digital data set. RESULTS: Mapping malaria incidence over the 10-year period in the study villages indicated the existence of a spatial pattern: the southern and western areas of Belize had consistently higher rates of malaria than northern areas. Examination of the seasonal distribution of malaria incidence by month over 10 years indicated that a statistically significant difference existed among districts and among months (p < 0.05). Spatial analysis of malaria incidence rates and of vegetation in Belize showed villages with high malaria rates having more broadleaf hill forests, agricultural land, and wetland vegetation types (i.e. SWF-seasonally waterlogged fire-induced shrubland of the plains). Statistical and spatial analyses of malaria incidence and of river distributions in Belize determined the high 10 percentile malaria incidence villages in western and southern Belize to have more rivers within two kilometers of the center of a village and a statistically significant correlation between proximity to rivers and villages (Spearman's gamma = -0.23; p < 0.05), especially in Stann Creek District (Spearman's gamma = -0.82; p < 0.05). CONCLUSIONS: Examination of the distribution of malaria during 10 years indicated transmission varied among geographic areas and among seasons. Additional studies are needed to examine, in more detail, the association between environmental and meteorological factors and malaria transmission. Furthermore, the role of An. darlingi in malaria transmission in Stann Creek needs further study since, of the three main vectors in Belize, An. darlingi has been found strongly associated with rivers.

Journal Article↗
PubMed1998

Unique expression pattern of a novel mosaic receptor in the developing cerebral cortex.

Recently, a new type of transmembrane protein with a unique combination of protein domains was characterized from human, rabbit and chicken. This protein exhibits features of the low-density lipoprotein receptor family and shows homology to the receptor of the neuropeptide head activator isolated from hydra. To study the temporal and spatial pattern of expression of this unusual new receptor we have isolated a murine homolog and, in accordance with its human counterpart, named it mSorLA. Northern blot analysis revealed the highest abundance of mSorLA transcripts in the adult brain, lower levels in a variety of other organs and expression during embryogenesis. In situ hybridization showed predominant localization in neurons of the cortex, the hippocampus and the cerebellum. During embryonic development mSorLA displayed a unique pattern of expression in the cerebral cortex, where a subpopulation of neurons was labeled before final differentiation. Transcripts of mSorLA were also detected outside the central nervous system in regions active in morphogenesis.

Animals↗
PubMed2006

Sex work in Tallinn, Estonia: the sociospatial penetration of sex work into society.

BACKGROUND: It is important to describe and understand the underlying patterns and dynamics that govern sex work in societies undergoing rapid political and social changes, its heterogeneity across populations, and its evolution through time in order to inform future research, sound policy formation, and programme delivery. OBJECTIVES: To describe the socioeconomic and cultural determinants, organisational structure, distinct categories, and spatial patterning of sex work in Tallinn, Estonia, and identify recent temporal changes in sex work patterns. METHODS: In-depth interviews with key informants; naturalistic observations of sex work and drug use venues, geo-mapping of sex work sites, review of media, public policy, and commissioned reports, and analyses of existing data. RESULTS: Sex work takes place in a hierarchy of locations in Tallinn ranging from elite brothels and "love flats" to truck stops. These sites vary in terms of their public health importance and social organisation. There are full time, part time, and intermittent male and female sex workers. Among others, the taxi driver, madam and the bartender are central roles in the organisation of sex work in Tallinn. Cell phone and internet technology enable sex work to be highly dispersed and spatially mobile. CONCLUSION: Future research and programmatic service delivery or outreach efforts should respond to the changing profile of sex work in Tallinn and its implications for STD/HIV epidemiology.

Adolescent↗
PubMed1996

Expression of Hoxb genes in the developing mouse foregut and lung.

Lung development in the mouse begins at embryonic day 9.5 (E9.5) when lung buds form in the foregut. Subsequently, there is extensive branching and cellular differentiation that depends upon specific epithelial-mesenchymal interactions. Homeobox genes are expressed in specific temporo-spatial patterns in the developing embryo and are known to be involved in axial patterning and specification of regional identity. Using whole mount in situ hybridization and immunohistochemistry, we studied the expression of Hoxb-1, b-2, b-3, b-4 and b-5 in the E9.5-E14.5 foreguts and lungs. Our results show that in E9.5 branchial arches and foregut, Hoxb genes are expressed in overlapping spatial domains and the anterior boundaries of these domains correspond to the position of a particular gene in the cluster-genes on the 3' end of the cluster are expressed more anteriorly in the branchial arches and foregut and those on the 5' end are expressed more posteriorly. Three of the genes, Hoxb-3, b-4, and b-5, are highly expressed in the foregut where the lung buds form. In contrast, in E10.5-E14.5 lung, there are two patterns of Hoxb gene expression. Hoxb-3 and b-4 are expressed in the mesenchyme of the trachea, mainstem bronchi, and distal lung, whereas Hoxb-2 and b-5 mRNA are present only in the mesenchyme of the distal lung buds. These results suggest that specific combinations of Hoxb gene expression are important in lung development and that Hoxb genes may be involved in specifying the differences between proximal (trachea and main bronchi) and distal (lung bud) mesenchyme.

Animals↗
PubMed1994

Meristem-specific gene expression directed by the promoter of the S-phase-specific gene, cyc07, in transgenic Arabidopsis.

A genomic clone for the cyc07 gene, which is expressed specifically at the S phase during the cell cycle in synchronous cultures of periwinkle (Catharanthus roseus) cells, was isolated. Determination of the nucleotide sequence of the clone revealed that the cyc07 gene consists of seven exons separated by six introns. Genomic Southern analysis indicated that the cyc07 gene is present as a single copy per haploid genome in periwinkle. Expression of related genes was detected in a wide range of other plants. Transgenic Arabidopsis plants were generated that expressed the gene for beta-glucuronidase (GUS) under the control of the promoter of the cyc07 gene. The tissue-specific pattern of expression directed by the promoter was investigated by analysis of GUS activity. Histochemical tests demonstrated that 589 bp of the 5'-upstream sequence of the cyc07 gene could direct specifical expression of the GUS reporter gene in meristematic tissues in transgenic plants. The spatial pattern of expression directed by the promoter was closely correlated with meristematic activity and cell proliferation, suggesting an association between the function of the cyc07 gene and cell proliferation.

Amino Acid Sequence↗
PubMed2004

On 'Analytical models for the patchy spread of plant disease'.

Epidemiologists are interested in using models that incorporate the effects of clustering in the spatial pattern of disease on epidemic dynamics. Bolker (1999, Bull. Math. Biol. 61, 849-874) has developed an approach to study such models based on a moment closure assumption. We show that the assumption works above a threshold initial level of disease that depends on the spatial dispersal of the pathogen. We test an alternative assumption and show that it does not have this limitation. We examine the relation between lattice and continuous-medium implementations of the approach.

Disease Outbreaks↗
PubMed2002

A Bayesian model for spatial wildlife disease prevalence data.

The analysis of the geographical distribution of disease on the scale of geographic areas such as administrative boundaries plays an important role in veterinary epidemiology. Prevalence estimates of wildlife population surveys are often based on regional count data generated by sampling animals shot by hunters. The observed disease rate per spatial unit is not an useful estimate of the underlying disease prevalence due to different sample sizes and spatial dependencies between neighbouring areas. Therefore, it is necessary to account for extra-sample variation and spatial correlations in the data to produce more accurate maps of disease incidence. The detection of spatial patterns is complicated by missing data in many of the geographical areas as the complete coverage of all areas is nearly impossible in wildlife surveys. For this purpose a hierarchical Bayesian model in which structured and unstructured over dispersion is modelled explicitly in terms of spatial and non-spatial components was implemented by Markov chain Monte Carlo methods. The model was empirically compared with the results of a non-spatial beta-binomial model using surveillance data of pseudorabies virus infections of European wild boars (Sus scrofa scrofa L.) in the Federal State of Brandenburg, Germany.

Animal Diseases↗
PubMedDate not supplied

Phenotypic and dynamical transitions in model genetic networks. I. Emergence of patterns and genotype-phenotype relationships.

Genotype-phenotype interactions during the evolution of form in multicellular organisms is a complex problem but one that can be aided by computational approaches. We present here a framework within which developmental patterns and their underlying genetic networks can be simulated. Gene networks were chosen to reflect realistic regulatory circuits, including positive and negative feedback control, and the exchange of a subset of gene products between cells, or within a syncytium. Some of these networks generate stable spatial patterns of a subset of their molecular constituents, and can be assigned to categories (e.g., "emergent" or "hierarchic") based on the topology of molecular circuitry. These categories roughly correspond to what has been discussed in the literature as "self-organizing" and "programmed" processes of development. The capability of such networks to form patterns of repeating stripes was studied in network ensembles in which parameters of gene-gene interaction were caused to vary in a manner analogous to genetic mutation. The evolution under mutational change of individual representative networks of each category was also simulated. We have found that patterns with few stripes (< or =3) are most likely to originate in the form of a hierarchic network, whereas those with greater numbers of stripes (> or =4) originate most readily as emergent networks. However, regardless of how many stripes it contains, once a pattern is established, there appears to be an evolutionary tendency for emergent mechanisms to be replaced by hierarchic mechanisms. These results have potential significance for the understanding of genotype-phenotype relationships in the evolution of metazoan form.

Biological Evolution↗
PubMed1996

A modular misexpression screen in Drosophila detecting tissue-specific phenotypes.

Genetic screens in Drosophila have lead to the discovery of many genes important for patterning and signal transduction in diverse organisms. Traditionally, the phenotypic effects of loss-of-function mutations are analyzed. As an alternative way to link genes and function, I have developed a versatile misexpression screen in Drosophila, the first such screen in higher eukaryotes. The screen identifies genes that, when over- or misexpressed in a pattern of interest, give a specific phenotype or modulate an existing mutant phenotype. It is based on Gal4 transactivation of a mobile enhancer and promoter that "targets" random endogenous genes for expression. The modular design of the screen allows directed expression in any temporal or spatial pattern. When activated in the developing eye, 4% of target inserts gave dominant phenotypes. One insertion was in the gene encoding Ras GTPase-activating protein; its overexpression phenotype was strongly enhanced by a mutation in Ras1. Thus, biologically relevant phenotypes and genetic interactions are identified using this method. The screen is a powerful new tool for developmental genetics; similar approaches can also be developed for other organisms.

Animals↗
PubMed2005

Estimation of leaf water potential by thermal imagery and spatial analysis.

Canopy temperature has long been recognized as an indicator of plant water status and as a potential tool for irrigation scheduling. In the present study, the potential of using thermal images for an in-field estimation of the water status of cotton under a range of irrigation regimes was investigated. Thermal images were taken with a radiometric infrared video camera. Specific leaves that appeared in the camera field of view were sampled, their LWP was measured and their temperature was calculated from the images. Regression models were built in order to predict LWP according to the crop canopy temperature and to the empirical formulation of the crop water stress index (CWSI). Statistical analysis revealed that the relationship between CWSI and LWP was more stable and had slightly higher correlation coefficients than that between canopy temperature and LWP. The regression models of LWP against CWSI and against leaf temperatures were used to create LWP maps. The classified LWP maps showed that there was spatial variability in each treatment, some of which may be attributed to the difference between sunlit and shaded leaves. The distribution of LWP in the maps showed that irrigation treatments were better distinguished from each other when the maps were calculated from CWSI than from leaf temperature alone. Furthermore, the inclusion of the spatial pattern in the classification enhanced the differences between the treatments and was better matched to irrigation amounts. Optimal determination of the water status from thermal images should be based on an overall view of the physical status as well as on the analysis of the spatial structure. Future study will involve investigating the robustness of the models and the potential of using water status maps, derived from aerial thermal images, for irrigation scheduling and variable management in commercial fields.

Agriculture↗
PubMed1986

Pattern sensitivity to boundary and initial conditions in reaction-diffusion models.

We consider Turing-type reaction-diffusion equations and study (via computer simulations) how the relationship between initial conditions and the asymptotic steady state solutions varies as a function of the boundary conditions. The results indicate that boundary conditions which are nonhomogeneous with respect to the kinetic steady state give rise to spatial patterns which are much less sensitive to variations in the initial conditions than those obtained with homogeneous boundary conditions, such as zero flux conditions. We also compare linear pattern predictions with the numerical solutions of the full nonlinear problem.

Animals↗
PubMedDate not supplied

The spatial relationship between infant mortality and birth defect rates in a U.S. city.

The spatial patterns of infant mortality and birth defect rates in the Des Moines, Iowa, urban region are described as a contoured surface based on the punctual kriging of address-matched vital statistics records from The Iowa Department of Public Health. Areas defined as having high rates are shown to be sensitive to the size of the spatial filtering units. There is no correlation between infant mortality and birth defect rates in the region. The significance of areas with high rates is determined by a Monte Carlo simulation procedure. One area of high infant mortality is found in the region, which contrasts with many smaller areas with high birth defect rates in the region. The observed birth defect rate pattern is consistent with the hypothesis that each birth was equally likely to result in a birth defect, while the infant mortality pattern is unlikely to be the result of such an equal likelihood process.

Congenital Abnormalities↗
PubMed1993

Cell-autonomous expression and position-dependent repression by Li+ of two zygotic genes during sea urchin early development.

The expression of two zygotic genes (HE and BP10) during sea urchin embryogenesis was previously found to be early, transient, spatially restricted and controlled at the transcriptional level. Here we studied how the expression of these genes is affected when cell interactions are abolished by dissociating blastomeres and when development is perturbed by treatment with Li+. We found that in isolated blastomeres, transient transcriptional activity (HE) is unchanged and both genes apparently function in the appropriate cell type. Thus HE/BP10 expression is largely cell-autonomous and should rely on maternal factors unevenly distributed in the egg. Treatment with lithium does not affect the temporal control but decreases the transcriptional activity and the size of the domain of expression of the HE/BP10 genes. As the Li+ concentration increases, the border of the domain is progressively shifted towards the animal pole. This alteration of the spatial pattern is the earliest molecular evidence of a change in cell fate detectable only much later by morphological criteria, and reveals a gradient of sensitivity to Li+ along the animal--vegetal axis. These results suggest that the activity of the HE/BP10 genes is strongly dependent on spatially organized maternal information controlling early development.

Animals↗
PubMed2003

Difference in the retinal cone mosaic pattern between zebrafish and medaka: cell-rearrangement model.

In fish retina, four kinds of photoreceptor cells (or cones) are two-dimensionally arranged in a very regular manner, forming cone mosaics. Mosaic pattern differs between species--two typical patterns are "row mosaic" and "square mosaic", exemplified by the cone mosaics in zebrafish and in medaka, respectively. In this paper, we study a cell-rearrangement model. Cells with pre-fixed fate exchange their locations between nearest neighbors and form regular mosaic patterns spontaneously, if the adhesive force between nearest neighbors and between next-nearest neighbors depend on their cell types in an appropriate manner. The same model can produce both row and square mosaic patterns. However, if the cell-cell interaction is restricted to nearest neighbors only, the square mosaic (medaka pattern) cannot be generated, showing the importance of interaction between next-nearest neighbors. In determining whether row mosaic (zebrafish pattern) or square mosaic (medaka pattern) is to be formed, two shape factors are very important, which control the way adhesions in different geometric relations are combined. We also developed theoretical analysis of the parameter ranges for the row mosaic and the square mosaic to have higher total adhesion than alternative spatial patterns.

Animals↗
PubMed1992

Spatiotemporal changes in Ha-ras p21 expression through the hepatocyte cell cycle during liver regeneration.

The protein product of the ras oncogene, Ha-ras (p21), is thought to be an important regulator of cell growth. The cytoplasmic relocalization of p21 in the cell during the cell cycle suggests a transient signaling role for this protein in association with its signal transduction function. Because of the importance of this role we examined spatial patterns in vivo of p21 expression at the protein and mRNA levels in hepatocytes during compensatory growth in rat liver following partial hepatectomy. A low level of p21 was immunolocalized on the cytoplasmic membrane of nonregenerating hepatocytes. The level of hepatic p21 increased significantly and without spatial restriction within the liver from 36 to 60 hr after partial hepatectomy (PH). p21 was localized in the cytoplasm of dividing hepatocytes and on the hepatic cytoplasmic membrane. The elevated p21 level decreased and was found mainly on hepatocyte plasma membranes by 96 hr after PH. Immunogold electron microscopy showed p21 localized over mitochondrial membranes and nuclei in nondividing regenerating hepatocytes. Approximately 50% of nonregenerating hepatocytes show nuclear localization of p21. This percentage changes with time following PH. The decrease in nuclear localization was accompanied with an increase in the low number of hepatocytes which demonstrated cytoplasmic localization in nondividing hepatocytes in regenerating liver. Flow cytometric analysis revealed a significant increase of p21 at 36 hr after PH which was 12 hr after the initial induction of ras mRNA. ras mRNA level increased 1.5-fold at 24 hr after PH and a maximum twofold induction was observed at 48 hr. Cell-cycle analysis of regenerating hepatocytes indicated a synchronized first peak of cell division 36-40 hr after PH. Dual parameter flow cytometry revealed that the level of p21 in hepatocytes in S phase and G2/M phase of the cell cycle was significantly higher than that in G0/G1 phase during regeneration. These findings suggest that p21 is important for the progression of regenerating hepatocytes to S phase and then to G2/M phase.

Amino Acid Sequence↗