Статті в журналах з теми "Looping construct"

At its most fundamental, entrepreneurship involves discovering, evaluating, and exploiting opportunities to create future goods and services (Shane and Venkataraman, 2000). Acknowledging that characteristics, emotions, cognitive biases, and past experiences influence entrepreneurial activity and decision making (Shepherd and Patzelt, 2017; De Winnaar and Scholtz, 2019), many propose that active reflection is an important facet of the entrepreneurial curricula (Nabi et al., 2016; Santos et al., 2016). For the entrepreneurial student, learning to make rationale decisions is paramount, and this is vitally linked to their ability to reflect and be self-aware. This study recounts the use of the eportfolio within a new large class (over 600 students) module in enterprise education. The module utilised an eportfolio reflective assignment to allow students express their feelings and knowledge about a series of attended entrepreneurial events and guest speaker seminars. In this paper, we present a novel insight into the efficacy of this curricular approach. External stakeholders who acted as mentors and speakers were asked to review a number of these student portfolios, and provide their thoughts on the assignments themselves, and the e-portfolio construct more generally. As such, this study highlights the multiple feedback loops of reflection which can be obtained from the eportfolio when used as a carefully considered pedagogical tool. It also highlights the integral role that industry stakeholders play in the enterprise curriculum.

Launched in 2013, Vine is a popular microblogging service that allows users to record, edit, and share six-second videos that loop ad libitum, until another video is selected. At this time, the communicative, expressive, and semiotic affordances of Vine and similar services have still to be fully explored by users and scholars alike. Through a multimodal analysis approach drawing on New London Group’s (1996) work, this paper investigates how people construct humour on Vine by artfully arranging different modes of expression. The analysis focused on user-enacted humour, as opposed to captured comical scenes or bare samples taken from TV shows or movies. The study hypothesises the social construction of a novel humorous language that draws on extant forms of humour and a variety of modes and techniques derived from audio-visual media and computer-mediated communication, as users inventively exploit the framework provided by the Vine platform. Findings show that users create instant characters to amplify the impact of their solo video recordings, use Vine as a “humorous confessional”, explore the potential of hand-held media by relying on “one hand and face” expressivity (the other hand holding the device for the video “selfie”), and use technology, internet slang, internet acronyms, emoticons/emojis, and hashtags to convey humour and complement the messages of the videos they post on Vine. The goal of this study is an exploratory analysis of humour and its discursive functions in an emergent social medium by considering its affordances, as users find new and creative ways to harness its expressive potential.

Abstract This paper examines aspects of strategic interaction and the construction of the social actor in a neo-Austinian framework of illocutionary acts. The basic premise of the neo-Austinian framework is conventionality, according to which illocutionary acts depend on social agreement. An important part of the framework is the felicity condition of entitlement, directly related to the hearer’s understanding of the conventions that should hold for an act performance. Two strategies of challenging and/or rejecting illocutionary acts are then identified tentatively dubbed looping and backfiring, related to the hearer’s perception of when the entitlement felicity condition is flouted. Both strategies can be overtly or covertly confrontational and demonstrate that in their social quality illocutionary acts serve to construct the social actor and build up interpersonal relations.

Abstract: This paper analyzes a clock using Arduino without Real time clock (RTC). The development of the digital clock in Arduino is to provide its own time without RTC. Generally, electronic circuit designers use RTC to construct a clock. Such a circuit requires an extra circuit and power. The CMOS battery supplies power to the RTC, once the CMOS battery power is discharged. It automatically erases the date and time and requires an update from an external device. Considering these facts, RTC is avoided and code with nested looping is used to maintain timing in Arduino. It enables us to modify the time using a keypad and LCD without an external device. Continuous power is supplied to the Arduino using a battery backup. Such a circuit is simple with reduced code. All features available in RTC are available in this circuit. Keywords: Arduino Atmega 328P, RTC, LCD16 x 2, Keypad 4x3.

Abstract Abstract 1469 Poster Board I-492 The LIM domain binding protein LDB1 is an essential cofactor of LIM-homeodomain (LIM-HD) and LIM-only (LMO) proteins in hematopoiesis and other developmental programs. We have shown that LDB1 and its LIM-HD and LMO interaction partners are protected from ubiquitylation by a small family of SSBPs. Recently, we demonstrated that these SSBPs bind to LDB1 homodimers to promote formation of a ternary complex containing two molecules of LDB1 and SSBP. This was dependent on an intact LDB1 dimerization domain (DD) and produced a shift, directly or indirectly, in the equilibrium between LDB1 monomer and dimer. In the present study, we introduced a 24-glycine linker between two full-length LDB1 peptide-coding sequences and expressed this forced or tethered LDB1 dimer (TD-LDB1) in vitro and in vivo. First, both TD-LDB1, introduced into cells by transfection, and endogenous LDB1 were found to have the same turnover rate, indicating that protection from ubiquitylation was independent of dimerization status. Second, TD-LDB1 was fused to the DNA binding domain of GAL4 (GAL4DBD) and in a second construct to the activation domain of herpesvirus VP16 (VP16AD) and these constructs were expressed in cells with a GAL4 reporter plasmid. Co-expression of GAL4-LDB1 and VP16-LDB1 significantly increased reporter luciferase activity as a result of dimerization, while co-expression of GAL4-TD-LDB1 with VP16-LDB1 did not, ruling out formation of LDB1 trimers. Likewise, co-expression of GAL4-TD-LDB1 with VP16-TD-LDB1 did not significantly affect luciferase activity, indicating that LDB1 also cannot form protein tetramers. In contrast, TD-LDB1 was able to bind SSBP2 and SSBP3 in both chemical cross-linking and mammalian two-hybrid assays, consistent with the SSBP interacting with preformed LDB1 dimers. Finally, the complete 200-amino acid DD of LDB1, reported as necessary and sufficient for protein dimerization, was confirmed in cross-linking analysis to act as a dominant negative inhibitor of LDB1 dimerization. When the DD was introduced into Lhx2-, Ldb1-, and Ssbp3-expressing cells, application of a modified electrophoretic mobility assay that can detect linking of two DNA probes in solution revealed that the DD reduced formation of a ‘looped’ complex containing two DNA probes and led to the appearance of a new complex containing Lhx2, Ssbp3, and Ldb1, apparently in a monomeric form. In summary, this work elucidates a novel function of SSBPs in enhancing LDB1 dimerization and, ultimately, long-range communication between cis regulatory regions in genes. In addition, it suggests that SSBPs bind dimeric LDB1 and induce an allosteric change in the adjacent SSBP interaction domain rather than vice versa. Finally, these results lead to the prediction that an SSBP- and LDB1-containing complex could promote looping between promoter-proximal and promoter-distal LIM-HD binding elements. Disclosures: No relevant conflicts of interest to declare.

Chronos versus Thanatos in a post-apocalyptic world as presented in Tatyana Tolstaya’s The Slynx and Oleh Shynkarenko’s KaharlykThe article explores issues related to images and concepts used by the writers of dystopias imagining a post-apocalyptic world to diagnose the condition of their national communities. The author discusses, among other things, the means of presenting time such as metaphors of looping or stopping time, which help construct the messages conveyed in the novels. Particular attention is paid to the contrast between Chronos and Thanatos — figures representing time and death, which are widely used in culture. In the analyzed novels, this contrast, which is characteristic of post-Enlightenment secular consciousness, is reflected in the creation of an antithesis between the historic and the archaic. Хронос contra Танатос в постапокалітичному світіна матеріалі романів Кись Тетяни Толстої і Кагарлик Олега ШинкаренкаВ статті розглядаються питання, пов’язані з образами і концепціями, за допомогою яких автори дистопій, зображуючи постапокаліптичний світ, діагностують кондицію власних національних спільнот. Аналізуються, зокрема, засоби представлення часу, як-то метафора часової петлі чи зупиненого часу, що допомагають будувати ідейне наповнення даних творів. Особлива увага звертається на характерне для післяпросвітницької світської свідомості протиставлення Хроноса і Танатоса, тобто культурних фігур смерті й часу, що в обраних творах знаходить своє віддзеркалення в побудові антитези історія — архаїка.

Protein–DNA loops are essential for efficient transcriptional repression and activation. The geometry and stability of the archetypal Lac repressor tetramer (LacI)–DNA loop were investigated using designed hyperstable loops containing lac operators bracketing a sequence-directed bend. Electrophoretic mobility shift assays, DNA cyclization, and bulk and single-molecule fluorescence resonance energy transfer (FRET) demonstrate that the DNA sequence controls whether the LacI–DNA loop forms a compact loop with positive writhe or an open loop with little writhe. Monte Carlo methods for simulation of DNA ring closure were extended to DNA loops, including treatment of variable protein hinge angles. The observed distribution of topoisomer products upon cyclization provides a strong constraint on possible models. The experiments and modeling imply that LacI–DNA can adopt a wide range of geometries but has a strong intrinsic preference for an open form. The flexibility of LacI helps explain in vivo observations that DNA looping is less sensitive to DNA length and shape than that expected from the physical properties of DNA. While DNA cyclization suggests two pools of precursor loops for the 9C14 construct, single-molecule FRET demonstrates a single population. This discrepancy suggests that the LacI–DNA structure is strongly influenced by flanking DNA.

Abstract Ewing sarcoma is an aggressive bone-associated tumor currently treated with dose-intense chemotherapy, radiation, and surgery. Ewing sarcoma affects adolescents and young adults with incidence rate of 3 per million. The hallmark of Ewing sarcoma is a translocated fusion transcription factor named EWS FLI that drives the oncogenic process. The lack of efficient and targeted treatment in Ewing sarcoma directly arises from the poor understanding of how EWS/FLI regulates expression of thousands of genes as well as the current lack of effective targeting of transcription factors. We hypothesize that FLI portion of EWS::FLI containing a crucial alpha-helix plays a novel role in transcription regulation of thousands of genes by modulating chromatin looping. We postulated this hypothesis based on recent evidence from our lab that established an alpha-helix immediately downstream of DNA binding domain as an important player in regulating transcriptional activity in A673 cell line. Structure-function mapping revealed that ETS domain alone was insufficient for full transcriptional regulation. The purpose of this study is to elucidate the mechanism underlying transcriptional regulation by FLI and it is likely to guide future studies on novel therapeutics for Ewing sarcoma patients as transcriptional factors remain elusive targets of therapy. We utilized knockdown/rescue experiments in which EWS FLI was depleted with shRNA and replaced with constructs containing the ETS DNA Binding Domain (DBD) alone or DBD+ (ETS DNA Binding Domain and 4th alpha-helix). The knockdown/rescue experiments were confirmed at mRNA level using qPCR, and at protein level using immunoblots. The binding pattern and transcriptional regulation of DBD and DBD+ were assessed with RNA-Seq respectively. The extent of roles each of these construct play in organization, structure, and function of chromatin are being assessed using Micro-C technique, a variation of Hi-C with improved resolution, higher signal-to-noise ratio and more information on chromatin domain boundaries and chromatin looping. The DNA binding and genomic localization of EWS FLI was unaltered by the deletion surrounding the DNA binding domain (which contains a 4th alpha-helix) in A673 cells. Despite this similarity in genomic localization and binding, the transcriptional output driven by EWS FLI was significantly diminished by the deletion. In addition, we observed this differential transcriptional output by the EWS FLI DBD and DBD+ in another Ewing sarcoma cell line called TTC466. Current work in our lab shows that EWS FLI has a substantial role in shaping the chromatin landscape of Ewing sarcoma cells. We observed this result with Hi-C technique upon the depleting EWS FLI from A673 cells. With this current study, we hope to understand if the flanking region neighboring the DNA binding domain contributes to the chromatin architecture remodeling function of EWS FLI and whether this function could be attributed to the transcriptional output differences in the DBD and DBD+ conditions. Citation Format: Ariunaa Bayanjargal. Studying the role of FLI portion of EWS FLI in transcription regulation via modulation of chromatin 3D landscape in Ewing sarcoma. [abstract]. In: Proceedings of the AACR Special Conference: Cancer Epigenomics; 2022 Oct 6-8; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2022;82(23 Suppl_2):Abstract nr B018.

Establishment of left-right (L-R) asymmetry is fundamental to vertebrate development. Several genes involved in L-R asymmetry have been described. In the Xenopus embryo, Vg1/activin signals are implicated upstream of asymmetric nodal related 1 (Xnr1) and Pitx2 expression in L-R patterning. We report here that Zic3 carries the left-sided signal from the initial activin-like signal to determinative factors such as Pitx2. Overexpression of Zic3 on the right side of the embryo altered the orientation of heart and gut looping, concomitant with disturbed laterality of expression of Xnr1 and Pitx2, both of which are normally expressed in the left lateral plate mesoderm. The results indicate that Zic3 participates in the left-sided signaling upstream of Xnr1 and Pitx2. At early gastrula, Zic3 was expressed not only in presumptive neuroectoderm but also in mesoderm. Correspondingly, overexpression of Zic3 was effective in the L-R specification at the early gastrula stage, as revealed by a hormone-inducible Zic3 construct. The Zic3 expression in the mesoderm is induced by activin (beta) or Vg1, which are also involved in the left-sided signal in L-R specification. These findings suggest that an activin-like signal is a potent upstream activator of Zic3 that establishes the L-R axis. Furthermore, overexpression of the zinc-finger domain of Zic3 on the right side is sufficient to disturb the L-R axis, while overexpression of the N-terminal domain on the left side affects the laterality. These results suggest that Zic3 has at least two functionally important domains that play different roles and provide a molecular basis for human heterotaxy, which is an L-R pattern anomaly caused by a mutation in human ZIC3.

ABSTRACT The lldPRD operon of Escherichia coli, involved in l-lactate metabolism, is induced by growth in this compound. We experimentally identified that this system is transcribed from a single promoter with an initiation site located 110 nucleotides upstream of the ATG start codon. On the basis of computational data, it had been proposed that LldR and its homologue PdhR act as regulators of the lldPRD operon. Nevertheless, no experimental data on the function of these regulators have been reported so far. Here we show that induction of an lldP-lacZ fusion by l-lactate is lost in an ΔlldR mutant, indicating the role of LldR in this induction. Expression analysis of this construct in a pdhR mutant ruled out the participation of PdhR in the control of lldPRD. Gel shift experiments showed that LldR binds to two operator sites, O1 (positions −105 to −89) and O2 (positions +22 to +38), with O1 being filled at a lower concentration of LldR. l-Lactate induced a conformational change in LldR that did not modify its DNA binding activity. Mutations in O1 and O2 enhanced the basal transcriptional level. However, only mutations in O1 abolished induction by l-lactate. Mutants with a change in helical phasing between O1 and O2 behaved like O2 mutants. These results were consistent with the hypothesis that LldR has a dual role, acting as a repressor or an activator of lldPRD. We propose that in the absence of l-lactate, LldR binds to both O1 and O2, probably leading to DNA looping and the repression of transcription. Binding of l-lactate to LldR promotes a conformational change that may disrupt the DNA loop, allowing the formation of the transcription open complex.

Abstract Polyadenylation is a post-transcriptional modification where the 3' end of an mRNA is cleaved and 250-300 adenines are added. It is predicted that 70-75% of human genes have more than one polyadenylation sequence (PAS) and are subject to alternative polyadenylation (APA). APA events affect the coding sequence of a gene when a proximal PAS is located within an intron, constitutive exon, or alternative exon. Gene expression is also affected if there are multiple PAS within the distal 3' untranslated region (UTR); proximal PAS usage shortens the 3'UTR, which can remove cis-regulatory regions such as miRNA and RNA-binding protein (RBP) sites. Furthermore, global changes in APA are linked to cellular state-proximal PAS usage is associated with immature developmental phases, cell proliferation, and cancerous phenotypes. Consequently, APA is a pertinent post-transcriptional modification that regulates gene expression and isoform generation across developmental stages and tissue types. Despite its significance, there are few APA studies in the hematology field, and those that exist have focused on global shifts in PAS usage. In this study, we uniquely focus on the APA mechanism of a single gene, RUNX1, and how this event can alter hematopoietic stem cell (HSC) homeostasis and hematopoiesis. There are three main isoforms of RUNX1 that differ in promoter and/or PAS usage. RUNX1b/c use different promoters, but have identical C-terminal regions. RUNX1a utilizes the same promoter as RUNX1b, but differs from both RUNX1b/c due to usage of a proximal PAS located in alternative exon 7a. RUNX1b/c are robustly expressed in most progenitor populations and differentiated blood cell lineages, whereas RUNX1a is restricted to human CD34+ HSCs. Functionally, RUNX1b/c promote HSC differentiation and lineage commitment, whereas RUNX1a expands HSCs and their engraftment potential, a property with therapeutic advantages but leukemic potential. Due to the difference in expression pattern and distinct functionality of RUNX1a compared to RUNX1b/c, it is relevant to study the APA event that dictates isoform generation. Elucidating this mechanism could provide valuable insight into the transient control of the HSC population for therapeutic benefit and illuminate new leukemogenic pathways. To study RUNX1 APA, we cloned alternative terminal exon 7a (RUNX1a) and constitutive exon 7b (RUNX1b/c) in between the two exons of a split GFP minigene reporter, along with 500 bp of their upstream and downstream flanking introns. We hypothesized that exon 7a would be skipped during processing of the minigene construct because the proximal PAS is rarely used in vivo. Conversely, exon 7b, the penultimate exon in RUNX1b/c, would be spliced in between the GFP exons, disrupting the GFP protein. These constructs were tested in KG-1a and U937 cells. Flow cytometry for GFP fluorescence supported our hypothesis as the exon 7a minigene produced a robust GFP signal and the exon 7b minigene produced no GFP signal. We confirmed that the GFP changes were due to the hypothesized mRNA processing events by performing RT-PCR using primers specific to the two GFP exons. These data show that important cis-regulatory elements that determine RUNX1 APA are located within exon 7a, 7b, and the cloned intronic regions. Next, we altered these minigenes by strategically making chimeric constructs that consist of either exon 7a or 7b with all combinations of upstream/downstream flanking introns. We discovered that replacing the intron upstream of exon 7a confers 2-5 fold greater splicing and polyadenylation of exon 7a, indicative of RUNX1a isoform generation. Therefore, a suppressor cis-element is located in this upstream intronic region. However, placing this intron upstream of exon 7b is not sufficient to reduce its inclusion between the GFP exons. Instead, both the upstream and downstream intronic regions flanking exon 7a are required. This suggests an RNA-looping mechanism that prevents splicing and usage of the exon 7a proximal PAS. Cleavage factor (CFIm) and Polypyrimidine-tract binding protein 1 (PTBP1) are RBPs involved in splicing and polyadenylation that alter mRNA processing by RNA-looping. We aim to narrow down the suppressor region upstream of exon 7a to identify a consensus sequence and the respective RBP that diminishes RUNX1 proximal PAS usage. This knowledge can be leveraged to enhance RUNX1a production and expand HSCs for therapeutic benefit. Disclosures No relevant conflicts of interest to declare.

To identify the molecular pathways that guide cardiac ventricular chamber specification, maturation and morphogenesis, we have sought to characterize factors that regulate the expression of the ventricular myosin light chain-2 gene, one of the earliest markers of ventricular regionalization during mammalian cardiogenesis. Previously, our laboratory identified a 28 bp HF-la/MEF-2 element in the MLC-2v promoter region, which confers cardiac ventricular chamber-specific gene expression during murine cardiogenesis, and showed that the ubiquitous transcription factor YB-1 binds to the HF-la site in conjunction with a co-factor. In a search for interacting co-factors, a nuclear ankyrin-like repeat protein CARP (cardiac ankyrin repeat protein) was isolated from a rat neonatal heart cDNA library by yeast two-hybrid screening, using YB-1 as the bait. Co-immunoprecipitation and GST-CARP pulldown studies reveal that CARP forms a physical complex with YB-1 in cardiac myocytes and immunostaining shows that endogenous CARP is localized in the cardiac myocyte nucleus. Co-transfection assays indicate that CARP can negatively regulate an HF-1-TK minimal promoter in an HF-1 sequence-dependent manner in cardiac myocytes, and CARP displays a transcriptional inhibitory activity when fused to a GAL4 DNA-binding domain in both cardiac and noncardiac cell context. Northern analysis revealed that carp mRNA is highly enriched in the adult heart, with only trace levels in skeletal muscle. During murine embryogenesis, endogenous carp expression was first clearly detected as early as E8.5 specifically in heart and is regulated temporally and spatially in the myocardium. Nkx2-5, the murine homologue of Drosophila gene tinman was previously shown to be required for heart tube looping morphogenesis and ventricular chamber-specific myosin light chain-2 expression during mammalian heart development. In Nkx2-5(−/−)embryos, carp expression was found to be significantly and selectively reduced as assessed by both whole-mount in situ hybridizations and RNase protection assays, suggesting that carp is downstream of the homeobox gene Nkx2-5 in the cardiac regulatory network. Co-transfection assays using a dominant negative mutant Nkx2-5 construct with CARP promoter-luciferase reporter constructs in cardiac myocytes confirms that Nkx2-5 either directly or indirectly regulates carp at the transcriptional level. Finally, a carp promoter-lacZ transgene, which displays cardiac-specific expression in wild-type and Nkx2-5(+/−) background, was also significantly reduced in Nkx2-5(−/−) embryos, indicating that Nkx2-5 either directly or indirectly regulates carp promoter activity during in vivo cardiogenesis as well as in cultured cardiac myocytes. Thus, CARP is a YB-1 associated factor and represents the first identified cardiac-restricted downstream regulatory gene in the homeobox gene Nkx2-5 pathway and may serve as a negative regulator of HF-1-dependent pathways for ventricular muscle gene expression.

Stretched out from end-to-end, the human genome -a sequence of 3 billion chemical letters inscribed in a molecule called DNA -is over 2 meters long. Famously, short stretches of DNA fold into a double helix, which wind around histone proteins to form the 10nm fiber. But what about longer pieces? Does the genome's fold influence function? How does the information contained in such an ultra-dense packing even remain accessible?In this talk, I describe our work developing 'Hi-C' (Lieberman-Aiden et al., Science, 2009; Aiden, Science, 2011) and more recently 'in-situHi-C' (Rao & Huntley et al., Cell, 2014), which use proximity ligation to transform pairs of physically adjacent DNA loci into chimeric DNA sequences. Sequencing a library of such chimeras makes it possible to create genome-wide maps of physical contacts between pairs of loci, revealing features ofgenome folding in 3D. Next, I will describe recent work using in situ Hi-C to construct haploid and diploid maps of nine cell types. The densest, in human lymphoblastoid cells, contains 4.9 billion contacts, achieving 1 kb resolution. We find that genomes are partitioned into contact domains (median length, 185 kb), which are associated with distinct patterns of histone marks and segregate into six subcompartments. We identify ∼10,000 loops. These loops frequently link promoters and enhancers, correlate with gene activation, and show conservation across cell types and species. Loop anchors typically occur at domain boundaries and bind the protein CTCF. The CTCF motifs at loop anchors occur predominantly (>90%) in a convergent orientation, with the asymmetric motifs "facing" one another. Next, I will discuss the biophysical mechanism that underlies chromatin looping. Specifically, our data is consistent with the formation of loops by extrusion (Sanborn & Rao et al., PNAS, 2015). In fact, in many cases, the local structure of Hi-C maps may be predicted in silicobased on patterns of CTCF binding and an extrusion-based model. Finally, I will show that by modifying CTCF motifs using CRISPR, we can reliably add, move, and delete loops and domains. Thus, it possible not only to "read" the genome's 3D architecture, but also to write it. Disclosures No relevant conflicts of interest to declare.

In this study of the learning of programming, two computer-based simulations (manipulative models) of program loops were compared with a computer-based tutorial combined with paper-and-pencil exercises. For the treatment group, one simulation was used prior to and one following classroom instruction on the WHILE-DO and REPEAT-UNTIL looping constructs. For the control group, the tutorial preceded classroom instruction, which was followed by the paper-and-pencil exercises. Students using the manipulative models were more successful in applying their knowledge of loops to a situation requiring transfer, but were no more successful on problems requiring interpretation or direct application. Previous programming experience did not produce a measurable effect on student performance on looping problems.

The Force parallel programming language designed for large-scale shared-memory multiprocessors is presented. The language provides a number of parallel constructs as extensions to the ordinary Fortran language and is implemented as a two-level macro preprocessor to support portability across shared memory multiprocessors. The global parallelism model on which the Force is based provides a powerful parallel language. The parallel constructs, generic synchronization, and freedom from process management supported by the Force has resulted in structured parallel programs that are ported to the many multiprocessors on which the Force is implemented. Two new parallel constructs for looping and functional decomposition are discussed. Several programming examples to illustrate some parallel programming approaches using the Force are also presented.

RUSH/SMARCA3 (SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily A, member 3) is capable of sequence-selective DNA binding and ATP-dependent DNA unwinding. In rabbit uterine epithelial cells, RUSH-1α (113 kDa) is the progesterone-dependent splice variant and RUSH-1β (95 kDa) is the oestrogen-dependent splice variant. Rabbit RUSH/SMARCA3 mRNA is primarily regulated at the proximal promoter (−162/+90) via a PRE (progesterone-response element) half-site/overlapping Y-box domain (−38/−26) and two Sp (specificity protein) 3 sites centred at −128 and −58. We investigated hormone regulation by exploring binding of transcription factors to a putative RUSH/SMARCA3 site (−616/−611) and the distal Sp3 (−131/−126) site. In response to progesterone, RUSH-1α binds the RUSH site and the Sp3 site becomes a functional binding site for Egr-1 (early growth-response gene product 1)/Sp (specificity protein)1/3/MAZ (Myc-associated zinc-finger protein)/MZF1 (myeloid zinc finger 1)/c-Rel. TransSignal TF–TF Interaction Arrays, supershift assays and ChIP (chromatin immunoprecipitation) analyses confirmed strong physical interactions between RUSH and Egr-1/c-Rel. Higher-order long-range interactions between RUSH and the Egr-1/c-Rel derivative of the anisotropic flexibility of the intervening DNA sequence were shown with 3C (chromosome conformation capture) assays. Transient transfection assays with mutant constructs showed the co-operative interaction between RUSH and Egr-1 mediates repression by c-Rel. Thus DNA-bound RUSH/SMARCA3 communicates with its own proximal promoter by looping the intervening DNA. Moreover, progesterone-dependent DNA looping is an adjunct to progesterone induction of the RUSH/SMARCA3 gene because the availability of RUSH isoforms and relevant binding partners is progesterone-regulated.

The DNA-binding protein CCCTC-binding factor (CTCF) and the cohesin complex function together to shape chromatin architecture in mammalian cells, but the molecular details of this process remain unclear. Here, we demonstrate that a 79-aa region within the CTCF N terminus is essential for cohesin positioning at CTCF binding sites and chromatin loop formation. However, the N terminus of CTCF fused to artificial zinc fingers was not sufficient to redirect cohesin to non-CTCF binding sites, indicating a lack of an autonomously functioning domain in CTCF responsible for cohesin positioning. BORIS (CTCFL), a germline-specific paralog of CTCF, was unable to anchor cohesin to CTCF DNA binding sites. Furthermore, CTCF–BORIS chimeric constructs provided evidence that, besides the N terminus of CTCF, the first two CTCF zinc fingers, and likely the 3D geometry of CTCF–DNA complexes, are also involved in cohesin retention. Based on this knowledge, we were able to convert BORIS into CTCF with respect to cohesin positioning, thus providing additional molecular details of the ability of CTCF to retain cohesin. Taken together, our data provide insight into the process by which DNA-bound CTCF constrains cohesin movement to shape spatiotemporal genome organization.

Abstract Approximately one-third of individuals with acquired aplastic anemia (AA) have abnormally short leukocyte telomere lengths. Mutations in genes responsible for maintaining telomere length and stability have been identified in constitutional AA (DKC1) and acquired AA (TERC and TERT). Telomerase, a reverse transcriptase, uses an RNA template (TERC) to extend nucleotide repeats. Telomeric repeat binding factor 1 (TERF1) binds to telomeric DNA, inhibits telomerase and induces bending, looping and pairing of duplex telomeric DNA. Telomeric repeat binding factor 2 (TERF2) binds to the telomere and protects it from degradation and fusion. There are few single nucleotide polymorphisms (SNPs) in TERF1 and TERF2 and nucleotide diversity in these genes is limited. We hypothesized that mutations and/or SNPs in TERF1 and/or TERF2 may be associated with acquired AA. Bi-directional sequence analysis was performed in 47 AA patients who failed to respond to immunosuppressive therapy, had family history of hematologic abnormalities without physical characteristics of dyskeratosis congenita, or short telomeres in leukocytes, on all exons and proximal promoter regions of TERF1 and TERF2. Regions with variation in the first 47 patients were sequenced in an additional 95 patients and 289 healthy controls. SNP frequencies were determined and case-control genotype data analyzed using additive and mutation dominant (MD) genetic models and SAS v8.02. Haplotypes were estimated and a case-control permutation test performed using PHASEv2.1. HaploStats was used to construct haplotypes, determine a global score p value, haplotype frequencies and odds ratios. 5244 base pairs (bp) were sequenced in TERF1 in 47 patients. SNPs were not present in exons 1–8. The proximal promoter, exon 9 and exon 10 were sequenced in the additional 95 patients and 289 controls. One patient had a mutation resulting in a conservative amino acid change (exon 9 Ala-Val). One SNP in intron 9 (C>T) was significant in the additive (p=0.049) and in the MD model, odds ratio 1.59 (p=0.033, 95% CI 1.06-2.39, 37.3% case and 31.8% control allele frequency [AF]). Two synonymous SNPs in exon 10 did not occur in patients but each had an AF of 1.4% in controls (p=0.044 for each). The MD model for these two SNPs suggested an association (p=0.044) but the low frequencies precluded additional calculations. Haplotype analysis showed statistically significant differences between cases and controls in global tests (HaploStats p=0.03 and PHASEv2.1 p=0.01). One haplotype had borderline significance in ordinal trait analysis and comparison to the most common haplotype (p= 0.057 and 0.079, respectively, 35.6 % case and 28.9% control frequency). These data suggest that genetic variation in TERF1 may be associated with aplastic anemia. Sequence analysis of TERF2 consisted of 4515 bp. The proximal promoter, exons 1–2, 4–5 and 7–10 did not have SNPs or mutations. A mutation (Ala-Ser, exon 6) in TERF2 was seen in one patient and no controls. SNPs seen in exons 3 and 6 were also sequenced in 95 patients and 289 controls. SNPs and haplotypes in TERF2 were not associated with AA. This pilot study identified a SNP and haplotype in TERF1 that may be associated with increased risk for AA. Variation at this site in TERF1 in combination with variation at other sites in other genes of the telomere complex could be risk factors for aplastic anemia. Highly penetrant mutations in TERF1 and TERF2 were not identified. These results should be confirmed in a larger study.

ABSTRACT Boundary elements have been found in the Abd-B 3′ cis-regulatory region, which is subdivided into a series of iab domains. Previously, a 340-bp insulator-like element, M340, was identified in one such 755-bp Mcp fragment linked to the PcG-dependent silencer. In this study, we identified a 210-bp core that was sufficient for pairing of sequence-remote Mcp elements. In two-gene transgenic constructs with two Mcp insulators (or their cores) surrounding yellow, the upstream yeast GAL4 sites were able to activate the distal white only if the insulators were in the opposite orientations (head-to-head or tail-to-tail), which is consistent with the looping/bypass model. The same was true for the efficiency of the cognate eye enhancer, while yellow thus isolated in the loop from its enhancers was blocked more strongly. These results indicate that the relative placement and orientation of insulator-like elements can determine proper enhancer-promoter communication.

Isabel Burton, née Arundell, was a model cosmopolitan wife to her famous explorer husband Richard Francis Burton for thirty years. Yet, two pieces of twenty-first-century neo-Victorian Burton-biofiction – The Collector of Worlds by Iliya Troyanov and The Strange Affair of Spring-Heeled Jack by Mark Hodder – write her clean out of their core narratives. Having explored these conspicuous absences, my article turns to the historical Lady Burton's life writing, focussing on The Life of Captain Sir Richard F. Burton, in which she narrativizes the couple's transnational lives. Two of its stylistic devices are discussed in detail – paratexts and the peculiar use of first-person narration – in order to trace in them a double-gesture by which Isabel Burton combines self-erasure with self-empowerment. The detailed analysis of six paratexts, as theorized by Philippe Lejeune and Gérard Genette, supports the claim that Isabel, from the text's fringe, constructs for herself a role that combines elements of the scribe, editor/curator, narrator and author. Looping back to the neo-Victorian biofictional texts, I propose that, in Life, the historical Isabel Burton fuses self-erasure (which Troyanov picks up) with self-empowerment (which Hodder picks up), to forge a female writerly identity compatible with her self-fashioning as the wife/widow of a Victorian transnational cosmopolitan.

Processing of most eukaryotic mRNAs includes and polyadenylation of the nascent transcript. Until now there has been no method for the reliable measurement of these processes in vivo. Therefore, in the present work a new technique was developed for measuring precisely the rate of cleavage and polyadenylation in vivo. The method uses a cis-antisense element targeted to an upstream poly (A) signal. Duplex formation of the antisense element with the poly (A) signal region prevents polyadenylation. In a series of expression vector constructs the antisense element was moved increasing distances downstream of its target poly (A) site, reasoning that if it takes the polymerase longer to reach the antisense element, polyadenylation would have more time to occur. Using this method the half time for commitment to cleavage and polyadenylation at the SV40 early poly (A) site and SPA (synthetic poly A site) was found to be 5 seconds. It was found that strong sites (SV 40 late poly (A) site) were processed faster. Commitment to cleavage and polyadenylation was found to be a multistep process. The expression results were confirmed with the help of RNase protection assay. Relationship between polyadenylation and transcription termination was also studied by using G-free assay and found to be positively correlated. Present data support looping moded suggesting some communication exists between polyadenylation complex and RNA pol. II for transcription termination.

During heart development, cells of the primary and secondary heart field give rise to the myocardial component of the heart. The neural crest and epicardium provide the heart with a considerable amount of nonmyocardial cells that are indispensable for correct heart development. During the past 2 decades, the importance of epicardium-derived cells (EPDCs) in heart formation became increasingly clear. The epicardium is embryologically formed by the outgrowth of proepicardial cells over the naked heart tube. Following epithelial-mesenchymal transformation, EPDCs form the subepicardial mesenchyme and subsequently migrate into the myocardium, and differentiate into smooth muscle cells and fibroblasts. They contribute to the media of the coronary arteries, to the atrioventricular valves, and the fibrous heart skeleton. Furthermore, they are important for the myocardial architecture of the ventricular walls and for the induction of Purkinje fiber formation.Whereas the exact signaling cascades in EPDC migration and function still need to be elucidated, recent research has revealed several factors that are involved in EPDC migration and specialization, and in the cross-talk between EPDCs and other cells during heart development. Among these factors are the Ets transcription factors Ets-1 and Ets-2. New data obtained with lentiviral antisense constructs targeting Ets-1 and Ets-2 specifically in the epicardium indicate that both factors are independently involved in the migratory behavior of EPDCs. Ets-2 seems to be especially important for the migration of EPDCs into the myocardial wall, and to subendocardial positions in the atrioventricular cushions and the trabeculae.With respect to the clinical importance of correct EPDC development, the relation with coronary arteriogenesis has been noted well before. In this review, we also propose a role for EPDCs in cardiac looping, and emphasize their contribution to the development of the valves and myocardial architecture. Lastly, we focus on the congenital heart anomalies that might be caused primarily by an epicardial developmental defect.

ABSTRACT During adeno-associated virus (AAV) type 2 productive infections, the p19 promoter of AAV is activated by the AAV Rep78 and Rep68 proteins. Rep-induced activation of p19 depends on the presence of one of several redundant Rep binding elements (RBEs) within the p5 promoter or within the terminal repeats (TR). In the absence of the TR, the p5 RBE and the p19 Sp1 site at position −50 are essential for p19 transactivation. To determine how a Rep complex bound at p5 induces transcription at p19, we made a series of p19 promoter chloramphenicol acetyltransferase constructs in which the p5 RBE was inserted at different locations upstream or downstream of the p19 mRNA start site. The RBE acted like a repressor element at most positions in the presence of both Rep and adenovirus (Ad), and the level of repression increased dramatically as the RBE was inserted closer to the p19 promoter. We concluded that the RBE by itself was not a conventional upstream activation signal and instead behaved like a repressor. To understand how the Rep-RBE complex within p5 activated p19, we considered the possibility that its role was to function as an architectural protein whose purpose was to bring other p5 transcriptional elements to the p19 promoter. In order to address this possibility, we replaced both the p5 RBE and the p19 Sp1 site with GAL4 binding sites. The modified GAL4-containing constructs were cotransfected with plasmids that expressed GAL4 fusion proteins capable of interacting through p53 and T-antigen (T-ag) protein domains. In the presence of Ad and the GAL4 fusion proteins, the p19 promoter exhibited strong transcriptional activation that was dependent on both the GAL4 fusion proteins and Ad infection. This suggested that the primary role of the p5 RBE and the p19 Sp1 sites was to act as a scaffold for bringing transcription complexes in the p5 promoter into close proximity with the p19 promoter. Since Rep and Sp1 themselves were not essential for transactivation, we tested mutants within the other p5 transcriptional elements in the context of GAL4-induced looping to determine which of the other p5 elements was necessary for p19 induction. Mutation of the p5 major late-transcription factor site reduced p19 activity but did not eliminate induction in the presence of the GAL4 fusion proteins. However, mutation of the p5 YY1 site at position −60 (YY1-60) eliminated GAL4-induced transactivation. This implicated the YY1-60 protein complexes in p19 induction by Rep. In addition, both basal p19 activity and activity in the presence of Ad increased when the YY1-60 site was mutated even in the absence of Rep or GAL4 fusion proteins. Therefore, there are likely to be alternative p5-p19 interactions that are Rep independent in which the YY1-60 complex inhibits p19 transcription. We concluded that transcriptional control of the p19 promoter was dependent on the formation of complexes between the p5 and p19 promoters and that activation of the p19 promoter depends largely on the ability of Rep and Sp1 to form a scaffold that positions the p5 YY1 complex near the p19 promoter.

Interdisciplinary research is devoted to the reflexive-deed reconstruction of the methodological theory of activity, created by G.P. Shchedrovitsky (1929 – 1994) in the frameworks of the collective thinking activity of the Moscow Methodological Circle representatives half a century ago as a complex organization of ontological representations, categorical means, integration-active capabilities and schematizations of methodological thinking, concentrated in the step-by-step formation of ever-growing methodological reflection. With the help of the author’s constructed metamethodological optics this theory is substantiated in its ontological, architectonical, epistemological and philosophic-methodological aspects, highlighted in the polysystemic architectonic mosaic of its ideas and themes, concepts and conceptions, foundations and principles, ideas and constructs, notions and categories, thought-schemes and models, matrices and paradigms, and the most importantly – in conscious thought-actional operation it can appear as a perfect system-actional approach, which is a harbinger of the latest methodology as an original organism or a unique field of TA (thought activity). The essential horizon of such a methodology primarily orients the thinker not on scientific-subject ideas, pictures and knowledge, but on the transformation and development of means, methods and structures of interpenetrating thinking and activity, ie on their reflection in the central link – methodological organization of thinking. Significantly enriching the normative (particularly, sign-symbolic) space of culture, the world of methodology is constructed as one of a kind, original and unique, layer or dimension of human existence – enabling thought-activity, thought-action, professional methodologization. Considering the subject field of the current research, firstly, reflection is highlighted as an attributive center of methodological thinking and methodology in general, secondly, the stages of development of methodological reflection as components of an action are analyzed, thirdly, it has been created for the first time the matrix of genesis of the indicated reflection as an act of methodology creation, fourthly, it has been revealed the method of constructing metamethodological optics of detailed study of the activity theory on the material of the reinterpreted reflexive-deed scenario of the inspirer’s action of system-actional methodology. In the author’s version of composing, metamethodological optics is a complex mechanism for selecting and specifying lenses-modules of interdependent thinking, understanding, activity, reflection, organized by the quintet scheme of philosophical categories as the basis for distinguishing systemic reflexive knowledge: the role of u n i- v e r s a l is performed by the vitacultural methodology developed by us, the place of g e n e r a l is occupied by a sphere of professional methodologization advocated by us, the position of s p e c i a l is occupied by a cyclic-deed approach that we have proposed, the position of i n i v i d u a l is taken by the author’s scheme-model of thought-deed, the place of s p e c i f i c is occupied by such a minimal fractal integrity of a special, conceptually diverse, idea of an object, which finds a graphic expression in the form of a thought scheme (a square wrapped in a circle) and four or five categorical definitions of this object. The main part of the study is devoted to a comprehensive content filling of each of the five reasoned stages of the methodological reflection becoming as components of full-fledged deed: propaedeutic stage – p r e – s i t u a t i o n, the quintessence of which is the idea of actional approach and methodological organization of thinking, the first stage – s i t u a t i o n, the core of which forms the construction of ontological schemes of activity and organization of the process of its reproduction, the second stage – m o t i v a t i- o n, where the most important acts are reflexive immersion of thinking into the world of activity and the emergence of ontology of thought-activity, the third stage – d e e d a c t i o n the main essence of which is reduced to reflexive immersion of TA ontology into a substantial horizon of thinking and to a compliance with the requirements of the multiple knowledge principle, the fourth stage – a f t e r a c t i o n, the most important in which is the reflexive closure of methodological thinking through various reflexive identifications. Thus, the thematically and substantively detailed horizontal of the newly created matrix of the methodological reflection genesis, which is the essence of self-thoughtful – philosophical and therefore methodological – thinking and alpha and omega of methodology in general. At the same time, among the most significant step-by-step creative products of the author’s performance it is worth noting: a) the concept of metamethodologization, that enabled the creation of the latest metamethodological optics of scrupulous elaboration of the activity theory according to the logic of a deed scenarioing of a creative way of the STA-methodology’s founder; b) the thought-scheme of component-tacts of the methodological turn of thinking as a reflexive-canonical deed; c) the cyclic-deed reconstruction of the method of systemic analysis in the unity of different procedures and stages of its implementation; d) the four-stage scheme of the evolution of a scientific subject in two orthogonal dichotomous dimensions of the implementation of methodological work: “empirical (sensory) – logical” and “specific – abstract”; e) the abstract ontological scheme of activity as a full-fledged deed of its reproduction in the cyclical complementarity of ontologems of situation, actualization, translation and reflexive practice and in the context of vitaculture; f) the biquater organizations of the corpus of epistemological units (idea, ontology, universum, STA-approach and scheme, category, theory, STA-methodology) in the polysystem substantiation of thought-actional representations; g) the thought-schemes of connection of ontogenetic stages of reflexive-deed closure of methodological thinking with different reflexive identifications, etc. As a result of the study conducted it is concluded that the model of reflexive-scenario looping of methodology, gained for the first time, for today is a holistic ontological picture of methodological TA, and in the long run – of the universe of thought-activity and thought-deed in general. A clear confirmation of this yet is the unique step-by-step spectrum of competent methodological practice – from the creation, development and usage of various technologies of methodological work and reflective metamethodologization to canon-oriented methodological seminars and sessions, organization-actional and organization-deed games.

This study was conducted to verify the construct model for organizational commitment among technical teachers at the Vocational College under the Ministry of Education. A study involving 25 vocational colleges with a total sample of 493 people consisting of teachers in mechanical engineering, electrical and electronic engineering fields, and engineering. This study uses Structural Equation Modeling (SEM) approach with AMOS 20 software. Researchers used the Organizational Commitment Questionnaire (OCQ) developed by Meyer, Allen, and Smith (1993) and Meyer and Allen (1997), comprising three substructures namely affective commitment, normative commitment and continuous commitment. The number of items is 24, 8 items per sub construct and after exploratory factor analysis is carried out there were only 22 items left. In order to ensure that the organization's commitment measuring model is verified, three factors have been evaluated as unidimensionality (looking at the factor loading ≥ 0.6), Validity (convergent validity AVE ≥ 0.5, and construct validity refers to fitness indexes) and Reliability (composite reliability, CR ≥ 0.6 and Average Variance Extracted ≥ 0.5). After the CFA analysis is carried out no items were dropped because all items have loading factors> 0.6, AVE ≥ 0.5 values for all sub constructs. While fitness indexes for Absolute Fit (RMESA = 0.062), Incremental Fit (CFI = 0.953, TLI = 0.945) and Parsimonious Fit (Chisq/df = 2.918) met the requirements set for all CR values ≥ 0.6. It can be concluded that after the measurement model was modified by looping between items, the model was fit to be used for structural model analysis process.

This time last year we proposed the theme of the 'loop' issue to the M/C collective because it sounded deeply cool, satisfying our poststructuralist posturings about reflexivity and representation, while also tapping into everyday cultural objects and practices. We expected that the 'loop' issue would generate some interesting and varied responses, and it did. We received submissions about music, visual art, language, child development, pop-cultural artefacts, mathematics and culture in general. These explorations of disparate fields seemed, however, to be tied together by a common thread: the concept of "generation" itself. Each article in the 'loop' issue describes a loop that does not simply repeat an original operation, but that through iteration creates new possibilities and new meanings. More recently, when we began the process of putting the 'loop' issue together we found ourselves faced with a problem we had not envisaged: how might we structure the issue to reflect our guiding metaphor? How might we make articles about loops read like a loop? For all its decentredness, the web is ultimately still a linear medium, and one that constructs fairly rigorous and striated hierarchies. Even M/C, which has tried to set itself apart from traditional printed academic journals, is still structurally reminiscent of them. We toyed with the idea of hyperlinking articles or changing the table of contents from a descending list to a circular one. Apart from the inconceivably difficult task of convincing our web-designers to change the table of contents template for a one-off issue, such strategies would not necessarily have the desired effect. Readers could choose not to make use of our designed loop—hypertext would still work against us because readers might jump around in any order they desired. Moreover, the very necessity for the issue to have a "feature article" would straighten out any loop we might construct. An editorially enforced structure, it seemed, was not the answer. So we turned to our contributions. Strangely enough (or perhaps not strangely at all), we found that, read together, they could be perceived as generating their own internal spiralling system of interconnected loops. We as editors have therefore sought to place the contributions in an order that brings out this cyclical reading. The issue's feature article is Laurie Johnson's "Agency, Beyond Strange Cultural Loops"; a highly accessible, gently amusing, and deeply thought-out meditation on the production and analysis of culture. Johnson works his way from the simple concept of infinite loops in computer programs, though the strange loops described by Douglas Hofstader and drawn by M.C Escher in his work Drawing Hands, to what he calls "strange cultural loops", in which the reception of original signs is shaped by what has been seen before. He contends that such loops constitute culture, because the very notion of cultural re-production is impossible without considering agency. Infinite loops are infinite only if we decide them to be, just as culture is recognisable if and how we choose it to be. What makes culture easily graspable in everyday life, yet so difficult to analyse, is that reception is as productive of culture as creation. The first of our two special visual art features is Vince Dziekan's "The Synthetic Image", which is at once an exhibition of 13 digital artists and an exploration of the curatorial process. "The Synthetic Image" is an astounding interactive exhibition/installation which the user is invited to explore via a kind of looped nodal map. Indeed, "The Synthetic Image" immerses the user in three kinds of loops. First, art and critique are drawn together into interlinking loops. Second, a centrifugal hypertextual structure is used to both create and display a narrative space. Third, the metaphor of the loop is used to discuss the synthetic nature of digital art that explores the relationship between the real and the virtual. M/C is extremely proud to present this exhibition in conjunction with the Department of Multimedia and Digital Arts at Monash University, Melbourne. Simone Murray explores cultural production through corporate loops in "Harry Potter, Inc.: Content Recycling for Corporate Strategy". Murray investigates not the first-wave success of the books but the second-wave take up of Harry Potter as a franchisable cultural product with substantial multi-demographic appeal. This is a detailed examination of how America Online-Time Warner (AOL-TW) has linked its corporate strategy to the characteristics of the Harry Potter brand. More than a simple sequential marketing operation, AOL-TW recycles content of the books, film, and soundtracks, in three ways: reusing digital content to sell its own products, licencing significant portions of content to secondary manufacturers, and, finally, using Harry Potter content to stimulate interest in non-Harry Potter AOL-TW products. Content recycling exemplifies current corporate drives toward synergy. In "Mastering the 'Visual Groove': Animated Electric Light Bulb Signs, Locations, and Loops", Margaret Weigel reminds that looping media are not new phenomena, dating from the Victorian era and coming to particular prominence in the looping electric light bulb signs of the late 1800s and early 1900s. Our reactions to electric signs were, she argues, strangely similar to those of many new media: moral panic and debate, leading to acceptance and even fondness. Weigel describes many of these whimsical modernist spectacles, particularly those of Broadway (the "Great White Way") in the early 1900s, and then describes their reception by tourists and locals. While tourists were drawn to the nightly spectacles, locals mastered and integrated the cyclical marketing systems into their daily lives. Greg Hainge's "Platonic Relations: The Problem of the Loop in Contemporary Electronic Music" proposes a way in which looping in electronic music may avoid the banal "Platonic" mode of repetition maligned by Deleuze in Difference and Repetition. In electronic music, Hainge argues, this passive approach to the loop conceives of the sampled element as "constitut[ing] an originary identity," the repetition of which constructs "an absolute internal resemblance". Moreover, Platonic looping itself forms a kind of technologically-determinist feedback loop within which the electronic artist finds him- or herself caught. By analysing the way in which electronic artist Kaffe Matthews breaks free of the Platonic mode, Hainge identifies a more "improvisational and dynamic aesthetic" of looping. In "Making Data Flow: On the Implications of Code Loops", Adrian Mackenzie explores and typologises loops in computer code. He argues that computer code has become an object of intense interest in cultural life, perhaps because computer code is at least as generative of meaning as content is supposed to be. Loops are an integral part of the coding process, and are also an interesting way to investigate the generation of meaning through information flows. Mackenzie finds the distinguishing feature of code loops to be their bounding conditions. Different bounding conditions, of course, generate different information flows. Given this, flows can be adapted to different cultural purposes by writers, artists, and hackers interested in exploring different spatio-temporal manifolds. Andrew T. Jacobs, in "Appropriating a Slur: Semantic Looping in the African-American Usage of 'Nigga'", unravels the fascinating rhetorical process by which the highly charged epithet 'nigger' has been reclaimed as 'nigga' by African-Americans. Drawing on rhetorical analysis and African-American sociology, Jacobs argues that co-opting the slur has involved three looping mechanisms—agnominatio, semantic inversion, and chiastic slayingthemselves combined into a looping process which he calls "semantic looping". He concludes that the use of "nigga" is a resistance strategy that functions through both recalling and refuting racism. In "Loops and Fakes and Illusions", Keith Russell investigates the role loops play in the childhood development of social understanding. Not only do loops figure in development, but, as Russell's reading of D.W. Winnicott demonstrates, childhood development itself is a sustaining loop. Following John Dewey, Russell contends that perplexity is the source of intellectual development, and that children exercise their perplexity by puzzling over illusions based around loops. Russell explores how these illusions and fakes demonstrate the tensions and dynamics of social reality, concluding that playing with loops is a lifelong process. Cameron Brown's "Rep-tiles with Woven Horns" is the second of our special visual art features. The title of Brown's article is also the title of the image gracing the cover of this issue. The image itself is particularly suited to the "loop" issue because it is a fractal created by recursion. Brown's article describes the geometry and mathematics behind the image, providing a step-by step demonstration of its creation. We think that even non-mathematicians will follow the logic of the steps involved, and gain a deeper appreciation of both the power and elegance of recursion. As we must, we conclude the 'loop' issue much as we began it, exploring the links between agency and the generative power of the loop. Like Laurie Johnson, Luis O. Arata's "Creation by looping Interactions" questions the creative process involved in M.C. Escher's Drawing Hands, but imagines it as an animated process with two different outcomes. While one outcome is a closed loop—akin to the Platonic looping described by Hainge—generating only itself, the other is an open loop. Open loops, Arata contends, are a form of interaction, a powerful reflexive dialogue of participatory creation. He shows that cutting-edge science is finding the reflexive creativity of open loops to be increasingly important both to practice and theory, concluding that innovation is all the richer for it. Thus, one might say that Johnson and Arata each takes the role of one of the hands in the artwork they both analyse: Escher's Drawing Hands. Within that larger loop, smaller loops are described, and so, like Nietzsche, we find ourselves "insatiably calling out da capo" (56). References Nietzsche, Friedrich. Beyond Good and Evil. Trans. R.J. Hollingdale. Harmondsworth: Penguin, 1973. Citation reference for this article MLA Style Mitchell, Peta and Rintel, E. Sean. "Editorial" M/C: A Journal of Media and Culture 5.4 (2002). [your date of access] < http://www.media-culture.org.au/mc/0208/editorial.php>. Chicago Style Mitchell, Peta and Rintel, E. Sean, "Editorial" M/C: A Journal of Media and Culture 5, no. 4 (2002), < http://www.media-culture.org.au/mc/0208/editorial.php> ([your date of access]). APA Style Mitchell, Peta and Rintel, E. Sean. (2002) Editorial. M/C: A Journal of Media and Culture 5(4). < http://www.media-culture.org.au/mc/0208/editorial.php> ([your date of access]).

This paper is concerned with deciding whether or not assertions are valid of a parallel process using methods which are directed by the way in which the process has been composed. The assertions are drawn from a modal logic with recursion, capable of expressing a great many properties of interest. The processes are described by a language inspired by Milner's CCS and Hoare's CSP, though with some modifications. The choice of constructors allows us to handle a range of synchronisation disciplines and ensures that the processes denoted are finite state. The operations are prefixing, a non-deterministic sum, product, restriction, relabelling and a looping construct. Arbitrary parallel compositions are obtained by using a combination of product, restriction and relabelling. We are interested in deciding whether or not an assertion A is valid of a process t. If it is valid, in the sense that every reachable state of t satisfies A, we write |= A:t. This paper investigates the extent to which the composition of t can guide methods for deciding |= A:t. For instance if t were a sum t_0 + t_1 we can ask what assertions A_0 and A_1 should be valid of t_0 and t_1 respectively to ensure that A is valid of t_0 + t_1. The paper formulates new compositional methods for deciding validity, and exposes some fundamental difficulties. Algorithms are provided to reduce validity problems for prefixing, sum, relabelling, restriction and looping to validity problems for their immediate components --- all these reductions depend only on the top-level structure of terms. The existence of these reductions rests on being able to 'embed' the properties of a term in the properties, or products of properties, of its immediate subterms. Because there is not such a simple embedding for the product construction of terms, as might be expected, similar reductions become much more complicated for products; although there are general results, and the reductions can be simple in special cases, the general treatment for products meets with fundamental difficulties. Whereas reductions for products always exist for this finite state language, they demonstrably no longer just depend on the top-level (product structure) of the term; in particular, a simple assertion is exhibited for which the size of the reduction must be quadratic in the number of states of the process. An attempt is thus made to explain what makes product different from the other operations with respect to compositional reasoning, and to delimit the obstacles to automated compositional checking of validity on parallel processes.

Why has software code become an object of intense interest in several different domains of cultural life? In art (.net art or software art), in Open source software (Linux, Perl, Apache, et cetera (Moody; Himanen)), in tactical media actions (hacking of WEF Melbourne and Nike websites), and more generally, in the significance attributed to coding as work at the pinnacle of contemporary production of information (Negri and Hardt 298), code itself has somehow recently become significant, at least for some subcultures. Why has that happened? At one level, we could say that this happened because informatic interaction (websites, email, chat, online gaming, ecommerce, etc) has become mainstream to media production, organisational practice and indeed, quotidian life in developed and developing countries. As information production moves into the mainstream, working against mainstream control of flows of information means going upstream. For artists, tactical media groups and hackers, code seems to provide a way to, so to speak, reach over the shoulder of mainstream media channels and contest their control of information flows.1 A basic question is: does it? What code does We all see content flowing through the networks. Yet the expressive traits of the flows themselves are harder to grapple with, partly because they are largely infrastructural. When media and cultural theory discuss information-network society, cyberculture or new media, questions of flow specificity are usually downplayed in favour of high-level engagement with information as content. Arguably, the heightened attention to code attests to an increasing awareness that power relations are embedded in the generation and control of flow rather than just the meanings or contents that might be transported by flow. In this context, loops provide a really elementary and concrete way to explore how code participates in information flows. Loops structure almost every code object at a basic level. The programmed loop, a very mundane construct, can be found in any new media artist's or software engineer's coding toolkit. All programming languages have them. In popular programming and scripting languages such as FORTRAN, C, Pascal, C++, Java, Visual Basic, Perl, Python, JavaScript, ActionScript, etc, an almost identical set of looping constructs are found.2 Working with loops as material and as instrument constitutes an indispensable part of producing code-based objects. On the one hand, the loop is the most basic technical element of code as written text. On the other hand, as process executed by CPUs, and in ways that are not immediately obvious even to programmers themselves, loops of various kinds underpin the generative potential of code.3 Crucially, code is concerned with operationality rather than meaning (Lash 203). Code does not directly create meaning. It circulates, transforms, and reproduces messages and patterns of widely varying semantic and contextual richness. By definition, flow is something continuous. In the case of information, what flows are not things but patterns which can be rendered perceptible in different ways—as image, text, sound—on screen, display, and speaker. While the patterns become perceptible in a range of different spatio-temporal modes, their circulation is serialised. They are, as we know, composed of sequences of modulations (bits). Loops control the flow of patterns. Lev Manovich writes: programming involves altering the linear flow of data through control structures, such as 'if/then' and 'repeat/while'; the loop is the most elementary of these control structures (Manovich 189). Drawing on these constructs, programming or coding work gain traction in flows. Interactive looping Loops also generate flows by multiplying events. The most obvious example of how code loops generate and control flows comes from the graphic user interfaces (GUIs) provided by typical operating systems such as Windows, MacOs or one of the Linux desktop environments. These operating systems configure the visual space of millions of desktop screen according to heavily branded designs. Basically they all divide the screen into different framing areas—panels, dividing lines, toolbars, frames, windows—and then populate those areas with controls and indicators—buttons, icons, checkboxes, dropdown lists, menus, popup menus. Framing areas hold content—text, tables, images, video. Controls, usually clustered around the edge of the frame, transform the content displayed in the framed areas in many different ways. Visual controls are themselves hooked up via code to physical input devices such as keyboard, mouse, joystick, buttons and trackpad. The highly habituated and embodied experience of interacting with contemporary GUIs consists of moving in and out, within and between different framing areas, using visual controls that respond either to pointing (with the mouse) or keyboard command to change what is displayed, how it is displayed or indeed to move that content elsewhere (onto disk, across a network). Beneath the highly organised visual space of the GUI, lie hundreds if not thousands of loops. The work of coding these interfaces involves making loops, splicing loops together, and nesting loops within loops. At base, the so-called event loop means that the GUI in principle stands ready at any time to accept input from the physical interface devices. Depending on what that input is, it may translate into direct changes within the framed areas (for instance, keystrokes appear in a text field as letters) or changes affecting the controls (for instance, Control-Enter might signal send the text as an email). What we usually understand by interactivity stems from the way that a loop constantly accepts signals from the physical inputs, queues the signals as events, and deals with them one by one as discrete changes in what appears on screen. Within the GUI's basic event loop, many other loops are constantly starting and finishing. They are nested and unnested. They often affect some or other of the dozens of processes running at any one time within the operating system. Sometimes a command coming from the keyboard or a signal arriving from some other peripheral interface (the network interface card, the printer, a scanner, etc) will trigger the execution of a new process, itself composed of manifold loops. Hence loops often transiently interact with each other during execution of code. At base, the GUI shows something important, something that extends well beyond the domain of the GUI per se: the event loop generates and controls informations flows at the same time. People type on keyboards or manipulate game controllers. A single keypress or mouse click itself hardly constitutes a flow. Yet the event loop can amplify it into a cascade of thousands of events because it sets other loops in process. What we call information flow springs from the multiplicatory effect of loops. A typology of looping Information flows don't come from nowhere. They always go somewhere. Perhaps we could generalise a little from the mundane example of the GUI and say that the generation and control of information flows through loops is itself regulated by bounding conditions. A bounding condition determines the number of times and the sequence of operations carried out by a loop. They often come from outside the machine (interfaces of many different kinds) and from within it (other processes running at the same time, dependent on the operating system architecture and the hardware platform). Their regulatory role suggests the possibility of classifying loops according to boundary conditions.4 The following table classifies loops based on bounding conditions: Type of loop Bounding condition Typical location Simple & indefinite No bounding conditions Event loops in GUIs, servers ... Simple & definite Bounding conditions determined by a finite set of elements Counting, sorting, input and output Nested & definite Multiple bounding conditions Transforming grid and table structures Recursive Depth of possible recursion (memory or time) Searching and sorting of tree or network structures Result controlled Loop ends when some goal has been reached Goal-seeking algorithms Interactive and indefinite Bounding conditions change during the course of the loop User interfaces or interaction Although it risks simplifying something that is quite intricate in any actually executing process, this classification does stress that the distinguishing feature of loops may well be their bounding conditions. In practical terms, within program code, a bounding condition takes the form of some test carried out before, during or after each iteration of a loop. The bounding conditions for some loops relate to data that the code expects to come from other places—across networks, from the user interface, or some other devices. For other loops, the bounding conditions continually emerge in the course of the loop itself—the result of a calculation, finding some result in the course of searching a collection or receiving some new input in a flow of data from an interface or network connection. Based on the classification, we could suggest that loops not only generate flows, but they generate those flows within particular spatio-temporal manifolds. Put less abstractly, if we accept that flows don't come from nowhere, we then need to say what kind of places they do come from. The classification shows that they do not come from homogeneous spaces. In fact they relate to different topologies, to the hugely diverse orderings of signs and gestures within mediatic cultures. To take a mundane example, why has the table become such an important element in the HTML coding of webpages? Clearly tables provide an easy way to organise a page. Tables as classifying and visual ordering devices are nothing new. Along with lists, they have been used for centuries. However, the table as onscreen spatial entity also maps very directly onto a nested loop: the inner loop generates the horizontal row contents; the outer loop places the output of the inner loop in vertical order. As web-designers quickly discovered during the 1990s, HTML tables are rendered quickly by browsers and can easily position different contents—images, headings, text, lines, spaces—in proximity. In shorts, nested loops can quickly turn a table into a serial flow or quickly render a table out of a serial flow. Implications We started with the observation that artists, writers, hackers and media activists are working with code in order to reposition themselves in relation to information flows. Through technical elements such as loops, they reappropriate certain facets of the production of information and communication. Working with these and other elements, they look for different points of entry into the flows, attempting to move upstream of the heavily capitalised sites of mainstream production such as the Windows GUI, eCommerce websites or blockbuster game titles. The proliferation of information objects in music, in visual culture, in database and net-centred forms of interactivity ranging from computer games to chat protocols, suggests that the coding work can trigger powerful shifts in the cultures of circulation. Analysis of loops also suggests that the notion of data or information flow, understood as the continuous gliding of bits through systems of communication, needs revision. Rather than code simply controlling flow, code generates flows as well. What might warrant further thought is just how different kinds of bounding conditions generate different spatio-temporal patterns and modes of inclusion within flows. The diversity of loops within information objects imply a variety of topologically complicated places. It would be possible to work through the classification describing how each kind of loop maps into different spatial and temporal orderings. In particular, we might want to focus on how more complicated loops—result controlled, recursive, or interactive and indefinite types—map out more topologically complicated spaces and times. For my purposes, the important point is that bounding conditions not only regulate loops, they bring different kinds of spatio-temporal manifold into the seriality of flow. They imprint spatial and temporal ordering. Here the operationality of code begins to display a generative dimension that goes well beyond merely transporting or communicating content. Notes 1. At a more theoretical level, for a decade or so fairly abstract notions of virtuality have dominated media and cultural studies approaches to new media. While that domination has been increasingly contested by more fine grained studies of how the Internet is enmeshed with different places (Miller and Slater), attention to code is justified on the grounds that it constitutes an increasingly important form of expression within information flows. 2. Detailed discussion of these looping constructs can be found in any programming textbook or introductory computer science course, so I will not be going through them in any detail. 3. For instance, the cycles of the clock chip are absolutely irreducible. Virtually all programs implicitly rely on a clock chip to regulate execution of their instructions. 4. A classification can act as a symptomatology, that is, as something that sets out the various signs of the existence of a particular condition (Deleuze 368), in this case, the operationality of code. References Appadurai, Arjun. Modernity at Large: Cultural Dimensions of Globalization. Minneapolis: U of Minnesota P, 1996. Deleuze, Gilles. The Brain is the Screen. An Interview with Gilles Deleuze. The Brain is the Screen. Deleuze and the Philosophy of Cinema. Ed Gregory Flaxman. Minneapolis: U of Minnesota P, 2000. 365-68. Hardt, Michael and Antonio Negri. Empire. Cambridge, MA: Harvard U P, 2000. Himanen, Pekka. The Hacker Ethic and the Spirit of the Information Age. London: Secker and Warburg, 2001. Lash, Scott. Critique of Information. London: Sage, 2002. Manovich, Lev. What is Digital Cinema? Ed. Peter Lunenfeld. The Digital Dialectic: New Essays on New Media. Cambridge, MA: MIT, 1999. 172-92. Miller, Daniel, and Don Slater. The Internet: An Ethnographic Approach. Oxford: Berg, 2000. Moody, Glyn. Rebel Code: Linux and the Open Source Revolution. Middlesworth: Penguin, 2001. Citation reference for this article MLA Style Mackenzie, Adrian. "Making Data Flow" M/C: A Journal of Media and Culture 5.4 (2002). [your date of access] < http://www.media-culture.org.au/mc/0208/data.php>. Chicago Style Mackenzie, Adrian, "Making Data Flow" M/C: A Journal of Media and Culture 5, no. 4 (2002), < http://www.media-culture.org.au/mc/0208/data.php> ([your date of access]). APA Style Mackenzie, Adrian. (2002) Making Data Flow. M/C: A Journal of Media and Culture 5(4). < http://www.media-culture.org.au/mc/0208/data.php> ([your date of access]).

An evolutionarily conserved feature of introns is their ability to enhance expression of genes that harbor them. Introns have been shown to regulate gene expression at the transcription and post-transcription level. The general perception is that a promoter-proximal intron is most efficient in enhancing gene expression and the effect diminishes with the increase in distance from the promoter. Here we show that the intron regains its positive influence on gene expression when in proximity to the terminator. We inserted ACT1 intron into different positions within IMD4 and INO1 genes. Transcription Run-On (TRO) analysis revealed that the transcription of both IMD4 and INO1 was maximal in constructs with a promoter-proximal intron and decreased with the increase in distance of the intron from the promoter. However, activation was partially restored when the intron was placed close to the terminator. We previously demonstrated that the promoter-proximal intron stimulates transcription by affecting promoter directionality through gene looping-mediated recruitment of termination factors in the vicinity of the promoter region. Here we show that the terminator-proximal intron also enhances promoter directionality and results in compact gene architecture with the promoter and terminator regions in close physical proximity. Furthermore, we show that both the promoter and terminator-proximal introns facilitate assembly or stabilization of the preinitiation complex (PIC) on the promoter. On the basis of these findings, we propose that proximity to both the promoter and the terminator regions affects the transcription regulatory potential of an intron, and the terminator-proximal intron enhances transcription by affecting both the assembly of preinitiation complex and promoter directionality.

Castration resistant prostate cancer (CRPC) continues to be androgen receptor (AR) driven. Inhibition of AR signaling in CRPC could be advanced using state-of-the-art biophysical and biochemical techniques. Structural characterization of AR and its complexes by cryo-electron microscopy would advance the development of N-terminal domain (NTD) and ligand binding domain (LBD) antagonists. The structural basis of AR function is unlikely to be determined by any single structure due to the intrinsic disorder of its NTD which not only interacts with coregulators but likely accounts for the constitutive activity of AR-splice variants (SV) which lack the LBD and emerge in CRPC. Using different AR constructs lacking the LBD their effects on protein folding, DNA binding, and transcriptional activity could reveal how interdomain coupling explains the activity of AR-SVs. The AR also interacts with co-regulators that promote chromatin looping. Elucidating the mechanisms involved can identify vulnerabilities to treat CRPC which do not involve targeting the AR. Phosphorylation of the AR coactivator MED-1 by CDK7 is one mechanism that can be blocked by the use of CDK7 inhibitors. CRPC gains resistance to AR signaling inhibitors (ARSI). Drug resistance may involve AR-SVs but their role requires their reliable quantification by SILAC-mass spectrometry during disease progression. ARSI drug resistance also occurs by intratumoral androgen biosynthesis catalyzed by AKR1C3 (type 5 17β-hydroxysteroid dehydrogenase), which is unique in that its acts as a coactivator of AR. Novel bifunctional inhibitors that competitively inhibit AKR1C3 and block its coactivator function could be developed using reverse-micelle-NMR and fragment-based drug discovery.

The fields of epigenetics and neuroscience have come to occupy a significant place in individual and public life in biomedicalized societies. Social scientists have argued that the primacy and popularization of the “neuro” has begun to shape how patients and other lay people experience themselves and their lifeworlds in increasingly neurological and genetic terms. Pregnant women and new mothers have become an important new target for cutting edge neuroscientific and epigenetic research, with the Internet constituting a highly active space for engagement with knowledge translations. In this paper, we analyze the reception by women in North America of translations of nascent epigenetic and neuroscientific research. We conducted three focus groups with pregnant women and new mothers. The study was informed by a prior scoping investigation of online content. Our focus group findings record how engagement with translations of epigenetic and neuroscientific research impact women's perinatal experience, wellbeing, and self-construal. Three themes emerged in our analysis: (1) A kind of brain; (2) The looping effects of biomedical narratives; (3) Imprints of past experience and the management of the future. This data reveals how mothers engage with the neurobiological style-of-thought increasingly characteristic of public health and popular science messaging around pregnancy and motherhood. Through the molecularization of pregnancy and child development, a typical passage of life becomes saturated with “susceptibility,” “risk,” and the imperative to preemptively make “healthy' choices.” This, in turn, redefines and shapes the experience of what it is to be a “good,” “healthy,” or “responsible” mother/to-be.

IntroductionWithin social media visibility campaigns, selfie captions usually work to produce coherent identity categories, linking disparate selfies together through hashtags. Furthering visibility politics, such selfie captions claim that authentic identities can be made visible through selfies and can be described and defined by these captions. However, selfie captions by the trans artist Alok Vaid-Menon challenge the assumption that selfies and their captions can make authentic identity legible. Through hashtags, emojis, and punning text, Vaid-Menon’s selfie captions interrogate visibility politics from within one of visibility politics most popular contemporary tools, demonstrating how social media can be used to theorize representation. Coherence, Visibility, and Authenticity through HashtagsMobilising and organising identitarian counterpublics through hashtags—from #DisabledAndCute (Wade) to #GirlsLikeUs (Jackson, Bailey, and Welles 2)—these captions operate as hyperlinks that lead users to collections of all the images similarly tagged or captioned. This draws attention to certain aspects of the images, and produces coherence and similarity, despite the actual diversity of the individuals participating in these projects of visibility. These captions also question the over-determination of visibility with authenticity in dominant discourse, and the assumption that visibility can guarantee authenticity. For example, this is apparent in the Human Rights Campaign’s 2014 publication Transgender Visibility: A Guide to Being You, which offers visibility as a critical strategy for “living as authentically as possible” (quoted in David 28).Further, as images that seem to enable direct, unmediated, and hence “authentic,” self-expression (Lorbinger and Brantner 1848), selfies are described as ideally designed for visibility politics (Duguay 4). Visibility politics relies on aesthetic representation to expand the boundaries of commonsense to include those who were previously excluded—all without challenging the underlying logic that produces the inclusion of some through the exclusion of others (Rancière 141–3). In social media visibility campaigns, selfie captions are therefore a critical tool, for they not only use hashtags to create webs of interconnected selfies that produce a coherent, visible identity category, but through doing so, they reinforce the illusion that selfies—as photographs—exhibit an unmediated relationship between sign and signified, offering a visual authentication of identity. Thus, social media visibility campaigns presume that the authentic self can be made legible through selfies and their captions, reiterating, as C. Riley Snorton writes, the “popular, long-held myth—that both the truth of race and the truth of sex are obvious, transparent, and written on the body” (3).Because visible markers of gender and race are assumed to offer access to the “truth” of identity (Rightler-McDaniels and Hendrickson 178), visibility politics are usually heavily invested in this idea of visible authenticity—they also, ultimately, provide a critical avenue for commodification, branding, and consumerism (Banet-Weiser 35; David 30). However, in direct contrast to this, the trans artist Alok Vaid-Menon—a non-binary South Asian performance artist whose pronouns are they/them—uses selfie captions to expose and explode the insufficiency of visibility politics, albeit while promoting their personal brand.Vaid-Menon: Captions, Hashtags, and Intersectional IdentitiesIn Instagram posts that include both still and video selfies, their punning captions undermine any direct relationship between sign and signified, and use playful language to challenge the logic that selfies can transparently communicate authentic identity. Instead of producing coherence, Vaid-Menon uses hashtags to insert charged, political posts within supposedly apolitical series, disrupting any claims to similarity. For example, although Vaid-Menon’s selfie captions draw attention to particular elements within the image, they highlight those aspects of the visual field that make it impossible to identify a single, unified identity.It is also worth discussing here how this plays out in a specifically visual medium such as Instagram. Drawing on the resources of this platform, these selfie captions include emojis, thereby doubling the elements of the visual field within the space of the caption, and emphasising the symbolic function of cultural signifiers of identity. Thus, Vaid-Menon’s selfie captions demonstrate that social media platforms are not merely conduits for visibility politics, but instead offer rich resources for interrogating and contesting the politics of representation.Throughout Vaid-Menon’s Instagram selfies, punning captions appear—examples include “beach the change you want to see in the world” and “fifty shades of gay.” In these captions, puns not only draw attention to the texture and flexibility of language—a linguistic playfulness that is always already present in social media platforms through ludic hashtags (Rightler-McDaniels and Hendrickson 187)—but also highlight elements within the image that put pressure on the idea of coherent and unified identity. By doing this, these captions explicitly declare that identity work is self-consciously performative, producing identities that are not a question of authenticity—even within the framework of “branded authenticity” (Banet-Weiser 11)—but that might instead be read through the more ambivalent notion of “sincerity” (Jackson 15).An example of this can be seen accompanying a slow-motion video selfie of Vaid-Menon in a blonde wig (AlokVMenon, 9 January 2016a). The significance of body hair for South Asian women and femmes is a reoccurring theme throughout Vaid-Menon’s selfie captions. They are vocal about the political significance of body hair, and use hashtags and text captions to address how body hair complicates their ability to communicate the truth(s) of their identity. In the video, brightly painted lips parted, Vaid-Menon twirls the blonde curls around their fingers, while the slow-motion effect emphasises the movement of each lock of hair. Simultaneously, Vaid-Menon’s dark body hair is prominent and visible, including chest hair, the shadow of a beard, and thick eyebrows.The image is accompanied by a caption which asserts punningly “gender is racial construct: blondes have more funding”, thereby transforming the gender studies dogma that “gender is a social construct” and the popular culture slogan that “blondes have more fun.” The caption uses this wording to point out that the gendering of body hair as masculine delimits femininity as whiteness, and also privileges white (cis) femininity within capitalism. Like the caption, the image also reveals how “gender is a racial construct,” staging the tensions between Vaid-Menon’s “natural” dark body hair (gendered masculine) and the bright, blonde wig they wear (gendered feminine, but racialised as white). Further, within late capitalism, the caption “blondes have more funding” lays claim to a possibility that the image forecloses—because “gender is a racial construct,” this increased funding is likely to be out of reach for brown trans femmes who look like Vaid-Menon. Together, the caption and the image suggest that hair is both the solution and the problem for Vaid-Menon—although “blondes have more funding,” the blondes who get funded are white, and definitely not covered in thick, dark body hair.Posting selfies that show off their body hair, Vaid-Menon regularly captions these images with the hashtag #TGIF (AlokVMenon, 19 August 2016) thereby taking advantage of the cross-platform utility and democratising function of hashtags (Rightler-McDaniels and Hendrickson 176) to insert these images into a space that is not usually one of critical race and gender analysis. Popular on Fridays, the hashtag #TGIF usually stands for “thank God it’s Friday,” but Vaid-Menon uses the ubiquitous hashtag to mean “thank goddess I’m femme.” As a result, the “thank god it’s Friday” hashtag introduces unsuspecting users to Vaid-Menon’s #TGIF selfies and their interrogation of the racialised politics of hair. Through inserting critical analysis of race and gender within such a light-hearted, non-serious hashtag, that is, by capitalising on the popularity of #TGIF, Vaid-Menon appears to defy the norms of discursive consistency within social media discourse (Rightler-McDaniels and Hendrickson 187) while simultaneously enhancing their personal brand (Banet-Weiser 59). Beyond hashtags, Vaid-Menon’s captions elaborate on the distinct pressures they experience around body hair, discussing how their body hair simultaneously obscures their ability to be recognised as femme and makes their race hyper-visible. In the caption on one #TGIF post, Vaid-Menon writes that, when they began shaving at age 13, it was an attempt at “becoming white.” Now, they write, they face pressure to authenticate their transfemininity by shaving, noting that, in this case, authenticity requires “invisibilization” (AlokVMenon, 15 November 2016).Vaid-Menon continues this theme in another selfie post, again problematising the supposedly direct relationship between authenticity and visibility. This example—in which Vaid-Menon poses against a violet background wearing a curly, blonde wig (AlokVMenon, 9 January 2016 b) their thick, dark hair contrasting strongly with the wig’s light gold—aims to critique the signifying power of the blonde wig.From the hyper-saturated colours, to the bright gold nose rings, to Vaid-Menon’s body hair, the selfie combines—and emphasises—markers of artifice and authenticity, femininity and masculinity. Reinforcing these contradictions, the caption interrogates the relationship between authenticity and visibility, stating “authenticity is a fraught project in a world that ritualizes your invisibilization.” Bringing together weighty concepts that occur in time, the caption speaks of ritual, the project of authenticity, and the process of invisibilisation, yet the selfie itself is a frozen instant, with nothing in the post clarifying what point of these processes, if any, it captures. In the selfie, the hyper-saturated colours highlight the wealth of information that the visual field makes available, but the image itself cannot answer the question of what visible markers, if any, communicate the truth of Vaid-Menon’s authentic identity. As the caption states, also foreclosing any answers, “authenticity is a fraught project,” and, moreover, that authenticity is threatened by what is not visible. While authenticity discourse presumes that the visual field offers the firmest epistemological grounds for assessing and legitimating identity, the visible may not convey the full reality of identity nor experience (Jackson 159). Furthermore, within selfie conventions, visual imperfection usually signifies authenticity (Lobinger and Brantner 1849), but this selfie has characteristics of professional photography, including the studio background, further marking it as a hybrid of authenticity and artifice.Through the intersection of the caption and the selfie, Vaid-Menon therefore casts into question the ability of the visual to successfully signify authentic identity. Thus, the caption reinforces and extends the work that the selfie does to trouble the coherence of Vaid-Menon’s identity. It should be noted, however, that this caption simultaneously participates in the production of Vaid-Menon’s personal brand, investing in a distinct mode of authenticity that Sarah Banet-Weiser has dubbed “AuthenticityTM,” an authenticity that is available to artists precisely through their creative and performative rejection of social norms (119–20). Refusing such normative assumptions about the relationship between hair, race, and gender, the caption and the selfie therefore position the blonde wig as simultaneously artificial and authentic.The tension between artifice and authenticity is explored further by Vaid-Menon in a set of two videos exploring the symbolism of the blonde wig, both captioned with an emoji of a blonde, white woman (AlokVMenon, 11 January 2016; 12 January 2016). By doubling the image of blonde hair within the caption—through the emoji that operates, rebus-like, as a substitute for language—these two captions shift the function of the blonde wig from a tactile, experiential object to an abstracted symbol of white womanhood. In the videos, Vaid-Menon, in character as “Becky” (Kelly) plays with the wig while delivering a monologue full of stereotypes about white women, a monologue that is summed up by the static, cartoonish emoji. As the visual spreads from the photograph into the space of the caption, the caption emphasises the symbolic—as opposed to the tactile or realist—function of the photographed wig.Across the series with the blonde wig, this shift from experiential object to abstract symbol happens primarily through the captions, although it also extends to the images. For example, accompanying the slow-motion video, the first caption puns “blondes have more funding” as the slow-motion video shows Vaid-Menon enjoying the physical sensation of the blonde curls. The slow-motion video creates an endless, looping present as its 7-second runtime repeats over and over, drawing our attention to the materiality of time and touch through the slow-motion effect. In the close, frontal framing of the video, the viewer does not see the pleasure of Vaid-Menon’s hand touching the wig itself, but rather its effect, as the curls fall slowly against Vaid-Menon’s cheek. Meanwhile, the punning caption is also concerned with texture, experience, and effect, drawing the viewer in to the texture of language. While the video stages an intimate, haptic pleasure, the selfie, posted later that same day, displays the wig, stressing what it might represent, rather than how it moves or how it feels. In the selfie, Vaid-Menon poses with one hand raised, caught in the act of twirling a curl, and the caption moves away from the pleasures of wordplay to a more overt political stance—“authenticity is fraught.” Here, their hand seems to pull the hair away from Vaid-Menon’s face, interrupting the sensuous intimacy of curls against their face.These selfie captions assert not only that cultural constructs make authentic visibility fraught for minoritised subjects, but, through the “transparent and economical” emoji (Bloom 248), these selfie videos and their emoji captions also serve to mediate blonde, white womanhood. As the image of the blonde wig proliferates, moving into the space of the caption, the final video selfie also introduces a second character, a white woman, presumably cisgender, wearing a different blonde wig, who appears suddenly behind Vaid-Menon.This tall, skinny woman with corkscrew blonde curls approaches the viewer with curiosity, swaying her body as she walks forward, with her eyes fixed on the camera. Pursing her lips, she produces the facial expression commonly described as “duckface,” a feminised facial expression that is common in selfies and marks the performative—rather than unmediated—self-expression they make possible. As she approaches Vaid-Menon and the camera, she ends up half-in and half-out of frame, lingering at the edge of our vision. Her presence has a disquieting and jarring effect, as Vaid-Menon continues their monologue without acknowledging her, despite the fact that she must be visible to Vaid-Menon on their cell phone screen. Then, because the video is a loop, the monologue ends abruptly, and the video restarts. As Vaid-Menon performs the role of Becky, the white woman who hovers eerily behind Vaid-Menon in the final video is pushed to the edge of the frame and ultimately vanishes at the moment of the loop. The structure of the loop is a provocative approach to questions of visibility, given that visibility politics asserts that visibility is teleologically directed toward future change, while in fact visibility politics reproduces the status quo that it makes visible (Keeling 33). Here, since Vaid-Menon only manages to displace “Becky” by enacting her (over and over), the final result is not (yet) an uncomplicated or uncompromised brown trans femme visibility.By staging the incoherence of claims to visible authenticity, Vaid-Menon’s selfie captions foreclose the possibility of successfully “passing” into coherent identity categories. In the series of posts with the blonde wig, Vaid-Menon never succeeds in seamlessly embodying any single identity category, and these tensions appear within the images as well as in the relationship between image and caption. This failure to “pass” is political, and as J. Jack Halberstam writes, there is a queer art to failure, for “under certain circumstances failing, losing, forgetting, unmaking, undoing, unbecoming, not knowing may in fact offer more creative, more cooperative, more surprising ways of being in the world” (2–3).Failure is also a critical aesthetic element in social media humour, with the hashtag #fail curating posts that ironically celebrate mistakes and failures (Zappavigna 152). In selfies and selfie captions, Vaid-Menon revels in the queer art of social media failure. For example, in a selfie posted on 23 December 2016, Vaid-Menon stares solemnly past the camera, wearing vibrant, contrasting colours, including a bobbed purple wig, bright yellow lipstick, and a dress covered with bright, multi-coloured polka dots. The caption on this colourful, clearly queer, photograph proclaims that Vaid-Menon is “str8 acting looking for same #discrete” (AlokVMenon, 23 December 2016).Everything in the caption operates as a promise that will never be fulfilled, as even the hashtag—#discrete—fails to connect the selfie to other, similar images, as this hashtag is populated by a wildly heterogenous mix of images ranging from sexual images, to landscape photography, to images of fashionable, modern homes. Here, Vaid-Menon participates in a common social media practice, subverting the utility of hashtags and using them as paratextual commentary rather than as tools for networked cataloguing. In this post, Vaid-Menon’s failure to conform to the standards of homonormativity—which would require Vaid-Menon to appear “straight-acting” and to be able to promise discretion to a lover—is pushed to excess, producing a glorious rainbow of queer failure. Similarly, in the series of posts featuring the blonde wig, Vaid-Menon’s campy, parodic version of blonde, white womanhood does not simply demonstrate soberly that the standards imposed by white supremacy and heterocispatriarchy are unreachable. Instead, the series produces this attempt to pass into acceptable white femininity as a strange, delirious failure, accompanied by brilliant colours, strobing slow-motion, and punning, incisive captions.ConclusionIn Vaid-Menon’s Instagram posts, selfies and their captions interrogate and challenge the assumption that authentic identity can transparently be made legible through selfies. Through hashtags, Vaid-Menon’s captions draw upon the resources of the social media platform to connect their selfies to a network of other—not necessarily similar—images, inserting their “thank goddess I’m femme” selfies amid the wealth of “thank god it’s Friday” Instagram posts. And, by using emojis as captions, Vaid-Menon undermines the ability of the caption to anchor the visual to coherent meaning by substituting images for language.Through images and captions, Vaid-Menon’s Instagram selfies restage the act of direct, immediate self-expression as a complicated negotiation of the mediating pressures of language, social media platforms, digital photography, and, ultimately, culture. Furthermore, although selfies are celebrated in popular culture and online activism for the “visibility” they seem to make possible, Vaid-Menon’s selfie captions indicate that social media can do far more than simply promulgate visibility politics. This is necessary, for, despite its compelling lure, visibility politics not only neglects to imagine alternative futures, but actually limits future possibilities through its focus on the present, which is inevitably shaped by the past (Keeling 23). Instead, while building their personal brand on Instagram, Vaid-Menon simultaneously uses selfies and selfie captions to interrogate visibility politics from within one of its most popular contemporary tools, exposing the limitations and compromises of “visibility.” Rather than merely a tool for representation, Vaid-Menon’s work demonstrates how selfies and selfie captions can produce theories of, and about, representation.ReferencesAlokVMenon. Instagram post. 9 January 2016 a. <https://www.instagram.com/p/BAVG-Q3Olqs>.AlokVMenon. Instagram post. 9 January 2016 b. <https://www.instagram.com/p/BAVSkAoOlmF>.AlokVMenon. Instagram post. 11 January 2016. <https://www.instagram.com/p/BAa-CXnOlla>.AlokVMenon. Instagram post. 12 January 2016. <https://www.instagram.com/p/BAdDLiCulhe>.AlokVMenon. Instagram post. 19 August 2016. <https://www.instagram.com/p/BJTp9cNhBPI>.AlokVMenon. Instagram post. 15 November 2016. <https://www.instagram.com/p/BM27QEFAmBu>.AlokVMenon. Instagram post. 23 December 2016. <https://www.instagram.com/p/BOWp_S3glq7>.Banet-Weiser, Sarah. Authentic TM: The Politics and Ambivalence in a Brand Culture. New York: New York UP, 2012.Bloom, Lynn Z. “Critical Emoticons.” Symplokē 18.1-2 (2010): 247–249.David, Emmanuel. “Trans Visibility, Corporate Capitalism, and Commodity Culture.” TSQ: Transgender Studies Quarterly 4.1 (2017): 28–44.Duguay, Stephanie. “Lesbian, Gay, Bisexual, Trans, and Queer Visibility through Selfies: Comparing Platform Mediators across Ruby Rose’s Instagram and Vine Presence.” Social Media + Society 2.2 (2016): 1–12.Halberstam, J. Jack. The Queer Art of Failure. Durham: Duke UP, 2011.Jackson, John L. Real Black: Adventures in Racial Sincerity. Chicago: U of Chicago P, 2005.Jackson, Sarah J., Moya Bailey, and Brooke Foucault Welles. “#GirlsLikeUs: Trans Advocacy and Community Building Online.” New Media & Society (2017), 1–21. DOI: 10.1177/1461444817709276.Keeling, Kara. The Witch's Flight: The Cinematic, the Black Femme, and the Image of Common Sense. Durham: Duke UP, 2007.Kelly, Cara. “What Does Becky Mean? Here's the History behind Beyoncé's 'Lemonade' Lyric That Sparked a Firestorm.” USA Today 27 April 2016. <https://www.usatoday.com/story/life/entertainthis/2016/04/27/what-does-becky-mean-heres-history-behind-beyoncs-lemonade-lyric-sparked-firestorm/83555996>.Lobinger, K., and C. Brantner. “In the Eye of the Beholder: Subjective Views on the Authenticity of Selfies.” International Journal of Communication 9 (2015): 1848–1860.Rancière, Jacques. Dissensus: On Politics and Aesthetics. London: Bloomsbury Academic, 2013.Rightler-McDaniels, Jodi L., and Elizabeth M. Hendrickson. “Hoes and Hashtags: Constructions of Gender and Race in Trending Topics.” Social Semiotics 24.2 (2013): 175-190.Snorton, C. Riley. Nobody Is Supposed to Know: Black Sexuality on the Down Low. Minneapolis: U of Minnesota P, 2014.Wade, Carrie. “’I Want to Be Visible’: A Queer #DisabledAndCute Photo Gallery.” Autostraddle.com. 20 Feb. 2017 <https://www.autostraddle.com/i-want-to-be-visible-a-queer-disabledandcute-photo-gallery-369532>Zappavigna, Michele. Discourse of Twitter and Social Media: How We Use Language to Create Affiliation on the Web. London: Continuum, 2012.

1. A Minor Philosopher According to Guattari and Deleuze's definition, a 'minor literature' is the literature of a minority that makes use of a major language, a literature which deterritorialises that language and interconnects meanings of the most disparate levels, inseparably mixing and implicating poetic, psychological, social and political issues with each other. In analogy, the Japanese media theorist Toshiya Ueno has refered to Félix Guattari as a 'minor philosopher'. Himself a practicing psychoanalyst, Guattari was a foreigner to the Grand Nation of Philosophy, whose natives mostly treat him like an unworthy bastard. And yet he has established a garden of minor flowers, of mongrel weeds and rhizomes that are as polluting to philosophy as Kafka's writing has been to German literature (cf. Deleuze & Guattari, Kafka). The strategies of 'being minor' are, as exemplified by Guattari's writings (with and without Deleuze), deployed in multiple contexts: intensification, re-functionalisation, estrangement, transgression. The following offers a brief overview over the way in which Guattari conceptualises media, new technologies and art, as well as descriptions of several media art projects that may help to illustrate the potentials of such 'minor machines'. Without wanting to pin these projects down as 'Guattarian' artworks, I suggest that the specific practices of contemporary media artists can point us in the direction of the re-singularising, deterritorialising and subjectifying forces which Guattari indicated as being germane to media technologies. Many artists who work with media technologies do so through strategies of appropriation and from a position of 'being minor': whenever a marginality, a minority, becomes active, takes the word power (puissance de verbe), transforms itself into becoming, and not merely submitting to it, identical with its condition, but in active, processual becoming, it engenders a singular trajectory that is necessarily deterritorialising because, precisely, it's a minority that begins to subvert a majority, a consensus, a great aggregate. As long as a minority, a cloud, is on a border, a limit, an exteriority of a great whole, it's something that is, by definition, marginalised. But here, this point, this object, begins to proliferate ..., begins to amplify, to recompose something that is no longer a totality, but that makes a former totality shift, detotalises, deterritorialises an entity.' (Guattari, "Pragmatic/Machinic") In the context of media art, 'becoming minor' is a strategy of turning major technologies into minor machines. a. Krzysztof Wodiczko (PL/USA): Alien Staff Krzysztof Wodiczko's Alien Staff is a mobile communication system and prosthetic instrument which facilitates the communication of migrants in their new countries of residence, where they have insufficient command of the local language for communicating on a par with the native inhabitants. Alien Staff consists of a hand-held staff with a small video monitor and a loudspeaker at the top. The operator can adjust the height of the staff's head to be at a level with his or her own head. Via the video monitor, the operator can replay pre-recorded elements of an interview or a narration of him- or herself. The recorded material may contain biographical information when people have difficulties constructing coherent narratives in the foreign language, or it may include the description of feelings and impressions which the operator normally doesn't get a chance to talk about. The Staff is used in public places where passers-by are attracted to listen to the recording and engage in a conversation with the operator. Special transparent segments of the staff contain memorabilia, photographs or other objects which indicate a part of the personal history of the operator and which are intended to instigate a conversation. The Alien Staff offers individuals an opportunity to remember and retell their own story and to confront people in the country of immigration with this particular story. The Staff reaffirms the migrant's own subjectivity and re-singularises individuals who are often perceived as representative of a homogenous group. The instrument displaces expectations of the majority audience by articulating unformulated aspects of the migrant's subjectivity through a medium that appears as the attractive double of an apparently 'invisible' person. 2. Mass Media, New Technologies and 'Planetary Computerisation' Guattari's comments about media are mostly made in passing and display a clearly outlined opinion about the role of media in contemporary society: a staunch critique of mass media is coupled with an optimistic outlook to the potentials of a post-medial age in which new technologies can develop their singularising, heterogenic forces. The latter development is, as Guattari suggests, already discernible in the field of art and other cultural practices making use of electronic networks, and can lead to a state of 'planetary computerisation' in which multiple new subject-groups can emerge. Guattari consistently refers to the mass media with contempt, qualifying them as a stupefying machinery that is closely wedded to the forces of global capitalism, and that is co-responsible for much of the reactionary hyper-individualism, the desperation and the "state of emergency" that currently dominates "four-fifth of humanity" (Guattari, Chaosmosis 97; cf. Guattari, Drei Ökologien 16, 21). Guattari makes a passionate plea for a new social ecology and formulates, as one step towards this goal, the necessity, "to guide these capitalist societies of the age of mass media into a post-mass medial age; by this I mean that the mass media have to be reappropriated by a multiplicity of subject-groups who are able to administer them on a path of singularisation" (Guattari, "Regimes" 64). Guattari consistently refers to the mass media with contempt, qualifying them as a stupefying machinery that is closely wedded to the forces of global capitalism, and that is co-responsible for much of the reactionary hyper-individualism, the desperation and the "state of emergency" that currently dominates "four-fifth of humanity" (Guattari, Chaosmosis 97; cf. Guattari, Drei Ökologien 16, 21). Guattari makes a passionate plea for a new social ecology and formulates, as one step towards this goal, the necessity, "to guide these capitalist societies of the age of mass media into a post-mass medial age; by this I mean that the mass media have to be reappropriated by a multiplicity of subject-groups who are able to administer them on a path of singularisation" (Guattari, "Regimes" 64). b. Seiko Mikami (J/USA): World, Membrane and the Dismembered Body An art project that deals with the cut between the human subject and the body, and with the deterritorialisation of the sense of self, is Seiko Mikami's World, Membrane and the Dismembered Body. It uses the visitor's heart and lung sounds which are amplified and transformed within the space of the installation. These sounds create a gap between the internal and external sounds of the body. The project is presented in an-echoic room where sound does not reverberate. Upon entering this room, it is as though your ears are no longer living while paradoxically you also feel as though all of your nerves are concentrated in your ears. The sounds of the heart, lungs, and pulse beat are digitised by the computer system and act as parameters to form a continuously transforming 3-d polygonal mesh of body sounds moving through the room. Two situations are effected in real time: the slight sounds produced by the body itself resonate in the body's internal membranes, and the transfigured resonance of those sounds is amplified in the space. A time-lag separates both perceptual events. The visitor is overcome by the feeling that a part of his or her corporeality is under erasure. The body exists as abstract data, only the perceptual sense is aroused. The visitor is made conscious of the disappearance of the physical contours of his or her subjectivity and thereby experiences being turned into a fragmented body. The ears mediate the space that exists between the self and the body. Mikami's work fragments the body and its perceptual apparatus into data, employing them as interfaces and thus folding the body's horizon back onto itself. The project elucidates the difference between an actual and a virtual body, the actual body being deterritorialised and projected outwards towards a number of potential, virtual bodies that can, in the installation, be experienced as maybe even more 'real' than the actual body. 3. Artistic Practice Guattari's conception of post-media implies criss-crossing intersections of aesthetic, ethical, political and technological planes, among which the aesthetic, and with it artistic creativity, are ascribed a position of special prominence. This special role of art is a trope that recurs quite frequently in Guattari's writings, even though he is rarely specific about the artistic practices he has in mind. In A Thousand Plateaus, Deleuze and Guattari give some detailled attention to the works of artists like Debussy, Boulez, Beckett, Artaud, Kafka, Kleist, Proust, and Klee, and Chaosmosis includes longer passages and concrete examples for the relevance of the aesthetic paradigm. These examples come almost exclusively from the fields of performing arts, music and literature, while visual arts are all but absent. One reason for this could be that the performing arts are time-based and processual and thus lend themselves much better to theorisation of flows, transformations and differentiations. The visual arts can be related to the abstract machine of faciality (visageité) which produces unified, molar, identical entities out of a multiplicity of different singularities, assigning them to a specific category and associating them with particular social fields (cf. Deleuze & Guattari, Tausend Plateaus 167-91) This semiotic territorialisation is much more likely to happen in the case of static images, whether two- or three-dimensional, than in time-based art forms. An interesting question, then, would be whether media art projects, many of which are time-based, processual and open-ended, can be considered as potential post-medial art practices. Moreover, given the status of computer software as the central motor of the digital age, and the crucial role it plays in aesthetic productions like those discussed here, software may have to be viewed as the epitome of post-medial machines. Guattari seems to have been largely unaware of the beginnings of digital media art as it developed in the 1980s. In generalistic terms he suggests that the artist is particularly well-equipped to conceptualise the necessary steps for this work because, unlike engineers, he or she is not tied to a particular programme or plan for a product, and can change the course of a project at any point if an unexpected event or accident intrudes (cf. Guattari, Drei Ökologien 50). The significance of art for Guattari's thinking comes primarily from its close relation with processes of subjectivation. "Just as scientific machines constantly modify our cosmic frontiers, so do the machines of desire and aesthetic creation. As such, they hold an eminent place within assemblages of subjectivation, themselves called to relieve our old social machines which are incapable of keeping up with the efflorescence of machinic revolutions that shatter our epoch' (Guattari, Chaosmosis 54). The aesthetic paradigm facilitates the development of new, virtual forms of subjectivity, and of liberation, which will be adequate to these machinic revolutions. c. Knowbotic Research + cF: IO_Dencies The Alien Staff project was mentioned as an example for the re-singularisation and the virtualisation of identity, and World, Membrane and the Dismembered Body as an instance of the deterritorialisation and virtualisation of the human body through an artistic interface. The recent project by Knowbotic Research, IO_Dencies -- Questioning Urbanity, deals with the possibilities of agency, collaboration and construction in translocal and networked environments. It points in the direction of what Guattari has called the formation of 'group subjects' through connective interfaces. The project looks at urban settings in different megacities like Tokyo, São Paulo or the Ruhr Area, analyses the forces present in particular local urban situations, and offers experimental interfaces for dealing with these local force fields. IO_Dencies São Paulo enables the articulation of subjective experiences of the city through a collaborative process. Over a period of several months, a group of young architects and urbanists from São Paulo, the 'editors', provided the content and dynamic input for a database. The editors collected material (texts, images, sounds) based on their current situation and on their personal urban experience. A specially designed editor tool allowed the editors to build individual conceptual 'maps' in which to construct the relations between the different materials in the data-pool according to the subjective perception of the city. On the computational level, connectivities are created between the different maps of the editors, a process that is driven by algorithmic self-organisation whose rules are determined by the choices that the editors make. In the process, the collaborative editorial work in the database generates zones of intensities and zones of tension which are visualised as force fields and turbulences and which can be experienced through interfaces on the Internet and at physical exhibition sites. Participants on the Net and in the exhibition can modify and influence these electronic urban movements, force fields and intensities on an abstract, visual level, as well as on a content-based, textual level. This engagement with the project and its material is fed back into the database and influences the relational forces within the project's digital environment. Characteristic of the forms of agency as they evolve in networked environments is that they are neither individualistic nor collective, but rather connective. Whereas the collective is determined by an intentional and empathetic relation between agents within an assemblage, the connective rests on any kind of machinic relation and is therefore more versatile, more open, and based on the heterogeneity of its components or members. In the IO_Dencies interfaces, the different networked participants become visible for each other, creating a trans-local zone of connective agency. The inter-connectedness of their activities can be experienced visually, acoustically, and through the constant reconfiguration of the data sets, an experience which can become the basis of the formation of a specific, heterogeneous group subject. 4. Guattari's Concept of the Machinic An important notion underlying these analyses is that of the machine which, for Guattari, relates not so much to particular technological or mechanical objects, to the technical infrastructure or the physical flows of the urban environment. 'Machines' can be social bodies, industrial complexes, psychological or cultural formations, they are assemblages of heterogeneous parts, aggregations which transform forces, articulate and propel their elements, and force them into a continuous state of transformation and becoming. An important notion underlying these analyses is that of the machine which, for Guattari, relates not so much to particular technological or mechanical objects, to the technical infrastructure or the physical flows of the urban environment. 'Machines' can be social bodies, industrial complexes, psychological or cultural formations, they are assemblages of heterogeneous parts, aggregations which transform forces, articulate and propel their elements, and force them into a continuous state of transformation and becoming. d. Xchange Network My final example is possibly the most evocative in relation to Guattari's notions of the polyvocity and heterogenesis that new media technologies can trigger. It also links up closely with Guattari's own engagement with the minor community radio movement. In late 1997, the E-Lab in Riga initiated the Xchange network for audio experiments on the Internet. The participating groups in London, Ljubljana, Sydney, Berlin, and many other minor and major places, use the Net for distributing their original sound programmes. The Xchange network is "streaming via encoders to remote servers, picking up the stream and re-broadcasting it purely or re-mixed, looping the streams" (Rasa Smite). Xchange is a distributed group, a connective, that builds creative cooperation in live-audio streaming on the communication channels that connect them. They explore the Net as a sound-scape with particular qualities regarding data transmission, delay, feedback, and open, distributed collaborations. Moreover, they connect the network with a variety of other fields. Instead of defining an 'authentic' place of their artistic work, they play in the transversal post-medial zone of media labs in different countries, mailing lists, net-casting and FM broadcasting, clubs, magazines, stickers, etc., in which 'real' spaces and media continuously overlap and fuse (cf. Slater). 5. Heterogenic Practices If we want to understand the technological and the political implications of the machinic environment of the digital networks, and if we want to see the emergence of the group subjects of the post-media age Guattari talks about, we have to look at connectives like Xchange and the editor-participant assemblages of IO_Dencies. The far-reaching machinic transformations which they articulate, hold the potential of what Guattari refers to as the 'molecular revolution'. To realise this revolution, it is vital to "forge new analytical instruments, new concepts, because it is ... the transversality, the crossing of abstract machines that constitute a subjectivity and that are incarnated, that live in very different regions and domains and ... that can be contradictory and antagonistic". For Guattari, this is not a mere theoretical question, but one of experimentation, "of new forms of interactions, of movement construction that respects the diversity, the sensitivities, the particularities of interventions, and that is nonetheless capable of constituting antagonistic machines of struggle to intervene in power relations" (Guattari, "Pragmatic/Machinic" 4-5). The implication here is that some of the minor media practices pursued by artists using digital technologies point us in the direction of the positive potentials of post media. The line of flight of such experimentation is the construction of new and strong forms of subjectivity, "an individual and/or collective reconstitution of the self" (Guattari, Drei Ökologien 21), which can strengthen the process of what Guattari calls "heterogenesis, that is a continuous process of resingularisation. The individuals must, at the same time, become solidary and ever more different" (Guattari, Drei Ökologien 76). References Deleuze, Gilles, and Felix Guattari. Kafka: Pour une Litterature Mineur. Paris: Ed. de Minuit, 1975. ---. Tausend Plateaus. (1980) Berlin: Merve, 1992. Guattari, Félix. Cartographies Schizoanalytiques. Paris: Ed. Galilée, 1989. ---. Chaosmosis: An Ethico-Aesthetic Paradigm. (1992) Sydney: Power Publications, 1995. ---. Die drei Ökologien. (1989) Wien: Passagen Verlag, 1994. ---. "Pragmatic/Machinic." Discussion with Guattari, conducted and transcribed by Charles J. Stivale. (1985) Pre/Text 14.3-4 (1995). ---. "Regimes, Pathways, Subjects." Die drei Ökologien. (1989) Wien: Passagen Verlag, 1994. 95-108. ---. "Über Maschinen." (1990) Schmidgen, 115-32. Knowbotic Research. IO_Dencies. 1997-8. 11 Sep. 1999 <http://io.khm.de/>. De Landa, Manuel. "The Machinic Phylum." Technomorphica. Eds. V2_Organisation. Rotterdam: V2_Organisation, 1997. Mikami, Seiko. World, Membrane and the Dismembered Body. 1997. 11 Sep. 1999 <http://www.ntticc.or.jp/permanent/mikami/mikami_e.php>. Schmidgen, Henning, ed. Ästhetik und Maschinismus: Texte zu und von Félix Guattari. Berlin: Merve, 1995. ---. Das Unbewußte der Maschinen: Konzeptionen des Psychischen bei Guattari, Deleuze und Lacan. München: Fink, 1997. Slater, Howard. "Post-Media Operators." Nettime, 10 June 1998. 11 Sep. 1999 <http://www.factory.org>. Wodiczko, Krzysztof. 11 Sep. 1999 <http://cavs.mit.edu/people/kw.htm>. Xchange. 11 Sep. 1999 <http://xchange.re-lab.net>. (Note: An extended, Dutch version of this text was published in: Oosterling/Thissen, eds. Chaos ex Machina: Het ecosofisch Werk van Félix Guattari op de Kaart Gezet. Rotterdam: CFK, 1998. Citation reference for this article MLA style: Andreas Broeckmann. "Minor Media -- Heterogenic Machines: Notes on Félix Guattari's Conceptions of Art and New Media." M/C: A Journal of Media and Culture 2.6 (1999). [your date of access] <http://www.uq.edu.au/mc/9909/minor.php>. Chicago style: Andreas Broeckmann, "Minor Media -- Heterogenic Machines: Notes on Félix Guattari's Conceptions of Art and New Media," M/C: A Journal of Media and Culture 2, no. 6 (1999), <http://www.uq.edu.au/mc/9909/minor.php> ([your date of access]). APA style: Andreas Broeckmann. (1999) Minor Media -- Heterogenic Machines: Notes on Félix Guattari's Conceptions of Art and New Media. M/C: A Journal of Media and Culture 2(6). <http://www.uq.edu.au/mc/9909/minor.php> ([your date of access]).