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Type IV secretion systems (T4SSs) mediate horizontal gene transfer, thus contributing to genome plasticity, evolution of infectious pathogens, and dissemination of antibiotic resistance and other virulence traits. A gene cluster of the Haemophilus influenzae genomic island ICEHin1056 has been identified as a T4SS involved in the propagation of genomic islands. This T4SS is novel and evolutionarily distant from the previously described systems. Mutation analysis showed that inactivation of key genes of this system resulted in a loss of phenotypic traits provided by a T4SS. Seven of 10 mutants with a mutation in this T4SS did not express the type IV secretion pilus. Correspondingly, disruption of the genes resulted in up to 100,000-fold reductions in conjugation frequencies compared to those of the parent strain. Moreover, the expression of this T4SS was found to be positively regulated by one of its components, the tfc24 gene. We concluded that this gene cluster represents a novel family of T4SSs involved in propagation of genomic islands.
The horizontal gene pool contributes to the diversification and adaptation of microorganisms. A major part of the horizontal gene pool consists of genomic islands. Genomic islands are well-characterized segments of bacterial genomes, often inserted at tRNA genes, that contain homologues of genes encoding integrases and other genes associated with conjugative plasmids or phages (17).
Prior to the 1970s Haemophilus influenzae was universally susceptible to ampicillin. In 1972, the first ampicillin-resistant isolate was detected, and soon after this strains resistant to tetracycline, chloramphenicol, erythromycin, and multiple antibiotics were identified and spread rapidly around the globe. Work over the past few years has provided evidence that horizontal transfer of genes in bacteria, including transfer of antibiotic resistance, is facilitated by genomic islands.
Genomic islands of many bacterial plant and animal pathogens encode type IV secretion systems (T4SSs) which are preferentially used for delivery of bacterial effector proteins across the bacterial membrane and the plasmatic membrane into the eukaryotic host cells (4, 18, 24, 28). T4SSs also mediate horizontal gene transfer, thus contributing to genome plasticity, the evolution of infectious pathogens, and dissemination of antibiotic resistance and other virulence traits (9, 22). The structures of the genetic determinants of T4SSs vary and consist of multiple genes organized into a single functional unit. These structures have been classified into major types based on a combination of gene content and shared homology. Hitherto, two different grouping schemes and nomenclatures have been used by investigators, as described in recent reviews. In one classification there are three major types, referred to as types F, P, and I, and these types associate with model conjugation systems described for plasmids F, RP4, and R64, respectively. In the other classification, types F and P are grouped together as type IVA and type I is type IVB. A third group in this classification is composed of "other" T4SS representatives (9, 10, 13, 32).
Hitherto, a major unresolved feature of genomic islands was the mechanism by which they are transferred between bacteria. The most widely held view is that genomic islands represent mobile elements, such as phage or conjugative plasmids that have either lysogenized or cointegrated with the chromosome, and that their transfer functions have become degenerate (17). This hypothesis has been favored by a number of investigators; however, observations of a family of syntenic genomic islands with deep evolutionary relationships have challenged this hypothesis based on findings for two members of this family, ICEHin1056 and the clc element. It is recognized that both ICEHin1056 and the clc element are capable of integration into the chromosome of the host, excision, and self-transfer to a new host and reintegration (15, 16, 49).
Preliminary analysis of in silico data acquired from sequencing projects suggested that there is a highly conserved module of genes that is responsible for horizontal transfer of these genomic islands; however, no functional analysis of the components of this putative transfer module was performed, and there was no in-depth description. A better understanding of genes involved in conjugative transfer and their relationship to well-characterized conjugative systems should provide a better understanding of how at least one family of genomic islands is mobilized in bacteria and may suggest that many more genomic islands than currently recognized contain genes for self-mobilization.
Here we describe identification and functional analysis of a cluster of genes encoding an uncharacterized T4SS present in ICEHin1056, an example of a diverse family of genomic islands. Within this gene cluster are homologues of genes in previously defined T4SSs. We show that this novel T4SS is responsible for formation of the conjugative pilus and the resulting conjugative transfer of ICEHin1056. In this study we not only identified a new lineage of T4SSs but also demonstrated that a highly evolved and conserved cluster of genes is involved in the mobilization of genomic islands.
Bacterial strains, plasmids, and growth conditions.All bacterial strains and plasmids used in this study are listed in Table 1. In most cases H. influenzae was grown on HIB medium (Columbia agar containing 15 μg/ml NAD and 15 μg/ml hemin). When required, this medium was supplemented with kanamycin (10 μg/ml), tetracycline (2 μg/ml), or ampicillin (4 μg/ml). All Haemophilus plate cultures were grown for 24 to 48 h at 37°C in an atmosphere containing 5% CO2. Liquid cultures of H. influenzae were grown in brain heart infusion broth (BHI) supplemented with NAD (10 μg/ml), hemin (15 μg/ml), and, when necessary, antibiotics at the concentrations described above and incubated at 200 rpm on a rotatory shaker at 37°C. Luria-Bertani broth was routinely used for growing Escherichia coli strains. When appropriate, Luria-Bertani medium was supplemented with ampicillin (50 μg/ml), kanamycin (50 μg/ml), or gentamicin (5 μg/ml).
PCR amplification and recombinant DNA methodology.Restriction endonucleases and DNA-modifying enzymes were obtained from New England Biolabs and were used according to the manufacturer's instructions. Recombinant DNA techniques and plasmid DNA preparation were performed as described by Sambrook et al. (45). Oligonucleotide primers were synthesized by Operon. Standard PCR amplifications were performed in 10-μl mixtures using Taq DNA polymerase or ProofStart DNA polymerase (QIAGEN) according to the supplier's instructions. Four-kilobase DNA fragments comprising the putative transfer region of ICEHin1056 were cloned into pGEM-T Easy (Promega). Recombinant plasmids were confirmed by PCR amplification and restriction endonuclease digestion. Genes of the putative type IV secretion system (tfc1, tfc2, tfc4, tfc6, tfc12, tfc14, tfc16, tfc22, tfc23, and tfc24) were inactivated by inserting a kanamycin resistance cassette and the H. influenzae DNA uptake signal sequence (47). These genes were chosen primarily because they exhibit homology to genes of other T4SSs; however, one of these genes, tfc19, a homologue of traG, lacked suitable restriction sites for insertional mutagenesis. An additional four genes in the module were inactivated because of well-placed unique restriction sites.
Generation of H. influenzae mutant strains.For construction of mutant strains by reciprocal recombination, 2 to 3 μg of the appropriate linearized plasmid DNA construct was used to transform H. influenzae strain Rd11 (Table 1), and transformants were selected with kanamycin (27). All transformants were checked by reculturing on HIB medium containing kanamycin and were subsequently confirmed to be mutants by PCR amplification. The PCR results were consistent with the hypothesis that only one copy of ICEHin1056 was present in each bacterium.
Conjugal transfer of the ICEHin1056 genomic island.The transfer efficiency of ICEHin1056 was determined using a modification of the method of Stuy (48). Donor or recipient cells were grown for 48 h on HIB agar, and approximately 108 cells were scraped off each plate and resuspended in 1 ml of BHI broth. Ten microliters of the suspension of donor cells and 100 μl of the suspension of recipient cells were gently mixed to obtain a ratio of donor cells to recipient cells of 1:10. Subsequently, this mixture was spread in the center of antibiotic-free HIB agar plates. The inoculum was allowed to dry prior to incubation for 6 h. The cells were then harvested by flooding the plate with 1 ml of BHI broth and resuspending them using a spreading paddle. Serial dilutions were plated to determine the viable counts and to determine the number of transconjugants, donors, and recipients using agar containing the appropriate selective antibiotic. Recipients and transconjugants were purified on agar plates containing tetracycline (2 μg/ml) and on agar plates containing kanamycin (10 μg/ml) plus tetracycline (2 μg/ml), respectively. Experiments were carried out in triplicate, and the mean and standard error for each strain were calculated.
Electron microscopy.To examine H. influenzae strains for the presence of the type IV secretion-dependent pilus, bacteria were grown overnight and then allowed to conjugate on antibiotic-free HIB agar plates for 6 h. The conjugating cells were harvested by flooding each plate with 1 ml of distilled water and subsequently resuspended them using a spreading paddle. A copper grid coated with Formvar and carbon was floated in the resulting suspension for 2 min. Excess fluid from the copper grid was removed prior to negative staining with 1% methyl tungstate. The cells were examined to determine the presence of a pilus by transmission electron microscopy.
Reverse transcriptase PCR (RT-PCR).Following growth for 12 h on solid medium and growth to an optical density at 600 nm of 0.6 in liquid medium, total RNA was prepared from harvested bacteria using the SV total RNA isolation system (Promega); the strains used were H. influenzae strains Rd and Rd11, as well as Rd11tfc1, Rd11tfc2, Rd11tfc4, Rd11tfc6, Rd11tfc12, Rd11tfc14, Rd11tfc16, Rd11tfc22, Rd11tfc23, Rd11tfc24, which represented mutants with mutations in the putative T4SS. Following RNA isolation, the yield was quantified spectrophotometrically. Subsequently, 550 ng of total RNA was reverse transcribed to produce cDNA using Superscript II reverse transcriptase (Invitrogen) and random primers (Promega). Coamplification of the cDNA transcripts for genes of the putative T4SS with an internal control allowed comparison of the levels of expression in the different RNA samples. PCR amplification of the cDNA of the putative T4SS genes was performed with oligonucleotides and probes listed in Table 2. Primers specific for the constitutively expressed housekeeping gene encoding beta-lactamase were also included in each PCR amplification as an internal control. To control for chromosomal DNA contamination, RNA samples were also directly used for PCR amplification.
Quantification of the tfc16 and tfc17 transcripts in strains Rd11 and Rd11tfc24 was performed using an ABI Prism 7000 sequence detection system (Applied Biosystems) according to the manufacturer's recommendations. Probes were labeled with 6-carboxyfluorescein. Plasmid standards were obtained by preparing serial 10-fold dilutions in 100 μl (final volume) of nuclease-free water such that 5 μl of standard added to a 50-μl real-time quantitative PCR mixture (1× TaqMan Universal PCR Master Mix [Applied Biosystems], each primer [Applied Biosystems] at a concentration of 500 nM, 100 nM TaqMan 6-carboxyfluorescein-labeled probe [Applied Biosystems]) contained from 3 × 105 to 3 × 101 copies. The reporter fluorescence was normalized to the fluorescence of the passive reference dye ROX. Amplification data, expressed as increases in reporter fluorescence, were collected in real time and were analyzed using the sequence detection system. Experiments were repeated in triplicate with two batches of cDNA made from RNA samples obtained from two independent preparations.
DNA sequence analysis.DNA sequence similarity searches using the BLASTN and BLASTX algorithms (1) and position-specific iterated BLAST (PSI-BLAST) (2) were performed by interrogating the National Center for Biotechnology Information (NCBI) website (http://ncbi.nlm.nih.gov/). The Smith-Waterman algorithm, using the Blosum65 score matrix, a gap opening penalty of 11, and a gap extension penalty of 1, was used to align low-homology open reading frames (ORFs) with the ORFs of known T4SSs and to compute P values for homology (29), using a Bonferroni correction to account for multiple testing. The Artemis comparison tool (ACT) (42) was used to visually compare T4SSs of various bacteria pairwise for homology by using the TBLASTX algorithm. The presence and locations of signal peptide cleavage sites in amino acid sequences were predicted with SignalP 3.0 Server (http://www.cbs.dtu.dk/services/SignalP/), using a combination of several artificial neural networks and hidden Markov models (6).
Evolutionary analysis.The evolutionary relationships of a set of genomic islands that carry putative T4SSs were investigated using three representative essential genes (tfc6, tfc14, and tfc16). Homologues of only these genes were present in all the genomic islands represented in the comparative analysis. It has been claimed elsewhere that tfc14 and tfc16 encode the archetypal T4SS proteins (32). The relationships were estimated using ClustalX based on either the amino acid sequences encoded by each gene individually or the amino acid sequences encoded by the concatenated sequences of tfc14 and tfc16. For these analyses, the following settings were used: correction for multiple substitutions and exclusion of positions with gaps. The output from the phylogenetic analysis was visualized with the TreeView software (38).
Identification of the conserved type IV secretion system.Genomic island ICEHin1056, which is an archetype of a genomic island that accounts for pandemic spread of antibiotic resistance among H. influenzae, is 59.4 kb long and consists of 64 ORFs. Based on preliminary bioinformatic analyses, 24 of the 64 ORFs of ICEHin1056, designated tfc1 to tfc24 (Fig. 1a), were thought to encode an uncharacterized T4SS. Seven of the 24 gene products of the ICEHin1056 putative T4SS have transmembrane helices, suggesting membrane-bound localization of the relevant proteins (Fig. 1a). Moreover, sequence analysis with SignalP 3.0 Server (6) revealed that 10 of the 24 genes of this gene cluster have signal peptide sequences that are typical for genes involved in T4SSs (Fig. 1a) (34). Characteristically, T4SSs are encoded by many genes expressing transmembrane domains and signal peptide sequences (32). DNA sequence similarity searches using the BLASTN and BLASTX algorithms (1) and position-specific iterated BLAST (2) together with the Smith-Waterman algorithm (29) were employed to identify components of this putative T4SS. This bioinformatic analysis revealed homology between tfc1 and the known T4SS gene traX (E = 1 × 10−60 in PSI-BLAST iteration 2), encoding a protein responsible for acetylation of F plasmid prepilin (35). Moreover, tfc6 was homologous to traD (E = 2 × 10−166 in PSI-BLAST iteration 1), a gene encoding a T4SS conjugative coupling protein involved in synchronization of mating pair formation with DNA transfer and "pumping" of DNA into recipient cells (22, 25). tfc14, tfc16, and tfc19 are traB (E = 9 × 10−06 in PSI-BLAST iteration 2), traC/virB4 (E < 3 × 10−159 in PSI-BLAST iteration 1), and traG (E = 2 × 10−42 in PSI-BLAST iteration 3) homologues, respectively, all of which are involved in pilus assembly (3, 32, 40). tfc2 and tfc22 exhibit homology with pilL (E = 4 × 10−19 in PSI-BLAST iteration 1) and pilT (E = 1 × 10−21 in PSI-BLAST iteration 1), respectively, whose products are required for type IV secretion pilus biogenesis (43, 44). Altogether, these in silico findings (Fig. 1a) suggest that the gene cluster which we investigated encodes components of a T4SS. Moreover, this gene cluster has been recognized in related genomic islands of other bacterial species, including Legionella pneumophila, Pseudomonas aeruginosa, Pseudomonas fluorescens, Photorabdus luminescens, Erwinia carotovora, and Salmonella enterica serovar Typhi.
ICEHin1056 type IV secretion system is evolutionarily distant from previously described T4SSs.Work over the past few years has led to classification of three distinct clusters of T4SSs: F-like (21, 22, 32, 33, 37), P-like (12, 13), and I-like (31, 46, 50). Relationships within and among clusters are based mainly on three genes, identified as either tfc6, tfc14, and tfc16 or traD, traB, and traC/virB4, respectively (named for the E. coli F plasmid T4SS). The tfc14 and tfc16 genes encode the quintessential T4SS proteins (32) and as such are ideal for comparative amino acid alignment analysis. The trees for these three genes showed different patterns of clustering across genomic islands, but substantial compatibility between trees was evident based on homologies to tfc14 and tfc16 (Fig. 2a to c). Combining the data for both sets of homologues of tfc14 and tfc16 produced a tree with strong bootstrap support for four major clusters (Fig. 3). The fourth type of T4SS is carried by a wide variety of genomic islands, including the clc element, pKLC102, PAPI, and Salmonella pathogenicity island 7, as well as other genomic islands derived from the following bacterial strains: L. pneumophila, P. aeruginosa, P. fluorescens, E. carotovora, and S. enterica serovar Typhi (Fig. 3).
Role of the ICEHin1056 type IV secretion system in pilus biogenesis.T4SSs are multisubunit cell envelope-spanning structures comprising a secretion channel and often a pilus or other surface filament or protein (13, 19, 32, 41, 51, 52). Despite the presence of a vestigial pilus gene cluster in the genome, H. influenzae strain Rd produces neither fimbriae nor type IV pili (5, 20). However, electron microscopic examination of strain Rd harboring genomic island ICEHin1056 (strain Rd11) revealed long thin pili (Fig. 4a to d), which is consistent with the hypothesis that the genes responsible for formation of pili are located on ICEHin1056. Only a single pilus approximately 7 to 7.5 nm in diameter and 1.5 μm long was attached to bacteria (Fig. 4a). In order to demonstrate the essential role of this putative T4SS for the formation of pili, 10 genes in this gene cluster, tfc1, tfc2, tfc4, tfc6, tfc12, tfc14, tfc16, tfc22, tfc23, and tfc24, were disrupted by insertion of a kanamycin resistance cassette, and mutant strains were examined for expression of pili by electron microscopy. These genes were chosen primarily because they exhibit homology to genes of other T4SSs or because of well-placed unique restriction sites. As shown in Table 3, disruption of these genes had a dramatic effect on the abilities of the mutants tested to produce pili. Pili were not detected in 7 of the 10 mutants analyzed, confirming the crucial role of this gene module in T4SS-dependent pilus expression. Not all genes associated with known T4SSs are involved in pilus biogenesis (13, 33). Disruption of the remaining three genes, tfc1, tfc6 and tfc24, did not diminish the production of pili (Fig. 4b to d and Table 3). The pilus-positive phenotype of strains Rd11tfc1 and Rd11tfc24 provided an explanation for the observed slight changes in the conjugation frequencies described below and suggest that tfc1 and tfc24 do not play a significant role in the formation of the conjugative type IV pili under our in vitro conditions. By contrast, despite its massive reduction in conjugative transfer, the mutant with a mutation in the tfc6 gene, encoding the conjugative coupling protein, produced pili (Table 3), confirming the hypothesis described by Panicker and Minkley (39) and verified by several other research groups that conjugative coupling proteins are essential for conjugation but are not required for T4SS pilus synthesis. However, most of the mutants examined were not able to express pili, revealing the essential role of this gene module in the expression of type IV secretion-dependent pili. None of the mutants has been complemented in trans as there is not a complementation system in H. influenzae that is easy to use; however, our experimental data show that the mutations had no polar effects on at least two of the downstream genes (tfc6 and tfc24 mutants produced pili in spite of pilus-negative phenotypes of upstream genes).
ICEHin1056 type IV secretion system is important for conjugation.Conjugation systems, assembled by a subfamily of the T4SSs, are usually used by bacteria in the process of conjugative transfer of DNA from donor cells to recipient cells (11). Genomic island ICEHin1056 in the parent strain was transferred from the donor to the recipient strain of H. influenzae at a frequency of 3 × 10−2. To obtain evidence for the function of the ICEHin1056 T4SS, all T4SS mutant strains were tested to determine their abilities to transfer the genomic island by conjugation to the same recipient. As shown in Table 3, the conjugation frequencies of the mutants were significantly lower than the conjugation frequency of the parent strain and ranged from 6 × 10−3 to 1 × 10−7. In agreement with our expectations, the lowest conjugation frequency, 1 × 10−7, was observed for the mutant with the disrupted tfc6 gene, encoding a putative conjugative coupling protein (8). Disruption of three genes, tfc1, tfc4, and tfc24, reduced the conjugal transfer frequencies to approximately 10−3, whereas disruption of the remaining seven genes, tfc2, tfc6, tfc12, tfc14, tfc16, tfc22, and tfc23, led to reduction of the conjugal transfer frequencies to approximately 10−7. In summary, the abilities of the majority of the mutants tested to conjugate were critically impaired, and the frequencies of transfer were reduced up to 100,000-fold compared to the frequencies of transfer observed for the parent strain, providing functional evidence of the importance of the T4SS for conjugal transfer of ICEHin1056 DNA; hence, we used the designation tfc (type four conjugation) for all components of this gene cluster.
Transcriptional analysis of the ICEHin1056 type IV secretion system.RT-PCR analysis of gene expression revealed that the ICEHin1056 T4SS gene cluster is expressed in the form of polycistronic transcripts (Fig. 1b and c). No mRNA transcripts were detected between tfc19 and tfc20 and between the first gene (tfc1) and the last gene (tfc24) of the ICEHin1056 type IV secretion cluster analyzed and the neighboring genes in the rest of the ICEHin1056 sequence (Fig. 1b and c). As Fig. 1b and c show, the RT-PCR analysis results suggest that the module consists of two transcripts. Transcriptional analysis by RT-PCR (Fig. 1b and c) revealed amplicons for the gene downstream of the gene interrupted by insertion of an antibiotic resistance cassette in each of the 10 T4SS mutant strains tested (data not shown). This suggests that there are no polar effects on genes downstream of insertional mutations.
Interestingly, RT-PCR analysis of gene expression of the ICEHin1056 T4SS indicated that there were no substantial changes in the pattern of expression in the mutants tested compared to the wild-type pattern, except for mutant tfc24. As shown in Fig. 5a, mutation of tfc24 greatly reduced expression of many of the type IV secretion genes. Quantification of tfc16 and tfc17 mRNA transcripts in strains Rd11 and Rd11tfc24 using the ABI Prism 7000 sequence detection system revealed that the number of copies of the tfc16 gene transcript was reduced from 530 × 102 in strain Rd11 to 263 × 102 in mutant Rd11tfc24 (Fig. 5b). Moreover, the number of copies of the tfc17 gene transcript was reduced from 537 × 102 in strain Rd11 to 411 × 102 in mutant Rd11tfc24 (Fig. 5b). The tfc24 product exhibits strong homology with conserved proteins having unknown functions found in a wide variety of bacterial species, including Haemophilus somnus, Haemophilus ducreyi, P. aeruginosa, P. fluorescens, Pseudomonas syringae, P. luminescens, E. carotovora, S. enterica serovar Typhi, and Yersinia pseudotuberculosis, and it contains a protein kinase C phosphorylation site. This suggests that tfc24 might encode a protein which plays a crucial role in regulation of the T4SS, whose function might be conserved between species. The role of this gene in regulation remains to be elucidated.
A set of genes present in ICEHin1056, a vector of antibiotic resistance in H. influenzae, was characterized, and we concluded that this vector represents a new family of T4SSs involved in DNA translocation among gram-negative bacteria. This interpretation was based on bioinformatic, evolutionary, and functional analyses. First, in the bioinformatic analysis, both protein motifs typical of T4SS gene clusters were found, and homologies to genes known to be associated with T4SSs were identified. Second, the evolutionary analysis of the two archetypal T4SS proteins revealed high levels of similarity between amino acid sequences for the three previously identified T4SS types plus a novel fourth type. Third, mutagenesis of genes indicated that the genes are involved in pilus formation and the resulting transfer of DNA by conjugation. Lastly, a transcriptional analysis suggested that this gene cluster is probably part of a regulated operon typical of other T4SSs. The putative origin of transfer has been localized between ORFs 60 and 61 of ICEHin1056, a position that is distinct from the position of the putative T4SS module investigated.
The genes that we analyzed are members of a set of core genes found in a diverse family of syntenic genomic islands (36). Well-recognized members of this family of genomic islands are widely distributed among proteobacteria, and additional variants have been identified among completed genome sequences present in NCBI databases.
Analysis of mRNA by RT-PCR suggested that transcription of the T4SS genes is polycistronic. The gene disruptions in the T4SS mutants described here have not been complemented in trans as no reliable complementation system is available for H. influenzae; however, our experimental data show that the mutations did not have polar effects on the downstream genes. Both tfc6 and tfc24 mutants produce pili in spite of the pilus-negative phenotype of upstream genes. Similarly, the conjugation frequencies of tfc4 and tfc24 mutants are only slightly reduced compared to those of mutants with mutations in upstream genes. Furthermore, transcriptional analysis suggested that there was no clear polar effect of gene disruption on downstream transcripts. This might be explained by the fact that the kanamycin resistance cassette used harbors an endogenous promoter which, when oriented in the same direction as the target gene, does not interfere with downstream gene expression.
Recent reviews of T4SSs have described the relationship between the recognized T4SSs (9, 13, 32). A simple consensus view is that T4SSs share two features: they encode bacterial protein and nucleoprotein export systems, and members of each type exhibit evidence of common ancestry. This definition is broad, and its interpretation is flexible; therefore, it is not surprising that the T4SS gene clusters described previously are diverse in a number of respects. They vary in gene content, the number of homologues that they share, and the gene order. This heterogeneity is a challenge to a simple classification. Therefore, it is not surprising that two approaches have been used for classification. Both approaches were founded on a combination of the genetic organization of the cluster and the evolutionary relationships among homologous genes. The first classification recognized three types based on the incompatibility group of the representative and archetypal DNA conjugative plasmid, namely, type F for IncF (plasmid F), type P for IncP (plasmid RP4), and type I for IncI (plasmid R64) (32). There is greater conservation of gene content and order within each type than there is between types. Alignment of amino acid sequences shared by proteins encoded by homologous genes supports the subdivision into these three groups. The alternative classification recognizes two major groups, type IVA and type IVB, and a third, "other" group (9, 10, 13). Type IVA includes the F and P types referred to above. Type IVB consists of type I, whose component modules are considered to be highly divergent from those of members of both type F and type P. The third group, labeled the "other" group, has not been well characterized.
The T4SS gene cluster reported here is distinct from previously recognized types whichever typing scheme is used. It represents a fourth lineage with a genetic distance as great as that between the F, P, and I lineages. The component genes of this module exhibit homology to a limited number of genes in the other lineages; only four homologues, tfc6 (traD), tfc14 (traB), tfc16 (traC/virB4), and tfc19 (traG) (Fig. 1a), are shared with the 12-gene VirB/D4 system found in the Ti plasmid of Agrobacterium tumefaciens and with the more numerous genes contributing to the T4SS gene clusters of the F plasmid and plasmid R64. Thus, using the alternative classification system, this new T4SS would be classified as a member of neither type IVA or type IVB but as a member of the "other" group. Despite the involvement in DNA conjugation, there is no recognized incompatibility group for the elements containing the new T4SS; consequently, it does not readily fit into the typing scheme consisting of types F, P, and I. We designated this lineage the GI type, as this T4SS is associated exclusively with genomic islands, as explained further below. The GI type T4SS gene cluster is a core component of a highly diverse family of syntenic genomic islands that presumably evolved in proteobacteria. Remarkably, the G+C contents of the GI type gene clusters found in this family of genomic islands range from 40% to approximately 70%, indicating that there is substantial divergence and evolutionary separation; however, the gene content and order have remained largely intact (36). This suggests that there is a particular fitness advantage to this coherent set of genes acting together, and intriguingly, so far these genes appear to be uniquely associated with a specific family of genomic islands.
The observations reported here describing the GI type of T4SS module from the archetypal genomic island ICEHin1056 provide direct evidence of how genomic islands may conjugally transfer. Hitherto, there was doubt about how such widely distributed structures could rapidly mobilize and spread through bacterial populations. Here we present evidence that one related, diverse, and widely distributed family of genomic islands possesses the genetic wherewithal to conjugate. Only two examples of this family of genomic islands have been shown to be conjugally transferable, namely, ICEHin1056 and the clc element (23, 36). It can be inferred that other members of this family of genomic islands mobilize intact from the distribution of the following genomic islands among their host bacteria: Salmonella pathogenicity island 7 (S. enterica serovar Typhi) (7), the Yersinia adhesion pathogenicity island (Y. pseudotuberculosis) (14), and PAP1 and pKLC102-like genomic islands (P. aeruginosa) (26, 30). The presence of a highly evolved and efficient conjugation system for mobilizing genomic islands provides a better explanation of how one family of genomic islands is able to propagate and efficiently enable bacterial populations to adapt to rapidly changing environments. What remains to be seen is whether the many diverse genomic islands not related to ICEHin1056 have dedicated conjugation systems that enable them to propagate through conjugation. Only recently have bioinformatic techniques become powerful enough to recognize and make predictions allowing recognition of the ICEHin1056 family of genomic islands and their related T4SS gene clusters. It is quite plausible that other distinct families of genomic islands will be identified and that new and divergent lineages (families) of T4SS gene clusters will be recognized.
Potentially, 24 genes were found in the T4SS module of ICEHin1056, and 7 of these genes are homologous to genes directly or indirectly associated with other T4SS gene clusters. Four of these seven genes are homologues to VirB/D4 genes found in the Ti plasmid of A. tumefaciens, a gene cluster considered by many workers to be the minimal essential set of T4SS genes; therefore, there are at least 18 genes with no homology to any of the remaining genes in the other type F, P, and I T4SS lineages, as determined by currently available techniques. Similarly, there are genes in types F, P, and I that are unique to each type, and the number may be more than 10 for type I and for type F. This heterogeneity between types for genes forming a T4SS module is consistent with the hypothesis that these genes were assembled through recruitment by a mixture of convergent (horizontal transfer) and divergent evolutionary processes. Curiously, within each type, there is marked preservation of coherence in each T4SS gene cluster. Once assembled, despite opportunities for recombination and reordering of component genes, the cluster apparently remains coherent due to a particular fitness advantage for a coordinated set of genes. Intriguingly, at least two genes, traC/virB4 and traB, appear to be sufficient to define membership in their respective lineages and may be essential for a cluster to function as a T4SS export system. If these two genes provide the key signature identifying other poorly related T4SSs among genomes and genomic islands in particular, searching for homologues of these genes may be the way to identify other distinct T4SS lineages.
We cordially thank E. Richard Moxon and Sarah L. Turner for critical reviews and comments on the manuscript and Kate Dingle and Gaynor Jenkins for their technical assistance with RT-PCR expression analysis.
This work was supported by a grant from the Medical Research Council. D.J.P.F. was supported by an equipment grant from the Wellcome Trust.
- Received 21 August 2006.
- Accepted 10 November 2006.
↵▿ Published ahead of print on 22 November 2006.
- American Society for Microbiology


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What You Need To Know About Progesterone Hormones
Progesterone is one of the many hormones produced that regulates and stimulates various functions in our bodies. While most of us are aware of estrogen, little attention is focused on progesterone. Progesterone is produced by the ovaries, the placenta when a woman is pregnant and also by the adrenal glands. It's primary function is to prepare the uterus for implantation of the fertilized egg and to maintain the pregnancy.

subset: commoncrawl


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Allied House Mortgage is suspended from originating or underwriting mortgages insured by the Federal Housing Administration.
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At the 2017 Istanbul Biennial, Mahmoud Khaled proposed a future in which private actions are protected from public scrutiny.
The day after Mahmoud Khaled left Istanbul following the unveiling of his new commission at this year's biennial, he saw the news that police in Cairo had arrested seven concert-goers for raising a rainbow flag during a performance by Lebanese rock band Mashrou' Leila.
Khaled, an Egyptian who lives and works in Norway, addresses topics of gender and sexuality in his practice and the Istanbul Biennial commission tackles the issues head on.
For Proposal for a House Museum of an Unknown Crying Man (2017), Khaled transformed Ark Kültür, an art centre in a 1930s' residential property, into the elaborately fictionalised home of a protagonist who has fled Cairo to live in exile in Istanbul. Khaled took as his starting point the trial of 52 men who were arrested in 2001 aboard a floating gay nightclub in Cairo. During this highly publicised ordeal, one of the men was photographed crying and covering his face with a white cloth. Until today, that image still remains a symbol for the homosexual community in Egypt, whilst the man's identity and humanity have been sidelined.
Khaled's installation delves deeply into this symbolic usurpation of individuality and evokes great sympathy for what that crying man represented: a personification of the LGBTQI community, not just in Egypt, but in Turkey and across the rest of the world.
Through a tightly curated audio guide, visitors to Khaled's installation found themselves moved from room to room with their gaze directed at the poignant details of the crying man's life, which conveyed a profound sense of melancholy, but in a poetic and elegant way.
His shelves were full of uninscribed trophies longing for accomplishments yet to come and, in his shower, a monitor continuously played a seven-minute scene from Maher Sabry's movie Toul Omry / All My Life (2008) – the first Egyptian film to openly address homosexuality, including the case of the Cairo 52. The table was laid with a single place setting, while upon the crockery, and in several other places around the house, a replicated image of The Crying Boy, a mass-produced print of a painting by Italian painter Giovanni Bragolin, could be found. Part of his own childhood, Khaled remembers this kitsch image hanging in the family home and in those of most of his friends. Just as nobody could ever tell him why the boy was crying or indeed, who he was, when Khaled realised that the image of the man under arrest covering his own tears was also becoming commodified, he was moved to try to construct a world in which a person's actions in private were not a matter for public scrutiny.
This is, of course, as idealised as the proposed house museum itself, which suggests that the legacy of someone as faceless and as publically shunned as the original crying man would be given weight.
What must be celebrated about this installation is the provocation to viewers to reconsider their own judgement, question patriarchal systems and, also, reflect on the fact that deep seated prejudices continue to fester, even within societies championing notions of progress.
Featured image: Mahmoud Khaled, Proposal for a House Museum of an Unknown Crying Man, 2017, mixed media, dimensions variable, image courtesy of the artist and Gypsum Gallery, produced with the support of Arts Council Norway, Ari Meşulam and Office for Contemporary Art Norway, photo by Sahir Uğur Eren.
A version of this article appeared in print in Selections, Letters From The Past #43 pages 62-63.


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A guide to cultural and recreational events in the Hudson Valley. Items for the calendar should be sent at least three weeks in advance to firstname.lastname@example.com.
PELHAM Rockwells Kevin Meaney and Brian Jian. Aug. 22 at 9 p.m. $10. Rockwells, 105 Wolfs Lane. rockwellsusa.net; 914-738-5881.
WEST NYACK Levity Live Steve Byrne. Through Aug. 16. $20. Chris D'Elia. Aug. 19 at 7:30 and 9:45 p.m. $30. Artie Lange. Aug. 21 at 7:30 p.m. and Aug. 22 at 7 and 9:30 p.m. $40. Levity Live, 4210 Palisades Center Drive. 845-353-5400; levitylive.com.
BETHEL Bethel Woods Center for the Arts Moonlit Movie Mondays: "The Rocky Horror Picture Show" (1975), directed by Jim Sharman. Aug. 17 at 8:30 p.m. $5 and $8. Bethel Woods Center for the Arts, 200 Hurd Road. bethelwoodscenter.org; 866-781-2922.
HUDSON Time & Space Limited "Love & Mercy" (2014), directed by Bill Pohlad. "Court" (2014), directed by Chaitanya Tamhane. "Tangerine" (2015), directed by Sean S. Baker. Through Aug. 23. $6 and $8. "A Midsummer Night's Dream" (2014), directed by Julie Taymor. Aug. 23 and 30 at 3 p.m. $7 to $12.50. Time & Space Limited, 434 Columbia Street. 518-822-8448; timeandspace.org.
NYACK Memorial Park An outdoor screening of "The Princess Bride" (1987), directed by Rob Reiner. Aug. 17 at 8 p.m. Free. Memorial Park, Piermont and Depew Avenues. rivertownfilm.org; 845-353-2568.
OSSINING Ossining Public Library "Selma" (2014), directed by. Aug. 19 at 6:30 p.m. Free. Ossining Public Library, 53 Croton Avenue. ossininglibrary.org; 914-941-2416.
PLEASANTVILLE Jacob Burns Film Center "Meru " (2015), screening and discussion with the filmmakers. Aug. 17 at 7:30 p.m. $10 and $15. "I Am What I Play"(2015), screening and discussion with Meg Griffin. Aug. 18 at 7:30 p.m. $7 and $12. "Lambert and Stamp" (2014), directed by James D. Cooper. Aug. 19 at 7:30 p.m. $7 to $12. "East Jerusalem/West Jerusalem" (2014), directed by Erez Miller and Henrique Cymerman. Aug. 20 at 7:30 p.m. $7 to $12. Jacob Burns Film Center, 364 Manville Road. 914-747-5555; burnsfilmcenter.org.
ROSENDALE Rosendale Theater "Me and Earl and the Dying Girl" (2015), directed by Alfonso Gomez-Rejon. Through Aug. 20. $5 and $7. "National Theater Live: 'A View From the Bridge,' " screening of the play by Arthur Miller. Aug. 23 at 3 p.m. $10 and $12. Rosendale Theater, 408 Main Street. rosendaletheatre.org; 845-658-8989.
For Children
BRONX Wave Hill "Family Art Project: Green Bugs Gathering." Aug. 16, 10 a.m. to 1 p.m. "Stories in the Garden: Animal Antics." Aug. 19 at 2 p.m. "Family Art Project: Delicate Dragonfly Wings." Aug. 22 and 23, 10 a.m. to 1 p.m. Free with admission to the grounds. $2 to $8; members and children under 6, free. Admission is free until noon on Saturdays. Wave Hill, 675 West 252nd Street. 718-549-3200; wavehill.org.
KATONAH Muscoot Farm Fairy Walk, activities in the woods. Costumes encouraged. Aug. 23, 1 to 3 p.m. Free. Muscoot Farm, 51 Route 100. 914-864-7282; muscootfarm.org.
PEEKSKILL 12 Grapes Music and Wine Bar Open mike for musicians ages 6 to 17. Aug. 16 at 6 p.m. Free. 12 Grapes Music and Wine Bar, 12 North Division Street. 914-737-6624; 12grapes.com.
PEEKSKILL The Field Library Children's Room Library Live: Hudson Valley Llamas, animal encounters. Aug. 20 at 6:30 p.m. Free. The Field Library Children's Room, 4 Nelson Avenue. fieldkids.wordpress.com; 914-737-0847.
PLEASANTVILLE Jacob Burns Film Center "The Muppets Take Manhattan" (1984), directed by Frank Oz. Aug. 22 and 23 at noon. $7 to $12. Jacob Burns Film Center, 364 Manville Road. burnsfilmcenter.org; 914-747-5555.
ROSENDALE Rosendale Theater Youth Arts: Puppet and Performing Arts Immersion Week. Ages 9 to 14. Aug. 17 to 24, 9 a.m. to 3 p.m. $145. "Outbounders and Dreamers," puppet theater. Aug. 21 at 5:30 p.m. $5 and $8. $20 for families. Rosendale Theater, 408 Main Street. 845-658-8989; rosendaletheatre.org.
SCARSDALE Greenburgh Nature Center "What's the Buzz With Honeybees?" Educational workshop. Aug. 16 at 2 p.m. $5 and $8. Greenburgh Nature Center, 99 Dromore Road. greenburghnaturecenter.org; 914-723-3470.
WEST NYACK Levity Live Mark Calabrese, comedy and magic. Aug. 16 at 2 p.m. $14. Levity Live, 4210 Palisades Center Drive. 845-353-5400; levitylive.com.
YONKERS Alamo Drafthouse Cinema "Kung Fu Panda 2," animated film. Aug. 17 through 20. $1 to $3. Alamo Drafthouse Cinema, 2548 Central Park Avenue. 914-226-3082; drafthouse.com/yonkers.
Music and Dance
ANNANDALE-ON-HUDSON The Richard B. Fisher Center for the Performing Arts, at Bard College Program Nine: "Sacred and Secular Choral Music From Five Centuries." Aug. 16 at 10 a.m. $30. Program 10: "Post-World War II Latin America." Aug. 16 at 1:30 p.m. $35. Program 11: "Musical Culture of the Hemisphere." Aug. 16 at 4:30 p.m. $25 to $75. The Richard B. Fisher Center for the Performing Arts, at Bard College. 845-758-7900; fishercenter.bard.edu.
BEACON Towne Crier Cafe Compton and Newberry, country. Aug. 16 at 7:30 p.m. $20. Chris Hillman and Herb Pedersen, country and rock. Aug. 20 at 7:30 p.m. $30 and $35. Red Molly, folk. Aug. 21 at 8:30 p.m. $25 and $30. Bruce Cockburn, folk and rock. Aug. 22 at 8:30 p.m. $45 and $50. The Revelers, country and blues. Aug. 23 at 7:30 p.m. $20 and $25. Towne Crier Cafe, 379 Main Street. 845-855-1300; townecrier.com.
BRONXVILLE Leonard Morange Park That Duo Show and Antique Car Show, classic rock. Aug. 20 at 6:30 p.m. Free. Leonard Morange Park, Parkway Road. 914-337-6040; bronxvillechamber.com.
CHATHAM PS21 Chatham "So You Think You Don't Understand (Modern) Dance?" Works in progress by Dance Omi Alumni. Aug. 21 at 2 p.m. Free. Keigwin + Company, contemporary. Aug. 21 and 22 at 8 p.m. $18 to $30. PS21 Chatham, 2980 Route 66. 518-392-6121; ps21chatham.org.
COLD SPRING Riverfront Bandstand Melissa Ferrick, folk. Aug. 16 at 6 p.m. Free. Chris O'Leary Band, blues. Aug. 23 at 6 p.m. Free. Riverfront Bandstand, Main Street. explorecoldspringny.com.
COLD SPRING The Chapel Restoration John Abercrombie, guitar, and Marc Copland, piano. Aug. 16 at 4 p.m. Donations accepted. The Chapel Restoration, 45 Market Street. chapelrestoration.org; 845-265-5537.
MARLBORO The Falcon Alexis P. Suter and the Ministers of Sound, soul and blues. Aug. 16, 10 a.m. to 2 p.m. E. J. Strickland and Transient Beings, jazz. Aug. 16 at 7 p.m. Jocelyn Arndt, soul. Aug. 17 at 7 p.m. Jeremy Baum's JB3 Trio, blues and jazz. Aug. 19 at 7 p.m. Eric Harland's Voyager, jazz. Aug. 21 at 7 p.m. Blue Chicken, rock. Aug. 22 at 8 p.m. Big Joe Fitz and the Lo-Fis, jazz. Aug. 23, 10 a.m. to 2 p.m. Donations accepted. The Falcon, 1348 Route 9W. 845-236-7970; liveatthefalcon.com.
NEW ROCHELLE Alvin & Friends Restaurant Mayte Vicens and Eduardo Goytia, tango. Aug. 21. Dance lesson at 8 p.m. followed by dancing until midnight. $25. The Chip Shelton Peace Time Ensemble, jazz. Aug. 23 at 5 and 7 p.m. $20. Alvin & Friends Restaurant, 14 Memorial Highway. alvinandfriendsrestaurant.com; 914-654-6549.
OSSINING Louis Engel Waterfront Park The Old Mill Singers, choral group. Aug. 21 at 7 p.m. Free. Louis Engel Waterfront Park, 160 Westerly Road. villageofossining.org; 914-941-3189.
PAWLING Daryl's House Adam Ezra Group, folk and rock. Aug. 16 at 2 p.m. No cover. Jason Gisser Band and the Differents, rock and soul. Aug. 16 at 7 p.m. $10 and $15. Dick Dale and Mishti, surf rock. Aug. 19 at 8 p.m. $35 and $50. "Satisfaction: the International Rolling Stones Show," a tribute concert. Aug. 20 at 8 p.m. $20 and $25. Albert Lee and Cindy Cashdollar, folk and rock. Aug. 21 at 8 p.m. $25 and $35. Kuf Knotz, hip-hop and reggae. Aug. 23 at 2 p.m. No cover. BeauSoleil avec Michael Doucet, Cajun and folk. Aug. 23 at 8 p.m. $20 and $25. Daryl's House, 130 Route 22. 845-289-0185; darylshouseclub.com.
PEEKSKILL 12 Grapes Music and Wine Bar Live Band Karaoke, hosted by J P Patrick. Aug. 20 at 8:30 p.m. No cover. The Bookends Band, classic rock. Aug. 21 at 9:30 p.m. $5 and $10. The Jon Bates Band, rock and soul. Aug. 22 at 9:30 p.m. $5 and $10. 12 Grapes Music and Wine Bar, 12 North Division Street. 914-737-6624; 12grapes.com.
PEEKSKILL Bean Runner Cafe The Black Dirt Band, blues. Aug. 21 at 8 p.m. $10. Joe Ferry and the Big Ska Band. Aug. 22 at 8 p.m. $10. Bean Runner Cafe, 201 South Division Street. 914-737-1701; beanrunnercafe.com.
PIERMONT The Turning Point James Maddock Trio, rock. Aug. 16 at 4 p.m. $30. The Turning Point, 468 Piermont Avenue. 845-359-1089; turningpointcafe.com.
POUGHKEEPSIE Mid-Hudson Civic Center The second annual Hudson Valley Freestyle Jam, with the Sugar Hill Gang, Cover Girls, Lisette Melendez and others. Aug. 22 at 8 p.m. $41 to $71. Mid-Hudson Civic Center, 14 Civic Center Plaza. 845-454-5800; midhudsonciviccenter.org.
POUGHKEEPSIE Victor C. Waryas Park Jazz in the Valley, with Ron Carter, Javon Jackson, Arturo O'Farrill Octet, Tia Fuller, Billy Drummond, Bill Charlap, Steve Kroon, Kenny Washington and Peter Washington. Aug. 16 at noon. $50 and $60. Victor C. Waryas Park, Main Street. 845-384-6350; jazzinthevalleyny.org.
TARRYTOWN Tarrytown Music Hall Jazz Is Dead, a Grateful Dead tribute band. Aug. 21 at 8 p.m. $35 to $55. Tarrytown Music Hall, 13 Main Street. tarrytownmusichall.org; 877-840-0457.
WOODSTOCK Bearsville Theater Kamasi Washington, jazz and funk. Aug. 19 at 8 p.m. $15. Chogyi Lama, Calvin Alfaro and Friends, rock. Aug. 20 at 7:30 p.m. $20. Anders Osborne, blues. Aug. 21 at 9 p.m. $25. Julianna Barwick and Wet, R&B. Aug. 22 at 9 p.m. $15. Bearsville Theater, 291 Tinker Street. 845-679-4406; bearsvilletheater.com.
WOODSTOCK Byrdcliffe Barn Kate Pierson, rock. Aug. 28 at 7:30 p.m. $75. $150 for concert and cocktail reception at 6:30 p.m. Byrdcliffe Barn, 485 Upper Byrdcliffe Road. woodstockguild.org; 845-679-2079.
WOODSTOCK Maverick Concert Hall Trio Solisti, classical. Aug. 16 at 4 p.m. $25 and $40. The Maverick Chamber Players. Aug. 22 at 6 p.m. $25 and $50. Ariel Quartet with Thomas Storm, baritone. Aug. 23 at 4 p.m. $25 and $40. Maverick Concert Hall, 120 Maverick Road. 845-679-8217; maverickconcerts.org.
YONKERS Empire City Casino, at Yonkers Raceway Lynyrd Skynyrd, Southern rock. Aug. 16 at 7 p.m. $55 to $250. Empire City Casino, at Yonkers Raceway, 810 Yonkers Avenue. 914-968-4200; empirecitycasino.com.
YONKERS Hudson River Museum Yonkers Community Music Showcase, song, dance, drama and spoken word. Aug. 21 at 8 p.m. Free. Hudson River Museum, 511 Warburton Avenue. 914-963-4550; hrm.org.
BRONX Bronx Museum of the Arts Boogie on the Boulevard, music, activities and local artists. The center lanes of the Grand Concourse from 161st to 167th Street will be closed to cars. Aug. 16, noon to 4 p.m. Free. Bronx Museum of the Arts, 1040 Grand Concourse. 718-681-6000; bronxmuseum.org.
BRONX Wave Hill Garden Highlights Walk, tour of seasonal highlights. Aug. 16 and 23 at 2 p.m. Free with admission to the grounds. $2 to $8; members and children under 6, free. Wave Hill, 675 West 252nd Street. 718-549-3200; wavehill.org.
CARMEL Putnam County Veterans Memorial Park The 15th annual Daniel Nimham Intertribal Pow Wow, Native American festival. Aug. 16, 10 a.m. to 5 p.m. Putnam County Veterans Memorial Park, 201 Gipsy Trail Road. 845-225-8154; nimham.com.
OSSINING Teatown Lake Reservation Field Day at Cliffdale, exploration and observation of insects. Aug. 29, 10:30 a.m. to noon. $7; members, free. Teatown Lake Reservation, 1600 Spring Valley Road. 914-762-2912; teatown.org.
YONKERS Untermyer Gardens Conservancy Tours of the Persian garden and estate restoration. Sundays at 11 a.m., Aug. 2 through 30. $10. Untermyer Gardens Conservancy, 945 North Broadway. 914-512-0436; untermyergardens.org.
Spoken Word
BRONX Bronx Museum of the Arts Lloyd Ultan and Shelley Olson discuss their book "The Bronx: The Ultimate Guide to New York City's Beautiful Borough." Aug. 22, noon to 1:30 p.m. Lunch and talk is $10; members, free. Bronx Museum of the Arts, 1040 Grand Concourse. 718-681-6000; bronxmuseum.org.
CROTON FALLS Schoolhouse Theater "Theater Finds an Audience on the Autism Spectrum," discussion. Aug. 18 at 7 p.m. Free. Schoolhouse Theater, 3 Owens Road. 914-277-8477; schoolhousetheater.org.
MOUNT VERNON St. Paul's Church "The Authors of St. Paul's," lecture. Aug. 19 at 1 p.m. Free. St. Paul's Church, 897 South Columbus Avenue. 914-667-4116; nps.gov/sapa.
NEW PALTZ Historic Huguenot Street, DuBois Fort "The Huguenots in France: What Came Before They Converted or Fled," lecture by Barbara D. Diefendorf. Aug. 22 at 5 p.m. $20 to $25. Historic Huguenot Street, DuBois Fort, 81 Huguenot Street. 845-255-1889; huguenotstreet.org.
VALHALLA Westchester Photographic Society, Westchester Community College "Post-Processing for Beginners," lecture by Arnold Breisblatt. Aug. 21 at 8 p.m. Free. Westchester Photographic Society, Westchester Community College, Technology Building, Room 108. 914-827-5353; wpsphoto.org.
ELMSFORD Westchester Broadway Theater "Backwards in High Heels," musical by Lynette Barkley and Christopher McGovern. Through Sept. 20. $54 to $84 for meal and show. Westchester Broadway Theater, 1 Broadway Plaza. broadwaytheatre.com; 914-592-2222.
GARRISON The Hudson Valley Shakespeare Festival, "The Winter's Tale" and "A Midsummer Night's Dream" by Shakespeare, with "An Iliad" by Lisa Peterson and Denis O'Hare, and "The Arabian Nights"by Mary Zimmerman, in repertory. (845) 265-9575; hvshakespeare.org. Through Aug. 30. $32 to $84. Wednesdays through Mondays, 9:30 a.m. to 5 p.m. Boscobel House and Gardens, 1601 Route 9D. 845-265-3638; boscobel.org.
GARRISON The Hudson Valley Shakespeare Festival, "Vanity Fair," by Kate Hamill. Aug. 20 at 7:30 p.m. $25. The Philipstown Depot Theater, 10 Garrison Landing. 845-424-3900; philipstowndepottheatre.org.
STONY POINT Penguin Rep Theater "Becoming Dr. Ruth," comedy by Mark St. Germain. Through Sept. 6. $40. Penguin Rep Theater, 7 Crickettown Road. 845-786-2873; penguinrep.org.
WOODSTOCK Woodstock Playhouse "Sweeney Todd," musical by Stephen Sondheim and Hugh Wheeler. Through Aug. 23. $32 to $40. Woodstock Playhouse, Routes 212 and 375. woodstockplayhouse.org; 845-679-6900.
YONKERS Hudson River Museum "The Tempest," Hudson Valley Shakespeare Festival's Conservatory Company. Aug. 22 at 8 p.m. Free. Hudson River Museum, 511 Warburton Avenue. 914-963-4550; hrm.org.
Museums and Galleries
BEACON Beacon "Beacon 3-D," outdoor public art exhibition. Through Oct. 15. Dawn to dusk. Beacon, various locations on Main Street. beacon3d.org; 917-318-2239.
BEACON Howland Public Library "Beacon Lens," Beacon Photographers group show. Through Sept. 6. Mondays, Wednesdays and Fridays, 9:30 a.m. to 5:30 p.m.; Tuesdays and Thursdays, 9:30 a.m. to 8 p.m.; Saturdays, 10 a.m. to 4 p.m.; Sundays, noon to 4 p.m. Howland Public Library, 313 Main Street. 845-831-1134; beaconlibrary.org.
BEACON Mount Gulian Historic Site Tours of the home, Dutch barn and restored garden. Sundays, Wednesdays, Thursdays and Fridays, 1 to 5 p.m. through Oct. 25. $4 to $8; members, free. Mount Gulian Historic Site, 145 Sterling Street. mountgulian.org; 845-831-8172.
BEACON RiverWinds Gallery "Selected Breed," oil paintings by Mary Smoot Souter. Through Sept. 6. Wednesdays through Mondays, noon to 6 p.m.; second Saturdays, noon to 9 p.m. RiverWinds Gallery, 172 Main Street. 845-838-2880; riverwindsgallery.com.
BETHEL Bethel Woods Center for the Arts "Written in Stone," sculptures by Harry Gordon. Through Oct. 12. "Threads: Connecting '60s and Modern Rockwear," from the collection of Andy Hilfiger. Through Dec. 31. $6 to $15; children under 3, free. Daily, 10 a.m. to 7 p.m. through Sept. 7. Bethel Woods Center for the Arts, 200 Hurd Road. bethelwoodscenter.org; 866-781-2922.
BRONX Bronx Museum of the Arts "Bronx Calling: The Third AIM Biennial," group show. Through Sept. 20. Thursdays through Sundays, 11 a.m. to 6 p.m.; Fridays, 11 a.m. to 8 p.m. Bronx Museum of the Arts, 1040 Grand Concourse. 718-681-6000; bronxmuseum.org.
BRONX The New York Botanical Garden "Frida Kahlo: Art, Garden, Life," paintings, works on paper and a garden. Through Nov. 1. $10 to $25; children under 2, free. Tuesdays through Sundays, 10 a.m. to 6 p.m. The New York Botanical Garden, Southern Boulevard. 718-817-8700; nybg.org.
COLD SPRING Buster Levi Gallery Works by Grace Knowlton, Barbara Smith Gioia and Maria Pia Marrella. Through Aug. 30. Fridays through Sundays, noon to 6 p.m.; and by appointment. Buster Levi Gallery, 121 Main Street. 845-809-5810; busterlevigallery.com.
COLD SPRING Gallery 66NY "Francesco Mastalia: Organic" and "Wilfredo Morel: Farm Art." Through Aug. 30. Thursdays through Sundays, noon to 6 p.m.; and by appointment. Gallery 66NY, 66 Main Street. gallery66ny.com; 845-809-5838.
GARRISON Garrison Art Center Riverside Crafts Fair, handmade items by 75 regional artists. Aug. 16, 10 a.m. to 5 p.m. $5; children under 13, free. Garrison Art Center, 23 Garrison's Landing. 845-424-3960; garrisonartcenter.org.
GARRISON Manitoga/The Russel Wright Design Center House, studio and landscape tours. Through Nov. 9. $10 to $20. Manitoga/The Russel Wright Design Center, 584 Route 9D. 845-424-3812; russelwrightcenter.org.
GREENBURGH Madeleine Gutman Gallery "Captivation Through Visual Inspiration," photographs by Monica Marrow. Through Sept. 15. Mondays through Fridays, 9 a.m. to 5 p.m. Madeleine Gutman Gallery, 177 Hillside Avenue. 914-682-1574; greenburghartsandculture.org;
HASTINGS-ON-HUDSON Upstream Gallery "Points of View From the Circle," Red Circle Photo Arts Collective. Through Aug. 30. Thursdays through Sundays, 12:30 to 5:30 p.m. Upstream Gallery, 8 Main Street. upstreamgallery.com; 914-674-8548.
HUDSON 510 Warren Street Gallery "Sea, Sky and Shore," paintings by Eleanor Lord. Through Aug. 30. Fridays and Saturdays, noon to 6 p.m.; Sundays, noon to 5 p.m. 510 Warren Street Gallery, 510 Warren Street. 510warrenstgallery.com; 518-822-0510.
HUDSON BCB Art "Big Collars," paintings by Barbara Friedman. Through Aug. 16. "The M. Van Dyke Group: Monoprints of the 1980s." Aug. 22 through Sept. 13. Thursdays through Sundays, noon to 6 p.m.; and by appointment. BCB Art, 116 Warren Street. 518-828-4539; bcbart.com.
HUDSON Carrie Haddad Gallery "Abstraction," Adam Cohen, Elizabeth Coyne, Donise English and Chris Duncan. Through Aug. 23. Mondays through Saturdays, 11 a.m. to 5 p.m.; Sundays, noon to 5 p.m. Carrie Haddad Gallery, 622 Warren Street. carriehaddadgallery.com; 518-828-1915.
HUDSON Jeff Bailey Gallery "Joshua Marsh: Drawings" and "Michael Berryhill: Shrine Time." Through Aug. 30. Fridays through Sundays, noon to 6 p.m.; and by appointment. Jeff Bailey Gallery, 127 Warren Street. baileygallery.com; 518-828-6680.
HUDSON John Davis Gallery Paintings by Janice Nowinski. Through Aug. 16. Works by William Ransom. Aug. 22 through Sept. 13. Thursdays through Mondays, 11 a.m. to 5 p.m.; and by appointment. John Davis Gallery, 362 1/2 Warren Street. johndavisgallery.com; 518-828-5907.
HYDE PARK The Franklin D. Roosevelt Presidential Library and Museum "The Spirit of the Gift: Gifts of State and Affection." Through Dec. 31. $6 and $9; children under 15, free. Daily, 9 a.m. to 6 p.m. through Oct. 31. The Franklin D. Roosevelt Presidential Library and Museum, 4079 Albany Post Road. 845-486-7745; fdrlibrary.marist.edu.
IRVINGTON Irvington Public Library "Pre-Columbian Mexican Pottery," Sumi ink drawings by Jim Maciel. Through Aug. 29. Mondays, Wednesdays, Fridays and Saturdays, 10 a.m. to 5 p.m.; Tuesdays and Thursdays, 10 a.m. to 9 p.m. Irvington Public Library, 12 South Astor Street. irvingtonlibrary.org; 914-591-7840.
KATONAH Katonah Museum of Art "Inside the Outside: Five Self-Taught Artists From the William Louis-Dreyfus Foundation." Through Oct. 11. $5 and $10; members and children under 12, free. Tuesdays through Saturdays, 10 a.m. to 5 p.m.; Sundays, noon to 5 p.m. Katonah Museum of Art, 134 Jay Street. 914-232-9555; katonahmuseum.org.
LARCHMONT Kenise Barnes Fine Art "Cool and Collected '15," group show. Through Sept. 10. Tuesdays through Saturdays, 10 a.m. to 5:30 p.m.; and by appointment. Kenise Barnes Fine Art, 1947 Palmer Avenue. 914-834-8077; kbfa.com.
LIVINGSTON MANOR Catskill Art Society "Audubon and Friends Too 2015," group show. Through Aug. 23. Mondays and Thursdays through Saturdays, 11 a.m. to 6 p.m.; Sundays, 11 a.m. to 3 p.m. Catskill Art Society, 48 Main Street. catskillartsociety.org; 845-436-4227.
MAHOPAC A. Eric Arctander Gallery, Putnam Arts Council "Works on Paper," group show. Through Aug. 30. Mondays through Thursdays, 10 a.m. to 4 p.m.; Saturdays and Sundays, 1 to 4 p.m. A. Eric Arctander Gallery, Putnam Arts Council, 521 Kennicut Hill Road. 845-803-8594; putnamartscouncil.com.
MAMARONECK Bullseye New York Gallery "Dark Ecologies," group show. Aug. 25 through Nov. 14. Tuesdays through Fridays, 10 a.m. to 6 p.m.; Saturdays, 10 a.m. to 5 p.m. Bullseye New York Gallery, 115 Hoyt Avenue. bullseyeglass.com; 914-835-3794.
MOUNTAINVILLE Storm King Art Center "Lynda Benglis: Water Sources," large-scale sculptures. Through Nov. 8. "Outlooks: Luke Stettner," site-specific outdoor installation. Through Nov. 29. $8 to $12; members and children under 5, free. Wednesdays through Sundays, 10 a.m. to 5:30 p.m. Storm King Art Center, Old Pleasant Hill Road. 845-534-3115; stormkingartcenter.org.
NEW PALTZ Samuel Dorsky Museum of Art, at the State University of New York at New Paltz "The Stories We Tell: Hudson Valley Artists 2015," group show. Through Nov. 8. Suggested donation, $5. Wednesdays through Sundays, 11 a.m. to 5 p.m. Samuel Dorsky Museum of Art, at the State University of New York at New Paltz, 1 Hawk Drive. newpaltz.edu/museum; 845-257-3844.
NYACK Edward Hopper House Art Center "John F. Simon, Jr.: The Ever Present Sun." Through Aug. 16. "Behind Doors and Through Windows: Reflections on Contemporary Domestic Life," group show. Aug. 22 through Oct. 18. $2 to $7; members and children under 16, free. Wednesdays through Sundays, noon to 5 p.m.; and by appointment. Edward Hopper House Art Center, 82 North Broadway. edwardhopperhouse.org; 845-358-0774.
OLANA Olana State Historic Site "River Crossings: Contemporary Art Comes Home," group show. Also on display at the Thomas Cole National Historic Site in Catskill. Through Nov. 1. $10 and $12; children under 12, free. Children under 16 admitted free at Thomas Cole. Olana State Historic Site, 5720 State Route 9G. rivercrossings.org.
OSSINING Ossining Public Library "The Geology of Light: A Photographic Life in Light & Stone," photographs by Bob Pliskin. Through Aug. 28. Monday and Thursday, 9 a.m. to 9 p.m.; Tuesdays and Fridays, 10 a.m. to 6 p.m.; Wednesdays, 1 to 9 p.m.; Saturdays, 9 a.m. to 5 p.m. Ossining Public Library, 53 Croton Avenue. 914-941-2416; ossininglibrary.org.
PEEKSKILL Bean Runner Cafe "Recent Work," George Dubinsky. Through Aug. 16. Sundays through Wednesdays, 8 a.m. to 6 p.m.; Thursdays, 8 a.m. to 8 p.m.; Fridays and Saturdays, 8 a.m. to 11 p.m. Bean Runner Cafe, 201 South Division Street. beanrunnercafe.com; 914-737-1701.
PEEKSKILL Hudson Valley Center for Contemporary Art "Love: The First of the Seven Virtues," group show. Through Dec. 6. $2 to $5; members and children under 8, free. Fridays, 11 a.m. to 5 p.m.; Saturdays and Sundays, noon to 6 p.m.; and by appointment. Hudson Valley Center for Contemporary Art, 1701 Main Street. 914-788-0100; hvcca.org.
PELHAM Pelham Art Center "Knit Knit Bomb Bomb," outdoor yarn installation. Through Aug. 31. Tuesdays through Fridays, 10 a.m. to 5 p.m.; Saturdays, 10 a.m. to 4 p.m. Pelham Art Center, 155 Fifth Avenue. pelhamartcenter.org; 914-738-2525.
PORT CHESTER Clay Art Center "My Name Is Nobody," Matthew Smith. Through Aug. 24. "There's No Such Thing as the Future," Kelley Donahue. Through Sept. 10. "Tristate of Mind," group show. Through Sept. 16. Mondays through Saturdays, 10 a.m. to 4 p.m.; and by appointment. Clay Art Center, 40 Beech Street. 914-937-2047; clayartcenter.org.
PORT EWEN Duck Pond Gallery, at Town of Esopus Library Watercolors by Nathan Milgrim. Through Aug. 29. Mondays, Tuesdays and Thursdays, 10 a.m. to 5:30 p.m.; Wednesdays, 10 a.m. to 8 p.m.; Fridays, 10 a.m. to 7 p.m.; Saturdays, 10 a.m. to 4 p.m. Duck Pond Gallery, at Town of Esopus Library, 128 Canal Street. esopuslibrary.org; 845-338-5580.
POUGHKEEPSIE The Franklin Delano Roosevelt Mid-Hudson Bridge "Bridge Music," sound installation by Joseph Bertolozzi. Through Oct. 31. Dawn to dusk. The Franklin Delano Roosevelt Mid-Hudson Bridge. josephbertolozzi.com.
POUGHKEEPSIE Vassar College, Frances Lehman Loeb Art Center "Punctuating Space: The Prints and Multiples of Richard Artschwager." Through Sept. 6. Tuesdays, Wednesdays, Fridays and Saturdays, 10 a.m. to 5 p.m.; Thursdays, 10 a.m. to 9 p.m.; Sundays, 1 to 5 p.m. Vassar College, Frances Lehman Loeb Art Center, 124 Raymond Avenue. 845-437-5632; fllac.vassar.edu.
POUND RIDGE Pound Ridge Public Library The 40th annual Ground Glass Photographic Group Show. Through Aug. 22. Tuesdays and Thursdays, 10 a.m. to 8 p.m.; Wednesdays, Fridays and Saturdays; 10 a.m. to 5 p.m.; Sundays, noon to 5 p.m. Pound Ridge Public Library, 271 Westchester Avenue. 914-764-5085; poundridgelibrary.org.
PURCHASE Neuberger Museum of Art of Purchase College "Plastic: Art in an Era of Material Innovation." Through Aug. 23. "Gold," group show. "Teresa Margolles: We Have a Common Thread." Through Oct. 11. $3 and $5; members and children 12 and under, free. Tuesdays through Sundays, noon to 5 p.m. Neuberger Museum of Art of Purchase College, 735 Anderson Hill Road. www.neuberger.org; 914-251-6100.
RHINECLIFF Morton Memorial Library "The Art Show," Benjamin Clinton Partrick. Through Aug. 29. Tuesdays, 2 to 6 p.m.; Wednesdays, 2 to 7 p.m.; Thursdays and Fridays, 9 a.m. to 12:30 p.m. and 2 to 6 p.m.; Saturdays, 9:30 a.m. to 12:30 p.m. Morton Memorial Library, 82 Kelly Street. morton.rhinecliff.lib.ny.us; 845-876-2903.
RIVERDALE Gallery 18, Riverdale Y "A Path to Plenty," acrylic paintings by Joyce Dutka. Through Aug. 28. Sundays through Thursday, 9 a.m. to 9 p.m.; Fridays, 9 a.m. to 4 p.m. Gallery 18, Riverdale Y, 5625 Arlington Avenue. riverdaley.org; 718-548-8200.
SAUGERTIES Cross Contemporary Art "Domestic Bliss," photographs by Susan Copich. Through Aug. 23. Thursdays through Mondays, noon to 6 p.m.; and by appointment. Cross Contemporary Art, 81 Partition Street. 845-399-9751; crosscontemporaryart.com.
TIVOLI Tivoli Artists Gallery "Tivoli: People and Place," group show. Through Aug. 23. Fridays, 5 to 9 p.m.; Saturdays, 1 to 9 p.m.; Sundays, 1 to 5 p.m. Tivoli Artists Gallery, 60 Broadway. 845-757-2667; tivoliartistsco-op.com.
WASSAIC The Wassaic Project, Maxon Mills "Deep End," group show. Through Sept. 20. Fridays, 5 to 7 p.m.; Saturdays, noon to 7 p.m.; and Sundays, noon to 5 p.m. The Wassaic Project, Maxon Mills, 37 Furnace Bank Road. wassaicproject.org; 347-815-0783.
WHITE PLAINS ArtsWestchester "Revisiting Arcadia: Contemporary Landscapes," group show. Through Aug. 29. Tuesdays through Fridays, noon to 5 p.m.; Saturdays, noon to 6 p.m. ArtsWestchester, 31 Mamaroneck Avenue. 914-428-4220; artswestchester.org.
YONKERS Blue Door Gallery "The Art of Healing," group show. Through Sept. 5. Thursdays and Fridays, 3 to 8 p.m.; Saturdays, noon to 5 p.m. Blue Door Gallery, 13 Riverdale Avenue. 914-375-5100; bluedoorartcenter.org.
YONKERS Hudson River Museum "Envy: One Sin, Seven Stories," installation by Adrien Broom. "Carla Gannis: The Garden of Emoji Delights." Through Sept. 26. $3 to $6; members, free. Wednesdays through Sundays, noon to 5 p.m. Hudson River Museum, 511 Warburton Avenue. 914-963-4550; hrm.org.


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So, I'm debating the ethics right now of offering this "tape" (1) for download.
What seems most intuitive to me is to contact each band in some fashion and ask for permission. Anything else seems presumptuous in a way that I am just uncomfortable with, personally. That's not to say others are wrong in saying, "please contact me if you want anything taken down" and offering downloads till that point. I know I have most certainly discovered music that was downloaded off of an internet site where I didn't do the due diligence to make sure that the artist had given permission, but then went out after being blown away and bought some vinyl direct from the artist. But personally, I feel the most comfortable on my own site with insuring consent every step of the way.
tl;dr? I think I'm just going to offer a track listing and hopefully inspire you to go out and track down these gems yourself. Of course, if I'm feeling particularly ambitious & find myself with some of the most scarce resource known to humanity: free time, then I may actually send out some e-mails and see what I can do. However, this setlist just feels so damn perfect for this winter, that I submit it to you here, this strange vision of a winter's day in hopes you will grab these songs yourself.
Of course, I find it impossible *not* to offer my pretentious-but-heartfelt quasi-poetic annotations. The story that this imagined day is; the story in between the songs, because of the songs, yet having nothing really to do with the songs..
Due to A) my utter ridiculousness & B) my Joycean infatuation, I submit this to you with suggested drink pairings (I warned you it'd be pretentious!), and time of day where each song falls in my imagination (plus some occasional anachrony where one is reflecting back on a time past or in the future.)
Here's the drink pairing list and letter code:
C = Coava Coffee Roaster's Girasoles from Costa Rica (a pleasantly intricate yet easy-to-drink & buttery cup to wake up with!)
L = Homemade Lemonade with Meyer lemons (all sweet, no sour, sorta like the pop songs they go along with) and Pomegranate juice to add a bit of winter into that traditional summer potable.
T = Foxfire Earl Grey with honey and some milk instead of cream, as the Irish take it (dreamy and comforting, yet agitates ever-so-slightly, like a lovers' arms waking you up as they squeeze you tighter in their sleep; waking you up but making you feel safe all over at the same time.)
B = Blanton's single-barrel bourbon (vanilla sweetness giving way to late rye spice; bite with sophistication. Rowdy and civilized all at the same time.)
E = Stumptown's Guatemalan Finca El Injerto Bourbon espresso, balancing chocolate with citrusy brightness and a hint of sweet spice as a finish. (A great middle of the day pick-me-up: sorta like these corresponding songs)
-Drink Pairing
#Track Number
"Song Name"
^Song Duration
+Time of Day
♥Band Name
-C #01: "Springtime" ^2:53 +7:06 AM ♥The Torn ACLs
Waking up with hope in your heart, knowing that if you keep doing the right thing, that good things are gonna happen. Ready for work, but shaking off the drudgery; looking for the little bit of daylight to catch on your fingertip before the inevitable rains begin anew, and loving them both.
-C #02: "My Heart Is A Drummer" ^3:17 +9:23AM ♥Allo Darlin'
As the day continues to move from a slow but bright-eyed start into that full and real feeling of being awake, thoughts are turning to love & that sense of being both giddy with excitement, yet strong and secure, absent-mindedly toying with a phone cord, giving yourself over to love, but never forgetting or losing yourself…I never liked Graceland, but I *love* the metaphoric Graceland & actually want to go out and get the album just for what it represents. There's a Graceland inside us all, in that place where the things that you love make you happiest: in that place where your heart is a drummer…(and we're talking Keith Moon, here)
-C #03: "Please Do Not Lie" ^4:30 +10:03 AM ♥Boa Constrictor vs. Honeydrips
The day has really begun in earnest. You're aware of where you are raw and where you are polished. You are unafraid and ready to go. Handclaps could be cheering choruses for the lilt in your heart. Things aren't perfect, but they are good, and this makes them real and sustainable; it makes them great.
-L #04: "Summertime" ^3:13 +11:24 ♥Pocketbooks
It's the dead of winter, but this song is the memory of a summer breeze in the midst of a sweltering August, without romanticization. Enjoying the warmth of your cardigan, remembering laughs and the sweet smoke from corn stalks on the BBQ as you all stood on the grass somewhere unforgettable.
-L #05: "Your Way, Mr. Leary" ^1:55 +12:17 PM ♥The Rainyard
Lunch is lively conversation with old friends and some new ones, too. There is talk of the night's forthcoming adventures. A catchy hook falls into your ear, whispered like an infectious secret, but then is gone just as quickly.
-C #06: "Philadelphia" ^3:36 +12:31 PM ♥Standard Fare
Some seemingly innocuous glimpse of something unrelated brings your mind back to your beloved and the temporary distance between you. You'll see that person after work, today…why is work taking *forever*? It bloody well feels like a year!
-L #07: "Fine Day For Sailing" ^2:26 +12:49 PM ♥Go Sailor
That's it, you've decided to cast off the hardships of the day with a breezy smile. Laugh it off with a dance somewhere inside of yourself. Forgive everything ever done to you with a laugh, but never forget, all the while staying too busy dancing to worry about a damn thing else. Or maybe you're not there yet, but that's where you want to be.
-L #08: "This Love Is Not Wrong" ^3:22 +1:05 PM ♥The Field Mice
Let the crowd say whatever they want about you. This feeling you have in your heart, this furnace you carry with you, keeping you warm and causing you to look up with a smile when everyone else is looking down with a bothered sneer, it is a love that is shared, a silent bond that will never ask for nor need the approval of anyone else.
-B #09: "Original Arrogance" ^1:37 +9:40 PM ♥Comet Gain
Finding yourself someplace where you were left, realizing that somehow you've gotten over everything that used to hold you back, and burning all those bad memories with the fire of urgency. Onward brazenly to this next new phase with all of the self-assurance you've ever needed to do anything you want to do. You're ready to create, and in big strokes with fast movement. Thinking forward into the night, and that moment where the hope for it meets with what it really is. Some threshold has been crossed, but you're not yet sure what.
-B #10: "I'm So Unclean" ^2:18 +3:15 PM ♥Evans The Death
Urgency gives way to reverie. You pine for your beloved as you enter your last big push to finish work for the day, having endured and conquered being beset by the (after)noontime demon: Ennui.
-T #11: "Green Eyed" ^2:54 +8:02 PM ♥September Girls
Thinking on the night beginning in earnest and thoughts that echo in your head: moments of cherished time alone, thoughts of being lost in a crowd, thoughts of being alone with your beloved. The future and where that turns from ideal form into what it really is. Pleasant surprises and otherwise. Again, returning to the echo of your thoughts. The night is young, and still, so are you…there is hope in the silent spaces between the noise.
-B #12: "Rip" ^2:27 +7:30 PM ♥Ringo Deathstarr
Somehow, and you're not sure how or why, things have taken a turn for the strange. You retreat into comforting thoughts. You're not sure if you are lucid dreaming or if reality has taken on a strange shimmer. You are not worried, but things are not as they were. You're not sure if they ever will be again. Your simple and quietly beautiful life of work and home is transformed into something you don't recognize…you stride forward into it anyway, casting not a backwards glance.
-B #13: "Do You Think It'll Snow Tonight?" ^1:28 +6:39 PM ♥The Cat's Miaow
A quiet wish, noisily uttered? A noise-y wish, quietly uttered? From the flurry inside to the possible one out…maybe tonight is best spent by the fireplace.
-E #14: "You Make Two Weeks Two Days" ^2:47 +5:30 PM ♥Baffin Island
Thinking forward to the commute home. Warm thoughts of returning to your beloved. How can time escape so quickly when life is so glorious now? How did it trudge along like an ancient horse on a tenderfoot's trail-ride when times were sour? The comfort of the company is enough to sustain the irrevocable velocity of wonderful moments rendered perfect by their second glance. Toiling in the diamond mine of memory never works up a sweat.
-E #15: "Your New Boyfriend" ^1:49 +4:43 PM ♥Rocketship
Back to where you are. The darkest times are rendered manageable by the sonic-honey rimming the medicine cup of hardship's medicine. Yesterday's scars give today's hugs meaning and perspective. You love like you used to. You love like new.
-E #16: "Folded In Half" ^3:36 +5:15 PM ♥The Bank Holidays
Harmony steps into your life as you put your day away and prepare for what comes next. Your thoughts escape up the spiral staircase until you are on top of the bus, watching the world from a newly-caught perspective. On your way home, only to leave it again, only to return. Love is the anchor that still lets the boat keep moving, but never too far from what matters.
-L #17: "Infatuation Street" ^3:17 +2:45 PM ♥Sweater Girls
Loveletters written on paper folded and burned as a floating wish into the sky, hoping only to be forgotten as they turn to ash. The weather on your face as the leaves have turned, the comforting scratch of wool against your skin.
-L #18: "Sea Horses" ^2:55 +5:01 PM ♥Blueboy
Life is more beautiful and more complicated than anyone had imagined.
-B #19: "Your Doubting Heart" ^3:15 +10:15 PM ♥The Hobbes Fanclub
You want to cry. Your beloved is lost. You know not where or how. Your memories are all dreams; your dreams are all memories. Jonas had an easier go of it than this sudden nightmare you have woken into. Everything has changed. Only Lethe notes register in your unbelieving ears. The music pulls you forward when you find your will lacking.
-B #20: "You Can Hide Your Love Forever" ^3:43 +10:15 AM ♥Comet Gain
Back to the morning; those burned wishes are not forgotten; they mature into these feelings that are as mundane yet comforting as your morning Orange Juice. Your desires will carry you when nothing else will. Dreams of a France or Scotland you've never seen, the friends in literature you've made, those words that have never been said that you always wanted…the home in that person's eyes. You can find the song when you can't find the words.
-B #21: "English Cities" ^2:04 +10:44 PM ♥Brilliant Colors
Pushing over smokestacks, past thoughtlessly brilliant interactions to the place where everyone is gathered. Stumbling through, you would rue your current state, wide awake fearing all is lost, were it not for the chaos of noise you have somehow scrolled above, to where you can see the pattern from a distance. You know not where your beloved is, but you enter the room where all your new friends are. Nightmares are raindrops that fall off of your slicker into puddles in the street you do not stop to consider.
-B #22: "You're Beautiful" ^1:06 +11:09 PM ♥The Faintest Ideas
Running up the flights of stairs into the room where they all are; you throw open the door, boldly declaring what you know, the beauty that's pushing its way out of your chest, effortlessly, as if from someone else, but it is *you*, it is the essence of you, but you don't wait for a reaction, you've turned around and are bounding out the door, down the steps before anyone's had a word. You don't care what they think; that's for them to decide. You wanted only for them to know.
-B #23: "For Ex-Lovers Only" ^2:35 +12:58 AM ♥Black Tambourine
Wandering rocks. The reaction that could have been, instead of the Scylla & Charybdis you have chosen . The fear at the end of the night. The wrong "what if" suddenly become real for a moment. A nightmare Baudelaire prose poem. A Berlioz vision of paregoric delusion. Your beloved warps before you, turning into a manifestation of all of one's own insecurities, pointing a silent accusatory finger, questioning your own capacity to be loved. You slam the door on the fears, letting everyone else be who they are. Let anyone walk away without a second thought, even if it does cost a tear.
-T #24: "Poor Students Dream of Marx" ^3:12 +7:01 PM ♥Cats on Fire
The world shudders awake with the slam of that door. You are in your own skin, seeing the world as it really is. The myths flee from your mind like smoke from a chimney. Seemingly impoverished by materialism, you've actually made the grade. You are alone, having put all of those insecurities to bed. Others show up, ask you questions. You glibly dismiss them. You will not be distracted again.
-T #25: "You Can Have What You Want" ^2:52 +1:10 AM ♥Papercuts
From tears, alone, you've found something in yourself again. You've discovered everything you need, and have chased the real and fake images that have hounded you. You are alone. You wouldn't mind some company, but you are fine and happy by yourself. There is nothing bittersweet about this peace, but there is contemplation.
-T #26: "Rats Blossom Into Boys" ^3:52 +1:33 AM ♥Kookooo Kitchen
You walk into the last room of the night. Things are happening for others. Talk. Drinks. Other things you don't want to look too closely at. You have no desire now for any of the former pastimes that once beguiled you. You've made it somehow to the other end of the night. You're not yet sure what you have learned but you are certain of what to do. Leave
-T #27: "The City Limit" ^5:53 +2:31 AM ♥The Radio Dept.
Streetlights cast their long rays, glimmering in the brisk dark night, pulsating rhythmically as you move past them, your bed calling to you. Feeling that familiar sense of being alone, you find yourself longing again, wistful, but not needing. Mildly happy with who you are and where you are, but wanting more. The shadows in the car give way to the streetlight drumbeat, like a strobe light in slow motion, to reveal your beloved nearly asleep in the passenger seat. How? Why has this all happened? Your loved one tells you that you have grown as you've needed to. You've dream-lived your fears and fantasies, and have come to the other side. Safe. While you were preoccupied, they had done the same. Paths have separated, crossed, split and then reunited in one giant thoroughfare, leading home; leading to a place that stands still as it runs and anchors only when it changes. Your hand is warm in your beloved's hand. A sigh, on the verge of sleep. Where it's at is where you are now; where you will always be as long as you desire it.
1:In a typically Fecked-up state of affairs, I call my mixes "tapes", which I burn onto CDs made to look like vinyl records, which are usually burned to mp3s and played on ipods/iphones. I think the only thing that would make the circle complete is if the iphone had an 8-track case


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Greater use of technology in infrastructure processes is a stand-out change of the past decade, but is it being embraced enough? Jon Masters catches the latest industry views at the Topcon-sponsored round table on digital advances
The premise is that use of digital technology can drive greater efficiency; that entire supply chains can work more collaboratively if they share common design models. The BIM agenda presents an opportunity to digitalise the whole lifecycle of assets. This can include initial site surveying through design and construction processes via BIM models and setting- out and machine control on site, then with as-built data, the operation and management of infrastructure.
So how far has the industry got in realising this potential? That was the subject for discussion when industry leaders gathered for a special round table as 2015 drew to a close.
"There are significant pockets of understanding, depending on the client and the supply chain supporting them on particular projects," says Topcon's business manager David Bennett.
"We've been talking about this whole agenda for a number of years now, about how the industry can use data and geographic information to manage assets. We have clients here around the table that are using this technology, and we have worked with a number of contractors on projects on the strategic road network, for example: they are using data management in taking designs out on site. It's not restricted to the roads sector, but there is not yet a lot of use to its full potential."
Is the problem its reliability and usability on the ground? Mark Lawton, chief engineering surveyor for Skanska: "Three-dimensional machine control (3DMC) has been around for 18 years or more, but has evolved from older robotic instruments to modern GNSS and optical systems that perform much better. What's really changed is the way the data is delivered out to site," he says. "It's now frequent, cheap and small. It's become integral to our business."
Many tier-one contractors haven't yet realised all the benefits of digital machine control, says Darren James,
"The software has now truly caught up with the hardware as a management tool. It's got to be holistic, though. Everyone in the supply chain has to embrace it for it to be really effective"
Nick Richards, managing director, Walters Group
managing director for infrastructure at Costain. "It's about controlling and positioning more efficiently and safely, working quicker and avoiding having anyone near the machine interface. Clients encourage use of this technology, but routinely what we've failed to do is to convince them in best-value language why to use it."
Ground improvement, remediation and earthworks specialist McAuliffe Group works frequently with the top 10 housebuilders. According to managing director John McAuliffe, machine control technology has become a business imperative: "It's about making best-value decisions. We're involved at land-transaction phases with major housebuilders, where it's about assessing the risks in the ground, then translating that into delivery," McAuliffe says.
"Housebuilders are generally working with very old technology – with bricks and mortar. It's what's in the ground and how we extract that value that's the differentiator."
Efficiencies that can be gained from the latest in machine control are "fantastic", says Nick Richards, managing director of plant, demolition and civil engineering contractor Walters Group. "The software has now truly caught up with the hardware as a management tool," he says. "It's got to be a holistic approach, though. Everyone in the supply chain has to embrace it for it to be really effective."
There is a role for clients to take in helping embed technology throughout the supply chain. "Yes, we have an obligation to ensure it's embraced, but it's got to be fit for purpose, it's got to be practical, and we need to ensure our workforce are ready for it," says Transport for London's head of asset strategy and investment, Andy Jinks.
"There's a behavioural aspect. The whole technology debate can be a double-edged sword: on the one hand, it helps us drive efficiency and increase capacity; on the other, it can drive a level of uncertainty and even fear. We have an obligation to overcome this to ensure technology is introduced in the right manner. An area of concentration for us on the Underground at the moment is exploring predict-and-prevent technology to improve our future maintenance practice."
Clients generally have no problem adopting new technology, provided the benefits are clearly articulated, says Highways England senior manager Phil Ellis. "Once it's understood, if we see the benefit, we will put it on the ground. It's not necessarily just clients or the supply chain that should be driving greater use of technology. We need to do so as a collective," he says.
This is all very well, but who should be paying for it? "I think the answer is: it should pay for itself," says Ellis.
Costain's James: "Ultimately it should become part of the cost base of what we're trying to do, but it should be at a lower cost than what we've traditionally done. Highways England is now paying less for some items of construction work because we've invested in this technology. Of course, if we cannot see a significant cost reduction down the line then we will not make such an investment, but we're now way beyond any leap of faith with this technology."
Another aspect to greater use of technology, digitalisation of processes and data management is its demand for different skills and cultural change within organisations. Consultant CH2M has been studying these effects. "We've worked with many clients and found there is a cultural change happening; most are embracing new technology," says CH2M information management consultant Ilias Krystallis.
"Taking a step back, what clients are really focusing on is implementation of standards for use of BIM. We are doing an exercise now measuring the benefits for clients around the development of standards: so, forming a data dictionary and drawing up a picture of what areas of their business directly benefit from particular parts or layers of the technological changes occurring."
What about the cultural change down the supply chain? Is it working to good effect between interfaces, between suppliers and different processes? Yes, says Skanska's Lawton: "I sit in the middle as a main contractor. I supply data to earthworks teams, so they can have what they need. I interface with the design team to get what I need, and then I will always conduct a BIM fire drill to validate the data and whether we can use it. That is happening all the time, day in, day out, as normal activity.
"Anything we need from the flow of data, we've subtly changed so that the workflow goes from concept to construction and delivery without issues. The part about the data coming back, however, for asset management and the like – that sits in another realm at the moment."
The different cultures of the BIM design world and the site realm do not often meet, says Topcon's Bennett. "But they did at our demonstrations, where we had designers at the controls of high-tech machines" – in a safe and controlled manner, he adds. "This helped show them why surveyors and site engineers are asking for data to be supplied in particular formats, so they could make the connection between the design model and the output required.
"Collaboration may be becoming an overused word in the BIM space, but it genuinely is vital that we pull
"The part about the data coming back, however, for asset management and the like – that sits in another realm at the moment"
Mark Lawton, chief engineering surveyor at Skanska
in all stakeholders to achieve the full potential. We're not going to bring it about alone, knocking on the door as technology providers."
ACE can provide a good overview on divergence in levels of take-up. Policy and external affairs director Julian Francis says: "We see a regional divide, and one related to the scale of engineering organisations. Larger ACE member companies, particularly in the South-east, are right on top of it, pushing BIM forward as much as they can. The further north we go, the more of an issue it becomes – especially for SMEs – around who's going to pay for it, why and how should they push on with it. We're conducting a series of roadshows with our legal teams, speaking to regional firms, clients and contractors about BIM's importance."
Greater amounts of technology and data can be seen everywhere and throughout infrastructure. How can this industry really exploit the data opportunity for managing assets?
Topcon machine control business development manager Andy McCann: "We have systems where the exact position of the final pass of the cutting edge of a machine can be recorded, loaded to a web portal, and fed back to the engineering surveying team for reporting as-built information for operational and asset management purposes. This is the level of data clients are now asking for."
A good example of likely future use of "big data" in infrastructure could manifest in management of the Underground. TfL's Jinks says: "Everything's possible. How can we use our CCTV analytics to understand the wear and tear on stations, or wifi and bluetooth pings to monitor and respond to passenger loadings? How do we use intelligent train weight data to inform passengers where is best to board? We just have to join it all up."
At the Topcon digital technology round table were:
Julian Francis, policy and external affairs director of ACE
Ilias Krystallis, information management consultant at CH2M
Darren James, infrastructure managing director at Costain
Phil Ellis, senior manager of Highways England
Andy Jinks, head of asset strategy/ investment at Transport for London
John McAuliffe, managing director of McAuliffe Group
Mark Lawton, chief engineering surveyor at Skanska
Nick Richards, managing director of Walters Group
Andy McCann, machine control business development manager at Topcon
David Bennett, business manager at Topcon
Paul Keenan, machine control business development manager at Topcon