Design, development and characterization of cyst phantom for ultrasound elastography applications.

Polyacrylamide based tissue mimicking phantom with embedded cysts is designed and analyzed for better understanding of cyst elastograms. Cysts filled with different fluids of varying density and bulk moduli are embedded in the phantom. Characterization is done based on parameters measured from the ultrasound B mode and elastogram of the cystic lesions. Such a phantom can serve as tool for better understanding of the elastographic appearance of cysts. Thus simple and complex cysts can be easily distinguished. It can also be used to teach a complex procedure like ultrasound guided fine needle aspiration.

Prevalence of atypical chest pain descriptions in a population from the southern United States.

INTRODUCTION: The character of chest pain (CP) is a major factor determining triage and admission for patients presenting to the emergency department (ED). Previous studies have found atypical descriptions in as little as 10-15% of patients with true myocardial ischemic pain. Atypical descriptions may be more prevalent in the Deep South of the United States because of cultural differences in the semantic description of pain. METHODS: A retrospective study of patients presenting to the ED of a southern U.S. urban hospital with enzyme-documented myocardial infarction was conducted to determine the prevalence of atypical CP descriptions. A multivariate analysis of those patients with atypical pain descriptions was conducted to determine the independent demographic factors associated with these descriptions. RESULTS: In a total of 77 subjects (56% black; 44% white) meeting the study criteria, 43% were found to have atypical elements in the character of their CP descriptions. Only the black race demographic was found to be significantly correlated with the atypical descriptions. The use of the descriptive term "sharp" accounted for nearly half of the atypical presentations. CONCLUSION: Regional differences in the description of the character of CP may result in misleading portrayals of ischemic heart disease in southern U.S. populations. These differences are associated with a higher prevalence of atypical CP because of semantic distinctions, such as the use of the term "sharp" as a descriptor of acuity rather than character or quality.

Apextrin, a novel extracellular protein associated with larval ectoderm evolution in Heliocidaris erythrogramma.

During the evolution of direct development in the sea urchin Heliocidaris erythrogramma major modifications occurred, which allowed the precocious formation of adult-specific structures and led to a novel larval body that surrounds these structures. The HeET-1 gene was isolated in a differential screen for transcripts enriched in the early embryos of H. erythrogramma relative to those of its indirect-developing congener, H. tuberculata. HeET-1 was unique among the three genes found in that no homologous transcript was detected in H. tuberculata total embryonic RNA blots. To verify this apparently extreme differential expression of the HeET-1 genes in Heliocidaris, we isolated the HeET-1 homologue from H. tuberculata genomic DNA and used it to probe blots of poly(A)+ RNA prepared from H. tuberculata embryos. It is expressed in H. tuberculata embryos at levels undetectable by this technique. The predicted amino acid sequence of HeET-1 suggested that it encodes a novel secreted protein. To assess the function of HeET-1, we raised polyclonal antisera to the HeET-1-encoded protein. We find that it is present in eggs in a type of secretory vesicle and that this maternal pool is gradually secreted after fertilization. As cells acquire apical-basal polarity in the blastula the protein becomes localized to the apical extracellular matrix, leading us to name the protein apextrin. The apical extracellular localization of apextrin is maintained in the columnar cells of the larval ectoderm until their internalization at metamorphosis. Ingressing mesenchyme cells rapidly endocytose apextrin upon leaving the vegetal plate. Comparison with fibropellin III, an apical lamina component, suggests that apextrin is an extracellular protein that is in tighter association with the plasma membrane than is the hyalin layer or apical lamina. We propose that apextrin is involved in apical cell adhesion and that its high level of expression may represent an adaptive cooption necessary for strengthening the large H. erythrogramma embryo.

Sea urchin Hox genes: insights into the ancestral Hox cluster.

We describe the Hox cluster in the radially symmetric sea urchin and compare our findings to what is known from clusters in bilaterally symmetric animals. Several Hox genes from the direct-developing sea urchin Heliocidaris erythrogramma are described. CHEF gel analysis shows that the Hox genes are clustered on a < or = 300 kilobase (kb) fragment of DNA, and only a single cluster is present, as in lower chordates and other nonvertebrate metazoans. Phylogenetic analyses of sea urchin, amphioxus, Drosophila, and selected vertebrate Hox genes confirm that the H. erythrogramma genes, and others previously cloned from other sea urchins, belong to anterior, central, and posterior groups. Despite their radial body plan and lack of cephalization, echinoderms retain at least one of the anterior group Hox genes, an orthologue of Hox3. The structure of the echinoderm Hox cluster suggests that the ancestral deuterostome had a Hox cluster more similar to the current chordate cluster than was expected Sea urchins have at least three Abd-B type genes, suggesting that Abd-B expansion began before the radiation of deuterostomes.

A sea urchin homologue of ceh-19, an unusual homeobox-containing gene from a nematode.

When screening for homeobox-containing genes from the sea urchin Heliocidaris erythrogramma (He), we isolated an exon of a gene which appears to be a homologue of the homeobox-containing gene, ceh-19, of Caenorhabditis elegans (Ce). The predicted translation of the sea urchin sequence shows 77% identity and 92% similarity to the first 53 amino acids of the homeodomain of ceh-19. The ceh-19 gene exhibits an intron in an unusual location in the 3' end of the homeobox; the He gene shares this feature.

Evolution of the fibropellin gene family and patterns of fibropellin gene expression in sea urchin phylogeny.

This study documents evolutionary modifications in the expression patterns of the sea urchin EGF I and EGF III genes, which encode a family of extracellular matrix proteins, the fibropellins. We show that the sea urchin apical lamina, a macromolecular extracellular matrix that surrounds the sea urchin embryo and is made up of the fibropellins, has been conserved through at least 250 million years of echinoid evolution. The contribution of different fibropellin family members to this structure has, however, changed over the course of sea urchin phylogeny, and between two congeneric species that exhibit different developmental modes. Mapping the evolutionary history of the EGF genes on a cladogram of relationships among sea urchins reveals that EGF I is present in all echinoids examined, while EGF III appears to have arisen by duplication and divergence from EGF I during the radiation of a suborder of the camarodont sea urchins some 35-45 million years ago. Alterations in the temporal expression patterns of these genes as well as the loss of one of the two EGF I transcripts and encoded protein are coincident with the evolution of a direct-developing larval form in Heliocidaris erythrogramma. H. erythrogramma and its congener Heliocidaris tuberculata, which develops via a typical echinopluteus larva, shared a common ancestor about 10 million years ago. The differences in fibropellin representation within the apical lamina of the various taxa indicate that a homologous embryonic structure can undergo substantial changes in composition during its evolutionary history.

Protein-DNA interactions at putative regulatory regions of two coordinately expressed genes, msp130 and PM27, during skeletogenesis in sea urchin embryos.

Development of the primary mesenchyme cells (PMCs) of the sea urchin embryo, which give rise to the larval skeleton, involves the coordinate onset of expression of several structural genes. As part of an effort to identify cis-acting elements that might play a role in this regulatory event, co-regulated genes were examined by two approaches. First, they were compared for conserved sequence elements. Four conserved elements were found as a cluster in all three genes examined, suggesting a regulatory role. Second, as a test for potential function, the putative regulatory regions of two of these genes were examined for protein binding sites. DNase I protection and gel mobility shift assays were used to: 1) identify several nuclear protein binding sites in these regions, two of which correspond to conserved elements among the genes; 2) demonstrate that the developmental time of appearance of the proteins that interact with these sites corresponds to the time of activation of the genes; and 3) show that two of the conserved sequence elements shared by these genes compete for the same binding proteins. These data identify putative regulatory elements, whose specific roles in the coordinate regulation of PMC-expressed genes can now be addressed directly using appropriate transgenic expression constructs.

Fibropellins, products of an EGF repeat-containing gene, form a unique extracellular matrix structure that surrounds the sea urchin embryo.

The sea urchin SpEGF 1 gene belongs to a growing family of developmentally important genes which encode proteins that contain repeated epidermal growth factor-like motifs. To characterize the embryonic expression of the protein products of this gene from Strongylocentrotus purpuratus, we generated polyclonal antisera from SpEGF I fusion proteins. These antibodies recognize two glycoproteins of 145 and 185 kDa, which we have named fibropellins. These proteins are present in unfertilized oocytes and throughout early development. The fibropellins are stored in cytoplasmic vesicles in the oocyte and are released soon after fertilization in a distinct secretory event following the exocytosis of cortical granule contents. Following secretion the proteins are localized in the basal surface of the hyaline layer. At the blastula stage the fibropellins become organized into distinct fibers which form a mesh-like network over the surface of the embryo. During subsequent development to the pluteus larva stage this network increases in overall morphological complexity and becomes regionally distinct. The molecular weights of the fibropellins and their pattern of embryonic localization indicate that these proteins form a component of the hyaline layer previously described as the apical lamina.

Dependence of the bilayer to hexagonal phase transition on amphiphile chain length.

Several series of amphiphiles of increasing chain length were tested for their abilities to modify the L alpha-HII transition of dielaidoylphosphatidylethanolamine using differential scanning calorimetry. Acylcarnitines, alkyl sulfates, alkylsulfobetaines, and phosphatidylcholines, with chain lengths between about 6 and 12 carbon atoms, show an increasing capacity to raise the L alpha-HII phase transition temperature of phosphatidylethanolamine. This is ascribed to increased partitioning of the added amphiphile from water into the membrane as the chain length increases. Alkyl sulfates and alkyltrimethylammonium bromides have diminished capacities to raise the L alpha-HII transition temperature as the chain length is increased from 12 to 16. This is caused by an increase in the hydrophobic portion of the amphiphile leading to a change in the intrinsic radius of curvature and a decrease in the hydrocarbon packing constraints in the HII phase relative to the shorter chain amphiphiles. The L alpha-HII transition temperature of phosphatidylethanolamine with acylcarnitines of chain length 14-20 carbon atoms, alkylsulfobetaines above 14 carbon atoms, and phosphatidylcholines with acyl groups having above 10 carbon atoms is relatively insensitive to chain length. We suggest that this is caused by a balance between increasing hydrocarbon volume promoting the HII phase through decreased intrinsic radius of curvature and greater relief of hydrocarbon packing constraints vs greater intermolecular interactions favoring the more condensed L alpha phase. This latter effect is more important for amphiphiles with large headgroups which can pack more efficiently in the L alpha phase. The phosphatidylcholines show a gradual decrease in bilayer stabilization between 10 and 22 carbon atoms.(ABSTRACT TRUNCATED AT 250 WORDS)

A sea urchin gene encodes a polypeptide homologous to epidermal growth factor.

A sea urchin DNA clone complementary to an embryonic messenger RNA whose protein product bears striking homology to the epidermal growth factor family of proteins has been identified and characterized. The structure of the protein is similar to that of previously identified regulatory genes in Drosophila and Caenorhabditis. RNA gel blot hybridization showed a unique temporal pattern of expression of this gene during embryogenesis and transcript enrichment in the embryonic ectoderm. These results suggest that this member of the epidermal growth factor gene family plays a role in early development decisions in sea urchin embryos.