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Abstract Chromatin state is thought to impart regulatory function to the underlying DNA sequence. This can be established through histone modifications and chromatin organisation, but exactly how these factors relate to one another to regulate gene expression is unclear. In this study, we have used super-resolution microscopy to image the Y loops of Drosophila melanogaster primary spermatocytes, which are enormous transcriptionally active chromatin fibres, each representing single transcription units that are individually resolvable in the nuclear interior. We previously found that the Y loops consist of regular clusters of nucleosomes, with an estimated median of 54 nucleosomes per cluster with wide variation. In this study, we report that the histone modifications H3K4me3, H3K27me3, and H3K36me3 are also clustered along the Y loops, with H3K4me3 more associated with diffuse chromatin compared to H3K27me3. These histone modifications form domains that can be stretches of Y loop chromatin micrometres long, or can be in short alternating domains. The different histone modifications are associated with different sizes of chromatin clusters and unique morphologies. Strikingly, a single chromatin cluster almost always only contains only one type of the histone modifications that were labelled, suggesting exclusivity, and therefore regulation at the level of individual chromatin clusters. The active mark H3K36me3 is more associated with actively elongating RNA polymerase II than H3K27me3, with polymerase often appearing on what are assumed to be looping regions on the periphery of chromatin clusters. These results provide a foundation for understanding the relationship between chromatin state, chromatin organisation, and transcription regulation – with potential implications for pause-release dynamics, splicing complex organisation and chromatin dynamics during polymerase progression along a gene.

Abstract Children experiencing educational neglect are likely to experience the harm associated with adverse life experiences and a range of emotional and psychological challenges. Using attachment theory and psychosocial frameworks, we devised and implemented an intervention designed to ameliorate the deleterious effects of challenging behaviours in an elementary school situated in a deprived neighbourhood of Istanbul, Turkey. For a period of eight months, 160 pupils received a psychosocial intervention once a week. Children’s patterns of behaviour, emotions, movements, interactivity, socialisation and interpersonal communications were observed during this period. Core activities of the intervention included library visits, reading, writing and listening games, maths games, drawing, animal animation, leaf making, ball games, parachuting games, colouring, hula-hoop and driver-car role plays. At the end of the intervention, one group of children made significant improvements in behavioural changes while no improvements were observed for another group. Although further research is required to generalize beyond the reference group, the findings suggest that a robust collaboration between inter-agency community partnership and universities can play a crucial role in responding to the needs of marginalized children with psychological and emotional problems.

While the biochemistry of gene transcription has been well studied, our understanding of how this process is organised in 3D within the intact nucleus is less well understood. Here we investigate the structure of actively transcribed chromatin and the architecture of its interaction with active RNA polymerase. For this analysis, we have used super-resolution microscopy to image the Drosophila melanogaster Y loops which represent huge, several megabases long, single transcription units. The Y loops provide a particularly amenable model system for transcriptionally active chromatin. We find that, although these transcribed loops are decondensed they are not organised as extended 10nm fibres, but rather they largely consist of chains of nucleosome clusters. The average width of each cluster is around 50nm. We find that foci of active RNA polymerase are generally located off the main fibre axis on the periphery of the nucleosome clusters. Foci of RNA polymerase and nascent transcripts are distributed around the Y loops rather than being clustered in individual transcription factories. However, as the RNA polymerase foci are considerably less prevalent than the nucleosome clusters, the organisation of this active chromatin into chains of nucleosome clusters is unlikely to be determined by the activity of the polymerases transcribing the Y loops. These results provide a foundation for understanding the topological relationship between chromatin and the process of gene transcription.

A 9-year-old male neutered Cockapoo was presented with an acute and progressive history of exercise induced weakness involving all limbs, and bilateral decreased ability to blink. Investigations revealed generalized myasthenia gravis alongside the presence of a thymoma and a cholangiocellular carcinoma. Symptomatic treatment through pyridostigmine bromide was used to control clinical signs, and complete surgical removal of the thymoma and cholangiocellular carcinoma was performed. Serum acetylcholine receptor antibody concentration was measured serially. Clinical remission defined as resolution of clinical signs alongside discontinuation of treatment was achieved by day 251 (8.2 months). Immune remission defined as normalization of serum acetylcholine receptor antibody concentration alongside resolution of clinical signs and discontinuation of treatment was achieved by day 566 (18.5 months). Neurological examination was normal, and the owners did not report any clinical deterioration during the final follow-up appointment on day 752 (24 months), hence outcome was considered excellent. This is the first report describing the temporal evolution of serum acetylcholine receptor antibody concentration in a dog with thymoma-associated myasthenia gravis which achieved immune remission following thymectomy. Treatment was successfully discontinued without any evidence of clinical deterioration thereafter despite serum acetylcholine receptor antibody concentration not normalizing for another 315 days (10 months).

Change agents are individuals who can successfully transform aspects of how organisations operate. In education, teachers as change agents are increasingly seen as vital to the successful operation of schools and self-improving school systems. To date, however, there has been no systematic investigation of the nature and role of teacher change agents. To address this knowledge gap, we undertook a systematic review into five key areas regarding teachers as change agents. After reviewing 70 outputs we found that current literature predominantly positions teacher change agents as the deliverers of top-down change, with the possibility of bottom-up educational reform currently neglected.

Vaginal mucosal surfaces naturally offer some protection against sexually transmitted infections (STIs) including Human Immunodeficiency Virus-1, however topical preventative medications or vaccine designed to boost local immune responses can further enhance this protection. We previously developed a novel mucosal vaccine strategy using viral vectors integrated into mouse dermal epithelium to induce virus-specific humoral and cellular immune responses at the site of exposure. Since vaccine integration occurs at the site of cell replication (basal layer 100-400 micrometers below the surface), temporal epithelial thinning during vaccine application, confirmed with high resolution imaging, is desirable. In this study, strategies for vaginal mucosal thinning were evaluated noninvasively using optical coherence tomography (OCT) to map reproductive tract epithelial thickness (ET) in macaques to optimize basal layer access in preparation for future effective intravaginal mucosal vaccination studies. Twelve adolescent female rhesus macaques (5-7kg) were randomly assigned to interventions to induce vaginal mucosal thinning, including cytobrush mechanical abrasion, the chemical surfactant spermicide nonoxynol-9 (N9), the hormonal contraceptive depomedroxyprogesterone acetate (DMPA), or no intervention. Macaques were evaluated at baseline and after interventions using colposcopy, vaginal biopsies, and OCT imaging, which allowed for real-time in vivo visualization and measurement of ET of the mid-vagina, fornices, and cervix. P value ≤0.05 was considered significant. Colposcopy findings included pink, rugated tissue with variable degrees of white-tipped, thickened epithelium. Baseline ET of the fornices was thinner than the cervix and vagina (p

Abstract Background Hox transcription factors specify segmental diversity along the anterior-posterior body axis in metazoans. While the different Hox family members show clear functional specificity in vivo, they all show similar binding specificity in vitro and a satisfactory understanding of in vivo Hox target selectivity is still lacking. Results Using transient transfection in Kc167 cells, we systematically analyze the binding of all eight Drosophila Hox proteins. We find that Hox proteins show considerable binding selectivity in vivo even in the absence of canonical Hox cofactors Extradenticle and Homothorax. Hox binding selectivity is strongly associated with chromatin accessibility, being highest in less accessible chromatin. Individual Hox proteins exhibit different propensities to bind less accessible chromatin, and high binding selectivity is associated with high-affinity binding regions, leading to a model where Hox proteins derive binding selectivity through affinity-based competition with nucleosomes. Extradenticle/Homothorax cofactors generally facilitate Hox binding, promoting binding to regions in less accessible chromatin but with little effect on the overall selectivity of Hox targeting. These cofactors collaborate with Hox proteins in opening chromatin, in contrast to the pioneer factor, Glial cells missing, which facilitates Hox binding by independently generating accessible chromatin regions. Conclusions These studies indicate that chromatin accessibility plays a key role in Hox selectivity. We propose that relative chromatin accessibility provides a basis for subtle differences in binding specificity and affinity to generate significantly different sets of in vivo genomic targets for different Hox proteins.

The positive promotion of social and emotional wellbeing in schools can be seen as a means to lessen the impact of social difficulties and behavioral problems on educational engagement. This study employed a multicomponent, whole school approach to promoting positive social and emotional wellbeing. School personnel identified nine primary Key Stage 2 pupils (ages 8 - 10) who were at risk of exclusion due to their disruptive and aggressive behavior. These pupils attended five maintained primary schools located across England. Following analysis of four baseline data points for stability, the Building Schools of Character (BSC) program was commenced. During this period, classroom observations were conducted weekly over the course of one academic school year. All pupils showed a marked decrease in disruptive behavior and were no longer considered at risk of exclusion by staff. These findings suggest a whole school positive social and emotional development approach can decrease disruptive behavior in the classroom.

As large migratory caribou herds decline globally and regional climate trends point to a warmer future, there is a need and a legislative requirement to ensure impacts of industrial development are fully assessed, particularly with respect to cumulative effects. In this paper we use a current proposal, the potential leasing of the 1002 lands on the Alaskan Arctic coastal plain of the Arctic National Wildlife Refuge for hydrocarbon development, to project the potential cumulative effects on the international Porcupine Caribou Herd. Using the caribou cumulative effects model, an existing decision support tool, we evaluate six alternative development scenarios for the 1002 lands, ranging from no leasing to full leasing with standard mitigation conditions. Compared to the no leasing option, at the current population size (218,000 caribou), our analysis projected that the likelihood of a herd decline over a 10-year period would increase from 3% to 19% depending on the leasing scenarios analyzed. This compares to an increased probability of decline from 11% to 26% if the starting population was 100,000, indicative of population estimates in the early 1970s. Our approach accomplishes one of the main steps in a comprehensive cumulative effects assessment, namely the quantification of past, present, and foreseeable future projects on a valued ecosystem component, the Porcupine Caribou Herd. We suggest the testing of underlining assumptions and refinements of the model required to more fully estimate the impacts of development. The use of transparent, quantitative decision support tools in assessing industrial development impacts on Arctic wildlife becomes more critical as climatic changes to Arctic landscapes accelerate.

OBJECTIVES/GOALS: The new CLIC Education & Career Development Gateway aims to be a translational science workforce ecosystem for CTSAs to share learning and training resources and career opportunities. The Gateway also provides individualized assistance to identify and implement TS learning and training resources. METHODS/STUDY POPULATION: The CLIC Education & Career Development Gateway, located on the CLIC website, is an entry way to: 1) the Education Clearinghouse, a platform where CTSA Program hubs can find and share educational resources individually or as part of resource kits; 2) the Opportunities Board, which includes jobs and mini-sabbaticals from CTSA Program hubs; and 3) the Education & Training Navigator, a personalized approach to education and training requests. These approaches help empower and support a cooperative learning and training community that is inclusive and collaborative, facilitating and amplifying opportunities for the sharing of educational resources throughout the translational science workforce. RESULTS/ANTICIPATED RESULTS: Through a person-centered, direct engagement approach, the anticipated outcomes of these efforts are to promote increased collaboration across CTSA Program Hubs and partners, and the amplification of accessible, relevant existing resources. Another anticipated outcome is increased production of educational materials through the reduction of work duplication and identification of gaps in education and training resources. The Gateway also provides an opportunity to communicate the work and efforts that consortium-level special groups (working groups, special interest groups, etc.) produce. Ongoing evaluations and suggestions will help determine future improvements and functionalities. DISCUSSION/SIGNIFICANCE OF IMPACT: CLIC’s education and training ecosystem promotes education as a community space to facilitate opportunities for collaboration and partnerships, amplifying visibility of the work created by members of the CTSA community, and encouraging a transformative career trajectory for trainees and scholars.

Stakeholders in healthcare are increasingly turning to real world evidence (RWE) to inform their decisions, alongside evidence from randomized controlled trials. RWE is generated by analysing data gathered from routine clinical practice, and can be used across the product lifecycle, providing insights into areas including disease epidemiology, treatment effectiveness and safety, and health economic value and impact. Recently, the US Food and Drug Administration and the European Medicines Agency have stated their ambition for greater use of RWE to support applications for new indications, and are now consulting with their stakeholders to formalize standards and expected methods for generating RWE. Pharmaceutical companies are responding to the increasing demands for RWE by developing standards and processes for each stage of the evidence generation pathway. Some conventions are already in place for assuring quality, whereas other processes are specific to the research question and data sources available. As evidence generation increasingly becomes a core role of medical affairs divisions in large pharmaceutical companies, standards of rigour will continue to evolve and improve. Senior pharmaceutical leaders can drive this change by making RWE a core element of their corporate strategy, providing top-level direction on how their respective companies should approach RWE for maximum quality. Here, we describe the current and future areas of RWE application within the pharmaceutical industry, necessary access to data to generate RWE, and the challenges in communicating RWE. Supporting and building on viewpoints from industry and publicly funded research, our perspective is that at each stage of RWE generation, quality will be critical to the impact that RWE has on healthcare decision-makers; not only where RWE is an established and evolving tool, but also in new areas that have the potential to disrupt and to improve drug development pathways.

Stakeholders in healthcare are increasingly turning to real world evidence (RWE) to inform their decisions, alongside evidence from randomized controlled trials. RWE is generated by analysing data gathered from routine clinical practice, and can be used across the product lifecycle, providing insights into areas including disease epidemiology, treatment effectiveness and safety, and health economic value and impact. Recently, the US Food and Drug Administration and the European Medicines Agency have stated their ambition for greater use of RWE to support applications for new indications, and are now consulting with their stakeholders to formalize standards and expected methods for generating RWE. Pharmaceutical companies are responding to the increasing demands for RWE by developing standards and processes for each stage of the evidence generation pathway. Some conventions are already in place for assuring quality, whereas other processes are specific to the research question and data sources available. As evidence generation increasingly becomes a core role of medical affairs divisions in large pharmaceutical companies, standards of rigour will continue to evolve and improve. Senior pharmaceutical leaders can drive this change by making RWE a core element of their corporate strategy, providing top-level direction on how their respective companies should approach RWE for maximum quality. Here, we describe the current and future areas of RWE application within the pharmaceutical industry, necessary access to data to generate RWE, and the challenges in communicating RWE. Supporting and building on viewpoints from industry and publicly funded research, our perspective is that at each stage of RWE generation, quality will be critical to the impact that RWE has on healthcare decision-makers; not only where RWE is an established and evolving tool, but also in new areas that have the potential to disrupt and to improve drug development pathways.

Beginning in 1969, the Provisional Irish Republican Army conducted a paramilitary campaign designed to unite Northern Ireland with the Republic of Ireland, creating a 32 county democratic socialist republic. The Provisional IRA’s campaign officially ended in 2005, but former Provisionals and others who followed them continue to pursue armed struggle to this day. The Provisional IRA and its successors are part of the centuries old and highly documented “resistance” of Irish people to British interference in Ireland. Over those centuries, state authorities ‒ the British, Irish, and Northern Irish governments ‒ have “resisted” the dissent of Irish Republicans. This paper draws on three different research methodologies available to social scientists ‒ counts of events that inform quantitative analyses, intensive interviews/oral histories, and visual sociology ‒ and argues that a multi-method approach will provide a better understanding of the dynamics of “resistance” in Ireland and, more generally, social protest.

It is widely recognized that school-aged children in most European countries are watching an average of 2.5 to 3.0 hours of television every day. If we add the time spent with radio, newspapers, internet, magazines, and other leisure-time media, the total amount of time children aged 9-15 spend with media is estimated to be about four to six hours every day. This may be a matter of alarm to some teachers and parents who see students spending less time reading or doing school assignments. They fear that children will not be involved in a balanced set of activities that will develop social skills, commitment to family and Church, and creativity-building hobbies.

Powszechnie wiadomo, że dzieci w wieku szkolnym w większości krajów europejskich codziennie oglądają telewizję przez około 2.5 do 3 godzin. Jeśli do tego dodamy czas spędzony na słuchaniu radia, czytaniu gazet i kolorowych magazynów, „błądzeniu” po Internecie i korzystaniu z innych mediów „wolnego czasu”, to całkowity czas poświęcony mediom przez dzieci w wieku 9-15 lat należałoby oszacować na 4-6 godzin dziennie. Dla niektórych nauczycieli i rodziców, którzy zauważają, że uczniowie spędzają mniej czasu na czytaniu i odrabianiu lekcji, może to być wręcz sytuacja alarmowa. Obawiają się oni, że dzieci nie zostaną objęte zajęciami rozwijającymi sprawności społeczne, zaangażowanie w rodzinę i Kościół oraz pobudzające twórczo hobby.

It is now well established that eukaryote genomes have a common architectural organization into topologically associated domains (TADs) and evidence is accumulating that this organization plays an important role in gene regulation. However, the mechanisms that partition the genome into TADs and the nature of domain boundaries are still poorly understood. We have investigated boundary regions in the Drosophila genome and find that they can be identified as domains of very low H3K27me3. The genome-wide H3K27me3 profile partitions into two states; very low H3K27me3 identifies Depleted (D) domains that contain housekeeping genes and their regulators such as the histone acetyltransferase-containing NSL complex, whereas domains containing moderate-to-high levels of H3K27me3 (Enriched or E domains) are associated with regulated genes, irrespective of whether they are active or inactive. The D domains correlate with the boundaries of TADs and are enriched in a subset of architectural proteins, particularly Chromator, BEAF-32, and Z4/Putzig. However, rather than being clustered at the borders of these domains, these proteins bind throughout the H3K27me3-depleted regions and are much more strongly associated with the transcription start sites of housekeeping genes than with the H3K27me3 domain boundaries. While we have not demonstrated causality, we suggest that the D domain chromatin state, characterised by very low or absent H3K27me3 and established by housekeeping gene regulators, acts to separate topological domains thereby setting up the domain architecture of the genome.

True to one of the themes in this issue of our journal, my essay raises two broadly pedagogical questions. First, what did Shakespeare learn about writing sonnets from his most accomplished contemporaries, Sidney and Spenser, and secondly what does he teach his own readers about how to read his sonnets? A single answer to both is proposed: he learned and taught that the abiding and primary subject of the sonnet is the sonnet itself and the kind of beauty it offers to readers. For want of a better term, “autodidactic” is proffered as a clumsy word to describe this two-way process of self-learning and self-teaching.

An economic and environmental feasibility study were carried out on the thermochemical conversion of glycerol to medium methane content biological synthetic natural gas (bio-SNG). A plant that processed 497 kg·h−1 of glycerol to bio-SNG was modelled as an on-site addition to a soybean biodiesel plant based in Missouri (USA) that produced 30 million litres of soybean biodiesel per year. Assuming the glycerol contained only 80 wt% free glycerol, the bio-SNG could substitute up to 24% of the natural gas at the soybean biodiesel plant. The discounted cash flow analysis showed it was possible to generate positive NPVs and achieve internal rates of return within the hurdle rate (12%) for biomass gasification technologies. From the environmental analysis it was found that the bio-SNG could reduce global warming potential by 28% when compared to conventional natural gas in the USA and translates to roughly 7% reduction in biodiesel natural gas emissions, if the maximum 24% of natural gas were to be substituted by bio-SNG. The work highlights the potential to divert waste glycerol to an onsite energy vector at soybean biodiesel plants with minimal change to the main biodiesel production process and potential reductions to soybean biodiesel global warming potential.

BACKGROUND: The cost-effectiveness of initial strategies in managing Canadian patients with uninvestigated upper gastrointestinal symptoms remains controversial.

We describe the first reported transmission to a human of simian foamy virus (SFV) from a free-ranging population of nonhuman primates in Asia. The transmission of an exogenous retrovirus, SFV, from macaques (Macaca fascicularis) to a human at a monkey temple in Bali, Indonesia, was investigated with molecular and serologic techniques. Antibodies to SFV were detected by Western blotting of serum from 1 of 82 humans tested. SFV DNA was detected by nested polymerase chain reaction (PCR) from the blood of the same person. Cloning and sequencing of PCR products confirmed the virus's close phylogenetic relationship to SFV isolated from macaques at the same temple. This study raises concerns that persons who work at or live around monkey temples are at risk for infection with SFV.

In Drosophila, protein trap strategies provide powerful approaches for the generation of tagged proteins expressed under endogenous control. Here, we describe expression and functional analysis to evaluate new Ubx and hth protein trap lines generated by the Cambridge Protein Trap project. Both protein traps exhibit spatial and temporal expression patterns consistent with the reported endogenous pattern in the embryo. In imaginal discs, Ubx-YFP is expressed throughout the haltere and 3rd leg imaginal discs, while Hth-YFP is expressed in the proximal regions of haltere and wing discs but not in the pouch region. The UbxCPTI000601 line is semilethal as a homozygote. No T3/A1 to T2 transformations were observed in the embryonic cuticle or the developing midgut. The homozygous survivors, however, exhibit a weak haltere phenotype with a few wing-like marginal bristles on the haltere capitellum. Although hthCPTI000378 is completely lethal as a homozygote, the hthCPTI000378/hthC1 genotype is viable. Using a hth deletion (Df(3R)BSC479) we show that hthCPTI000378/Df(3R)BSC479 adults are phenotypically normal. No transformations were observed in hthCPTI000378, hthCPTI000378/hthC1, or hthCPTI000378/Df(3R)BSC479 embryonic cuticles. We have successfully characterised the Ubx-YFP and Hth-YFP protein trap lines demonstrating that the tagged proteins show appropriate expression patterns and produce at least partially functional proteins.

Hox genes encode a family of transcription factors that are key developmental regulators with a highly conserved role in specifying segmental diversity along the metazoan body axis. Although they have been shown to regulate a wide variety of downstream processes, direct transcriptional targets have been difficult to identify and this has been a major obstacle to our understanding of Hox gene function. We report the identification of genome-wide binding sites for the Hox protein Ultrabithorax (Ubx) using a YFP-tagged Drosophila protein-trap line together with chromatin immunoprecipitation and microarray analysis. We identify 1,147 genes bound by Ubx at high confidence in chromatin from the haltere imaginal disc, a prominent site of Ubx function where it specifies haltere versus wing development. The functional relevance of these genes is supported by their overlap with genes differentially expressed between wing and haltere imaginal discs. The Ubx-bound gene set is highly enriched in genes involved in developmental processes and contains both high-level regulators as well as genes involved in more basic cellular functions. Several signalling pathways are highly enriched in the Ubx target gene set and our analysis supports the view that Hox genes regulate many levels of developmental pathways and have targets distributed throughout the gene network. We also performed genome-wide analysis of the binding sites for the Hox cofactor Homothorax (Hth), revealing a striking similarity with the Ubx binding profile. We suggest that these binding profiles may be strongly influenced by chromatin accessibility and provide evidence of a link between Ubx/Hth binding and chromatin state at genes regulated by Polycomb silencing. Overall, we define a set of direct Ubx targets in the haltere imaginal disc and suggest that chromatin accessibility has important implications for Hox target selection and for transcription factor binding in general.

Polycomb-group (PcG) and Trithorax-group proteins together form a maintenance machinery that is responsible for stable heritable states of gene activity. While the best-studied target genes are the Hox genes of the Antennapedia and Bithorax complexes, a large number of key developmental genes are also Polycomb (Pc) targets, indicating a widespread role for this maintenance machinery in cell fate determination. We have studied the linkage between the binding of PcG proteins and the developmental regulation of gene expression using whole-genome mapping to identify sites bound by the PcG proteins, Pc and Pleiohomeotic (Pho), in the Drosophila embryo and in a more restricted tissue, the imaginal discs of the third thoracic segment. Our data provide support for the idea that Pho is a general component of the maintenance machinery, since the majority of Pc targets are also associated with Pho binding. We find, in general, considerable developmental stability of Pc and Pho binding at target genes and observe that Pc/Pho binding can be associated with both expressed and inactive genes. In particular, at the Hox complexes, both active and inactive genes have significant Pc and Pho binding. However, in comparison to inactive genes, the active Hox genes show reduced and altered binding profiles. During development, Pc target genes are not simply constantly associated with Pc/Pho binding, and we identify sets of genes with clear differential binding between embryo and imaginal disc. Using existing datasets, we show that for specific fate-determining genes of the haemocyte lineage, the active state is characterised by lack of Pc binding. Overall, our analysis suggests a dynamic relationship between Pc/Pho binding and gene transcription. Pc/Pho binding does not preclude transcription, but levels of Pc/Pho binding change during development, and loss of Pc/Pho binding can be associated with both stable gene activity and inactivity.

Insulator or enhancer-blocking elements are proposed to play an important role in the regulation of transcription by preventing inappropriate enhancer/promoter interaction. The zinc-finger protein CTCF is well studied in vertebrates as an enhancer blocking factor, but Drosophila CTCF has only been characterised recently. To date only one endogenous binding location for CTCF has been identified in the Drosophila genome, the Fab-8 insulator in the Abdominal-B locus in the Bithorax complex (BX-C). We carried out chromatin immunopurification coupled with genomic microarray analysis to identify CTCF binding sites within representative regions of the Drosophila genome, including the 3-Mb Adh region, the BX-C, and the Antennapedia complex. Location of in vivo CTCF binding within these regions enabled us to construct a robust CTCF binding-site consensus sequence. CTCF binding sites identified in the BX-C map precisely to the known insulator elements Mcp, Fab-6, and Fab-8. Other CTCF binding sites correlate with boundaries of regulatory domains allowing us to locate three additional presumptive insulator elements; "Fab-2," "Fab-3," and "Fab-4." With the exception of Fab-7, our data indicate that CTCF is directly associated with all known or predicted insulators in the BX-C, suggesting that the functioning of these insulators involves a common CTCF-dependent mechanism. Comparison of the locations of the CTCF sites with characterised Polycomb target sites and histone modification provides support for the domain model of BX-C regulation.

Effective ecological monitoring is central to the sustainability of subsistence resources of indigenous communities. For caribou, Arctic indigenous people's most important terrestrial subsistence resource, body condition is a useful measure because it integrates many ecological factors that influence caribou productivity and is recognized by biologists and hunters as meaningful. We draw on experience working with indigenous communities to develop a body condition monitoring protocol for harvested animals. Local indigenous knowledge provides a broad set of caribou health indicators and explanations of how environmental conditions may affect body condition. Scientific research on caribou body condition provides a basis to develop a simple dichotomous key that includes back fat, intestinal fat, kidney fat and marrow¬fat, as measures of body fat, which in autumn to early winter correlates with the likelihood of pregnancy. The dichotomous key was formulated on "expert knowledge" and validated against field estimates of body composition. We compare local indigenous knowledge indicators with hunter documented data based on the dichotomous key. The potential con¬tribution of community body condition monitoring can be realized through the continued comparative analysis of datasets. Better communication among hunters and scientists, and refinement of data collection and analysis methods are recommended. Results suggest that specific local knowledge may become generalized and integrated between regions if the dichotomous key is used as a generalized (semi-quantitative) index and complemented with other science and community-based assessments.