[Endovascular treatment strategies for distal entry tear of Stanford type B aortic dissection].

Currently, thoracic endovascular aortic repair (TEVAR) is the first-line treatment for patients with complicated Stanford type B aortic dissections. However, TEVAR does not occlude the distal entry tear of dissections, and blood flow persists in the false lumen. Dissections might progress in some patients. Studies showed that distal entry tear increased the possibility of late aortic events during follow-up. Thus, treatment of distal entry tear is necessary in some high-risk patients after TEVAR. In this article, the current treatment strategies of distal entry tear are summarized, which include PETTICOAT, STABILISE, covered stent, fenestrated and branched stent-grafts, false lumen embolization, vascular occluder, and Knickerbocker. However, the number of the cases of most approaches is so limited that the indications and effectiveness need to be further studied. Selecting the right treatment for the right patient is of great importance.

Characterization of the structure and transcriptional patterns of the gene encoding the late histone subtype H1-beta of the sea urchin Strongylocentrotus purpuratus.

We have cloned and characterized the gene encoding the late histone H1-beta subtype from the sea urchin Strongylocentrotus purpuratus. The gene contains all of the upstream sequence homologies previously seen in late H1-gamma genes. The expression of H1-beta mRNA is coordinated with that of H1-gamma mRNA, and like H1-gamma it is expressed in all adult somatic tissues tested.

Isolation, characterization, and expression of the gene encoding the late histone subtype H1-gamma of the sea urchin Strongylocentrotus purpuratus.

We cloned and characterized the gene encoding H1-gamma, a late histone subtype of the sea urchin species Strongylocentrotus purpuratus. The predicted primary sequence of H1-gamma is 216 amino acids in length and has a net charge of +70, which is high for a somatic H1 histone. The H1-gamma gene appears to be a unique sequence gene that is not tightly linked to the core histone genes. The 770-base-pair transcribed region of the H1-gamma gene is bordered on the 5' side by two previously described H1-specific sequence elements and on the 3' side by a hairpin loop structure and CAGA box sequences. We detected 3,900 stored maternal H1-gamma mRNA transcripts per egg. The number of H1-gamma transcripts per embryo rises by 9.5 h postfertilization, but the maximum rate of accumulation (4,300 molecules per min per embryo) occurs in the late-blastula-stage embryo between 14 and 21 h after fertilization. The number of H1-gamma mRNA molecules peaks 21 h after fertilization when there are 2.0 X 10(6) molecules per embryo (a 500-fold increase) and then decreases over the next 3.25 h to 1.3 million molecules per embryo. Between 24 and 82 h after fertilization the number of H1-gamma transcripts declines steadily (210 molecules per min per embryo) to reach approximately 5.4 X 10(5) H1-gamma mRNAs by 82 h postfertilization. Surprisingly, the number of late H1 mRNA molecules per embryo is greater than the number of late H2B mRNA molecules beginning at the early gastrula stage of development.

An embryonic enhancer determines the temporal activation of a sea urchin late H1 gene.

Normal development requires that individual genes be expressed in their correct temporal patterns, but the mechanisms regulating this process during early embryogenesis are poorly understood. We have studied the early and late sea urchin histone genes during embryogenesis to address the molecular mechanisms controlling temporal gene expression. By measuring the changes in expression of cloned H1-beta DNA constructs after microinjection into fertilized one-cell zygotes, we demonstrated that a highly conserved 30-base-pair segment of DNA between positions -288 and -317 (USE IV) is responsible for the transcriptional activation of this late histone gene at the late blastula stage. In this report, we demonstrate that an oligonucleotide corresponding to USE IV acts as an embryonic enhancer element capable of activating the simian virus 40 early promoter in a stage-specific manner. Using an in vivo competition assay and in vitro DNase I footprinting and mobility shift assays, we also identified a protein(s) that interacts with this enhancer. Results of the competition assay suggested that this factor acts to stimulate transcription of the H1-beta gene. The factor was found to be stored in mature eggs as well as in all embryonic stages examined. The mobility of the factor found in eggs, however, differed from that of the embryonic form, which suggested that posttranslational modification occurs after fertilization.

Tramtrack69 is positively and autonomously required for Drosophila photoreceptor development.

Cell-fate specification and cellular differentiation are tightly controlled by both positive and negative transcriptional factors during development. The Drosophila BTB/POZ (Bric-a-brac Tramtrack Broad complex/Pox virus and Zinc finger) domain-containing Tramtrack (Ttk) proteins have been previously shown to be transcriptional repressors and inhibitors of the neuronal fate of cells such as photoreceptors. Here we provide evidence that one of the Ttk proteins, Ttk69, also plays a positive and autonomous role in promoting or maintaining differentiation of photoreceptor neurons at the late stages of Drosophila eye development. Consistent with this notion, the Ttk69 protein, but not Ttk88, is expressed in all photoreceptor cells during pupal stage. Thus, Ttk69 appears to play a dual function by serving negative and positive regulatory roles at different stages of photoreceptor development.

Evolution of the hedgehog gene family.

Effective intercellular communication is an important feature in the development of multicellular organisms. Secreted hedgehog (hh) protein is essential for both long- and short-range cellular signaling required for body pattern formation in animals. In a molecular evolutionary study, we find that the vertebrate homologs of the Drosophila hh gene arose by two gene duplications: the first gave rise to Desert hh, whereas the second produced the Indian and Sonic hh genes. Both duplications occurred before the emergence of vertebrates and probably before the evolution of chordates. The amino-terminal fragment of the hh precursor, crucial in long- and short-range intercellular communication, evolves two to four times slower than the carboxyl-terminal fragment in both Drosophila hh and its vertebrate homologues, suggesting conservation of mechanism of hh action in animals. A majority of amino acid substitutions in the amino- and carboxyl-terminal fragments are conservative, but the carboxyl-terminal domain has undergone extensive insertion-deletion events while maintaining its autocleavage protease activity. Our results point to similarity of evolutionary constraints among sites of Drosophila and vertebrate hh homologs and suggest some future directions for understanding the role of hh genes in the evolution of developmental complexity in animals.

Repression of Drosophila photoreceptor cell fate through cooperative action of two transcriptional repressors Yan and Tramtrack.

The Drosophila yan gene encodes an E twenty-six (ETS) domain nuclear protein with a transcription repressor activity that can be downregulated through phosphorylation by mitogen-activated protein kinase (MAPK). Before photoreceptor precursor cells commit to a particular cell fate, Yan is required to maintain them in an undifferentiated state. We report here identification of tramtrack (ttk) mutations that act as dominant enhancers of yan. We show that ttk synergistically interacts with yan to inhibit the R7 photoreceptor cell fate. Since ttk products are nuclear proteins with zinc-finger DNA-binding motifs, yan and ttk represent two nuclear regulators essential for the control of cellular competence for neural differentiation. Reduction of either yan or ttk activity suppresses eye phenotypes of the kinase suppressor of ras (ksr) gene mutation, which is consistent with the involvement of yan and ttk in the Ras/MAPK pathway.

The N-terminal BTB/POZ domain and C-terminal sequences are essential for Tramtrack69 to specify cell fate in the developing Drosophila eye.

The BTB/POZ (broad complex Tramtrack bric-a-brac/Pox virus and zinc finger) domain is an evolutionarily conserved protein-protein interaction motif. Many BTB-containing proteins are transcriptional regulators involved in a wide range of developmental processes. However, the significance of the BTB domain in development has not been evaluated. Here we present evidence that overexpression of the Tramtrack69 (Ttk69) protein not only blocks neuronal photoreceptor differentiation but also promotes nonneuronal cone cell specification in early Drosophila eye development. We show that the BTB domain is essential for Ttk69 function and single amino acid changes in highly conserved residues in this domain abolish Ttk69 activity. Interestingly, the Ttk69 BTB can be substituted by the BTB of the human Bcl-6 protein, suggesting that BTB function has been conserved between Drosophila and humans. We found that the Ttk69 BTB domain is critical for mediating interaction with the Drosophila homolog of C-terminal-binding protein (dCtBP) in vitro, and dCtBP(-) mutations genetically interact with ttk69. Furthermore, the C-terminal region downstream of the DNA-binding zinc fingers is shown to be essential for Ttk69 function. A dCtBP consensus binding motif in the C terminus appears to contribute to Ttk69 activity, but it cannot be fully responsible for the function of the C terminus.

The Drosophila zfh-1 and zfh-2 genes encode novel proteins containing both zinc-finger and homeodomain motifs.

Two of the most common DNA-binding motifs found in eukaryotic transcriptional regulatory proteins are the homeodomain and the C2-H2 zinc finger. In Drosophila, homeodomain and zinc-finger proteins have been implicated in a wide variety of developmental processes. Until now, no proteins have been described in which both these DNA-binding motifs are present. We report here the isolation of genes encoding two such Drosophila proteins from a cDNA expression library. The product of the zfh-1 gene (zinc-finger homeodomain protein 1) contains one homeodomain and nine C2-H2 zinc fingers. The product of the zfh-2 gene possesses three homeodomains and sixteen C2-H2 zinc fingers. For zfh-1, antisera raised against nonoverlapping regions of the gene product all recognize a 145 kDa protein on protein immunoblots, suggesting that the different DNA-binding motifs are actually all present in the mature gene product. The novel arrangement of interspersed homeodomain and zinc-finger motifs in the primary sequences of the zfh-1 and zfh-2 gene products may signify an unusual mechanism of transcriptional regulation by these proteins.

The embryonic expression patterns of zfh-1 and zfh-2, two Drosophila genes encoding novel zinc-finger homeodomain proteins.

The zfh-1 and zfh-2 genes of D. melanogaster encode novel proteins containing both homeodomain and C2-H2 zinc-finger DNA-binding motifs. Antisera against these proteins were used to investigate their expression patterns during embryonic development. The zfh-1 gene is expressed in the mesoderm of early embryos and in a number of mesodermally-derived structures of late embryos, including the dorsal vessel, support cells of the gonads, and segment-specific arrays of adult muscle precursors. In addition, zfh-1 is expressed in the majority of identified motor neurons of the developing CNS. The mesodermal zfh-1 expression requires the products of the twist and snail genes. The zfh-2 gene displays a more limited expression pattern, largely restricted to the CNS of late embryos. Ubiquitous zfh-1 expression in transgenic flies bearing an hsp70-zfh-1 construct has specific developmental consequences, including embryonic CNS defects as well as adult eye and bristle abnormalities. The expression patterns of zfh-1 and zfh-2 suggest that both genes may be involved in Drosophila neurogenesis and that zfh-1 may have additional functions in mesoderm development.

Executive system dysfunction in the aged monkey: spatial and object reversal learning.

As part of the effort to characterize age-related cognitive changes in executive system function in a nonhuman primate model of human aging, the performance of seven rhesus monkeys, 20 to 28 years of age, was compared to that of five young adult monkeys, 6 to 11 years of age, on spatial and object reversal tasks. No differences in performance were found between the two groups in the initial learning of either task. On spatial reversals, aged monkeys were impaired relative to young adults, but there was no difference in overall performance between the groups on object reversals. Central to this article, a perseverative tendency was noted in the aged group on both spatial and object reversal tasks. Changes in executive system dysfunction may represent an important aspect of age-related cognitive decline.

Recognition memory span in rhesus monkeys of advanced age.

Assessment of recognition memory was performed on eight rhesus monkeys of advanced age (25 to 27 years of age) using the delayed recognition span test (DRST). Their performance was compared to that of five young adult animals (5 to 7 years of age) on two stimulus conditions of the DRST: spatial position and color. Both trial unique and repeating series were used for each of the two conditions. As a group, aged monkeys were impaired on both the spatial and color conditions of the DRST, achieving about two-thirds of the span of the young adult group in each condition. Error analyses revealed that monkeys in the aged group also produced more perseverative responses (i.e., displacing the previously correct disk) than did young adults. Together the findings suggest that monkeys of advanced age are impaired on tasks with memory loading demand characteristics.

Isolation and characterization of the gene encoding the testis specific histone protein H2B-2 from the sea urchin Lytechinus pictus.

We have cloned and characterized the gene encoding a sperm specific H2B-2 histone subtype from the sea urchin L. pictus. The gene is not clustered with any other histone genes. However, it resembles other histone genes in many respects. The gene contains no intervening sequences or polyadenylation signals. Like other histone genes it contains the conserved 3' hairpin loop sequence and CAAGAAAGA box necessary for the processing of the 3' end of the histone transcript. Upstream of the gene in addition to the TATA box, and two copies of inverted CCAAT Boxes is a conserved sequence element found in many H2B genes. Unlike most histone genes, the H2B-2 gene is expressed in a sex and tissue specific manner only in testis. The sperm H2B-2 subtype encoded by this gene is 142 amino acids in length. The larger size of this H2B protein is accounted for by residues in the N-terminus of the protein consisting of a series of pentapeptide repeats typical of this histone subtype.

Negative control of photoreceptor development in Drosophila by the product of the yan gene, an ETS domain protein.

Loss-of-function mutations in the yan gene result in the differentiation of supernumerary photoreceptors in the Drosophila eye. The yan gene encodes a protein with an ETS DNA-binding domain that accumulates in the nuclei of undifferentiated cells during the early stages of eye development. Our data suggest that yan functions as a cell-autonomous negative regulator of photoreceptor development; in the presumptive R7 and cone cells, yan appears to act antagonistically to the proneural signal mediated by sevenless and Ras1.

Evolutionary conservation of the leucine-rich repeat transmembrane protein Gp150 in Drosophila and Bombyx.

Gp150 is a transmembrane glycoprotein belonging to the leucine-rich repeat (LRR) superfamily. Here we report the molecular characterization of a Gp150 homolog in Drosophila virilis, which is separated from Drosophila melanogaster by about 60 million years. A silkworm Bombyx mori Gp150 homolog was identified through a sequence database search. Sequence analysis revealed high conservation in the LRRs and cysteine motifs flanking the LRR region in the extracellular domain of Gp150. Using an in vivo assay, we demonstrated that the extracellular domain is essential for Gp150 function. Moreover, structural features unique to the Gp150 proteins were identified that include an incomplete carboxy-flanking cysteine motif, acidic regions on both sides of the LRR region in the extracellular domain, and a short cytoplasmic domain with three putative tyrosine phosphorylation motifs, which might be involved in interaction with SH2 domains. Thus, Gp150 defines a new subfamily of LRR proteins and may be involved in signal transduction. Sequence comparison of the two Drosophila gp150 genes demonstrated a high degree of conservation of genome organization downstream of the gp150 gene. Interestingly, D. virilis gp150 coding region appears to have an additional intron, an uncommon feature in homologs of other genes. The expression pattern of gp150 during embryogenesis in D. melanogaster and D. virilis was found to be identical. gp150 transcripts were localized generally to regions where cells are undergoing dramatic morphogenetic movements. This is further corroborated by the localization of gp150 transcripts in eye imaginal discs in the region spanning the morphogenetic furrow.

Shortsighted acts in the decapentaplegic pathway in Drosophila eye development and has homology to a mouse TGF-beta-responsive gene.

Differentiation in the Drosophila eye imaginal disc traverses the disc as a wave moving from posterior to anterior. The propagation of this wave is driven by hedgehog protein secreted by the differentiated cells in the posterior region of the disc. Hedgehog induces decapentaplegic expression at the front of differentiation, in the morphogenetic furrow. We have identified a gene, shortsighted, which is expressed in a hedgehog-dependent stripe in the undifferentiated cells just anterior to the furrow and which appears to be involved in the transmission of the differentiation-inducing signal; a reduction in shortsighted function leads to a delay in differentiation and to a loss of photoreceptors in the adult. shortsighted is also required for a morphogenetic movement in the brain that reorients the second optic lobe relative to the first. shortsighted encodes a cytoplasmic leucine zipper protein with homology to a mouse gene, TSC-22, which is transcriptionally induced in response to TGF-beta.

Both basal and ontogenic promoter elements affect the timing and level of expression of a sea urchin H1 gene during early embryogenesis.

Late histone H1-beta mRNA accumulates with the correct ontogenic pattern following microinjection of the cloned gene into fertilized sea urchin eggs. Sequences upstream of the gene encoding the sea urchin H1-beta protein contain both basal and developmentally regulated elements. One late H1-specific activator sequence (USE IV) is required for the accumulation of mRNA following the blastula stage of development. All late H1 genes also contain a highly conserved GC-rich sequence resembling a low-affinity binding site for the mammalian transcription factor Sp1 that is required for basal expression of the H1-beta gene at all stages of embryogenesis. When this GC-rich sequence (GGGCTG) is converted to a perfect core Sp1 sequence (GGGCGG), the H1-beta transcripts accumulate to much greater levels and their peak accumulation is shifted to the early blastula stage rather than late blastula and gastrula stages of development. Coincidently, early H1 genes, whose peak expression is also at the early blastula stage, all contain the same core consensus sequence (GGGCGG). Thus, both gene-specific activator sequences, as well as sequences that resemble sites for general transcription factors, may play a major role in determining the temporal patterns of gene expression during early embryogenesis.

Evolution of functional diversification of the paired box (Pax) DNA-binding domains.

The Pax gene family consists of tissue-specific transcriptional regulators that always contain a highly conserved DNA-binding domain with six alpha-helices (paired domain), and, in many cases, a complete or residual homeodomain. Numerous genes of this family have been identified in animals, with the largest number found in vertebrates. Our evolutionary analyses indicate that the vertebrate Pax gene family consists of four well-defined and statistically supported groups: group I (Pax-1, 9), II (Pax-2, 5, 8), III (Pax-3, 7), and IV (Pax-4, 6). Group I paired domains share a most recent common ancestor with Drosophila Pox meso, group II with Pox neuro, group III with paired and gooseberry, and group IV with the eyeless gene. Two groups containing complete homeodomains (III and IV) are distantly related, and the intergroup relationships are (I,III), (II,IV). These four major groups arose before the divergence of Drosophila and vertebrates prior to the Cambrian radiation of triploblastic metazoan body plans. We conducted an analysis of fixed radical amino acid differences between groups in a phylogenetic context. We found that all four fixed radical amino acid differences between groups I and III are located exclusively in the N-terminal alpha-helices. Similarly, groups II and IV show three fixed radical differences in these alpha-helices but at positions different from those in groups I and III. Implications of such fixed amino acid differences in potentially generating sequence recognition specificities are discussed in the context of some recent experimental findings.

Developmental control of promoter-specific factors responsible for the embryonic activation and inactivation of the sea urchin early histone H3 gene.

We have begun an investigation of the molecular basis for the temporal embryonic expression of the early histone H3 gene of the sea urchin Strongylocentrotus purpuratus. Cloned constructs exhibit the proper temporal regulation following microinjection into one-cell zygotes of the related sea urchin species, Lytechinus pictus. Deletion analysis of the upstream promoter region of the H3 gene revealed several regions that are involved in both positive and negative control. DNase I footprinting, mobility shift, and methylation interference experiments reveal multiple sequence-specific DNA-binding proteins that interact with at least five distinct regions within 200 bp upstream of the RNA initiation site. Extracts prepared from staged embryos revealed that the ability of the factors to bind their target sequences was regulated. Proteins bound at four different sites were detected only at stages when the H3 gene was active transcriptionally. In addition, three different forms of a CCAAT-binding protein also are regulated temporally. The activity of these protein(s), however, correlates inversely with the transcriptional activity of the gene. The TATA box and CCAAT sequences are all that is required for expression of low levels of H3 transcripts with the proper temporal pattern. This approach should be useful in understanding the mechanisms used to regulate temporal patterns of gene expression during early embryogenesis.

Loss of tramtrack gene activity results in ectopic R7 cell formation, even in a sina mutant background.

We have screened a collection of transposable-element-induced mutations for those which dominantly modify the extra R7 phenotype of a hypomorphic yan mutation. The members of one of the identified complementation groups correspond to disruptions of the tramtrack (ttk) gene. As heterozygotes, ttk alleles increase the percentage of R7 cells in yan mutant eyes. Just as yan mutations increase ectopic R7 cell formation, homozygous ttk mutant eye clones also contain supernumerary R7 cells. However, in contrast to yan, the formation of these cells in ttk mutant eye tissue is not necessarily dependent on the activity of the sina gene. Furthermore, although yan mutations dominantly interact with mutations in the Ras1, Draf, Dsor1, and rolled (rl) genes to influence R7 cell development, ttk mutations only interact with yan and rl gene mutations to affect this signaling pathway. Our data suggest that yan and ttk both function to repress inappropriate R7 cell development but that their mechanisms of action differ. In particular, TTK activity appears to be autonomously required to regulate a sina-independent mechanism of R7 determination.