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The Saccharomyces cerevisiae Shu2 protein is an important regulator of Rad51, which promotes homologous recombination (HR). Shu2 functions in the Shu complex with Shu1 and the Rad51 paralogs Csm2 and Psy3. Shu2 belongs to the SWS1 protein family, which is characterized by its SWIM domain (CXC...Xn...CXH), a zinc-binding motif. In humans, SWS1 interacts with the Rad51 paralog SWSAP1. Using genetic and evolutionary analyses, we examined the role of the Shu complex in mitotic and meiotic processes across eukaryotic lineages. We provide evidence that the SWS1 protein family contains orthologous genes in early-branching eukaryote lineages (e.g., Giardia lamblia), as well as in multicellular eukaryotes including Caenorhabditis elegans and Drosophila melanogaster. Using sequence analysis, we expanded the SWIM domain to include an invariant alanine three residues after the terminal CXH motif (CXC…Xn…CXHXXA). We found that the SWIM domain is conserved in all eukaryotic orthologs, and accordingly, in vivo disruption of the invariant residues within the canonical SWIM domain inhibits DNA damage tolerance in yeast and protein-protein interactions in yeast and humans. Furthermore, using evolutionary analyses, we found that yeast and Drosophila Shu2 exhibit strong coevolutionary signatures with meiotic proteins, and in yeast, its disruption leads to decreased meiotic progeny. Together our data indicate that the SWS1 family is an ancient and highly conserved eukaryotic regulator of meiotic and mitotic HR.
HOMOLOGOUS recombination (HR) is an error-free DNA repair pathway that is essential both in mitotic cells to ensure DNA stability and in meiotic cells for faithful chromosome segregation. HR begins with double-strand break (DSB) formation and DNA end processing that give rise to 3′ single-stranded DNA (ssDNA) overhangs (Heyer et al. 2010). A key step in HR is the formation of RecA-like filaments on these ssDNA ends. During mitosis in the budding yeast Saccharomyces cerevisiae, the RecA-like protein Rad51 coats the ssDNA, whereas during meiosis, both Rad51 and another RecA-like protein, Dmc1, form on ssDNA ends (Lin et al. 2006; Heyer et al. 2010). Formation of RecA-like filaments on the DNA is essential for the homology search and strand-invasion steps that define HR. Therefore, regulation of RecA filament formation is critical for accurate repair of DNA damage and chromosome segregation. Given their importance, both Rad51 and Dmc1 are extremely well-conserved descendants of the archaeal protein RADA (Diruggiero et al. 1999; Komori et al. 2000; Lin et al. 2006; Chintapalli et al. 2013).
There are many proteins that both promote Rad51 filament formation and disassemble inappropriate filaments. Interestingly, in many organisms, the proteins that stabilize Rad51 filaments themselves share structural homology with Rad51 and evolved from the archaeal RADB homolog (Lin et al. 2006). In humans, these Rad51 paralogs include RAD51B, RAD51C, RAD51D, XRCC2, XRCC3, and SWSAP1, whereas in the budding yeast, they include RAD55, RAD57, CSM2, and PSY3 (Schild et al. 2000; Takata et al. 2001; Martin et al. 2006; She et al. 2012; Tao et al. 2012; Godin et al. 2013; Sasanuma et al. 2013b). It has been proposed that SHU1 is also a Rad51 paralog (Martin et al. 2006). A great deal of work has been done to characterize these proteins in vitro, in vivo, and phylogenetically (Kawabata et al. 2005; Lin et al. 2006; Heyer et al. 2010; Suwaki et al. 2011). Importantly, many human Rad51 paralogs are mutated in cancers and are associated with cancer predisposition (Vaz et al. 2010; Suwaki et al. 2011; Park et al. 2012; Pennington and Swisher 2012; Shamseldin et al. 2012; Somyajit et al. 2012; Filippini and Vega 2013). Consistent with a critical role in genome maintenance, disruption of the yeast Rad51 paralogs results in a mutator phenotype and in some cases increased chromosomal rearrangements, which are often observed in tumor cells (Huang et al. 2003; Shor et al. 2005). In budding yeast, Shu1 and the Rad51 paralogs Csm2 and Psy3 form an obligate heterotetramer called the Shu complex (also referred to as the PCSS complex) with a fourth member, Shu2, whose major defining feature is a SWIM domain (Shor et al. 2005; Martin et al. 2006). The association of divergent Rad51 paralogs with a SWIM domain–containing protein is conserved in fission yeast Schizosaccharomyces pombe, where the Rad51 paralogs Rlp1 and Rdl1 interact with a Shu2 homolog Sws1 (Martin et al. 2006). Similarly, the human homolog of Shu2, hSWS1, interacts with the Rad51 paralogs RAD51D and XRCC2 (Martin et al. 2006; Liu et al. 2011). Importantly, hSWS1 functions as an obligate heterodimer with hSWSAP1, which itself resembles a previously unidentified highly divergent hRad51 paralog (Liu et al. 2011).
The conserved association between Shu2-like proteins and the Rad51 paralogs promotes Rad51-dependent HR via a largely undetermined mechanism (Godin et al. 2013). In all species where Shu2-like proteins have been described, their disruption results in a reduction in Rad51 filament formation and a corresponding decrease in HR (Shor et al. 2005; Martin et al. 2006; Liu et al. 2011; Sasanuma et al. 2013b). Strikingly, these defects are similar to disruption of the Rad51 paralogs (Sasanuma et al. 2013b). It remains to be determined what the functional significance of the SWIM domain is and why Rad51 paralogs associate with SWIM domain–containing proteins to promote HR throughout several eukaryotic lineages.
To further our understanding of the Shu2/SWS1 protein family, we characterized its evolution across taxonomic groups with a special focus on conservation of the defining SWIM domain. During this process, we confirmed the orthologous relationship of Shu2 and hSWS1 and discovered previously unknown SWS1 orthologs in multiple species, including Caenorhabditis elegans and Drosophila melanogaster. Interestingly, evolutionary analysis implicates yeast Shu2 and its fly ortholog CG34314 in meiosis because they show strong rate covariation with meiotic proteins—specifically with those that contribute to HR. Through analysis of eukaryotic Shu2/SWS1 orthologs, we found that the SWIM domain is invariant and can be expanded to include an invariant alanine residue after the canonical SWIM domain. Furthermore, we identified in the literature a cancer patient from the COSMIC database who harbors a mutation in this invariant alanine (Shepherd et al. 2011). In vivo disruption of the invariant SWIM domain residues likely results in protein instability and loss of function. Together our work indicates that the SHU2 gene is found in all major eukaryotic lineages, where it promotes HR in both mitosis and meiosis.
Materials and Methods
Yeast strains, plasmids, and media
The strains used in this study are listed in Supporting Information, Table S1 and are isogenic to RAD5+ W303, except for the PJ69-4A and PJ69-4α yeast two-hybrid strains (Thomas and Rothstein 1989; James et al. 1996; Zhao et al. 1998). The primers used are listed in Table S2. Standard protocols were used for yeast culturing, transformation (LiOAc method), sporulation, and tetrad dissection. The medium was prepared as described previously with twice the amount of leucine (Sherman et al. 1986). The yeast two-hybrid (Y2H) plasmid for hSWS1 was created by PCR amplification using pJ636 (pcDNA3-3HA-hSWS1) from Paul Russell as a template with oligonucleotides hSWS1.F and hSWS1.R and subcloned into the EcoRI and SalI restriction sites of pGBD-C1 (Martin et al. 2006). hSWSAP1 was PCR amplified from the MYC-SWSAP1 vector from Jun Huang with oligonucleotides SWSAP1.F and SWSAP1.R and subcloned into the EcoRI and SalI restriction sites of pGAD-C1(Liu et al. 2011). Creation of pGBK-shu2-C114S (Shu2.C114S.F and Shu2.C114S.R), pGBK-shu2-C116S (Shu2.C116S.F and Shu2.C116S.R), pGBK-shu2-F119A (Shu2.F119A.F and Shu2.F119A.R), pGBK-shu2-C176S (Shu2.C176S.F and Shu2.C176S.R), pGBK-shu2-H178A (Shu2.H178A.F and Shu2.H178A.R), pGBK-shu2-A181T (Shu2.A181T.F and Shu2.A181T.R), pGBD-SWS1-C85S (hSWS1.C85S.F and hSWS1.C85S.R), pGBD-SWS1-C87S (hSWS1.C87S.F and hSWS1.C87S.R), pGBD-SWS1-F90A (hSWS1.F90A.F and hSWS1.F90A.R), pGBD-SWS1-C103S (hSWS1.C103S.F and hSWS1.C103S.R), pGBD-SWS1-H105A (hSWS1.H105A.F and hSWS1.H105A.R), and pGBD-SWS1-A108T (hSWS1.A108T.F and hSWS1.A108T.R) was accomplished using site-directed mutagenesis of the pGBK-SHU2 or pGBD-SWS1 plasmids.
Integration of a C114S mutation at the endogenous SHU2 locus was accomplished by creating an integration vector by subcloning with EcoRI and PstI from pGBK-shu2-C114S in the yiPLAC211 integration vector. Wild-type (WT) yiPLAC-SHU2 was created by reversing the C114S mutation by site-directed mutagenesis using Shu2.S114C.F and Shu2.S114C.R. yiPLAC211 was subsequently mutagenized to C116S, F119A, C176S, H178A, and A181T with the primers listed earlier (Figure S2). The WT and mutant versions of yiPLAC211-shu2 were linearized with BamHI and transformed into W9100-2D. Pop-outs were screened on 5-fluorooritic acid and verified via PCR. All inserts were verified by DNA sequence analysis.
Serial dilutions
The indicated strains were grown to an OD600 of 0.5 and then fivefold serially diluted onto rich medium (YPD) or YPD with either 0.006, 0.012, and 0.02% methyl methanesulfonate (MMS) or 50 mM hydroxyurea (HU). For strains harboring pGBK-shu2 vectors, serial dilutions were performed as earlier with the exception that cells were grown in SC-TRP medium prior to plating onto YPD medium or YPD with 0.02% MMS. Plates were incubated for 2 days at 30° prior to imaging.
Yeast two-hybrids
The GAL4 DNA-activating domain (pGAD) expressing plasmids was transformed into PJ69-4A, and the GAL4 DNA-binding domain (pGBK or pGBD) expressing plasmids was transformed into PJ69-4α. The SHU1 gene was disrupted with NatNT2 using pFA6A-NatNT2 as described by Janke et al. (2004) in both the PJ-694A or PJ69-4α strain backgrounds where indicated. The plasmid containing PJ69-4A and PJ69-4α haploid yeast cells was mated, and diploids were selected by growth on SC-LEU-TRP solid medium. Individual colonies were grown to early log phase to OD600 0.2, and then 5 µl was spotted onto medium to select for the plasmids (SC-LEU-TRP) or onto medium to select for expression of the reporter HIS3 gene (SC-LEU-TRP-HIS) indicating an interaction. Plates were incubated for 2 days at 30° and photographed. The experiments were done in triplicate.
Homology searching and phylogenetics
Yeast Shu2 homologs from fungi were retrieved from the nonredundant amino acid sequence database at NCBI using PSI-BLAST. Hits with E-values below 0.005 were used for subsequent iterations. An additional PSI-BLAST search beginning with the human SWS1 (ZSWIM7) protein led to hits across many eukaryotic taxa and in archaea after three iterations. While Shu2 homologs were found in hundreds of species, a selection representing major lineages and model organisms is shown in Figure 2. Protein-sequence phylogenies were inferred in PhyML using the LG substitution model with four rate classes (Guindon and Gascuel 2003). Branch support values were generated with the approximate likelihood ratio test (aLRT).
Evolutionary rate covariation of Shu2 and fly SWS1 with meiotic and mitotic proteins
Values of evolutionary rate covariation (ERC) were calculated using previously described methods (Clark et al. 2012). Briefly, orthologous protein sequences were collected from species with sequenced genomes and aligned in muscle (18 fungal species and 12 Drosophila species for their corresponding data sets) (Edgar 2004; Clark et al. 2012; Findlay et al. 2014). For each protein, we then estimated amino acid branch lengths using a fixed-tree topology and the aaml program of the PAML package. Branch lengths then were transformed to relative rates using a projection operator (Sato et al. 2005; Yang 2007). The ERC value between any two proteins was calculated as the correlation coefficient between their evolutionary rates (Table S3, Table S4, Table S5, Table S6, Table S7, Table S8, Table S9, and Table S10).
The elevation of Shu complex ERC values as a group was tested by comparison with 100,000 random sets of genes of the same size (N = 4 proteins). A P-value was estimated from the number of random protein sets with mean ERC values equal to or greater than the mean ERC between Shu complex proteins. Sets of mitotic, meiotic, and "recombinase activity" proteins were obtained from the Gene Ontology Annotation Database through the Yeast Mine and FlyBase Query Builder web tools for yeast and Drosophila, respectively (Wilson et al. 2008; Balakrishnan et al. 2012). Statistical significance for ERC between Shu2 and these functional groups was determined by comparing their ERC distributions with the whole-proteome background (all genes) through Wilcoxon rank-sum tests.
Spore viability assay
Diploid yeast strains where both copies of the indicated genotype had been disrupted or mutated at the endogenous locus were sporulated at 30°. The individual spores were tetrad dissected onto rich medium, and spore viability was ascertained. A plate of 22 individual tetrads was analyzed in triplicate, and standard deviations were calculated.
Mitotic recombination assays
Mitotic recombination rates were calculated from WT, shu1∆, and shu2∆ cells containing the leu2-∆EcoRI::URA3::leu2-∆BstEII direct-repeat recombination assay, as described by Alvaro et al. (2007). Gene conversion (GC) events were measured by Leu+ Ura+ colonies, and single-strand annealing (SSA) recombinants were measured by Leu+ Ura− colonies. Nine individual colonies were analyzed for each genotype, and the experiment was performed in triplicate. The mitotic recombination rate and standard deviation were calculated as described earlier (Lea and Coulson 1949).
Characterization of shu2∆ recombination phenotype and its physical interactions with the other Shu complex members
The Rad51 paralogs have a number of defining features that are related to promoting Rad51 filament formation, such as decreased rates of Rad51-mediated gene conversion on disruption (Sung 1997; Alvaro et al. 2007; Godin et al. 2013). Consistently, we previously found that deleting either CSM2 or PSY3 results in a decrease in gene conversion and a subsequent increase in Rad51-independent repair (Godin et al. 2013). To determine if Shu1 or Shu2 has a similar role in promoting Rad51-dependent repair, we performed a heteroallelic recombination assay in WT, shu1∆, and shu2∆ cells (Figure 1A). In this assay, a recombination event between two leu2 heteroalleles with an intervening URA3+ gene can generate a LEU2+ prototroph through repair by Rad51-dependent sister-chromatid GC (LEU2+ URA3+) or Rad51-independent intrachromosomal SSA (LEU2+ URA3−). Similar to csm2∆ and psy3∆ cells (Godin et al. 2013), we found that shu1∆ and shu2∆ cells have significantly decrease rates of Rad51-dependent GC (P ≤ 0.02 and P ≤ 0.005, respectively) with a corresponding increase in rates of error-prone SSA (Figure 1A). These results demonstrate that although Shu2 is not a Rad51 paralog, it exhibits many of the same phenotypes as the other Shu complex members.
The human Shu2 homolog hSWS1 interacts with multiple Rad51 paralogs either directly or indirectly through hSWSAP1 (Martin et al. 2006; Liu et al. 2011). We asked whether other yeast Shu members similarly bridge the protein-protein interactions between Shu2 and Shu1 or Psy3 (Shor et al. 2005; Ball et al. 2009). To address this question, we performed Y2H analysis of yeast expressing SHU2 in the GAL4 DNA-binding domain (pGBK-SHU2) and tested its interaction with either SHU1 or PSY3 expressed in the GAL4 DNA-activating domain (pGAD-SHU1, pGAD-PSY3) in a genetic background in which one of the four SHU genes is disrupted (Figure 1B and data not shown). We found that loss of SHU1 disrupts the Shu2-Psy3 Y2H interaction (Figure 1B). Therefore, similar to hSWS1, which interacts with the other hRad51 paralogs through hSWSAP1, the interaction of Shu2 with the Rad51 paralog Psy3 is likely stabilized by Shu1.
Shu2 orthologs are conserved across eukaryotes
Although Shu2 homologs were identified in fission yeast and humans (Shor et al. 2005; Martin et al. 2006), their phylogenetic orthology with budding yeast Shu2 has not been demonstrated, nor have Shu2 orthologs been identified in other major eukaryotic lineages, including important model organisms. To produce a complete picture of the SWS1 family, we searched for Shu2 homologs across eukaryotic and outgroup archaeal lineages. PSI-BLAST queried with yeast Shu2 yielded hits across fungal species and metazoans. PSI-BLAST queried with human SWS1 (ZSWIM7) led to hits across eukaryotes, including plants, and archaeal proteins containing a SWIM domain. In addition, we identified putative SWS1 orthologs in several early-branching eukaryote lineages, including Diplomonadida (Giardia lamblia), Euglenozoa (Leishmania and Phytomonas), green algae (Coccomyxa and Bathycoccus), Stramenopiles (Aphanomyces, Albugo, Phaeodactylum, and Ectocarpus), Alveolata (Paramecium, Plasmodium, and Oxytricha), and Ichthyosporea (Capsaspora) (sequences in File S1). These sequences clustered with known SWS1 orthologs in phylogenetic trees separated from archaea outgroup sequences. This deep diversity, in addition to known fungal, metazoan, and plant orthologs, suggests an ancient origin of the SWS1 protein family. PSI-BLAST recovered a single-protein sequence from each species, suggesting that there were few or no Shu2/SWS1 family duplications and that these orthologs are well conserved.
To define a comprehensive Shu2/SWS1 protein family, we constructed a phylogeny using Shu2 homologs from major eukaryotic and archaeal lineages (Figure 2A). In the resulting phylogeny, eukaryotic sequences were cleanly separated from the archaeal sequences with high branch support (aLRT = 0.98). Moving forward from the archaeal root, the well-supported branches between eukaryotic sequences were in agreement with accepted speciation events, thus supporting the orthology of these Shu2/SWS1 sequences. Although the short alignment (219 amino acids) of Shu2/SWS1 did not provide sufficient power to infer all branch nodes with strong support, these sequences appear to be true orthologs. Notably, this tree revealed Shu2 orthologs in D. melanogaster and C. elegans, which we will refer to as dmSws1 and ceSws-1 hereafter, as well as an Arabidopsis thaliana ortholog, AT4G33925. The archaeal SWIM domain–containing proteins served only to root the tree, and we are not able to comment on the orthology or specific function of those sequences.
A defining feature of the Shu2/SWS1 protein is the SWIM domain, a zinc-binding feature (Figure 2B) (Martin et al. 2006). To further ensure orthology between yeast Shu2 and human SWS1 (also known as ZSWIM7) in exclusion of other SWIM domain–containing proteins in humans (ZSWIM1–6 and ZSWIM8), C. elegans (pqn-55), and D. melanogaster (CG34401), we constructed a phylogeny with all SWIM domain proteins from these species and all putative eukaryotic SWS1 orthologs (Figure 2C). Homology between these proteins is limited to the SWIM domain, so their alignment is limited to a region of 30 highly conserved amino acids. As expected for a small alignment, many branching nodes were not well resolved; however, the branching pattern cleanly separates the SWS1 orthologs from the other SWIM domain proteins with moderate support (aLRT = 0.70). This topology further supports the putative orthology of the sequences in Figure 2A (i.e., they descended from a single common ancestor).
Shu2 and its orthologs have evolutionary histories strongly correlated with recombination and meiosis-related proteins
To better define the biologic function of the Shu complex, we performed coevolutionary analysis in both budding yeast and Drosophila. This analysis exploits the observation that functionally related proteins tend to have covarying rates of evolution because they experience shared evolutionary pressures. This property can be quantified as ERC, reflecting the degree to which two proteins have rates of sequence evolution that covary between species (Clark et al. 2009; Clark and Aquadro 2010; Clark et al. 2012). ERC for a protein pair is quantified as a correlation coefficient (ranging between −1 and 1) for which higher values reflect stronger rate covariation. ERC values are typically elevated between functionally related proteins genome-wide in yeasts and Drosophila, as well as between meiosis and DNA repair proteins in mammals (Clark et al. 2012, 2013; Findlay et al. 2014). Hence an elevated ERC value for a protein pair suggests cofunctionality between them.
We first demonstrated that members of the yeast Shu complex have significantly covarying rates with each other, as we might predict given their cofunctionality (Figure 3A); their pairwise ERC values were highly elevated as a group (mean ERC = 0.52, permutation test P < 0.00001), whereas the expected mean ERC for a random gene set is zero. These ERC values were calculated across a phylogeny of 18 fungal species, including S. cerevisiae. We also found elevated ERC values between the Shu complex members and Srs2 (Figure 3A), consistent with a conserved physical and/or genetic interaction between Shu2 and Srs2 (a DNA helicase that disassembles Rad51 filaments) in both budding and fission yeast (Ito et al. 2001; Martin et al. 2006; Bernstein et al. 2011).
To determine the coevolutionary relationship of the Shu complex with broader functional groups within fungi, we studied Shu2's ERC values with mitotic and meiotic proteins. Shu2 did not show significant rate covariation with mitotic proteins, but ERC values between Shu2 and meiotic proteins were significantly elevated, suggesting strong coevolution between them (P = 1.1 × 10−8) (Figure 3B and Table S3 and Table S4). Importantly, these results were unchanged when recombination-related genes and genes shared by the two sets were removed from analysis, so the association is not limited to HR proteins (mitosis P = 0.27; meiosis P = 1.1 × 10−6). Similar to Shu2, Psy3 also exhibits significantly increased ERC values with meiotic but not mitotic proteins (Figure S1A and Table S5 and Table S6). In contrast to Shu2 and Psy3, both Shu1 and Csm2 show significant rate covariation with both mitotic and meiotic proteins (Figure S1, B and C, and Table S7, Table S8, Table S9, and Table S10). In addition, the D. melanogaster Shu2 ortholog dmSws1 (CG34314) also showed rate covariation with meiotic proteins (P = 0.013) and with Rad51 paralogs (P = 0.0066) but not with mitotic proteins as a class (Figure 3D). The Drosophila dmSws1 results also remained unchanged after removing recombination-related and shared genes (mitosis P = 0.62; meiosis P = 0.0238). Most notably, Rad51C and Rad51D showed extremely high rates of covariation, whereas the other two Rad51 paralogs, Spn-A and Spn-B, showed modestly elevated levels (Figure 3C). Finally, we tested for coevolutionary associations of meiotic and mitotic genes with the mammalian Shu2 ortholog SWS1 (ZSWIM7). Contrary to results in fungi and Drosophila, ERC values between mammalian SWS1 and meiotic and mitotic genes were not generally elevated. Overall, results from fungi and D. melanogaster suggest that the Shu complex has conserved meiotic and mitotic roles in eukaryotic species.
Expansion of the SWIM domain to include an invariant alanine three amino acids downstream of the canonical CXC...Xn...CXH motif
The SWIM motif was originally defined as a zinc-binding motif that contains the canonical CXC...Xn...CXH sequence (Liu et al. 1995; Makarova et al. 2002; Banerjee et al. 2004). However, on analysis of our evolutionarily deep alignment, we identified an invariant alanine located three amino acids downstream from the CXH motif (Figure 4A and Figure S2). Interestingly, in humans, this invariant alanine in hSWS1 is mutated to a threonine (A108T) in a cancer patient from the COSMIC database (Shepherd et al. 2011). To determine whether alanine 108 may be functionally important, we constructed a Y2H vector containing hSWS1 mutagenized to include this mutation, SWS1-A108T, and mutations in the canonical SWIM domain residues (C85S, C87S, C103S, and H105A)(Figure 4B). Suggesting that these residues are functionally important, disruption of the canonical SWIM domain or the invariant alanine (A108) results in a reduced Y2H interaction with hSWS1's obligate binding partner hSWSAP1 (Figure 4B). Interestingly, another highly conserved residue, F90, does not result in reduced Y2H interaction when mutated to an alanine (Figure 4B, SWS1-F90A). These results suggest that the SWIM domain in hSWS1 including the invariant alanine is likely important for hSWS1 function.
The SWIM domain is important for Shu2's functionality in vivo
To investigate the role of the SWIM domain and the invariant alanine, we created Y2H vectors harboring the analogous mutations in the budding yeast SHU2 SWIM domain (Figure 5A). Via Y2H analysis, we found that mutating the canonical SWIM domain residues (C114S, C116S, C176S, and H178S) in shu2 results in an undetectable Y2H interaction with Shu2's binding partners Shu1 and Psy3 (Figure 5A). Interestingly, mutating alanine 181 to a threonine leads to a reduced Y2H interaction with Psy3 but not Shu1. Furthermore, while the hSWS1-F90A mutant maintains its Y2H interaction with hSWSAP1, we found that the corresponding mutation in yeast shu2, F119A, results in a reduced Y2H interaction with Psy3 but not Shu1 (Figure 5A). These results suggest that the canonical SWIM domain in yeast Shu2 is likely important for function and that the conserved alanine and phenylalanine also may be components of this domain.
To determine whether the impaired Y2H interactions of the SWIM domain mutants result in diminished DNA damage tolerance in yeast, we complemented shu2∆ cells with a WT SHU2- or a mutant shu2-expressing plasmid. We found that the WT SHU2 plasmid complements the MMS sensitivity of shu2∆ cells, while mutations in the canonical SWIM domain (C114S, C116S, C176S, and H178A) do not (Figure 5B). Interestingly, despite the altered protein-protein interactions observed in both shu2-F119A and shu2-A181T, both mutant plasmids complement the MMS sensitivity of a shu2∆ cell (Figure 5B). These results indicate that the interaction between Shu2 and Psy3 may be dispensable for Shu2's mitotic function. These findings were confirmed when we stably integrated the canonical SWIM domain mutants (C114S, C116S, C176S, and H178A) as well as F119A at the endogenous SHU2 locus (Figure S3). Unfortunately, we were unable to integrate A181T into our yeast strains. Because the Shu genes were originally characterized by their ability to suppress the sensitivity of a top3∆ or sgs1∆ strain to MMS or HU treatment, we also examined whether the integrated SWIM domain alleles also would rescue sgs1∆ DNA damage sensitivity such as disruption of SHU2 and found that the canonical SWIM domain mutants do (Figure 5C). Similar findings also were observed with a SWIM domain mutation in the fission yeast gene sws1-C152S (Martin et al. 2006). Thus our analysis of the four canonical SWIM domain mutants demonstrates that the conserved SWIM domain is important for this protein family's function in the repair of MMS-induced DNA lesions.
Given the conserved nature of the Shu complex in meiotically dividing yeast, we asked how disruption of the SWIM domain would affect meiotic outcome in S. cerevisiae. To test this, we sporulated diploids homozygous for deletion of a single Shu complex member and screened for viability of the resulting offspring. In agreement with two recent reports (Hong and Kim 2013; Sasanuma et al. 2013b), we found that loss of any member of the Shu complex causes a marked decrease in spore viability, with Csm2 and Psy3 resulting in a more severe defect (P ≤ 0.02 for all) (Figure 5D). Next, we tested whether mutations of the SWIM domain similarly would lead to decreased meiotic progeny. We found that the SWIM domain mutants C114S, C116S, C176S, and H178A are also defective for spore viability compared with WT yeast (P ≤ 0.05 for all) (Figure 5E). In contrast, the shu2-F119A allele spore viability was similar to that of WT yeast (Figure 5E). These results demonstrate that the canonical SWIM domain is necessary for Shu2's role during meiosis.
Here we describe how the SWS1 protein family has evolved throughout major eukaryotic lineages to interact with the Rad51 paralogs to promote HR. One defining feature of the SWS1 protein family is its invariable SWIM domain (consisting of the CXC…Xn…CXH motif, where X is any amino acid and n is a variable number of amino acids), and we show that this domain is important for protein functionality. Furthermore, our analysis indicates that the SWIM domain contains an invariant alanine two residues after the CXH motif that would expand the domain to CXC…Xn…CXHXXA. Specifically, in budding yeast, mutation of the canonical SWIM domain residues reduces Rad51-dependent repair, meiotic viability, and protein-protein Y2H interactions. Additionally, we show further evidence that the Shu complex has an important meiotic function and that this function is evolutionarily conserved. We demonstrate that the SWS1 family is found across eukaryotes and are the first to identify orthologs of SWS1 in important model organisms such as D. melanogaster and C. elegans, as well as in additional fungal and ancient eukaryotic lineages.
Regulation of meiotic recombination is critical for maintenance of chromosome copy number and genetic diversity. Consistent with a central role for the yeast Shu complex during meiosis, loss of Shu2 or its binding partners reduces the viability of meiotic offspring, implicating this complex as a critical meiotic regulator (Figure 5, D and E). It was demonstrated recently that the Shu complex has an important function in the recruitment of Rad51 to meiotic DSB sites and that Shu1 promotes homolog bias (Hong and Kim 2013; Sasanuma et al. 2013b). In agreement with these findings, patterns of coevolution also highlight the importance of the Shu complex in meiosis because we found that all the Shu complex members, including Shu2, are coevolving with other meiotic proteins, as shown by elevated ERC values (Figure 3 and Figure S1). Importantly, we did not observe elevated ERC values between Shu2 and Csm2 with mitotic proteins, unlike the other Shu complex members, Shu1 and Psy3 (Figure 3 and Figure S1).
On further analysis, we identified the D. melanogaster SWS1 ortholog, which is also likely to function during meiosis. dmSws1 is expressed primarily in the ovaries, which are the site of meiotic crossing over in flies (St. Pierre et al. 2014; B. R. Graveley, G. May, A. N. Brooks, J. W. Carlson, L. Cherbas, C. A. Davis, M. Duff, B. Eads, J. Landolin, J. Sandler, K. H. Wan, J. Andrews, S. E. Brenner, P. Cherbas, T. R. Gingeras, R. Hoskins, T. Kaufman, S. E. Celniker, personal communication). In contrast, dmSws1 is not significantly expressed in other adult tissues, including the testes, which do not produce recombinant gametes in Drosophila. Consistent with dmSws1's ovarian expression, we found that dmSws1 also exhibits strong coevolutionary signatures (ERC) with meiotic proteins, just as observed for its budding yeast ortholog Shu2 (Figure 3, B and D). Also similar to Shu2, which physically interacts with the Rad51 paralogs, we found that dmSws1 strongly coevolves with the Rad51 paralogs dmRad51D and dmRad51C (Figure 3, A and C), which are also expressed primarily in the ovaries. Despite this conserved evolutionary pattern, we were unable to detect a physical Y2H interaction between dmSws1 and dmRad51D or between ceSws-1 and Rfs-1. However, this may be due to the lack of a third unidentified binding partner, as is required for Y2H interaction between hSWS1 and its hRAD51 paralogs. For example, we only detected a Y2H interaction between hSWS1 and its obligate binding partner hSWSAP1 despite the physical interaction of hSWS1 with other Rad51 paralogs (Martin et al. 2006; Liu et al. 2011). Alternatively, the strong ERC values between dmSws1 and the Rad51 paralogs may be explained by a conserved genetic but not physical interaction. Altogether, our work suggests that the SWS1 protein family likely consists of conserved pro-recombinogenic factors for meiotic HR in multiple eukaryotic lineages. In the future, direct experimental evidence to examine the role of dmSws1 in meiotic and mitotic Rad51 filament formation will be necessary to confirm this hypothesis.
While our work indicates that the SWS1 family of proteins functions in meiosis, it is clear that these proteins also promote mitotic HR (Shor et al. 2005; Mankouri et al. 2007; Ball et al. 2009; Bernstein et al. 2011; Godin et al. 2013; Xu et al. 2013). Multiple groups including our own have demonstrated that loss of SHU2 confers sensitivity to replication fork–damaging MMS in various budding yeast strains, and consistent with these findings, previous reports have indicated that loss of SWS1 results in a sensitivity to replication fork–blocking agents in both fission yeast and human HeLa cells (Martin et al. 2006; Liu et al. 2011). Furthermore, we showed here that the SWIM domain in Shu2 is important for the resistance of the Shu complex to MMS-induced DNA damage. The role and importance of Shu2 and the Shu complex in promoting mitotic HR are not completely understood, although they appear to act in part by regulating the activity of Srs2.
Srs2 is a DNA helicase that destabilizes Rad51 filaments in vitro, which, in turn, regulate HR in vivo (Rong et al. 1991; Krejci et al. 2003; Veaute et al. 2003; Burgess et al. 2009). Previous reports have shown that Shu2 physically interacts with Srs2 (Ito et al. 2001; Martin et al. 2006) and that loss of Shu1 results in increased Srs2 occupancy at DSB sites (Bernstein et al. 2011). Together these findings support a model in which the Shu complex may promote Rad51 filament formation by inhibiting Srs2 in mitosis. However, a new report indicates that Srs2 has an additional important function in regulating Rad51-dependent repair during meiosis (Hong and Kim 2013; Sasanuma et al. 2013b). In contrast to its role in mitosis, the function of Srs2 in meiosis is pro-recombinogenic. Furthermore, disruption of SRS2 does not rescue or alter the meiotic defect observed in a Shu complex mutant, suggesting that the genetic interaction between the Shu complex and Srs2 is different during mitosis and meiosis (Sasanuma et al. 2013a, b). Despite the differing mitotic and meiotic roles of Srs2, the conserved physical interaction between Shu2 and Srs2 and the strong evolutionary covariation clearly indicate that these proteins have a functionally important relationship (Figure 3A). In the future, the strong ERC observed between Shu complex members could be exploited to identify additional protein modifiers of its HR function, as demonstrated in a recent ERC study (Findlay et al. 2014). Moreover, it remains uninvestigated whether hSWS1 or its binding partner, hSWSAP1, retains this physical interaction with the putative Srs2 homologs such as PARI or RTEL or whether they promote meiotic HR. In conclusion, the SWS1 protein family is an important factor in both mitotic and meiotic HR, and future work will shed light on its unique regulatory mechanisms for Rad51.
We thank Rodney Rothstein for providing materials and reagents, Paul Russell and Jun Huang for expression constructs of the human proteins used to make the yeast two-hybrid vectors, and Shahm Raslan for experimental assistance. This work was supported by grants from the National Institutes of Health (GM-088413 and ES-0244872), a V Foundation Scholar Award to K.A.B., and a Charles E. Kaufman New Investigator Award from the Pittsburgh Foundation and a CURE grant from the Commonwealth of Pennsylvania Department of Health to N.L.C.
Supporting information is available online at http://www.genetics.org/lookup/suppl/doi:10.1534/genetics.114.173518/-/DC1.
Communicating editor: J. Sekelsky
- Received December 9, 2014.
- Accepted February 2, 2015.
- Copyright © 2015 by the Genetics Society of America


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SAN FRANCISCO (MarketWatch) -- Vulcan Materials Co. VMC, +0.36% said Monday that it has acquired a crushed stone quarry in Polk County, Tenn., from Polk County Stone LLC. Terms of the transaction were not disclosed. The assets acquired include the stone quarry and related processing plant, located 40 miles east of Chattanooga, the Birmingham, Ala.-based construction materials company said. The facility will be integrated into Vulcan's mid-south division, which is headquartered in Knoxville, the company said.
Vulcan Materials buys Tenn. quarry from Polk County Stone
Quote References
+0.51 +0.36%

subset: commoncrawl


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FLINT (WJRT) (4/16/2019) - "It was sickening. I mean it just brought back all the memories of the fire here and how helpless you feel," explained David White, President of the Whaley Historical House Association.
Watching France's Notre Dame Cathedral burn has been difficult for him; because, he knows they now have a long road ahead of them.
In November 2015, they dealt with a similar devastating fire at the Whaley Historic House Museum.
"And, the Flint Fire Department, just as you saw in Paris, they removed items from the house," White explained. "We have the original loan book from Citizens Bank to Billy Durant for him to buy the road car company and bring it to Flint in 1885." That eventually led to General Motors.
The home is filled with countless pieces of Flint's history, dating back to the late 1800s.
White said only about a dozen artifacts were lost in the flames.
A little more than three years later, the restoration is finally about 99-percent complete.
"It probably should've happened faster," White said. "There was a lot of delay, a lot of delay from the insurance company - what they would pay, what they wouldn't pay, and contractors, you know."
For now, they're still waiting on a few of the furniture pieces to be returned and to restore much of the word work still suffering from water damage.
"The total cost of the renovation, which includes removing all the artifacts and taking care of them for the time period, was $2.3 million," he said. "We're still lacking $50,000 to bring us back into the black. And so, we're working very hard towards that."
Insurance money, grants and donations covered the rest.
He's grateful the community stepped up in more ways than one, an effort he's also seeing in Paris.
"Certainly people were taking a risk to go into the Cathedral to remove the historical documents and that. So, we have to be very thankful for those people," White said.


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Surface Pattern Designer Claudia Owen
Claudia Owen is a surface pattern designer who lives in Australia, a lover of creating patterns and applying them to a variety of products such as wallpapers, fabrics, rugs, stationery and homewares.
'I've always liked art and design so when it came time to decide what career path I wanted to follow a creative path was on the top of the list. My background is in graphic design but somewhere along the way I fell in love with the world of surface pattern design.'
'I love to create patterns and I'm particularly drawn to geometric and strong bold designs. Patterns are great in that they have a wide range of applications for home wares, fabrics, stationery products and the list goes on and on. I don't think the word 'love' can actually fully express how much I love and enjoy doing what I do!
This year I will have some of my work at the Hong Kong Houseware Fair, Canton Fair in China as well as Surtex in the United States which is one of the largest art licensing shows in the world. Exciting times ahead!'
'For the last couple of months I've been busy working on a collection of silk scarves that are GOTS approved (Global Organic Textile Standard) and eco friendly. The collection comprises of six different designs and each design is bright and they reflect my love of patterns, colours and textures. And to go with the scarves I designed a number of matching accessories such as phone covers, throw pillows, stationery items and lots of other fun products.'


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The Path To Finding Better Solutions
What You Need to Know About Netsuite Integrations With the ever evolving technology, more and more consumers are increasingly demanding for the products and services that they wish to purchase to be available online. This has, in turn, compelled many enterprises both small and large to keep abreast with this changing technology. In fact, your company will not only require to be accessible online. But will also need you to integrate it fully with software that will combine all the CRM, ERP, PSA and E-commerce data to enable your firm to run successfully. Hence the best integration platform as a service software that will fulfill this mission for your enterprise will be Net Suite. Net suite is billed as the best cloud ERP vendor in the world. What net suite does is that it ensures that all your client relationship management, the whole of your enterprise resource planning and your professional services automation is managed within a single software package together with your E-commerce needs. Therefore, helping you minimize not only your IT footprint that you will have previously required to manage all your ERP applications but will also allow you to focus all your time and energy to drive the creative side of your firm. Hence this saves you time that you would have used in dealing with some endless tasks of creating a mailing list, fumbling with records and dealing with tedious accounting details. Usually, net suite assimilates all these needs of your business into one easy to learn and follow package that allows for the management and pricing of all your products and services.
Incredible Lessons I've Learned About Software
Using net suite as the integrating software for your firm's website means that you can bring all your E-commerce needs to a simple database that is extremely user friendly and easily adaptable. Hence you will not be required to train your staff on programming or employ staff who have prior experience in using net suite. Basically, all they will need to do is to log in as an administrator of net suite for them to key in data and make any changes to your website without undergoing any difficulties.
Valuable Lessons I've Learned About Software
Once you download the net suite from the open source network and your login account has been opened correctly it will fully integrate into your enterprise that is from accounting, site management, e-commerce and client relations. Thus, enabling your enterprise to run smoothly, therefore, giving you plenty of time to put all your energies to improving your client's satisfaction. Which will often go a long way in creating sustainable growth of your company because of loyal customers. Apart from this, the net suite software will enable you to have more time to concentrate on building strategies that you will use to market your firm more. The final benefit of using net suite is that it creates a global platform for your business. Which you can use to sell your products and services to an international consumer base without having to worry about language barriers and different currencies as net suite can easily convert language and currency from one form to the other hence allowing for seamless transactions from one part of the world to the other. Lastly, it is imperative to know that the application programming interface permits for the installation of third party applications that are supported and maintained by net suite to allow its users with the upgrade process.


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Sports Titans: Magic Johnson
Laker legend key player in another L.A. sports renaissance
Earvin "magic" Johnson Jr. arrived here 39 years ago as the No. 1 overall draft pick of the Los Angeles Lakers, soon to be the leader of a basketball renaissance that produced five National Basketball Association titles for the city.
Nearly four decades later, Johnson is part of another sports renaissance in Los Angeles – as both a team owner and executive – this one spanning multiple sports and institutions.
"We as fans are really blessed and should be happy," Johnson said, citing the return of professional football to the city; Stan Kroenke's Inglewood stadium and the Los Angeles Football Club stadium projects; and Los Angeles' winning bid for the 2028 Olympics as pillars in a new era of sports.
And the sports renaissance is not just about the fan experience, Johnson pointed out. It's also a boon for the local business marketplace.
"Creating a building, a new stadium provides so many job opportunities, contract opportunities for different suppliers and companies in the city and then you think about tax revenue for the city itself," he said. "There's a lot of money. That's why cities recruit teams to come because they know what it means for the economy."
And Johnson is in the thick of much of this economic boom, attached in some form or another to at least five local sports franchises.
He's part of ownership groups that control the Los Angeles Dodgers, The Los Angeles Sparks, the Los Angeles Football Club and e-sports franchise Team Liquid/aXiomatic. The Laker great also is making a return to his former team as president of basketball operations, brought on by managing owner Jeanie Buss to help turn the team around after several down years.
All this comes on top of his more than 30-year run as chief executive of Magic Johnson Enterprises Inc. His eponymous operating company has franchised national brands such as Starbucks, Burger King and 24 Hour Fitness, and joined Ron Burkle's Yucaipa Co. of West Hollywood and Canyon Capital Partners of Century City in several investment funds.
Johnson's transition from the hardwood to the owner's box came with some hardship, however, and Johnson is candid in describing the difficulties players often face once their athletic careers wane.
"Business, when I first started, was hard because you knew pick-and- roll, you knew free throws, you knew how to throw a no-look pass," Johnson said. "But EBITDA and this and that. Oh my God. It's a steep learning curve."
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Punisher - Nerd Journal
Jan 24th, 2018 by wordswithnerds
John and Craig discuss #Netflix's #Marvel's #ThePunisher, share what they watched, try to figure out if #StarWars #Solo #AStarWarsStory is actually going to be released, and continue to fight #HashtagOveruse.
Words With Nerds is available on iTunes, Google Play, Stitcher, or through your podcast app of choice. Interact with the show on twitter at @nerdswords. You can reach the show hosts directly at @kesseljunkie (John) and @craigsorrell (Craig).
But why let Twitter have all the fun? Interact with the boys on Facebook at facebook.com/WordsWithNerds.
Help the show: Visit our sponsors LootCrate at www.lootcrate.com/wordswithnerds and enter wordswithnerds at checkout to get receive a discount on your purchase.
Want delicious pre-cooked meals delivered to your door (LIKE A KING!,) check out our sponsor www.mealtimebox.com . Delicious meals delivered to you, no cooking, no hassle, no trouble. Dinner is served! enter NERDS at checkout for a 15% discount for this amazing service.


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Feb. 27 Membership Evening
What's New for 2019 Sheridan Nurseries
Mar. 27 Seeds, Magazines, Books, etc. Recycle
Unearthed Alexandria Risen*
Apr. 24 The Narcissus Show
Raised Bed Gardening Tara Nolan*
(link to Narcissus Show Schedule)
Apr. 27 District 17 Annual General Meeting
Hosted by Georgina Brock Garden Club
"Muddy Boots"
Martin Galloway - Your Garden Wakes Up for Spring
Penny Beaudrow, Lauri Hoeg - Sacred Plants - Spiritual and Medicinal Uses
May BHS Garden Work Schedule begins
(link to BHS Garden dates)
May 11 Compost Giveaway
Whitby Operations Centre
May 22 The Spring Show
Drama in the Butterfly Garden Carol Pasternak*
(link to Spring Show Schedule)
May 25 Plant Sale
Grass Park, Brooklin 9:00am
(link to additional information)
May 30 -
June 2 Brooklin Spring Fair
June 12 The Iris & Peony Show
Geraniums Bill Grainger
(link to Iris & Peony Show Schedule)
June 26 The Rose Show
Edible Gardening Cathie Hooper
(link to Rose Show Schedule)
July 19-21 OHA Annual Convention
Windsor, ON
(link to OHA Annual Convention page)
July 20 Whitby in Bloom Garden Tour
(link to Town of Whitby Garden Tour details page)
Aug. 28 The Annuals Show
Square Foot Gardening Gini Sage
(link to Annuals Show Schedule)
Sep. 25 Autumn Show
An English Country Garden Marjorie Mason
(link to Autumn Show Schedule)
Oct. 23 Pumpkins, Pies & Pictures Show
Light up your Garden with Bulbs Jeff Mason
(link to Pumpkins, Pies & Pictures Show Schedule)
Nov. 27 BHS AGM & Social
Dessert, Entertainment, Awards, Elections
St. Thomas Anglican Church, 101 Winchester, East @ 6:30pm
(link to BHS AGM & Social Show Schedule)
* These speakers are sponsored by Whitby in Bloom


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« Happy New Year | Main | WMH »
Sunday, January 09, 2005
Mix and Share
For the past few years, I've put together a little holiday mix CD with songs. This year moved a bit quickly, though, and I didn't have time to send them out to anyone I knew. So, in the interest of fulfilling the needs of my readership (and post something substantial), I thought I'd share the tracklisting with some of my thoughts on the songs. Yes, you are extremely lucky to read this. More that actually pertains to real life soon enough!
"That Spirit of Christmas" by Ray Charles from Spirit of Christmas
This is always a good way to start off a holiday CD, especially with one of the best Christmas songs I can remember. Whether it's being played on National Lampoon's Christmas Vacation or Dexter Sy's notoriously rare Nintendojo Holiday movie, this one hits me ::points to heart:: right here. ::bites lip, lowers voice to a whisper:: Right here.
"Everything Must Go" by Steely Dan from Everything Must Go
Huang-sensei turned me on to Steely Dan, and they do have quite a few catchy songs. We got to see them perform at Roseland in NYC before he left for Japan, too, and they were a very good live band. All of that means nothing, though, when you play this song. The lyrics are weighty and tremendous, and anyone who has struggled (with anything really) will find something in this for them. Personally, it will always remind me of my first site and how things went with that.
Talk about the famous road not taken
In the end we never took it
And if somewhere on the way
We got a few good licks in
No one's ever gonna know
'Cause we're goin' out of business
Everything must go
"Miracle Drug" by U2 from How To Dismantle An Atomic Bomb
Not my favorite song on the album (that would easily be "Sometimes You Can't Make It On Your Own"... a killer), but the chorus is just so uplifting and really makes you feel like you can do anything. This one came at the right time, and brightens up my outlook on the new year.
"The Ocean's Breathes Salty" by Modest Mouse from Good News for People Who Love Bad News
Jen hooked me up with their CD, and while that single "Float On" is really good, this song has some of the best guitar playing I've heard in awhile. By the middle of the song, you'll find yourself belting it out, playing the steering wheel as if you're in front of the packed Foto Autodromo (better because all 100K will kick your ass if you're bad). When the song ends, though, you'll find yourself in front of that same steering wheel, wondering where your rock career took that fateful detour into video game journalism.
"Killing Me" by Robbie Williams from The Ego Has Landed
I like Robbie Williams, even though he seems to be an overrated, overexposed Brit pop singer with a fair share of bad habits. At least no one knows his name here, and I can put this song on the CD without fear. This is one of his better ones. A bit of gíc trivia: his song, "She's The One," was the soundtrack for the European launch of the Sega Dreamcast. See? That's why I get paid the big bucks!
"Pause For Porno" by Dr. Dre from 2001 (Instrumental)
Gotta love this one. Even the standard porn track on many rap albums gets the full treatment from Dr. Dre. It's the track that will kick off the infamous Cloudch... XenDis... GameDAIL... whatever DVD for E3 in Los Angeles. Just feels sunny and smoggy. This track is meant to cleanse the palate and ushers in the second act (as you can see, I don't play around with mix CDs).
"Let Me Know" by DJ Quik feat. Hi-C, unreleased
If there's ever a song that just gets people jumping, it's this one. Feel the West Coast vibe, let the rays hit you, and don't drive too fast. This is coupled with the next track to make a one-two punch of goodness...
"Getsu to Oji" by Kenji Niinuma from Katamari Damacy
This keeps it going, with a very colorful and light Japanese track that'll make you wish you knew what the hell he was saying. From Katamari Damacy, this is one of the songs that keeps you going through that game.
"What's Up Fatlip?" by Fatlip from What's Up Fatlip?
This starts a couple of biographical tracks that I've grown increasingly addicted to. An oldie but goodie, this also has a Spike Jonze video that is one of the most compelling music videos ever put to film.
I'm far from hard
Emotionally scarred
On Pico Boulevard
I was regarded as a retard
I make myself sick
Get on my own nerves
Immature, insecure,
Grown up nerd
Has-been MC
On a label that's unstable
Choppin' bliggy on a table
"Spaceship" by Kanye West feat. GLC & Consequence from College Dropout
One of the best albums of the year, I've been trying to play this one for everyone I know. This is one of the many great songs on it. Can't say anything but buy it.
I've been workin' this graveshift
and I ain't made shit
I wish I could
buy me a spaceship and fly
past the sky
"Spottieottiedopaliscious" by OutKast from Aquemini
Who woulda thought the fellas who made Southernplayalisticadillacmuzik, one of my first rap CDs, would go on to become huge superstars. Well, they have, and this is one of my favorite songs from them (thanks to Charles for playing this for me). Right up there with "Bombs Over Baghdad," you have to listen to the horns and realize you're listening to some of the best stuff you'll ever hear.
"Garden Grove" by Sublime from Sublime
Keeping it with the some of the older songs from the early 90s on the CD, this is from one of my favorite groups of the time. You'll find yourself singing along way too quickly, and you'll feel anger as you realize they were cut way too short.
"Slug" by Passengers from Original Soundtracks 1
This was a cool album as I remember it. U2 got together with Brian Eno (one of their longtime producers and a well-known artist himself) to create this album. "Slug" is one of my favorites from that album, with a fantastic beginning followed by a more impactful final ten seconds or so. Oh yeah, the middle is very good, too.
"Wataridori 2" by Cornelius from Wired Creative Commons
What's a mix CD without some funky electronic shiz? I grabbed this free CD sampler and found it was quickly the best track on the whole thing (which included Chuck D and the Beastie Boys). You should be able to find it on a lot of sites. I especially like the thunder-out-of-nowhere. More songs should have that.
"Blow, Gabriel, Blow" by Cole Porter and performed by Jonathan Pryce, Kevin Kline & Chorus from De-Lovely
We saw this movie on the way to London, and while it was pretty good, this song makes it a must-have. I didn't know where else to put this, and while it may sound out of place it really is sweet.
Once I was headed for hell,
Once I was headed for hell;
But when I got to Satan's door
I heard you blowin' on your horn once more,
So I said, "Satan, farewell!"
And now I'm all ready to fly,
Yes, to fly higher and higher and higher!
"This Is Your Life" by Billy Paul from Me and Mrs. Jones: The Best of Billy Paul
Couldn't think of a better way to end this one. As Justin and Patrick would say, this song just makes me go (ahem) "Oooof!"
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Did the track listing on my mix CD work on your iTunes? If not, then I'll just have to write a similarly informative post on my own blog.
Posted by: Huang-sensei at Jan 9, 2005 8:07:52 PM
Steely Dan rocks, and there are a few catchy tunes from Robbie W. Though my fav is that track from Dr. Dre, west coast stylez!
John, send me an email. And tony, if you there, send me one too.
Posted by: Plate at Jan 10, 2005 4:10:12 AM
As Justin and Patrick would say, this song just makes me go (ahem) "Oooof!"
True! You do make a really weird sound when listening to that song.
Posted by: Patrick at Jan 13, 2005 1:19:08 AM
I know. And I'm okay with that. Indeed!
Posted by: John at Jan 13, 2005 5:04:16 PM
I copy would be nice Christmas present! Considering how hard it is for John to copy a file. Looking forward to sample John's ear. Have tasted his ear before, waxy but good!
Posted by: charles aka answerman at Jan 18, 2005 10:57:12 AM


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Deactivation of user done by the shipper admin of the company. The shipper admin has the ability to deactivate the user through the 'Shipper Admin' section >> 'User overview'.
By pressing the 'Details' button the shipper admin can review the user information.
In order to deactivate the user, the shipper admin needs to press the 'Deactivate' button and confirm the action.
To confirm the deactivation, the shipper admin needs to insert 'DEACTIVATE' and press the 'Yes, I want to' button.
The process of user deactivation is immediate and irreversible.


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The growing role of institutional investors in financial markets.
In 2013, the primary institutional investors in the OECD – pension funds, public pension reserve funds (PPRFs), mutual funds, and insurance companies – held USD 92.6 trillion in assets, a number now well above pre-crisis levels. In that same year, the combined GDP of the OECD countries was USD 47.3 trillion. In 2001, OECD institutional assets represented 1.4x GDP; this number has since grown to 2.0x GDP, highlighting the growing role of institutions as financial intermediaries. Put another way, the accumulation of savings in such financial channels has never been so large, which underscores the important role that institutions can play as sources of productive long-term capital. This function has become increasingly important during a time period in which the role of traditional financial intermediaries such as banks and their capacity to provide long-term financing is changing in the post credit crisis period. The scope of this survey covers OECD countries, some non-OECD countries, and member countries of the G20.


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Calling all actors!
This is it! Your chance to experience what it is like on a film set.
This course is designed to bring actors and non-actors up-to-speed with the skills and techniques needed to successfully work as a screen actor.
The Saturday Screen Acting School will give you the experience of what it is like to work in front of a camera. You will also be given dozens of insider tips only the professionals know. Build your confidence and enhance your skills.
Participants encouraged to bring in a headshot and a favourite snapshot *recommended but not essential
Morning Workshops
Bring a good notebook. Wear comfortable clothes. There are lots of group demonstrations and exercises!
1.The Truth is not your friend
Most acting techniques teach 'truth', but nothing in movies is real.
Let tutor Patrick Tucker take you through a series of exercises and tutorials that will explain how screen acting is very different from stage acting and why.
Cheating of Positions and Vocal levels and Reactions.
a. SHOOT 3 test scenes and PLAYBACK
More details of actual shooting procedures and solutions given.
b. SHOOT more test scenes with half a dozen participants.
2.Screen Business
LIVE SHOOT! Participants put the ideas into practice.
What it is and how it works
The plusses as well as the minuses.
Headshots and what they mean
Act your headshot
SHOOT participants using Typecasting
SHOOT other exercises including head turns and over shoulder listening.
Lunch 13:00 – 14:00
Don't waste this hour!
Use the time to network with fellow participants and form new alliances and find collaborators.
If you've ever been to a casting session and not been booked, does it mean you weren't good enough? Or maybe you were taught the wrong way to approach a casting.
In this session, Patrick Tucker will explain how to read and play for an audition.
SHOOT some examples.
Commercial casting explained
SHOOT an audition (2 versions)
Everyone does a commercial casting, and the results played back.
Acting sequentially (not simultaneously)
5.Getting Work
Learn how to take the skills you have learned so far today and create valuable marketing assets you can use immediately to enhance the chances of your getting booked for a job.
Create examples
Check Lists – review the days work and prepare for the future
Who Should Attend
In short, anyone wishing to be an actor, or any actor wanting to hone up on their screen acting skills. This intensive one day workshop is also recommended for writers, directors and producers at any level. It is essential for those who have attended a Drama School, but feel that they were not prepared for the real world of Screen Acting in their course, and what to do in their auditions.
About Patrick Tucker
Patrick Tucker started directing for the stage in 1968, and for the screen in 1976, and has been doing both ever since. To date he has directed over 250 theatre productions and over 200 dramas for the screen (including one feature) at venues in the UK and all over the world, including America, Canada, Denmark, Germany, Ireland, Israel, Jordan, Kenya, Latvia, South Africa, South Korea, and three productions at Shakespeare's Globe London.
His last stage work was Measure, For Measure for the Blackfriars Theatre in Virginia, and on screen a Russian sit-com Olimpiada 80 filmed in Latvia (in Russian). He has lectured and run courses on the various aspects of acting and directing since the mid-1970s (presenting directing workshops for Raindance since 1997, and at Central Film School since it opened in 2009), and his books Secrets of Acting Shakespeare (Routledge 2nd Edition 2017) and Secrets of Screen Acting (3rd Edition Routledge 2014) contain many original insights – as do his workshops. He is currently preparing Secrets of Screen Directing – the Tricks of the Trade for publication in 2019.
Ask a Question about Saturday Screen Acting School


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Prototype of serial BMW X2 came back to the roads
Externally, the novelty will not significantly differ from the concept shown by Bavarians in the autumn at the motor show in the French capital. Recently a prototype was seen passing the road tests.
Precursor of BMW X2 debuted at the Paris Motor Show last year, and the camouflaged prototype of the production version came back to the roads to pass tests. Compared to the previous test vehicles, the car has received the serial rear lights. Unlike the show car presented in Paris, the car has received a pair of large side mirrors, standard elbows of the exhaust pipes and smaller wheels. BMW X2 differs from large X4, X6, and X2 in a less sloping roof line.
BMW X2 is based on UKL platform, which is also a basis for the 2-Series Active Tourer, Gran Tourer and Mini, so we can expect an all-wheel drive system. The range of power trains is presented by the 3- and 4-cylinder turbo engines. At the moment nothing is known about the release date, but according to the prototype, it will not be surprising if the public premiere will take place in the second half of 2017.


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WASHINGTON (1/15/15)--A current proposal from the Internal Revenue Service (IRS) to remove the 36-month non-payment testing period prong from cancellation of debt has the support of the Credit Union National Association. CUNA expressed support for the proposal in a comment letter filed with the IRS Tuesday.
"We believe the proposed change could result in reducing the regulatory compliance burden on reporting entities, including credit unions, in assessing whether a debt has been discharged," reads the letter, signed by Luke Martone, senior assistant general counsel at CUNA. "In addition, we believe the proposal may alleviate some taxpayer confusion that has resulted under the existing regulation."
The proposal would specifically remove a component of the IRS rule that a deemed discharge of indebtedness, for which a Form 1099-C, Cancellation of Debt, must be filed, occurs at the expiration of a 36-month non-payment period.
Credit unions and other covered financial institutions would no longer be required to file a 1099-C solely because a debtor failed to make payments for 36 months on an outstanding debt.
Under current regulation, the form must be used to cancel a debt income of $600 or more, if any of the right prongs mentioned in the rule exists. Seven of these prongs are specific instances resulting in a discharge of debt.
The eighth, the expiration of the non-payment testing period, does not result in a discharge of debt, but instead creates a "presumption of a deemed discharge of debt, nevertheless triggering the statue's information reporting requirements," the letter reads.


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When my guyz took me to the KW Knitters fair they each choose a pattern.
Mr Bunnstible was my DH's choice
I just love the construction of this Bunny
Waste yarn is knit in then removed and the stitches are picked up for the ears.
Supper simple recommend this pattern to bunny makers.
This Months Kal for the Carry along forum on Rav:
I put this on the list for the Kal, so I really should do it!
My Current Sock Project:
Foots first Socks 45% done started July/13
If you missed it "Basic Bra Making Part 1 & Part 2" are now up!
I have decided to reorder my wip list so the oldest ones are first and list my monthly Kal and my Sock Project separately, I always have these two on the go. So I think this will make more sense.
Current WIP List (9)
- Pine Forest Baby Blanket 50% done started Jan 11/12
- SPROUT – Growing Roots sweater 60% done started May 17/12
- Top Down Swing Pullover – 98% done started Jan 1/13
- Sea of Dreams Blanket 75% done started Mar 9/13
- Mile a minute afghan – crochet 50% done started Apr 24/13
- Isla lace Pillowcase edge 52% done started June 18/13
- Rose Harbour Shawl 75% done started Jun 28/13
- Fanflower Fling 50% done started Jul 12/13
- Mr Bunnstible started Sept 17/13
Ruffle ScarfDone started Dec 3/12 finished Jul 7/13 Cozy Winter ShawlDone started Dec 1/12 – finished Jul 11/13 Eileen DressDone started Jun 21/13 – finished Jul 11/13 Easy Peasy Shawl– Done started Jul 14/12 – finished Jul 17/13 All about love shawletteDone Started Aug 3/13 – finished August 17/13 CalimetryStarted Sept 6/13 finished Sept 6/13
FO's 2013 = 90
Used of 38 purchased patterns from Rav = 9 (29 to go)
Stash Used 2013 = 77
Sewing FO's = 4
Books 2013 = 30 + 2 = 32
Thanks again for visiting,
cya Friday:0)
I'm off to visit Tami's, then on to my blogwalk
come join me in the blogpond :0)
Find me on Ravelry.com and also my Etsy shop
Oh and FYI I have 50% off Back to School Sale at my Ravelry Shop, now we all have so much time with the kids in school. Sale lasts to the end of the month. 3 months till Christmas people!


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Allanton village and its hall is about to get a new logo and local people will have the final say in which design is chosen.
Last autumn villagers were asked to sketch designs for new signs outside the hall and for its Facebook and Twitter pages. These ideas were passed on to Kevin Shearer of Printspot who refined them and produced two logos.
At the village's Easter Saturday coffee morning on March 31, in Allanton Village Hall (complete with Mr Ford's hot-cross buns) residents will be asked to choose their favourite logo.
One design shows Stevenson's bridge (the designer of lighthouses) over the Whiteadder. The other design identifies the village as a place where lots of tailoring went on – that was the days when this part of Berwickshire was very much based on sheep farming. At one time, Allanton had seven bespoke tailors whose customers came from far and wide using the train to Chirnside station and onward by pony and trap. One tailor had 'branches' in Edinburgh and Newcastle.
This Saturday, March 24 at 7.30pm the Campbell Duo will be entertaining in the hall - an evening of guitar and flute duos - plus wine and beer from the Hetton Brewery.


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Regain Control Of Your Health With Dental Sedation [BLOG]
Most of the time, fear is a healthy emotion that keeps you from doing things that put you in harm's way. Rather than walk down a dark alley alone late at night, you think maybe it's a better idea to stay on a more public, well-lit path.
Thank goodness for instinct, right?
But when fear becomes irrational and keeps you from participating in life's important events or from taking proper care of yourself, that's when it's time to do something about it.
Your oral health is intimately linked to your total body health.
This is why our team at Times Square Dental wants to help you get over your dental anxiety and regain control of your health this year.
We're starting with today's blog that explains where your fear of the dentist may be coming from and our sedation dentistry in our Boise, ID office can help you overcome it!
Start By Figuring Out The Cause Of Your Anxiety
As important as it is to allow a dental professional to clean and exam your mouth twice a year, it feels like an impossible feat for someone with dental anxiety.
The sad thing is that millions of people feel this way.
Here are some common reasons why:
*Past Dental Trauma*
One of the most common reasons adults are too afraid to go to the dentist is because they're still traumatized by bad dental appointments from their childhood.
Who would want to go back to the dentist if the only memories they have of dental care are scary and painful?!
*Problems With Control*
Adults sometimes feel uneasy at the dentist because they're busy parents and professionals who find sitting still or not being the one in control really uncomfortable. It's not rooted in an actual fear of dental treatment so much as it is an unsettling feeling of handing over the reigns for a bit.
*Too Much Time Has Passed*
Maybe your fears about dental care didn't start out so bad, but you've stayed out of the dentist office long enough that now, it just feels like too much time has passed for you to feel comfortable going back.
Without knowing about all the ways dentistry has changed in the last few years, you're going to create a monster in your mind about what the dentist and hygienist have in store for you, which is perfectly understandable.
At Times Square Dental, though, you'll find you have no reason to feel this way.
*The Stories You Hear*
There's a reason we love going to the movies, watching TV shows, and reading books.
It's because stories have the power to pull us out of our own reality and let us exist in another time and place.
But that's not necessarily a good thing if what you've heard about the dentist office is always negative. Friends and family members sharing their own dental horror stories could be enough to keep you from making routine appointments.
Regain Control Of Your Health With Dental Sedation
If you're like most people, when you're afraid of something, you feel tempted to simply avoid it rather than face it.
While a lucky few might be able to convince themselves to tough things out and just do whatever it is they dread, you feel like that's just not possible.
Some level of fear can certainly be healthy, but fear that keeps you from doing things you know are good for you, like going to the dentist for routine cleanings and exams, surely isn't.
It controls you.
And as a result, it controls your health.
This is why our team at Times Square Dental is so dedicated to your comfort. We know how important it is that you feel safe and confident enough to get the professional dental care you need to keep your smile healthy and beautiful.
It's that reason that you'll find sedation dentistry and comfort options in our Boise, ID dental office.
*Your Options Are Inhaled, Oral, or IV Sedation*
You've probably heard of inhaled sedation, only it's commonly called laughing gas. This is a mild relaxation intervention that's safe and effective for all ages that starts working and wears off immediately.
For more intense anxiety, you can choose between oral or IV sedation, a decision Dr. Hastings or Dr. Dial can help you make depending on your treatment needs.
Of all the innovations in modern dentistry, the growing trend among dentists relying on sedation to help ease the nerves of fearful patients has been among the most significant.
That's because millions of people now have an opportunity to get the dental treatment they desperately need, and thus, completely turn their oral health around.
We also offer wall-mounted TV's in our rooms so you can distract yourself with your favorite programs!
All you have to do is ask one of our caring team members when you need a break or more help feeling comfortable, calm, and relaxed.
Choose Better Oral Health Today!
You have the power to turn your oral health around.
With sedation in our Boise, ID dental office, you don't have to let fear control you or your smile anymore!
Choose better oral health today. Call Times Square Dental at 208-278-1069 fill out our convenient online form to schedule an appointment.


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Hardcopy Research: last shipped December 4, 2019Online Research:Strategist updated a/o 12/04/2019Workbook updated a/o 12/06/2019File Downloads Weekly >> updated a/o 12/06/2019File Downloads Monthly >> updated a/o 12/01/2019
Media: 02/07/2012: 2011 Top Performers
08/16/2011: 6 stocks insiders are snapping up
The data contained in this report were taken from statistical services, reports in our possession, and from other sources. The opinions and estimates expressed are our own, and we make no representation either as to the accuracy or as to the existence of other facts or interpretations which might be significant. The information herein was gathered from responsible sources but we cannot guarantee its accuracy or completeness. We may from time to time have a position in the securities described in this report and may buy or sell such securities.


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We take pride in what our guests say about their expiriences staying here with us at Ivanhoe, please take a moment to read through our ratings or please leave a review of your own.
If you're looking for a hotel or guest house offering bed and breakfast, en-suite guest accommodation in Bridlington for a holiday, short weekend break or somewhere to stay for a local event then you've come to the right place!
Whether you are coming for the sea, sun and sandy beach, walking or bird watching in the Yorkshire Dales and Moors, or a more energetic pastime such as jet-skis, windsurfing or sailing, the Ivanhoe Guest House is well placed for all your needs.


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7 Solar Plants Set Up in Kyoto, Hyogo, Fukui
Sousuke Kudou, technical writer
Tango Solar Power Co Ltd announced Jan 24, 2019, that it has set up seven solar power plants in Miyazu and Maizuru cities (Kyoto Prefecture), Toyooka City (Hyogo Prefecture) and Wakasa-cho (Fukui Prefecture).
Tango Solar Power is a special purpose company (SPC) established and financed by The Kaneshita Construction Co Ltd, Omron Field Engineering Co Ltd (Meguro-ku, Tokyo) and Kyocera Corp.
The solar panel capacity and grid capacity of the seven plants are 3.228MW and 2.831MW, respectively, in total. They are expected to generate 3,481MWh of electricity, which is equivalent to the amount consumed by 700 households, per year, eliminating 1,885t of CO2 emissions.


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McConnell Says He Will Delay Start of Senate Recess
(CN) — Senate Majority Leader Mitch McConnell said Tuesday the Senate will delay the start of the August recess until the third week of the month, as he and his fellow Republicans prepare to unveil a revised version of their healthcare bill before the end of the week.
In a statement Tuesday, Majority Leader Mitch McConnell said the delay is necessary to complete work on legislation and deal with Trump administration nominees.
He later told reporters GOP lawmakers plan to unveil their revised health care bill Thursday and begin voting on it next week. Once that's accomplished, he said, the Senate can move on to work on other issues.
But even as he described his plans to reporters, the prospects of his divided party actually repealing much of the Affordable Care Act seemed as gloomy as ever.
Earlier, one Republican senator said he is "very pessimistic" that the health care measure will be approved, while another warned leaders about retaliation by conservative voters should they react to a collapse of the measure by striking a deal with Democrats.
Underscoring the uncertainty, No. 3 Senate GOP leader John Thune of South Dakota told the Associated Press he would not make predictions about the health care vote.
"I think we have narrowed it down to where we know where the decision points are," he said.
The Senate had been scheduled to begin its five-week recess on July 31. The delay would push it to the week of Aug. 14.
McConnell said that once the Senate "completes its work on health care reform," it would deal with the defense policy bill and nominations.


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Disclosure: I'm not going to talk explicitly about ecofeminism in this week's column. This column is about a few things mainstream feminists often have difficulty acknowledging.
Feminism originated in a white-supremacist, xenophobic, homophobic, transphobic society. As I said in my first column, people who don't identify as feminists because they don't know what the term means deeply frustrate me. But as history shows, feminism excluded and continues to exclude — be it intentionally or unintentionally — women who aren't white, educated, straight, cisgender, American, etc. How can we expect everyone to identify as feminists if the feminist movement does not incorporate an end to the oppression of all female-identifying people?
To illustrate how mainstream feminism has been ignorant of these issues and exclusive towards certain women, here's a brief history of it:
"First-wave" feminism began in the 19th century, driven by white, middle-and-upper-class Protestant women bent on achieving women's suffrage in the United States. While suffrage theoretically came with the 19th amendment in 1920, female minorities continued to face discrimination at the polls, such as black women in the Jim Crow South. Today, minorities still face difficulties when voting in certain (red) states due to recent voter ID laws, restrictions on voting hours, gerrymandering and other (primarily race-based) discriminatory policies. So, though first-wave feminism did achieve one important goal, this goal wasn't extended to all women for decades, and still isn't enjoyed equally by all women in the United States.
Second-wave feminism began in the 1960s, leading to the legalization of abortion and the passage of other laws that impacted women everywhere in the United States. But white, well-off women still led the movement, and many of their goals focused on equal job opportunities for (college-educated) women like themselves, which did not have the same impact for lower class women, immigrant women and women of color who faced discrimination and fewer job and education opportunities.
Third-wave feminism began in the 1990s and continues today. Many third-wave feminists have become more aware of issues related to race, class and gender identity, and many feminists from different backgrounds have called out mainstream feminism's problems, but there are still improvements to be made.
Many mainstream feminists continue to ignore and marginalize women of color. Lena Dunham's show "Girls" (2012 – present), for example, features straight, white, college-educated, well-off girls who live in diverse New York City but somehow never interact with anyone from backgrounds different from their own. Despite the fact that Dunham's show only represents a small portion of the female population, Dunham has implied in the past that she speaks for "all" women of our generation. Similarly, Sheryl Sandberg's book "Lean In" (2013) instructs women how they can "have it all:" a corporate job that probably requires eight years of costly higher-level education, a happy home life and an awesome husband. But the book doesn't address underlying structural barriers that women — especially women marginalized by race, class, sexual orientation and gender identity — face. Still, mainstream feminists have called both Sandberg and Dunham "feminist icons."
Finally, some feminists today refuse to include transgender women. Even women's colleges in the United States — including Barnard, Bryn Mawr, Smith and Wellesley — deny admission to transgender women, who are disproportionately at risk for sexual violence and therefore may need the safe space of an all-women's college more than anyone.
Throughout most of its duration, mainstream feminism has, implicitly and explicitly, excluded marginalized groups of people. Feminism, by definition, strives for equality regardless of gender, but this goal loses its credibility if it does not incorporate the need for equality based on social, racial, sexual-orientation and gender-identity differences as well.
Simply put, if the feminist movement does not become more sensitive and actively involved in these issues, it will not and should not succeed.


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Job D. (Darby) Turner Appointed to University of Kentucky College of Law Visiting Committee
LEXINGTON, KY: Greenebaum Doll & McDonald PLLC is pleased to announce that Job D. (Darby) Turner, a Member in the firm's Lexington office, has been appointed to the University of Kentucky College of Law Visiting Committee (Visiting Committee).
The Visiting Committee is comprised of 30 to 40 of the most distinguished graduates of the College of Law, including United States Congresspersons, Governors, judges, corporate CEO's and law partners. The Visiting Committee meets regularly with the Dean, faculty and students of the College of Law, and also consults with the President, Provost and other top administrators of the University.
Mr. Turner is a member of Greenebaum's Corporate and Commercial Group and the Tax and Finance Group. His practice focuses on general corporate representation, tax increment financing, real estate law, governmental affairs, including representation before the Lexington-Fayette Urban County Planning and Zoning Commission, and business and estate planning matters.
In addition to his role with the Visiting Committee, Mr. Turner serves on the Board of Directors for the University of Kentucky Athletic Association. He received his bachelor's degree from the University of Kentucky and his law degree from the University of Kentucky College of Law.
About Greenebaum Doll & McDonald PLLC
Greenebaum Doll & McDonald PLLC is a widely-respected business law firm with approximately 150 professionals in five offices, serving local, national and international clients in virtually every industry. For more information, visit www.greenebaum.com.


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Automation undeniably, has helped industrial communication market trends to thrive and has advanced the operational work process across numerous sectors
Selbyville, United States - July 21, 2019 /MarketersMedia/ —
The global industrial communication market has thrived successfully in the recent decade as growing adoption of these components in Industry 4.0 has effectively helped increased productivity in major industries. Labor reduction, waste reduction, repeatability, enhanced quality control, and integration with existing business systems have given companies an added advantage by lowering long-term costs, which could indirectly increase product output and revenue.
Communication networks like LAN (Local Area Network) or WAN (Wide Area Network) can communicate large amounts of data via limited number of channels. Whereas industrial networking enable implementing various communication protocols between field devices, digital controllers and several automation related software tools and external systems. Therefore modern industries have demonstrated increased adoption of industrial communication networks over traditional LAN and WAN. This has led to industrial communication market size depicting a substantial increase lately.
Sample Copy of This Report @ https://www.gminsights.com/request-sample/detail/3367
For instance, according to renowned automation conglomerate ABB, and one of the notable players in industrial communication market, the firm's revenue increased by 5% to $7,395 million in 2018. This has been credited to the strong growth in robotics & motion, robust performance achieved from electrification products and a steady result owing to industrial automation.
Industrial communication market is being driven by robotization trends in industries in the emerging economies of the APAC region. Moreover, the increasing adoption of the Industry 4.0 revolution further augments the prospects for the industrial communication industry. Asia Pacific industrial communication industry is forecast to grow at a CAGR of more than 12% over 2019-2025.
Besides APAC region, robot sales in the U.S. has also significantly helped to expand industrial communication market. Robot sales in the United States has been recorded to hit a new peak of almost 38,000 units, setting a record for the eighth year in a row from 2010 to 2018. Robot density in the U.S. manufacturing industry reached 200 robots per 10,000 employees by 2018. Increasing rate of robotization and robot density will drive the industrial communication market substantially.
Make an Inquiry for purchasing this Report @ https://www.gminsights.com/inquiry-before-buying/3367
Among numerous sectors deploying industrial robots, automobile and electronics sectors have been credited with highest share in the year 2018. Automotive sector had accounted for the deployment of 116,000 units of industrial robots whereas, electrical and electronics sector has been credited for the use of 113,000 units of industrial robots. Growing demand of industrial robots is among the key drivers of industrial communication market growth.
Industrial communication market footprint is seemingly strengthened by the industry players in manufacturing, automobile and electronics sectors. For instance, an automation company, ABB, has reportedly announced that it will introduce collaborative robots to medical laboratories as it opens a new healthcare hub at the Texas Medical Center (TMC) innovation campus in Houston, Texas. This new facility will focus on developing non-surgical medical robotics systems. ABB already has its robots operational in food and beverage sector and is playing a significant role to propel industrial communication market growth.
In a nutshell, it would not be incorrect to quote that industrial communication industry is likely to come up as a highly profitable vertical of the overall electronics cosmos. As per estimates, global industrial communication market is anticipated to exceed USD 160 billion by 2025.
Browse Complete Report Summary @ https://www.gminsights.com/industry-analysis/industrial-communication-market
Table of Contents (TOC) of the report:
Chapter 4. Competitive Landscape, 2018
4.1. Introduction
4.2. Major market players, 2018
4.2.2. Cisco Systems
4.2.3. Rockwell Automation
4.2.4. Schneider Electric
4.2.5. Siemens AG
4.3. Market challengers, 2018
4.3.1. Belden Inc.
4.3.2. Eaton Corporation
4.3.3. HMS Industrial Network
4.4. Other prominent vendors
Chapter 5. Industrial Communication Market, By Component
5.1. Key trends, by component
5.2. Hardware
5.2.1. Market estimates and forecast, 2014 – 2025
5.2.2. Switches
188.8.131.52. Market estimates and forecast, 2014 – 2025
5.2.3. Hubs, routers & gateways
188.8.131.52. Market estimates and forecast, 2014 – 2025
5.2.4. Connectors
22.214.171.124. Market estimates and forecast, 2014 – 2025
5.2.5. Communication interfaces
126.96.36.199. Market estimates and forecast, 2014 – 2025
5.2.6. Controllers & processors
22.214.171.124. Market estimates and forecast, 2014 – 2025
5.2.7. Others
126.96.36.199. Market estimates and forecast, 2014 – 2025
5.3. Software
5.3.1. Market estimates and forecast, 2014 – 2025
5.4. Services
5.4.1. Market estimates and forecast, 2014 -- 2025
5.4.2. Installation service
188.8.131.52. Market estimates and forecast, 2014 – 2025
5.4.3. Maintenance service
184.108.40.206. Market estimates and forecast, 2014 – 2025
Chapter 7. Industrial Communication Market, By Application
7.1. Key trends, by application
7.2. Aerospace & defense
7.2.1. Market estimates and forecast, 2014 – 2025
7.3. Automotive & transportation
7.3.1. Market estimates and forecast, 2014 – 2025
7.4. Chemical
7.4.1. Market estimates and forecast, 2014 – 2025
7.5. Electrical & electronics
7.5.1. Market estimates and forecast, 2014 – 2025
7.6. Energy & power
7.6.1. Market estimates and forecast, 2014 – 2025
7.7. Food & beverages
7.7.1. Market estimates and forecast, 2014 – 2025
7.8. Industrial manufacturing
7.8.1. Market estimates and forecast, 2014 – 2025
7.9. Oil & gas
7.9.1. Market estimates and forecast, 2014 – 2025
7.10. Pharmaceutical
7.10.1. Market estimates and forecast, 2014 – 2025
7.11. Others
7.11.1. Market estimates and forecast, 2014 – 2025
Browse Complete Table of Content @ https://www.gminsights.com/toc/detail/industrial-communication-market
About Global Market Insights:
Global Market Insights, Inc., headquartered in Delaware, U.S., is a global market research and consulting service provider; offering syndicated and custom research reports along with growth consulting services. Our business intelligence and industry research reports offer clients with penetrative insights and actionable market data specially designed and presented to aid strategic decision making. These exhaustive reports are designed via a proprietary research methodology and are available for key industries such as chemicals, advanced materials, technology, renewable energy and biotechnology.
Contact Info:
Name: Arun Hegde
Email: Send Email
Organization: Global Market Insights, Inc.
Address: Global Market Insights, Inc., 4 North Main Street,
Phone: 1-888-689-0688
Website: https://www.gminsights.com/pressrelease/industrial-communication-market
Source URL: https://marketersmedia.com/global-industrial-communication-market-size-to-penetrate-the-160-billion-usd-business-realm-by-2025/88898811
Source: MarketersMedia
Release ID: 88898811


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Wall Street may be taking profits on Walmart's (WMT) - Get Report better than planned second quarter earnings, but they very easily have reason to quickly return to the stock.
Here are several positive things gleaned from Walmart's results that could serve as fodder for the bulls.
Online Sales Crushed It, Again
Walmart's online sales surged 60% in the quarter, following a 63% pop in the first quarter. Bottom line: by offering more stuff on its website (more than 67 million items and counting...) and better linking web operations with stores, Walmart is fighting back hard against ruthless Amazon (AMZN) - Get Report . This isn't pie in the sky stuff, the data actually supports it.
While the Rest of Retail Is Dying...
Walmart, and even Target (TGT) - Get Report , had solid second quarters against a backdrop of death in the mall. Last week, department-store retailers including Macy's Inc. (M) - Get Report , J.C. Penney Co. Inc. (JCP) - Get Report and Kohl's Corp. (KSS) - Get Report released a wave of disappointing earnings reports as the decline of bricks and mortar heightens, as TheStreet's Lindsay Rittenhouse reports.
Discounters are winning because they sell food, but also as their websites are just getting integrated with stores better.
The Economy Is Just Fine
Despite everything that is happening out in the political landscape, Walmart's traffic increases at both its U.S. namesake stores and Sam's Club suggest the economy is chugging along. Credit rising wages, especially among Walmart workers.
Bring on those Sears (SHLD) earnings...
Wondering what else Walmart is up to?
Don't miss these top stories on TheStreet:


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"Once you realize your government's sole purpose is to reserve murder of you and your family as its sacred task depending on your lack of fealty and obedience, everything sort of falls into place on why governments throughout the ages have been so murderous at home and abroad". William Buppert
Wednesday, December 27, 2017
Theft, Government Style
By Porter
As almost certainly you know already, the dollars in your bank [or wallet] are being stolen every day, no matter what your account balance may indicate. Just because the explicit numbers haven't declined, doesn't mean the implicit value hasn't. Inflation is absolutely the most palatable tax for politicians, because it does its work in such professional silence. The government doesn't even have to overtly tax at all to operate. It could simply print the necessary cash for its operations, as was precisely the case with Quantitative Easing. But do you think that would be getting Government for free?
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Villa Lagoon Tile -- our own line of encaustic mosaic cement floor tile. The tile is hand made to order and has our own designs. This type of tile is also called Cuban tile and "mosaicos hidraulicos". www.VillaLagoonTile.com
DAC-ART Building System The DAC-ART Corporation PO Box 6130, Mobile AL 36660, Ph 800-803-2601, 251-490-1954
General Contractor--Dan Zehentner-- Zehentner
Inc., 7355 Coopers Landing, Foley, AL 36535
Ph. 251-752-4878
Architextural Millwork & Cabinetry
2515 Hwy 4 West., Century, FL 32535
ph 850-256-4420
Billy and Valerie Dunsford--Custom kitchen cabinets, bathroom cabinetry.
Hurd Windows from Builder Resource in Mobile, AL. Wood windows, distinctive designs, limitless options. Builder Resource, 3450 Armour Avenue, PO Box 7326, Mobile, AL 36670, ph (251) 479-5412
ACP -- Evoba Wood Ceiling System -- Acoustic Ceiling Products, LLC., PO Box 1581, Appleton, WI 54912-1581 Ph Product Info: 800.434.3750 / Technical Support: 800.558.0615
Swift Supply, Inc. 24300 Canal Rd., Orange Beach, AL 36561, 251-974-5675, Fax 251-974-5677
Fastenal 1214 Hwy 59 South, Summerdale, AL 36580, Ph (251) 947-6002
Air Conditioning--American Standard® 780 Commerce Drive
P.O. Box 6265, Gulf Shores, AL 36547, Ph (251) 968-7600, (850) 492-9500
YardEnvy -- Fine decor for home and garden including patio furniture, fountains, arbors, bird feeders, fire pits, weather vanes and more. www.YardEnvy.com
Scott's Concrete, PO Box 851, Magnolia Springs, AL 36555
Concrete Acid Stain--Xtreme Concrete Designs
Chad Blackwell .--- 251-269-4368
Concrete Acid Stain--Reynolds Redi-Mix --David Trull at Reynolds Ready Mix has been very helpful. He is the local DIY concrete acid stain supplier, 251-747-4238 or 1-800-239-3879. Call David for orders of Concrete Acid Stain product.
Engineer--Anil Badve, P.E., P.O. Box 554, Foley, AL 36536 ph (251) 943-5052
New Stone Age Builders -Mike Rode-P.O. Box 4732, Gulf Shores ,AL 36542,
Vent cover grilles from Decor and Restore, Inc. who also sell specialized rugs.


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STOCKHOLM (Reuters) - Sweden's government has dropped plans for a bank tax after a consultation process drew largely negative feedback and will instead propose that banks pay more into the industry's resolution fund, the finance ministry said on Friday.
The centre-left minority government has long sought to impose a financial services tax, saying the sector - which is exempt from value-added tax - should pay more to the state.
But critics say a tax would hit the financial services industry, which is a major contributor to the Nordic country's economy and could push some financial firms to move jobs abroad.
Banks currently pay an annual fee to the resolution fund, which is a part of the system for crisis management in the financial sector. The fees add about 7 billion crowns to state coffers, the ministry said in Friday's statement.
The new proposal, which is backed by the government and the Left Party would strengthen public finances by more than 3 billion Swedish crowns (266.35 million pounds) in 2018 and by more than 6 billion crowns in 2019 due to a growing banking sector, the ministry said.
Reporting by Bjorn Rundstrom; Editing by Catherine Evans