EPHA2 Is Associated with Age-Related Cortical Cataract in Mice and Humans

Age-related cataract is a major cause of blindness worldwide, and cortical cataract is the second most prevalent type of age-related cataract. Although a significant fraction of age-related cataract is heritable, the genetic basis remains to be elucidated. We report that homozygous deletion of Epha2 in two independent strains of mice developed progressive cortical cataract. Retroillumination revealed development of cortical vacuoles at one month of age; visible cataract appeared around three months, which progressed to mature cataract by six months. EPHA2 protein expression in the lens is spatially and temporally regulated. It is low in anterior epithelial cells, upregulated as the cells enter differentiation at the equator, strongly expressed in the cortical fiber cells, but absent in the nuclei. Deletion of Epha2 caused a significant increase in the expression of HSP25 (murine homologue of human HSP27) before the onset of cataract. The overexpressed HSP25 was in an underphosphorylated form, indicating excessive cellular stress and protein misfolding. The orthologous human EPHA2 gene on chromosome 1p36 was tested in three independent worldwide Caucasian populations for allelic association with cortical cataract. Common variants in EPHA2 were found that showed significant association with cortical cataract, and rs6678616 was the most significant in meta-analyses. In addition, we sequenced exons of EPHA2 in linked families and identified a new missense mutation, Arg721Gln, in the protein kinase domain that significantly alters EPHA2 functions in cellular and biochemical assays. Thus, converging evidence from humans and mice suggests that EPHA2 is important in maintaining lens clarity with age.     

Melatonin and melanocyte functions

The effect of melatonin on melanocyte functions was studied by incubating whole-skin organ cultures with melatonin, as well as by assessing melatonin positivity in melanocytes versus dendricity and pigmentation, when arrested in the G(2) phase. From this study, it was observed that melatonin positivity is inversely related to the length of UV exposure. Increasing melatonin levels are related to decreasing dendricity and pigment donation during photoresponse in the G(2) phase. Melanocyte melatonin positivity increases with dark incubation and is higher with a pulse of UV exposure after dark incubation with melatonin. This increase is associated with a doubling of melanocyte number after dark incubation and a further doubling upon exposure to a pulse of UV. The melanocytes directly take up melatonin, which results in a marked increase in their numbers. Thus, extreme caution should be exercised when using melatonin as an anticancer drug. This finding also simulates the melanocyte repopulation of the skin with repigmentation during summer in polar animals.     

Identification of Microsatellite Repeats in Turbot (Scophthalmus maximus) and Dover Sole (Solea solea) using a RAPD-Based Technique: Characterization of Microsatellite Markers in Dover Sole

We have used a RAPD-based technique to identify several microsatellite repeats in turbot (Scophthalmus maximus) and Dover sole (Solea solea) and report the characterization of six novel polymorphic microsatellite markers for Dover sole. These are the first such markers to be developed for this flatfish species. They exhibit much higher levels of heterozygosity than those previously observed with allozyme loci and should prove useful in addressing population genetic questions as well as more fundamental aquaculture-related questions. Received ; accepted July 13, 1999.     

AVIS: AJAX viewer of interactive signaling networks

MOTIVATION: Increasing complexity of cell signaling network maps requires sophisticated visualization technologies. Simple web-based visualization tools can allow for improved data presentation and collaboration. Researchers studying cell signaling would benefit from having the ability to embed dynamic cell signaling maps in web pages. SUMMARY: AVIS is a Google gadget compatible web-based viewer of interactive cell signaling networks. AVIS is an implementation of AJAX (Asynchronous JavaScript with XML) with the usage of the libraries GraphViz, ImageMagic (PerlMagic) and overLib. AVIS provides web-based visualization of text-based signaling networks with dynamical zooming, panning and linking capabilities. AVIS is a cross-platform web-based tool that can be used to visualize network maps as embedded objects in any web page. AVIS was implemented for visualization of PathwayGenerator, a tool that displays over 4000 automatically generated mammalian cell signaling maps; NodeNeighborhood a tool to visualize first and second interacting neighbors of yeast and mammalian proteins; and for Genes2Networks, a tool to connect lists of genes and protein using background protein interaction networks. AVAILABILITY: A demo page of AVIS and links to applications and distributions can be found at http://actin.pharm.mssm.edu/AVIS2. Detailed instructions for using and configuring AVIS can be found in the user manual at http://actin.pharm.mssm.edu/AVIS2/manual.pdf.     

The distribution of long range admixture linkage disequilibrium in an African-American population

OBJECTIVES: To better understand the effect of admixture on long range linkage disequilibrium (LD), we characterized extended LD in gene-rich regions of an African-American population. METHODS: Approximately 290 cM of chromosomes 1, 3, 6, 11-17, 20 and 22 were scanned using 109 polymorphic microsatellite markers spaced an average of 3 cM apart. Disequilibrium between loci (D') was based on maximum-likelihood estimates of haplotype frequencies computed for 200 unrelated African Americans. RESULTS: Mean D' values were highest on chromosomes 6p23-p21.3 (D' = 0.33) and 15p22.2-p25.3 (D' = 0.34), and lowest on chromosome 12p11.2-q14 (D' = 0.21). Overall, the variance in LD among chromosomes accounted for approximately two-thirds of the total LD variance. Of the 434 locus pairs spaced between 0.3 and 38.7 cM apart, there was no detectable correlation between LD and recombination distance and a weak negative correlation between LD and physical distance (r(s) = -0.12; p = 0.031). For the 192 intrachromosomal locus pairs where allele frequency data were available from the Centre d'Etude du Polymorphisme humain (CEPH), we found a statistically significant positive correlation between LD and the allelic frequency differences (delta) between the African-American study population and Caucasian reference CEPH population (r(s) = 0.53; p < 0.0001). The correlation between LD and both recombination and physical distance was markedly increased for locus pairs with high delta levels. CONCLUSIONS: Our results suggest that recent Caucasian admixture maintains a high level of long range LD in African Americans on a genomic scale, and selected markers with large African American/Caucasian delta levels may be useful in association studies.     

Effects of natural and anthropogenic gradients on native and exotic winter annuals in a southern California Desert

Native annual plant species constitute a large proportion of the plant diversity found in arid vegetation types within the southwestern United States; yet, little is known about controls on diversity patterns along natural and anthropogenic gradients. In this study we evaluated native species richness and exotic species cover across overlapping gradients of precipitation, wind, and N deposition in the Colorado Desert of southern California. Factors allowing native diversity to persist under high N deposition and high wind were also evaluated in a second, focused study at one end of the gradient. We found that gradients in precipitation, nitrogen deposition, and wind were the most important factors to native richness and exotic species cover across the landscape, while local heterogeneity in bare ground influenced richness and cover at the high deposition/windy, or high-disturbance, end of the gradient. Patterns of native diversity were evaluated across the gradients using non-metric multidimensional scaling, which showed diversity was split into two axes: one strongly correlated to precipitation and the other strongly correlated with disturbance factors. The disturbance factors were also positively associated with exotic grass and forb cover. In total, these results indicate that large-scale patterns in disturbance and exotic species cover negatively affect native annual plant species diversity but native species can also persist due to local heterogeneity.     

Rhamnocitrin-3-glucuronid in Verbesina myricephala

The structure of a new flavonol-glycoside isolated from the herb of Verbesina myricephala Sch. Bip. has been established as the 3-O-β-d-glucopyranuronide of 3,5,4′-trihydroxy-7-methoxy-flavone (rhamnocitrin-3- glucuronide.     

Actin-Dependent Receptor Colocalization Required for Human Immunodeficiency Virus Entry into Host Cells

Human immunodeficiency virus (HIV) envelope binds CD4 and a chemokine receptor in sequence, releasing hydrophobic viral gp41 residues into the target membrane. HIV entry required actin-dependent concentration of coreceptors, which could be disrupted by cytochalasin D (CytoD) without an effect on cell viability or mitosis. Pretreatment of peripheral blood mononuclear cells, but not virus, inhibited entry and infection. Immunofluorescent confocal microscopy of activated cells revealed CD4 and CXCR4 in nonoverlapping patterns. Addition of gp120 caused polarized cocapping of both molecules with subsequent pseudopod formation, while CytoD pretreatment blocked these membrane changes completely.     

Control of brown dog tick,Rhipicephalus sanguineus using assembly pheromone encapsulated in natural polymer,chitosan

In the current study, an attempt was made to encapsulate assembly pheromone using natural polymer, chitosan. Chitosan beads were prepared by incorporating assembly pheromone in conjunction with an acaricide, namely, deltamethrin. In the in vitro bioassay, the test beads attracted and killed 79 % of unfed larvae, 88 % of unfed nymphs and 61 % of unfed adults of the brown dog tick, Rhipicephalus sanguineus, in 24 h of exposure. Field trials were carried out to attract and kill the pre-parasitic environmental stages. The beads were dispersed onto specially designed devices and they were placed in infested kennels. The devices were observed after 10 days.     

A whole-genome screen of a quantitative trait of age-related maculopathy in sibships from the Beaver Dam Eye Study

Age-related maculopathy (ARM) is a leading cause of visual impairment among the elderly in Western populations. To identify ARM-susceptibility loci, we genotyped a subset of subjects from the Beaver Dam (WI) Eye Study and performed a model-free genomewide linkage analysis for markers linked to a quantitative measure of ARM. We initially genotyped 345 autosomal markers in 325 individuals (N=263 sib pairs) from 102 pedigrees. Ten regions suggestive of linkage with ARM were observed on chromosomes 3, 5, 6, 12, 15, and 16. Prior to fine mapping, the most significant regions were an 18-cM region on chromosome 12, near D12S1300 (P=.0159); a region on chromosome 3, near D3S1763, with a P value of.0062; and a 6-cM region on chromosome 16, near D16S769, with a P value of.0086. After expanding our analysis to include 25 additional fine-mapping markers, we found that a 14-cM region on chromosome 12, near D12S346 (located at 106.89 cM), showed the strongest indication of linkage, with a P value of.004. Three other regions, on chromosomes 5, 6, and 15, that were nominally significant at P< or =.01 are also appropriate for fine mapping.