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Jazz Legends Legacy: Steve Brown 64, M.S. 68
Glenn Brown 36 and Marie Ward Brown 35 first met on the steps of the old Ithaca Conservatory of Music building in DeWitt Park. Years later, in 1960, their son Steve Brown came to his parents alma mater as a classical percussion student of Warren Benson. Benson was then the faculty sponsor of the student-run jazz lab, led by senior and founder Al Hamme 61. Steve Brown became the groups guitarist during his freshman year. When Hamme graduated, Brown took over, directing the band for most of his remaining undergraduate years, with Bob Levy 66 covering one semester.
After Browns 1964 graduation he taught public school music on Long Island for three years (during which time his youngest brother, Ray Brown 68, who now teaches jazz at Cabrillo College in California, led the jazz lab), before returning to IC in 1967 to continue his studies. He proposed his own program of study, becoming the first-ever recipient of a masters degree in classical guitar from IC. During the year he spent studying for it Brown returned to playing in the jazz lab, performing with his brother Ray.
Steve Brown had made an impression at Ithaca. As soon as he completed his masters degree, IC hired him to lead the big band, teach jazz courses, and direct the jazz lab, which hes been doing ever since. If you do the math youll see that there have been only four directors of the IC jazz lab since the bands founding in 1957: students Al Hamme, Steve Brown, Bob Levy, Ray Brown, and then Steve Brown again, this time as a faculty member.
Over the years the jazz bands and classes have included students majoring in performance, education, and jazz. Brown says that hes proud to serve the music education community, knowing that students equipped to teach jazz are in demand in the public schools. He incorporates pedagogy into all his classes so that his students leave IC with the ability to teach jazz "methodically and engagingly."
As the principal jazz studies professor, Brown has a variety of duties. He leads the big band and electric guitar ensemble, teaches jazz guitar and electric bass students, and teaches classes in jazz piano, jazz history, improvisation, and arranging. Even though he has been asked to join the faculty at the Berklee School of Music in Boston, which specializes in jazz and contemporary media, he has happily stayed in Ithaca, where he can "do it all."
Both part-time lecturers in jazz studies graduated from the School of Music. Paul Merrill, M.M. 01, was Browns student and is now his colleague. For him, the essence of Browns influence on Ithacas musical community is his dedication to "working with all levels and affecting musicians from every background."
Lauri Robinson-Keegan 89, M.M. 00, who is known in the jazz community as Cookie Coogan, leads the vocal jazz ensemble. She points out what an incredibly generous mentor Brown is: "Hes always willing to share with anyone who is interested. He also involves his students in the real jazz world --- he takes students to play gigs with him, and he plays on their gigs occasionally. Steve helps nurture young jazz musicians trying to find their niche in the world."
Brown also maintains an active schedule of performances as a guitarist and recording artist. He and his brother Ray were recently featured in a concert in Pittsburgh called "Brothers in Jazz." Steve Brown was also one of the all-stars who performed at the State University of New York at Binghamtons farewell concert celebrating Al Hammes retirement. After starting the IC jazz lab, Hamme had gone on to direct Binghamtons jazz program.
Browns writing has also garnered attention: Kendor recently published his original big band piece Catch Phrase, and he has written two books in widespread use: Jazz Solos for Guitar Vol. 1 and An Introduction to Jazz Improvisation, a collaboration with Ray.
During the School of Musics recent expansion, a chamber jazz room was added as a gift from Steve McCluski 74 and Kim Joslyn McCluski 74 in honor of the Brown family. A plaque there honors all seven members of the family who graduated from IC --- Steve; his wife, Barbara Katz Brown 74, M.S. 75; his parents (his mother passed away this winter); his brother Ray; Rays wife, Susan Cahill Brown 71; and his brother Glenn Brown 59 --- plus two who attended other colleges, Steves brother Roger and his sister, Jeanne Brown DAcquisto. The room is a place where classes and small chamber groups can get together to hone their skills.
Browns boundless energy and enthusiasm for jazz, his passion for sharing, and his inspiring personality all make him the venerable master for dozens of students every year. As Paul Merrill puts it, "Jazz has no greater mentor, torchbearer, player, teacher, and friend than Steve."Photo of Steve Brown rehearsing the big band by Richard Killen
A. Ozolins, Ithaca College Office of Publications, 6 August, 2002


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- Research article
- Open Access
Identification of multiple integrin β1 homologs in zebrafish (Danio rerio)
© Mould et al; licensee BioMed Central Ltd. 2006
- Received: 09 March 2006
- Accepted: 20 June 2006
- Published: 20 June 2006
Integrins comprise a large family of α,β heterodimeric, transmembrane cell adhesion receptors that mediate diverse essential biological functions. Higher vertebrates possess a single β1 gene, and the β1 subunit associates with a large number of α subunits to form the major class of extracellular matrix (ECM) receptors. Despite the fact that the zebrafish (Danio rerio) is a rapidly emerging model organism of choice for developmental biology and for models of human disease, little is currently known about β1 integrin sequences and functions in this organism.
Using RT-PCR, complete coding sequences of zebrafish β1 paralogs were obtained from zebrafish embryos or adult tissues. The results show that zebrafish possess two β1 paralogs (β1–1 and β1–2) that have a high degree of identity to other vertebrate β1 subunits. In addition, a third, more divergent, β1 paralog is present (β1–3), which may have altered ligand-binding properties. Zebrafish also have other divergent β1-like transcripts, which are C-terminally truncated forms lacking the transmembrane and cytoplasmic domains. Together with β1–3 these truncated forms comprise a novel group of β1 paralogs, all of which have a mutation in the ADMIDAS cation-binding site. Phylogenetic and genomic analyses indicate that the duplication that gave rise to β1–1 and β1–2 occurred after the divergence of the tetrapod and fish lineages, while a subsequent duplication of the ancestor of β1–2 may have given rise to β1–3 and an ancestral truncated paralog. A very recent tandem duplication of the truncated β1 paralogs appears to have taken place. The different zebrafish β1 paralogs have varied patterns of temporal expression during development. β1–1 and β1–2 are ubiquitously expressed in adult tissues, whereas the other β1 paralogs generally show more restricted patterns of expression.
Zebrafish have a large set of integrin β1 paralogs. β1–1 and β1–2 may share the roles of the solitary β1 subunit found in other vertebrates, whereas β1–3 and the truncated β1 paralogs may have acquired novel functions.
- Integrin
- Zebrafish Embryo
- Zebrafish Genome
- Knee Region
- Hybrid Domain
Integrins are a family of metazoan cell surface receptors that play critical roles in cell adhesion, migration, differentiation and survival [1]. Integrins are heterodimeric glycoproteins containing non-covalently associated α and β subunits, and are grouped into sub-families according to the identity of the β subunit. In humans, eight different β subunits combine with 18 different α subunits to form 24 functionally distinct heterodimers. Integrins have a large extracellular domain responsible for interacting with extracellular ligands, and a small intracellular domain that binds to cytoskeletal and signaling proteins. Integrins assimilate information from the extracellular and intracellular environments by acting as bi-directional transducers of signals across the cell membrane. Hence, the binding of an extracellular ligand can elicit activation of intracellular signaling pathways and reorganisation of the cytoskeleton [2, 3]. Conversely, changes in intracellular signaling can result in the stimulation or inhibition of ligand binding due to conformational changes in the extracellular domains of an integrin [4–6].
Integrin ligands include the major ECM components laminins, fibronectin and collagens. The β1 integrin sub-family contains 12 different heterodimers in mammals, which form the major group of cell-ECM receptors. An ECM-binding β chain is probably the most ancient, in evolutionary terms, of all the integrin β chains [7, 8]. β1 integrins have widespread essential functions both during development and in the adult organism [9–12].
The A domain is the portion of the β subunit that is involved in extracellular ligand binding; this domain also is critical for the interactions with the α subunit that lead to heterodimerisation. The A domain consists of a central β-sheet encircled by seven α helices, and has two large sequence insertions compared to other A domains [16]. These insertions, which are critical for ligand binding and for association with the α subunit, lie in the loops between β-strands B and C and between β-strand D and α helix 5 [13, 14]. The βC-βD loop (otherwise known the 'specificity-determining loop') contains a small disulfide-bonded segment that contributes to the differences in ligand-binding specificity between different integrin heterodimers [17–19]. The A domain also contains three cation-binding sites: MIDAS (metal-ion dependent adhesion site), ADMIDAS (adjacent to MIDAS) and LIMBS (ligand-associated metal binding site) [13, 14]. The MIDAS site is critical for ligand recognition, whereas the ADMIDAS and LIMBS sites appear to have regulatory roles [20, 21].
X-ray crystal structures [13–15] of the extracellular domains have revealed that the overall shape of the β subunit is that of a 'head' connected to a long 'leg' with an intervening 'genu' or 'knee' (Fig. 1). The A domain and hybrid domain form part of the integrin head region, whereas the PSI and EGF-1 and EGF-2 lie in the knee region; the remaining EGF repeats and βTD domain contribute to the leg region. The conversion of an integrin from an inactive to an active state is proposed to involve an unbending of the knee region, and a key hinge point in the knee region has recently been proposed to lie in EGF-2 [15]. A critical feature of this unbending is the release of the hybrid domain from interactions with other integrin domains, thereby allowing the hybrid to undergo an outward pivoting. In turn, this movement causes conformational changes in the A domain that promote ligand recognition [14, 22]. The β subunit cytoplasmic tail contains a HDRRE motif for interaction with the α subunit cytoplasmic domain [23], and NPXY sequences that are involved in binding to cytoskeletal proteins such as talin and filamin [24].
The zebrafish is an emerging model organism with a large number of attractive features for studying the function of genes, such as: (i) the external and rapid development of embryos, (ii) the transparency of embryos, which allows visualisation and tracking of individual cells or groups of cells, (iii) the high degree of conservation between zebrafish and human genes, (iv) the similarities between human and zebrafish embryonic development resulting in a large proportion of tissues and organs being grossly similar between the two species, (v) its genetic tractability, aided by the availability of powerful genetic tools such as gene knock-down by morpholino-modified antisense oligonucleotides (morpholinos) [25] and gene knockout by target-selected mutational inactivation of genes (TILLING)[26], and (vi) the partially complete genome sequence [59]. In addition, zebrafish is finding increasing use as a model organism to study human diseases [27, 28]; in many of these disorders integrin-ligand interactions play important roles in the initiation or progression of the disease [29].
To date, our knowledge of the biological roles of integrins in vertebrates has been derived mainly from knockout studies in mice [30, 31]. For the reasons described above, zebrafish provide a very attractive alternative model system for studying integrin function during development and disease. Currently, however, very little published information is available concerning the sequences and complement of integrin genes in this organism. Here we have investigated if β1-like genes and transcripts are found in zebrafish. We found that zebrafish have two β1 paralogs that are closely related to the single gene found in other vertebrates. Surprisingly, we also found multiple, more divergent, zebrafish β1 paralogs, many of which are truncated and entirely novel forms.
Zebrafish have two β1 paralogs (β1–1 and β1–2) that are closely related to human β1
Zebrafish have a third β1 paralog (β1–3), which is more divergent in sequence from human β1
Percentage amino acid identities between zebrafish and human β1 sequences
% id
Zebrafish have truncated β1 paralogs that are closely related to β1–3
Corresponding database entries for zebrafish β1 sequences
Accession Number
Gene Prediction
Genomic Location
Corresponding (or additional) EST
Source of EST
whole adult
gastrula stage
<72 h embryos
pooled gut
All the truncated paralogs are closely related to β1–3 (Fig. 5B), with ~80% sequence identity overall. Hence, together with β1–3 they form a novel group of β1 paralogs. In a surprising contrast to what is observed regarding sequence conservation between β1–1 β1–2 and human β1(Fig. 3), the PSI and hybrid domains are almost perfectly conserved between the truncated paralogs and β1–3, whereas the A domains are much less well conserved (Fig. 5B). In addition, the sequence of EGF-1 is highly conserved, and the first two EGF repeats of β1tr-3 are almost identical in sequence to those of β1–3 (Fig. 5B). The major region of sequence variation between the truncated β1 paralogs is the putative linker sequence in EGF repeat 2 [15], with some of the paralogs having a six amino acid deletion in this region (Fig. 4).
The truncated paralogs, like β1–3, have a substitution of two key aspartate residues by Ser/Asn at the ADMIDAS. In addition, these paralogs are also lacking an essential LIMBS residue (E189 in human β1 [E169 in the mature sequence]) due to a glutamate to arginine substitution. A similar mutation in human β1 (APM and J. Askari, unpublished data) or β3 [33] perturbs ligand recognition. Hence, the A domains of the truncated paralogs may not be functional for ligand binding, or may possibly bind to ligands in a different manner to other β subunit A domains. The MIDAS coordinating residues are conserved in the truncated paralogs; hence, it remains possible that they may bind ligands in a manner akin to the A domains found in some α subunits [6].
Overall, the sequences of the A domains in the truncated paralogs are highly divergent from other chordate sequences; indeed, they are only slightly more similar to human β1 A domain (57–58% identity) than the Ciona β1 A domain (56% identity). The loops important for ligand binding and association with the α subunit are poorly conserved. For example, the truncated paralogs all have a large deletion of nine amino acids in the β B-βC loop. A second part of the α/β subunit interface involves residues in the βD-α5 loop, especially those that form a short stretch of 310 helix (DGKL) [13]. This region is highly conserved between the full-length paralogs and human β1 (Fig. 2) but has several substitutions in the truncated paralogs. Taken together, these findings suggest that the truncated β1 paralogs may have weaker associations with α subunits, associate with different α subunits, or even may not associate with α subunits at all. However, even if the truncated paralogs are able to associate with α subunits, they would form non-signaling receptors, as they lack the transmembrane and cytoplasmic domains.
Genomic and phylogenetic analysis reveals evolutionary relationships between the β1 paralogs
Gene sequences corresponding to β1–1, β1–2 and β1–3 can be identified in the zebrafish genome (Table 2) (although there are a number of inaccuracies and gaps in the GENESCAN ab initio gene predictions). Currently, the gene for only one truncated paralog can be identified from the latest genome sequence (Zv6), corresponding almost exactly to ESTb (Table 2). β1–2, 'ESTb' and β1–3 sequences lie adjacent to one another in a tandem array on chromosome 2; β1–1 lies in a separate region of the genome (chromosome 24). The genomic sequences show almost perfect conservation of exon- intron boundaries (data not shown), suggesting that all the β1 paralogs arose from duplication of a single ancestral β1 gene.
The ML tree was supported by maximum parsimony and Bayesian analyses. Only neighbor joining analysis (not shown) produced two vertebrate clades where β1–1, β1–2 and human β1 grouped separately to the other sequences. However, the genomic locations of the different paralogs add support to the ML analysis. For example, the position of β1–2 and β1–3 in close proximity on the same scaffold suggests that β1–2 and β1–3 arose from a tandem duplication of a common ancestor. The presence of a truncated paralog in the same region of the genome adds weight to the proposal (supported by all the phylogenetic analyses) that the ancestor of the truncated paralogs arose from a duplication of the precursor of β1–3.
The divergent β1 paralogs are expressed later in development than β1–1 and β1–2
The divergent β1 paralogs are expressed less widely than β1–1 and β1–2
EST sequences corresponding to the different β1 paralogs can be identified in the NCBI databases (Table 2). The majority of these EST sequences correspond to those of truncated paralogs. Although no EST sequences that exactly matched β1tr-1 or β1tr-2 were found, ESTs corresponding to the 5' and 3' ends of β1tr-3 could be identified (Table 2). (Conversely, although there are many additional EST sequences that correspond exactly with ESTa (Table 2), we did not amplify any precisely matching sequences from zebrafish embryos.) EST sequences also provide evidence concerning the expression of the different β1 paralogs in different tissues or at different stages of development (Table 2). ESTs for the truncated paralogs (especially ESTs corresponding to ESTa) are found in cDNA from several different tissues, and appear to be particularly abundant in liver and kidney.
The novel findings of this report are: (i) Extensive expansion of the β1 integrin gene family has taken place in zebrafish, (ii) a group of divergent β1 paralogs with regulatory site mutations is present, some of which are novel truncated forms that lack the transmembrane and intracellular domains, (iii) the different β1 paralogs exhibit varied patterns of temporal and tissue expression. Our findings raise several important questions. For example: Why do zebrafish have multiple versions of the single β1 gene found in higher vertebrates? What is the function of the divergent paralogs? Which α subunits does each paralog pair with?
Gene duplication is an important mechanism for the generation of phenotypic complexity, diversity and innovation. Gene duplication can occur either through tandem duplication of an individual gene, segmental duplication of a portion of a chromosome, or through whole genome duplication. Recent analyses have provided strong support for a whole genome duplication in the lineage of ray-finned fishes approx. 350 million years ago [34–36]. Following this duplication, most of the gene duplicates were rapidly lost. However, a small proportion of these duplicates were retained. Hence, for approximately 20% of single-copy human genes, two corresponding orthologs are found in zebrafish [37]. It is possible that β1–1 and β1–2 arose from the fish-specific whole genome duplication. Consistent with this possibility, β1–1 and β1–2 have different chromosomal locations and furthermore, both gene loci show synteny with the p11.22 region of human chromosome 10, which contains the β1 (ITGB1) gene (APM, unpublished data). Two co-orthologs of β1 are also found in other teleost species such as fugu [38].
In the classical model of gene duplication [39], new gene duplicates face one of two fates: either one copy mutates to a pseudogene (called nonfunctionalization), or one copy preserves the original function, and the other copy mutates freely until by chance it obtains a sequence that confers a new, beneficial, positively selected function (called neofunctionalization). In a third possibility, called subfunctionalization or subfunction partitioning, the complementary partitioning of ancestral subfunctions is the mechanism that preserves gene duplicates [37]. Two classic examples of subfunction partitioning in zebrafish are the pair of sox9 genes, sox9a and sox9b [40], and the two mitf genes, mitfa and mitfb [41]. It is possible that ancestral β1 integrin functions have been partitioned between β1–1 and β1–2. Subfunction partitioning can occur through tissue-specific expression of gene duplicates and/or expression at different times of development. We found that there are differences in temporal expression of β1–1 and β1–2, with the expression of β1–2 peaking in early development, whereas expression of β1–1 was highest during later development. Also some differences in the tissue expression between these two paralogs were noted, with low expression of β1–1 in adult intestine. It is also possible that partitioning could take place if each β chain associates with a distinct subset of α chain partners. The possible subfunction partitioning of β1 roles by β1–1 and β1–2 may provide distinct advantages for functional genetic analyses in zebrafish. Whereas knockout of the single β1 gene in mice is very early embryonic lethal [9, 10], knockout (or knockdown) of only one of these genes in zebrafish may have a much less severe phenotype, making it possible to study the roles of β1 integrins during the later stages of development.
In contrast to β1–1 and β1–2, it seems likely that the divergent β1 paralogs arose through tandem duplications. Taking the genomic, phylogenetic and sequence identity data together, the most likely scenario is that the precursor of β1–2 underwent a tandem duplication that gave rise to the ancestor of β1–3; a subsequent duplication of this paralog gave rise to the progenitor of the truncated paralogs. It seems likely that the divergent paralogs have acquired novel functions. With the exception of β1tr-3, these paralogs are expressed at later times of development and in a more tissue-restricted manner than β1–1 and β1–2. It therefore appears that the divergent paralogs may be involved in tissue differentiation, maintenance or remodelling, rather than in development/organogenesis. The expression of many of these paralogs in tissues such as liver and kidney supports this view.
One clue to the function of the divergent paralogs is that they possess a double substitution (DD to SN) in the regulatory ADMIDAS cation-binding site in the A-domain. Preliminary results suggest that engineering an identical substitution in human β1 (in a heterodimer with α5 subunit) produces a constitutively inactive integrin, i.e., an integrin with very low affinity for ligand (J. Askari and A.P. Mould, unpublished data). It is therefore possible that β1–3 could act as a dominant-negative β1 subunit by competing with β1–1 and β1–2 for α subunit partners or intracellular binding factors. It is also proposed that expression of high levels of unoccupied integrins can trigger apoptosis [42]; hence, expression of β1-3 could also regulate cell survival. If the truncated paralogs are unable to associate with α subunits they could be secreted into the extracellular environment. There are now many examples of cell surface receptors that also have a secreted antagonistic counterpart, e.g. VEGFR, FGFR-1 and IL-1R [43–45]. It is therefore intriguing to speculate that truncated integrin paralogs could have a similar mode of action.
It is also possible that the divergent paralogs could have acquired new binding partners. The very high degree of sequence conservation of the PSI, hybrid domain and EGF-1 between the divergent paralogs suggest that these domains form a key functional region of these polypeptides, and furthermore that the function of these domains is common to all the divergent β1 paralogs. Such a degree of conservation suggests that these domains interact with protein ligands (even though there is currently no evidence in other integrins that these domains have extracellular ligands). The PSI, hybrid, and EGF-1 domains lie adjacent to each other at, or close to, the knee region of the integrin [15]. Proteins that bind to these domains could therefore alter function, either by constraining the integrin in a bent (low affinity) form or by perturbing the interactions between PSI/hybrid/EGF-1 domains and other integrin domains, thereby favouring the unbent (high affinity) form. A key hinge point in the β subunit knee region is thought to be the near the start of EGF-2 [15]. Interestingly, some of the truncated paralogs have a six amino acid deletion in this linker region (Fig. 4), which is likely to change the conformational state of the protein, and may therefore modify function. Intriguingly, the second EGF repeat in β1tr-3 is essentially identical to that of β1–3, implying that this repeat may also have a conserved role between these two paralogs.
It is currently unclear how many separate genes are represented by the truncated β1 paralogs and ESTs. Although the sequences β1tr-1 and β1tr-2 are 98% identical to each other they are probably distinct gene products because they have different 3'UTRs. In addition, although β1tr-2 and β1tr-3 may have identical 3'UTR sequences, it appears unlikely that β1tr-3 is an alternatively spliced form of β1tr-2 because the differences between the two coding sequences are spread throughout the primary structures (Fig. 4 and Additional file 1). Additionally, the truncated paralogs cannot simply represent different alleles of the same gene because at least three of these paralogs could be amplified from RNA derived from a single fish (APM and Paul Walker, unpublished results). Furthermore, the genomic sequence that corresponds to ESTb (which is 98% identical in coding sequence to β1tr-2) does not contain sequences that would allow the generation of different transcripts by alternative splicing (data not shown). In summary, the currently available evidence suggests that the truncated paralogs originate from separate (yet to be characterised) genes. Since the genomic scaffold is assembled from shotgun sequences it is possible that, due to the almost identical sequences of the truncated paralogs, the scaffold is incorrectly assembled and additional genes of truncated paralogs may be present in the same region. In the future, the availability of finished clones in the region of the genome that contains 'ESTb' for detailed analysis should help to clarify the precise number of truncated β1 paralogs in the zebrafish genome. A further puzzle is why none of the truncated β1 sequences we amplified from zebrafish RNA exactly matched the ESTa or ESTb sequences. A possible explanation is that the β1tr-1, β1tr-2 and β1tr-3 sequences were only the predominant sequences obtained by subcloning of the RT-PCR products. Other clones obtained have not yet been fully sequenced, some of these may match the EST sequences more precisely.
Truncated forms of β subunits have previously been reported in sponge and man [7, 46]. However, these polypeptides represent alternatively spliced forms of full-length β subunits that are truncated within the hybrid domain, and therefore probably result in non-functional proteins [47]. In contrast, the truncated β1 paralogs reported here appear to be unique gene products, and the position of the truncation would preserve the structure and function of all of the extracellular domains with the exception of the βTD.
A key subject of future investigations will be to identify the α subunit partners of the different paralogs. All twelve α chains that pair with β1 in higher vertebrates have orthologs in the zebrafish genome, and at least two of these α chains have two co-orthologs (APM, unpublished results). It seems very unlikely that all of the β1 paralogs identified here are able to associate with all of the >14 potential α chain partners. For example, if each of the six β1 paralogs has retained the ability to pair with all of the α chains then a minimum of 84 different heterodimers could form. This would present a huge increase in complexity over the twelve heterodimers found in higher vertebrates. It is likely, therefore that in this process of multimeric protein evolution, previously termed 'molecular incest' [48], that only a limited number of possible combinations was retained based on function.
It is not yet clear if divergent β1 paralogs are found in other teleosts. Currently only limited gene information is available on other fish species, with the exception of the pufferfish Takifugu rubripes and Tetraodon nigroviridis. Analysis of the pufferfish genomes suggests that only two β1 paralogs are present in fugu and tetraodon; these paralogs are closely related to zebrafish β1–1 and β1–2 (AP Mould, unpublished results). However, since β1–3 probably arose from an ancient duplication of the ancestor of β1–2 it would be surprising if the former paralog was retained only in the zebrafish lineage. Furthermore, although the divergent β1 paralogs have not been detected in pufferfish genomes, pufferfish may have retained fewer gene duplicates than zebrafish [49].
The presence of expanded gene families is hypothesized to have made a major contribution to the extraordinary diversity and evolutionary success of teleosts, which make up half of all vertebrate species. It is known that fish genomes are more 'plastic' than other vertebrate genomes, partly due to higher rates of gene duplication [50]. Fish-specific novel members of gene families may contribute to a large extent to the distinct physiology of fishes and mammals, while differential retention of duplicate genes may have facilitated the isolation of emerging species during the vast radiation of teleosts [51, 52]. Our findings show that integrins are a dynamic family of genes that have evolved in multiple ways after the divergence of the common ancestors of the mammalian and fish lineages. Expansion of the integrin family may also correlate with expansion of extracellular matrix gene families, e.g. collagens, in teleosts [34].
At least six paralogs of the integrin β1 gene have been identified in zebrafish, demonstrating that this species has a greatly expanded integrin repertoire. Two of these paralogs may share the functions of the single β1 subunit found in higher vertebrates, whereas the remaining paralogs may have acquired novel roles. This is the first description of truncated β1 chains, which we speculate could be secreted proteins that act as regulators of integrin functions.
Sequencing of zebrafish integrin β1 paralogs
The complete amino acid sequence of human β1 [Swiss-Prot: P05556] was used to probe the Danio rerio genome ([59, 61]) using TBLASTN [53] to identify genes with highest sequence identity to human β1. To amplify complete β1 sequences, primers were designed using putative 5' and 3' sequences (identified as described above) or from EST sequences on the NCBI database [60]. The following primers were used: β1–1, 5'-ATGGACCTGAAGCTACTTTTCATATC-3' and 5'-CTGATGGCCATTATTTGCCTTCG-3', β1–2, 5'-ATGGACGTAAGGCTGCTCCTG-3'and 5'-CACGTTCGTCCATTATTTGCCCTC-3', β1–3, 5'-ATGAAAATGAAGCTGCTGTTATTATC-3' and 5'-CACTTTCCCTCATATCTGGGATTC-3' β1tr-1, 5'-ATGGATATAACAGTTTTGTTATTATCAG-3' and 5'-ATGTATAACATGAGGTCATGATGTAC-3' β1tr-2 5'-ATGGATATAACAGTTTTGTTATTATCAG-3' and 5'-GTATAACATGTGTCTCAATATATGATG-3'
Total RNA was prepared from 4-day old embryos using TRI reagent (Sigma), and reverse transcription was performed using Superscript II (Invitrogen) according to the manufacturer's instructions. PCR reactions were performed using Phusion (New England Biolabs). Cycling parameters were 98°C for 30 s, followed by 40 cycles of 98°C for 10 s, 60°C for 20 s and 72°C for 60 s. An additional sequence, β1tr-3, was amplified from reverse transcribed RNA from adult kidney (prepared as described below) using the same primers as for β1tr-2. The PCR reactions generated products of ~2.4 kB for β1–1, β1–2 and β1–3, and ~1.9 kB for β1tr-1, β1tr-2 and β1tr-3. EST clones ESTa and ESTb were obtained from RZPD German Resource Center for Genome Research, product numbers IRAKp961B08165Q [IMAGE:6525557] from adult kidney cDNA library and IMAGp998I1214695Q3 [IMAGE:7001749] from whole adult body cDNA library, respectively.
PCR products were analysed on 1.5% agarose gels. Sequencing of PCR products or EST clones was performed using the BigDye cycle sequencing kit (Applied Biosystems). PCR products from reactions using the 5'-ATGGATATAACAGTTTTGTTATTATCAG-3' forward primer contained a mixture of sequences and therefore these products were subcloned into the Zero Blunt TOPO cloning kit (Invitrogen). Sequencing of individual clones revealed a single most abundant sequence (β1tr-1, β1tr-2 or β1tr-3). Alignment of zebrafish and human sequences was performed using CLUSTAL W [62].
Phylogenetic analysis
The β1 integrin sequences identified in Danio rerio were aligned with the human β1 sequence [Swiss-Prot: P05556] and the ancestral β1-like gene previously identified in Ciona intestinalis (JGI Ciona v1.0 ci0100141446) [8] using CLUSTAL X [54]. Gap-containing sites were removed from each alignment and Maximum Likelihood trees were inferred using PROML from the PHYLIP package [55]. The JTT model of amino acid substitutions was used with and without global rearrangements and correction for rate heterogeneity (α value obtained from TREEPUZZLE [56]). The topologies of the trees were tested using three independent methods. Neighbor-Joining and Maximum Parsimony bootstrap replicates were obtained using the PHYLIP package [55]. Bayesian tree inference values were produced from the MrBayes programme [57].
RT-PCR analysis
Embryos were harvested at different times post fertilization. Tissues from adult fish (~9 months old) were removed by dissection and flash frozen in liquid nitrogen. Total RNA was prepared using Trizol (QIAGEN). Reverse transcription was performed using M-MLV reverse transcriptase or Omniscript (Invitrogen) according to the manufacturer's instructions. PCR reactions were performed using recombinant Taq (a gift from P. Walker, University of Manchester, UK) or Advantage 2 polymerase (BD Biosciences). For the truncated β1 sequences, primers were designed to give a single product of approx. 600 bp that is unique to β1tr-1, β1tr-2 or β1tr-3. The following primers were used:
β1–1 5'-ATGGACCTGAAGCTACTTTTCATATC-3' and 5'-GTGACGTTTCTCCAGCCAATGTG β1–2 5'-GATGGTAATGAATGCACCAAGGC-3' and 5'-GGAGTCGGAGGTAAGCGTTCC-3' β1–3 5'-GTGTTGTTTGATATAGAAATCACGGCT-3' and 5'-CGTATCCCACTTGGCATTATTTTTCTC-3' β1tr-1 5'-CCAGGATCTGGATGCATACTG-3' and 5'-ATGTATAACATGAGGTCATGATGTAC-3' β1tr-2 5'-CCAGGATCTGGATGCATACTG-3' and 5'-GTATAACATGTGTCTCAATATATGATG-3' β1tr-3 5'-CATGATGAGGTGCTGGCGGATG-3' and 5'-GTATAACATGTGTCTCAATATATGATG-3'
Cycling parameters were 95°C for 2 min, followed by 30 cycles of 95°C for 30 s, 60°C for 20 s and 72°C for 60 s. PCR products were analysed on 1.5% agarose gels. The identity of selected products was confirmed by DNA sequencing. As a control, a fragment of β-actin was amplified using the primers 5'-CCACGAGACCACCTTCAACT-3' and 5'-CATTGTGAGGAGGGCAAAGT-3' for 28 cycles of 95°C for 30 s, 55°C for 40 s and 72°C for 60 s. Negative controls consisted of no cDNA template reactions.
JHJ and ACG are supported by BBSRC PhD studentships, AJLH by a fellowship from Cancer Research UK, and APM and MJH by The Wellcome Trust. We thank Paul Walker for the gift of recombinant Taq, and Drs Sally Stringer, Paul Walker and Patrick Buckley for helpful discussions.
Authors' Affiliations
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This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

subset: commoncrawl


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Tag: Mayan Ruins
Update #1: Time to Wail: The Western Wall of the Mayan Ruins is Gone
Over the last year, several people have sadly reported the legal destruction of the western wall of the "Mayan Ruins" in Garden City as a result of the building of two new homes in this Garden City area.
Femenias Findings: Exploring the Link Between Camp Mills and the "Mayan Ruins" Motor Parkway Bridge
Frank Femenias believes there may be a link between the short-life of the Camp Mills (1918-1919) and the reason the nearby "Mayan Ruins"Motor Parkway Bridge was never completed. Did Frank convince you with these amazing overlays?
Update #3: Mystery Foto #28 Reveals Another Piece of Long Island History in Garden City
Mystery Foto #28 has uncovered a piece of Long Island history. Garden City Village Historian Bill Bellmer notes "the aerial shows the Camp Mills Base Hospital remains, one of my local specialties. The image you posted is better than any other that I have."
Update #3: Mystery Foto #28 Solved: A 1931 Aerial of the Motor Parkway in Garden City and Mineola
Author Richard Panchyk challenged you to identify this spectacular Long Island aerial.
Mystery Foto #37 Solved: An Amazing Aerial of the Long Island Motor Parkway In Garden City
It was time for another amazing Mystery Foto.
Mystery Foto #32 Solved: A 1950 Image of the Motor Parkway Looking West to the LILCO Gas Tank
Art Kleiner, Paula S. and her son Jeff challenged you again to identify this 1950 image of our favorite parkway.
Exploring the "Mayan Ruins" Bridge in Garden City- Updated: 4/23/2017
With the permission of a surrounding neighbor, members of the Long Island Motor Parkway Preservation Society took an up-close look last Saturday of the "Mayan Ruins" Motor Parkway Bridge in Garden City.
Sam & Dave's Excellent Lost Motor Parkway Adventure III: The "Mayan Ruins" in Garden City
In the third post of the Russo's second series, Sam & Dave explore the amazing "Mayan Ruins" Motor Parkway Bridge in Garden City.
The 1938 U.S. Department of Agriculture Aerial Suvey of Nassau County #1: Searingtown to Garden City
On August 3, 1938, the United States Department of Agriculture conducted an aerial survey of Nassau County. These enlargeable aerials (NC_8_3_38 series) have been posted on the Stony Brook University Digital Research Collections.
William A. Bellmer Article: "The Long Island Motor Parkway in Garden City"
For the January 2016 newsletter of the Garden City Eastern Property Owners Association, village historian William A. Bellmer wrote this article on the Motor Parkway.


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Combing a light weight linen fabrication, understated front tie detail and stunning floral print, the Malibu Top is a must have for the Spring/Summer wardrobe. All design and wear elements considered, the top features elastane at the back and sleeve cuff. With endless styling options, complete the look with the Malibu Shorts, elevate with a tailored pant or for a casual look pair with a denim skirt.
Model height: 179cm | Model wears Size 36


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MOSCOW, May 22 (Reuters) - Igor Shuvalov, former first deputy prime minister, has been selected to become the new head of the Russian state development bank VEB, three sources familiar with the decision told Reuters.
Shuvalov is set to replace Sergei Gorkov, who has been in charge of VEB since Feb 2016.
It was not immediately clear why Gorkov was apparently being replaced. Sources said they did not know whether the document on Shuvalov's appointment has been already signed.
Spokespeople for Shuvalov and VEB declined to comment.
Kremlin spokesman Dmitry Peskov, asked about the issue during a daily conference call with reporters, said: "We never discuss staffing issues and won't do so this time either." (Reporting by Tatiana Voronova, Polina Nikolskaya and Darya Korsunskaya Writing by Katya Golubkova; Editing by Kevin Liffey)


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Gift Card and Gift Card Holder designs from the beginning of my career!
Monday, September 29, 2014
Throwing it back
Posted by Julia Warren at 11:38 PM
2 comments:
- Julie said...
Animal print balloons. You are my creative girl. Love it. xoxo Maman
- June 14, 2015 at 10:56 PM
- Julie said...
Julia is a creative genius.
- June 14, 2015 at 10:57 PM
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Cidade da Beira – Moçambique
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Edwin W. Slade, Jr., D.M.D., J.D.
Edwin W. Slade, Jr., D.M.D., J.D., graduated from the University of Connecticut and received his dental degree from the University of Pennsylvania School of Dental Medicine. He was a resident in Dental Medicine and Surgery at the Medical College of Pennsylvania and completed his residency in Oral and Maxillofacial Surgery at the University of Connecticut/Hartford Hospital. He received his law degree cum laude from Temple University School of Law and is a Fellow of the American College of Legal Medicine.
Dr. Slade is Board Certified by the American Board of Oral and Maxillofacial Surgery and the National Board of Dental Anesthesiology. He is the Past President of both the Pennsylvania and Delaware Valley Societies of Oral and Maxillofacial Surgeons. He is a Fellow of the American Association of Oral and Maxillofacial Surgeons (AAOMS) and served both as District 2 Trustee and the Treasurer of the AAOMS. He is presently the AAOMS representative on the Board of Directors of the Accreditation Association for Ambulatory Health Care (AAAHC). He is a Fellow of the American College of Dentistry and the International College of Dentistry.
Dr. Slade is an Adjunct Clinical Associate Professor in the Department of Oral and Maxillofacial Surgery and Pharmacology at the University of Pennsylvania School of Dental Medicine, having taught there for 16 years.


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I recently read a study that stated only 1 out of about 5 relationships are actually thriving. Tai Lopez
Learn how knowing these 26 love stories can help you understand your partner and save your relationship.
Learn what my mentor's taught me that allowed me to make money while traveling and having fun - https://www.TaiLopez.com/67youtube
For a behind the scenes look of a day in the life of an entrepreneur follow me on snapchat - @TaiLopez


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Other Reports
Logistics Recommendations for an Improved US Arctic Research Capability
This 1997 USARC report emphasizes the importance of a strong US arctic logistics capability to enhance and expand current US arctic research and addresses both general and specific arctic logistics needs. It contains recommendations developed through a broad survey of the US academic arctic research community regarding the logistics needed to improve the capability and safety of the US arctic research program.
Available Format: 4.89 MB pdf
July 1997


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The best:
The following paragraph is specifically for Carlos...Carlos, there's nothing like perpetuating racial stereotypes under the guise of "comedy." Granted, you're not the only comedian that does it, but you're the only one that misses the whole comedy part. Oh yeah, someone pass on that "yelling a joke doesn't make it funnier" advice to Joe Rogan too.
Yelling a joke doesn't make it funnier. Doing little twitches while telling a joke doesn't make it funnier. Pointing at the camera or various people in the audience doesn't make it funnier. Laughing hysterically at your own jokes doesn't make them funnier. Relying on stereotypes for 99.8% of your material doesn't make you funnier. Having your own show doesn't make you funny. Your not funny, nor are you by any means a respected comedian, but rather, you are a commercialized joke, a fat vulture feeding on the remains of the Chappelle show. Perhaps when you take the time to construct a joke coherently, rather than fall back on "in-yo-face"-it y, will you truly know what it means to be a comedian.


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When I'm in the process of outlining or writing my initial draft of a book, I often skip a lot of physical character descriptions in favor of completing my book. During this phase I focus on the action of the story and advancing the plot to my intended HEA. My second phase of writing involves building up character, location and emotional descriptions.
This method works well for me. It helps me actually get my but in a chair and write, without worrying about big details. However, once the "bones" of a book is complete I always go back and add the meat. Good (and hopefully great) books are built on the development of characters and the connections readers are able to make with either, the characters, subject and situations depicted.
When I flesh out my characters, I admit to not spending a lot of time repeating physical characteristics. I'm of a mind that if I describe a person once, I don't need to keep reminding readers every other page or chapter what color their eyes are. Plus I'd rather spend my thesaurus time on emotions and sex scenes ;). Although, I don't often repeat physical descriptions, I do include them– specifically skin tone, race or ethnicity. In a publishing world drowning in whitewashed characters this is very important to me.
Growing up and reading romance novels, I didn't realize how removed color and diversity was from the books I read until one day someone lent me my first Brenda Jackson novel. On the cover was a woman that looked like me. Admittedly the woman was more toned and hair wind swept than me, but for the first time in my romance reading lifetime, a heroine was described as being brown. That blew me away. In that moment I felt a connection with the romance world I didn't realize had been missing. My parents raised me to be confident, and I never doubted that my life could be filled with as much love and adventure as I dared. However, that day I felt as if the rest of the world finally knew that too.
It wasn't till I was much older that I realized how sad that moment in my life was. At that point in time I had read hundreds of romance novels and only one of them featured a person of color as hero or heroine that wasn't a stereotyped caricature. This is a horrible ratio.
The landscape is much different now. There are much more diverse writers and books featuring diverse characters available now. No doubt that there is plenty of room for more (much more). However, as more color and overall diversity is introduced to the books being published (by all kinds of authors and genres), I think it is important to remember what those first few lines of character description mean to a reader who has never had the opportunity to see themselves represented in a book before. That moment of recognition, that simple indelible link formed by a reader is priceless.
Abled, white heterosexual characters are so much the norm, that it can be almost jarring to read others described. That jarring moment for those readers not abled/heterosexual/white can be a strange mixture of euphoria, connection, and sadness. Because the truth is, it shouldn't be such a surprise to see yourself positively reflected in fiction. It should be part of the normal experience, like white-noise, much like the alabaster skinned heroines or steel-eyed heroes that still make up the bulk of romance stories today.
Laurel Cremant is an opinionated author and reader of romance with a wicked sense of humor. RNIC was smart (or crazy) to bring her on as a blogger. In 2016 she took over the management of this site and relishes her new title of "Overlord of Awesome"


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Mar 9, 2017|
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Yeah let me start off only give credit to this entire organization this is really. Entire team effort and look at the number of players we've been able to sign early in this process. That doesn't happen by itself and so you start with ownership and Terry Cain and and the via via allocation of resources and then dug his staff my staff you look at. Jim over north Kenton he yanked him in the hole the really the entire building and this is this an entire entirely a team effort from day one. And in the credit goes aside so. War woods wrecked oh here. Right. Or just blown. Up. Well you know this is this is a tough time a year as far as that goes this time a year and you know there's some players and those are good players and a and and we wish them the best in terms of their careers you know we're gonna do everything in our power. To continue to add pieces to this roster in guys that fit the DNA. That we're looking for and in competition at every position that's for missions against. Iran. Well as a football decision strictly football decision and that that's a talk about a tough are the business is a tough time a year. And and that's my toughest parts in my job as it is when guys leave this building and you know that. So last week when you look at aaron's career when he left Texas a great career and and he's a lot for this organization so we wish the wish Aaron well when. And we're moving forward it as the organizations of football team. Yeah Mike you know and you look at me look at the list of players we've been able to add to this point. You'll see a lot of overlap there with intangibles. Toughness. Versatility and in. In his case as is accused of Mike and when you look at his career he's played multiple positions he played at Iowa had a great career. We find good football players and he's played in nickel position the corner position in the safety positions so and we just felt like this is a great. And great value for us and good football player and he's played in big games and and so we're very fortunate to have Micah. Shaw worked pros won't get it Hillary Clinton at this point so when you say that Aaron Williams what football you know that he can. I try to try to go what does that exactly means if you could expand it was you know whether it's it's. Players as well so on work or what might be. Well irons good football player and and we just feel like at this time as an organization and our philosophy moving forward. That we're going in different directions this time and so when I say it's a football decision that's it's lemon. Three. And a life. I. Right you know again when you mean the question with my Canon and Jordan porn are different so. Versatility. Smarts intangibles. Toughness is restart and and then you look at the skills and on the field ball skills. The ability take the football away in score on defense or put her set up our offense and scoring position of the big for us. Moving forward in and just the ability to run the defense and be that quarterback in the secondary was exports. I know that crazy just aren't really never ends. I don't envision. What are your linebackers right now it seems like there's some that he's had some players who. It is sabres cellular in the secondary when those guys that he's not looking for specific skill sets to the ordinance. But I mean that's we have to be created we are gonna look to add at every position like I mentioned yesterday competition death at every position BT. Linebacker no different. We've played with two linebackers in my career before 7% of the time three linebackers. So you know another percent of time so. You know you you take what you having and you put the players in position be successful and so that's where the creativity comes in with in the system. What this signing. Sessions two fullbacks got. Hurtful but what if he's. Say much hope what does that say don't have those guys. Offensive. Plant. It's just not typical. Paula soon common reasons why this next statement seems. Before buck fullback is is a big part of this nonsense and in and let's let's say it's it's not limited to playing the full back position. There's versatility within that position that's and that's what might bring to the table as you mentioned Mike Holmgren. And I had a chance to be around Mike for a number of years in Carolina know what he brings that table in terms of leadership intangibles again. And what happens in that locker room on Monday through its through Saturday and so that's a big part of it and then. Mike's versatility on the field with the ability to play fullback slot up at fullback and also a transition to have that position he enters the football he's done extremely well so. I had a front row seat to it to observe what Mike Steele said is both on and off the field that was key force and you talk about it again it's been a probe formerly. No different with the mark on this case. We have Melissa there your cars. What all. And that's that's a question France. Where would it Hitler's you like. Over those days that things. The told those are I think you get a feel pretty early what what he's all about I mean he's. First of all as a family man and that person. And and ordered be greatly you've got to lead by example and you see them on a daily basis with with the man of a Mike's caliber off the field. And what he does in the communities. Is phenomenal as well so he's he's just got that it's that people gravitate towards. He's he's known as it is solid dancer around around the locker room. And when I think you'll enjoy it team if and when he gets in the and Sony who likes to showcase some of those town so now he's he's a good addition for us. Race culture change she's loved sports it sounds like that's the type of guy that he would come and make culture change. Critical for him not just a guy like that guys like can you maybe a little and say it's your philosophy. Changing cultures make sure this team which was. I don't mean culture is big and and and in order to have him resulting chemistry you've got to first have the culture and so you bland. High character individuals with with talent. And you had a chance to have that culture and so that is a big part of this and in moving this organization forward in the right direction. There's. Or did you know that he could influence and be okay. Classroom use. That is so when it was thinking. Well this is this is a business and we talk about you've heard me says before earning their right to win. And you know that's. That's we've got to stay focused on the task at hand and and I'm I'm all for as we mentioned chemistry earlier and and billing that chemistry in the locker that's a big part of it. Yet we've got to make sure that we stay focused on the task at hand and that means earned their right to went on a daily basis and and and so I just. No I don't I don't believe that playing video games. In the locker room is part of earned the right wing and and so. We're going to be focused disciplined and accountable football team and and the media you do it one day at a time. I'm so I don't lose hopefully it. Take our real heat cream but. What. Coke glory but his thoughts about receiving position and people's judgment. Going into this game and how you might. Think about dressing room. Knowing that there are always. Right thing you know we're gonna looked at every position wide receiver no difference we do have some house with the with the losses but the team mentioned earlier so we've gotten. Its merits we need to address them and a look at every position no different and wide receiver. Cameron looked at and methodical way and and put this team and position in football games. Because locals band OK go through what was. It. Was qualities. Or network may. Use this. There. I'm sorry to the quality of people are looking for. Work record isn't better. And again just. Guys that wanna compete and got intangibles and no different that the quarterback position I'm looking for certain type of an individual that's why are the right way. Loves to compete. Lost the process. Is gonna put this. Football team and positioning games and and and so. Whether it's as a starter that back up for the third stringer. Burnett good football players this team. And there we're looking every option. And versatility alive to speak feed. Inside just how important is versatility hybrids and rolls on. As soon as that term that we use called you know. Position flexibility for versatility known position flexibility same ads right and so that's important you look at. Micah Hyde Mike has been a return Ares played safety and mentioned corner and nickel so. That's important for us Mike Tolbert played both fullback and halfback. Extremely well so you're looking for. For that value in seeing guys in them when you when you add that component in the locker room. With with these individuals that's big for us. It's. Well you go back to the process and really since day one night when I entered an building in January it's it's been a total team effort and again credit that's got to go to the staff and Doug and his staff the coaching that coaches in just the whole process has been phenomenal. We've had very productive and healthy meetings and and and look when you had healthy means that means sometimes you have to get disagreements and those are those are good those are those are healthy and so. That's been part of the process and at the end of days is a decision we all agreed upon that this would put this organization. In the right direction this time. Certain groups with access. To. You. Murder. Why else would work I'll let you know later. It and stuck or. Process and well and you'd it to now as you go through it I mean you look at every option to say hey. We've got Tyrod and and you look at. The quarterbacks are roster than you you look at that freeagent coolest quarterbacks and and you look at the draft system quarterback so. Like I like I mentioned earlier we're gonna we looked at every option Marie continue to look at every option to add pieces to this to this roster. That are gonna allow us to compete have great competition and at depth and and no different in this case and in yet to look at it certainly from all different angles and you mention them. Really think. But Stephon Gilmore moving. You know focus your attention on you bouncy or to. Step up because it is start for Republicans. We got a lot of lack confidence and in both rounds are being. And keep on as well and some of the other players Russians that position so. Having said that you know we're gonna pursue every opportunity to improve their position as well on there's nothing off the table at this point. And. He says it and the fact that. It was the funds he's. We don't know if you're brave of you guys all important is it freed brings you guys and we're. Yours Leslie Frazier. Maybe some preconceptions from the book what it was like your lies. Well you know you gotta start somewhere certainly and so. You know it's important that we do bring guys in. Basically get him in the ball to learn from day one right and learn together and and that's a big part of the continuity. Builds that communication writes when guys are around one another they did though she communication and it did fuel from one another and in and you build the chemistry and and the communication on defense in particular in this case this is vital to our success though. That's important and as we continue to build this thing. That camaraderie. Just being around one another whether it's off the field in the locker room. And I deleted on table by the way. Pack so oh or you know whatever I mean that's that's that's important. I don't know anything about it. Ball skills. All. Does that still. Work. Yeah I'd again you know just like Gary position nothing's off the table and look to add to this roster in every way shape or form and were up there right. Came down earlier it's only a dozen and going right back up that dug in his staff for up there working as well as much to have so we're at pieces. To this roster nothing's off the table corporate officers. What's the current plan that. Mills. It's. You know we've got some pieces there and you mention a couple of them see Georgetown some good things for this for this organization. And I am looking at it hard just like the other positions. Thanks to us.


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Each edition of Tax Matters consists of free-flowing responses by three tax practitioners to a question regarding a current issue in tax law and policy. Tax Matters commentaries provide insightful perspectives on a broad range of topics, making important contributions to the dialogue within the tax bar about cutting-edge issues. Although the commentaries are certainly of interest to the academic community, they are primarily directed toward tax professionals and their clients.
United States taxation of worldwide income combined with a high corporate tax rate disadvantages US-headed multinational groups compared with groups with the same income mix but a non-US parent. The disadvantage has become more pronounced in recent years as more countries move to territorial systems and lower rates.
US groups have a strong tax incentive to restructure—including by foreign mergers—so their parent is non-US. They then will never owe US tax on foreign source income earned in subsidiaries of the non-US parent, and so may be able to outbid US competitors for foreign acquisitions. Corporate inversions also enable conversion of US-source income into untaxed foreign-source income through deductible payments to foreign affiliates. Moreover, US dividend withholding tax no longer applies.
Congress and the Treasury have reacted several times to limit inversions. Section 367(a) regulations finalized in 1996 tax US shareholders on gain if together they end up with more than half the shares of the new foreign parent. Enacted in 2004, section 7874 generally treats the new parent as domestic if at least 80% of its shares are held by former shareholders of the inverting domestic corporation by reason of having held its shares, unless the group has substantial business activities in the new foreign parent's home country. Last year temporary regulations narrowly construed this exception.
Foreign mergers continue to occur notwithstanding section 7874, however, including Perrigo/Elan (Ireland) (announced July 29) and Actavis/Warner Chilcott (Ireland) (announced May 20). Reports of expected tax savings have renewed focus on the policy issues inversions present. Why are they of more concern than decisions to incorporate a new business abroad? To what extent do expected savings arise from deductible related party payments? What do inversions say about whether the United States can sustain a system increasingly out of step with a world of territorial, low-rate systems? Why do inversions reduce effective tax rates if US groups already can permanently defer tax—and book tax expense—on reinvested business income of foreign subsidiaries? What practical effect has section 7874 had? Should tax planners advise more US-headed multinational groups to merge or restructure, planning around section 7874, so that they have a non-US parent? What if any non-US tax constraints limit inversions?
By Robert Scarborough, Lecturer in Law, Columbia Law School


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He helped a U.S. president avoid nuclear Armageddon. He married a woman who lived in a bottle. And the entire world reeled in shock when he was shot by an unknown assailant.
Actor Larry Hagman died Friday of complications from cancer. He was 81.
Hagman's breakthrough role was in film, opposite Henry Fonda's president in the 1964 political thriller Fail-Safe. But his real fame came from television, first in the surreal '60s sitcom I Dream of Jeannie and then, most significantly, in the 1970s as the iconically villainous J.R. Ewing in the internationally adored nighttime soap opera Dallas.
Hagman returned to his most infamous role in the sequel series to Dallas, which debuted earlier this year and became an instant hit. He announced his ongoing cancer treatment shortly after filming began in 2011. He died while still filming in the city for which the show is named.
He is survived by Maj, his wife of 50 years — a union almost unprecedented in showbiz circles — and son Preston and daughter Kristina.
Hagman was born into the business, in Fort Worth, Texas in 1931, the son of soon-to-be Broadway star Mary Martin, who would later originate the indelible Tony Award-winning roles of Nellie Forbush in South Pacific, Peter in Peter Pan and Maria in The Sound of Music.
Hagman's father was a lawyer whose clients included several rich Texas oil barons, memories of whom Hagman would later draw on to play the meanest oilman of them all, J.R.
His parents divorced in 1936, with the 5-year-old Hagman shuttled off to Los Angeles to live with his maternal grandmother. He spent most of his childhood and adolescence in private and military schools, none of them for very long. After a brief time with his mother in New York, he moved back to Texas to take a short-lived stab at college.
Failing that, and having inherited the Martin acting gene, the young Hagman landed several jobs in regional stock theatre. In 1951, his mother stepped back in and brought him to London for a minor role in the West End version of South Pacific. He stayed in Britain for another five years, serving in the U.S. air force, mostly in travelling USO shows.
Hagman returned to New York in 1956 and earned more minor roles in several off-Broadway and Broadway productions. At the same time, he was making inroads into television drama, which in the '50s was still drawing most of its talent from the New York stage.
He made his film debut in 1964 in Ensign Pulver, a sequel to 1955's Mr. Roberts, which had headlined Henry Fonda, the star of Hagman's next film, Fail-Safe.
TV's I Dream of Jeannie followed a year later. This time Hagman had the lead, as harried astronaut Maj. Tony Nelson who had to cope with the magical mishaps conjured up by his genie, played with a demure and, in retrospect, disturbing subservience by Barbara Eden.
Reacting to the news of Hagman's death, Eden wrote this weekend on her Facebook page: "I still cannot completely express the shock and impact from the news that Larry Hagman has passed. I can still remember, that first day on Zuma Beach with him, in the frigid cold. From that day for five more years, Larry was the centre of so many fun, wild, shocking ... and in retrospect, memorable moments that will remain in my heart forever.
"Amidst a whirlwind of big laughs, big smiles and unrestrained personality Larry was always, simply Larry. You couldn't fault him for it, it was just who he was."
Even before donning that huge Stetson in Dallas, Hagman was notoriously larger than life. He was opinionated, eccentric and an inveterate practical joker, known in some circles as "the Mad Monk of Malibu." He smoked pot with Jack Nicholson and dropped acid with rocker David Crosby. He also smoked cigarettes and drank extravagantly.
Though no less colourful in personality, Hagman put aside his unhealthy habits following the successful removal and replacement of a cancerous liver in 1995.
This was four years after the end of Dallas, a 13-season run which transformed Hagman into an international superstar: an estimated 350 million people in 57 countries tuned in to the pivotal 1980 cliffhanging episode "A Family Divided," which had everyone asking, "Who shot J.R.?"
Hagman seized the opportunity to bully his way to a better deal and ended up making $1 million a year. Not quite Charlie Sheen money — Sheen was making slightly more than that per episode when he left Two and a Half Men — but substantial enough at the time to make Hagman one of the highest-paid television actors ever.
While the network and production company frantically hatched contingency plans, commissioning scripts based on the premise that J.R. had died, Hagman calmly held out, whisking his family off to Europe to wait out the storm.
Hagman's fame at the time was such that his own mother resented it. There is an often-told story among veteran TV journalists of a Hagman-hosted press party at his Malibu home at which Mary Martin loudly complained to anyone who would listen that she was a much bigger star than her son.
In middle age, Hagman became an advocate for various causes, from organ donation to antismoking to clean energy, which would have appalled his oilman alter ego. He was not, despite his Texas background, a fan of George W. Bush, whom he referred to as "a sad figure, not too well educated, who doesn't get out of America much. He's leading the country towards fascism."
Hagman bounced back from his health woes in the late 1990s to take on a range of character roles in both movies and TV and to revive J.R. in several TV-movie specials. And then in the contemporary Dallas spinoff that he was still shooting last week at the time of his death.
In the first episode of the new series, J.R. initially appeared as a frail, barely aware old man, only to be proven hale and hearty and ornery as ever by the end of the episode. How doubly appropriate, given Hagman's penchant for gags, and J.R's Ewing's Machiavellian brilliance at faking out his unwitting opponents.
On the weekend, the New York Times quoted Hagman from a 1980 interview: "Life is terminal, death is not. I think death is just another stage of our development. I honestly believe that we don't just disappear. We don't go into a void. I think we're part of a big energy curtain, an energy wave, in which we are like molecules."
Molecule or not, Larry Hagman is not about to disappear. Not as long as there are reruns and DVD box sets.


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SAO PAULO, Sept 27 (Reuters) - Brazil's state-run oil company Petróleo Brasileiro SA said on Thursday in a securities filing it has reached an agreement with U.S. officials to settle investigations in the United States.
Petrobras, as the company is known, will provision $853.2 million in its third-quarter results to pay for the deal with the Department of Justice and the Securities and Exchange Commission. (Reporting by Carolina Mandl)


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My name is Maggie Raynard. After sixteen years being just plain me, suddenly, when I lose my temper, my fingers become weapons of mass destruction. Turns out I'm a semi-god, descended from Aphrodite. Sou nds cool in theory, but when I accidentally put my ex-boyfriend in a coma, things go downhill pretty fast. Now some new guy named Mac Finnegan has made it his mission in life to continually piss me off. I'm stuck learning how to use my new powers while also dealing with regular high school problems, and with this annoying—and super-hot—guy all up in my business, I'm about to flip out. But it gets worse. I just learned there's this watchdog council of semis who keeps an eye out for any bad apples. They think I'm the baddest of the bunch and want to take me out before I do any more damage. My nemesis Mac might turn out to be my salvation, only he's got secrets of his own...and they may just kill us both.
About The Author: Christine O'Neil is one half of the happiest couple in the world. She and her handsome hubby currently reside in Pennsylvania with a four-pack of teenage boys and their two dogs, Gimli and Pug. If she gets time off from her duties as maid, chef, chauffeur, or therapist, she can be found reading just about anything she can get her hands on, from Young Adult novels to books on poker theory. She doesn't like root beer, clowns or bugs (except ladybugs, on account of their cute outfits), but lurrves chocolate, going to the movies, the New York Giants and playing Texas Hold 'Em. Writing is her passion, but if she had to pick another occupation, she would be a pirate…or, like, a ninja maybe. She loves writing fun and adventure-filled romance stories, but also hopes to one day publish something her dad can read without wanting to dig his eyes out with rusty spoons. Christine loves to hear from readers, so please feel free to get in touch with her via the Contact Page. Christine also writes adult romance under the name Christine Bell.
- 1 sterling silver ring and a Team Mac pin as well as temporary tattoos of the magpie.
- 1 sterling silver ring and a Team Mac pin as well as temporary tattoos of the magpie.
- 1 fourteen karat gold ring, and a Team Mac pin as well as temporary tattoos of the magpie.
Week 1
9/2/2013- Crossroad Reviews- Guest Post
9/3/2013- The Book Belles- Review
9/4/2013- Manga Maniac Cafe- Interview
9/5/2013- A Backwards Story- Guest Post
9/6/2013- Coffee, Books and Me- Interview
Week 2
9/9/2013- Books Are Magic- Review
9/10/2013- K-Books- Guest Post
9/11/2013- Curling Up With A Good Book- Review
9/12/2013- Fantasy Book Addict- Interview
9/13/2013- Ticket to Anywhere - Review


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Dr Mike's Wellness Blog
Dr Mike's Wellness Blog, with the latest information and health advice, makes it extremely convenient for readers to learn better ways to improve their personal health. It provides a wealth of articles, editorials and helpful programs designed for increasing every user's health. Dr Mike's Wellness Blog represents a valuable opportunity for health devotees from across the world. By letting the renowned Dr Mike share his expertise in wellness through his regularly updated blog, the website…Read More


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After friday news and Richard's voiceover in beer comercial I consider to start drinking a beer. Bye Bye green tea! Thank You to Kings and MissCachou for finding this commercial. :) Now I become full RAcoholic! BTW Beer is very good for hair.
1. A beer NEVER leaves the toilet seat up.
2. A beer lasts longer than seven seconds.
3. A beer won't expect you to cook dinner when you're not hungry.
4. A beer will never expect you to sit in the wet spot IT makes.
5. A beer doesn't care if you go shopping.
6. A beer doesn't mind when your mother visits.
7. A beer does as many chores as a man, with a LOT less complaining.
8. Having a beer can't make you pregnant.
9. A beer won't tease you because you once liked Barry Manilow.
10. You don't have to laugh at a beer's jokes.
11. A beer won't mind at all if you're not in the mood for beer.
12. If a beer had a sports car, it wouldn't love it more than you.
13. A beer doesn't want to go out alone with the other beers.
14. A beer doesn't sulk.
15. A beer wouldn't waste its money on Playbeer magazine.
16. A beer won't switch the TV channel.
17. A beer doesn't have to sleep with the windows open.
18. A beer doesn't snore.
19. A beer can't interrupt.
20. A beer doesn't care that you can't find your car's carburetor.
21. A beer doesn't think black leather bikinis are neat.
22. A beer doesn't belch. Or fart.
23. A beer doesn't mind having pantyhose dry in the bathroom.
24. A beer doesn't care that you don't balance your checkbook.
25. A good beer is easy to find.
26. A beer can't pout.
27. A beer doesn't have hair in strange places.
28. A beer doesn't have friends who will drink your beer.
29. A beer wouldn't yell if you dented the car.
30. A beer won't get jealous if you enjoy another beer.
31. A beer won't care if you gain five pounds.
32. A beer will be there for anytime of the month.
33. A beer doesn't want children.
34. A beer doesn't think poetry is queer.
35. A beer isn't ready until you're ready.
36. If the beer is finished before you are, you can have another beer.
37. Hangovers go away.
38. A beer tastes good.
39. Having a beer doesn't make you want to take a shower.
40. A beer will never invite friends home for dinner without calling.
41. A beer's life does not revolve around the football.
42. A beer would never make fun of your new outfit.
43. A beer never needs a shave.
44. You don't have to let a beer win.
45. A beer doesn't care what toppings you get on the pizza.
46. Just because you have dinner with a beer doesn't mean you have to sleep with a beer too.
47. A beer doesn't have morning breath.
48. A beer is happy to go where ever you want to go.
49. A beer will never drink the last beer.
50. A beer will never take the newspaper apart before you've read it.
51. When a beer is finished, it doesn't roll over and go to sleep.
52. A beer wouldn't mind if you wanted it to wear a condom.
53. A beer is never temperamental.
54. A beer will never complain about your cooking.
55. A cold beer is a good beer.
56. A beer will never worry about losing its hair.
57. A big, fat beer is nice to have.
58. A beer won't steal the covers.
Emm...still problem is that I don't drink beer...


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Kmart Has A $35 Hand Held Steamer Better Than Your Iron
There are times when getting out the iron, putting up the ironing board and doing a whole load of clothes you've put off for weeks can be super painful.
Your ironing board may or may not stay up for days after, and you may forget to actually put the iron away because after use it's too hot.
Behold, your new best friend - the Kmart Hand Held Garment Steamer.
(Image: Kmart)
Not only is this product small and light, it's robust, tough and only $35 buckaroos!
The steamer is quick and easy to use.
Just hang your fave top, dress or pants up on a doorknob or rail, fill up the steamer's water tank, wait thirty seconds and then aim the steamer towards the fabric. Voila. Crease be gone.
You're welcome.


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Part-time receptionist needed for an award-winning studio
- Recruiter
- Location
- London
- Salary
- 24000-30000 per annum
- Posted
- 27 Oct 2017
- Closing date
- 23 Nov 2017
- Ref
- 38510
- Job function
- Secretarial
- Job level
- Experienced (non manager)
- Hours
- Part time
- Contract
- Permanent
- Listing type
- Job vacancy
An AJ100 rated architecture practice based in London's design hub is looking for a front of house receptionist to join their team on a part time basis. Spread between 2 studio locations, which are in close proximity, this person will be responsible for providing first class service to both internal and external clients. Key areas of responsibility will include maintaining the reception area, being key point of contact for clients, answering calls, keeping the office security up to date, booking couriers, cabs, train tickets and car hire, liaising with distributors, assisting with admin duties, and processing / monitoring expenses. The ideal candidate will be IT literate, professional, have good communication skills, and excellent attention to detail. If you would like the chance to work at a progressive, award-wining studio, which is one of the leading design-based architectural practices in the UK, then apply now. This is a 12 month fixed term contract. Salary is pro rata.
Similar jobs


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Feel like you're sitting on cotton candy or a cloud with this perfect mid-century bar chair. Gable features a shiny round base and a footrest in polished brass, with a fixed height of 1m. It is fully upholstered in cotton velvet and features a super stylish design, with an accent low back and a cylindrical shape.
Length: 55,50 cm | 21,85 in
Width: 63,50 cm | 25 in
Height: 100 cm | 39,37 in
Feet: Polished Brass; Cover: White Leather
These are our choice of products to pair your favorite with, the ones that will complement your decor in seconds.
Our very own list of products similar to the one you're looking for, available for you to choose your next favorite!


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Anti-BDS Legislation Passes Pennsylvania Senate and House
Read the full article at: http://jewishexponent.com/2016/10/26/anti-bds-legislation-passes-pennsylvania-senate-and-house/
An anti-BDS bill has passed the Pennsylvania Serante by a 47 to 1 vote.
Less than a month after California Gov. Jerry Brown signed a bill against the boycott, divestment and sanctions (BDS) movement, the Pennsylvania Senate has passed a similar bill by a vote of 47 to 1. After an affirmative concurrence vote in the House, Pennsylvania HB 2107 is now on Gov. Tom Wolf's desk for his signature.
Should the bill be enacted, Pennsylvania would become the 14th state to pass anti-BDS legislation.
The prime sponsor of the bill is state Rep. Matthew E. Baker, a Republican serving Tioga County and parts of Bradford and Potter counties.
The bill — which also addresses questions regarding public access to procurement records and awarding a contract to a single source — contains anti-BDS language within the context of prohibited contracts.
The bill asserts that "Israel is America's dependable, democratic ally in the Middle East, an area of paramount strategic importance to the United States" and that "it is in the interest of the United States and the Commonwealth to stand with Israel and other countries by promoting trade and commercial activities and to discourage policies that disregard that interest."
For that reason, a company contracted by the Commonwealth is not permitted to be involved in a boycott—defined in the bill as "to blacklist, divest from or otherwise refuse to deal with a person or firm when the action is based on race, color, religion, gender or national affiliation or origin of the targeted person or entity."
Jacob Millner, of the Israel Project — a national nonpartisan educational organization that has been instrumental in pursuing anti-BDS legislation across the country — said he was pleased about the passage of the bill.
"I feel good about it. I'm pretty confident, though I don't like to count my chickens before they hatch. But Pennsylvania has a lot of organization who have done a lot of good work on making this happen."
Millner says the Pennsylvania bill is noteworthy because "it's important that states make a statement on the right side of history, that they won't invest their money or spend state money on companies that would engage in anti-Israel and anti-Semitic [activities]."
Millner called BDS "a form of economic warfare against the only Jewish state," and added, "the state wouldn't condone a company that would discriminate against African-Americans, women or gays and lesbians, and discrimination based on national origin is similar. The state has to take a stand on that."


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Follow me as I travel around Ecuador gathering research for my books in the "Culture Smart!" series. Join me as I learn about the people, food, culture, history and fabulous natural treasures of this fascinating Andean country.
Monday, January 14, 2013
I'm Off to Discover Ecuador
I thought the best way to launch this blog was to post a new video from the Ecuador Tourism Ministry that captures the essence of the Ecuador I discovered many years ago when I traveled overland from Caracas through Colombia to Quito.
Far from the postcard images of stately churches and big-eyed llamas I found a land of contrasts; tiny mountain hamlets seemingly unchanged since the days of the Incas, steamy jungles full of colorful butterflies and birds, and crazy cities like Guayaquil where the parties last all night.
I also discovered remote beaches with salt-encrusted surf dudes just as cool and laid back as the ones I'd left behind in Caribbean Colombia and Venezuela.
Back then I had no guidebook or any real clue what I would find in Ecuador but the offer of a free bed and a few weeks of freedom to explore was enough to get me on the bus.
Once there I just followed my instincts and went with the flow: Quito, Otavalo, Mindo, San Rafael, San Vicente, Cuenca, Vilcambamba, Guayaquil, Galapagos - each place different to the one before, like a microcosm of South America squeezed into one tiny country.
That first adventure and the people I met on it has set the tone for all subsequent trips.
And now I'm setting off again for the land of volcanoes, beaches and jungles and all I can think about is chowing down on a cuy (roast guinea pig), sipping on a cool juice made from tomate de arbol, and meeting heaps of local peeps who can help me get a deeper understanding of this amazing country.
If you want to help me gather research for my new book "Culture Smart! Ecuador" send me an email at email@example.com or leave a comment below.
In the meantime, I'll be posting photos and videos and recipes and anything interesting that I stumble across on my travels.
Posted by Russell Maddicks at 11:28 AM
Labels: adventure, ama la vida, Cuenca, Culture Smart Ecuador, cuy chicas bellas, discover, Ecuador, Montanita, Quito, Quito Visitor's Bureau, Roast Guinea Pig, Russell Maddicks, tourism, travel
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Oh oh! Looks like the 'plus-size model' label is completely OUT when it comes to Crystal Renn, since the "Hunger" author appears to have lost even more weight, as these new pictures reveal. Let's see the explanation from NY Mag:
Ford posted photos of Crystal Renn posing in a Fashion for Passion tank, sales of which benefit photographer Nicholas Routzen's campaign for children and the arts. The blogosphere is shocked by how much weight she appears to have lost, especially after she looked to have lost weight before walking in the last Chanel cruise show, which was huge for her career.
Crystal's agent Gary Dakin was contacted and he said:
"She is still a 39-40 hip (she fluctuates like any woman), and a solid size 10. She has no urge [to lose weight]. She's an amazing model. She can make herself look bigger or smaller depending on what the client wants."
Definitely not looking like a 10 in the above pictures!
So what is it? Further (intentional or unintentional) weight loss? Angles? Photoshop? Crystal's slimming "talent" (as the agent says)? Maybe lens distortion?