HIV-1 Vpr Enhances Viral Burden by Facilitating Infection of Tissue Macrophages but Not Nondividing CD4+ T Cells

Prior experiments in explants of human lymphoid tissue have demonstrated that human immunodeficiency virus type 1 (HIV-1) productively infects diverse cellular targets including T cells and tissue macrophages. We sought to determine the specific contribution of macrophages and T cells to the overall viral burden within lymphoid tissue. To block infection of macrophages selectively while preserving infection of T cells, we used viruses deficient for viral protein R (Vpr) that exhibit profound replication defects in nondividing cells in vitro. We inoculated tonsil histocultures with matched pairs of congenic viruses that differed only by the presence of a wild-type or truncated vpr gene. Although these viruses exhibited no reduction in the infection or depletion of T cells, the ability of the Vpr-deficient R5 virus to infect tissue macrophages was severely impaired compared with matched wild-type R5 virus. Interestingly, the Vpr-deficient R5 virus also exhibited a 50% reduction in overall virus replication compared with its wild-type counterpart despite the fact that macrophages represent a small fraction of the potential targets of HIV-1 infection in these tissues. Collectively, these data highlight the importance of tissue macrophages in local viral burden and further implicate roles for CC chemokine receptor 5, macrophages, and Vpr in the life cycle and pathogenesis of HIV-1

CpG Island microarray probe sequences derived from a physical library are representative of CpG Islands annotated on the human genome

An effective tool for the global analysis of both DNA methylation status and protein–chromatin interactions is a microarray constructed with sequences containing regulatory elements. One type of array suited for this purpose takes advantage of the strong association between CpG Islands (CGIs) and gene regulatory regions. We have obtained 20 736 clones from a CGI Library and used these to construct CGI arrays. The utility of this library requires proper annotation and assessment of the clones, including CpG content, genomic origin and proximity to neighboring genes. Alignment of clone sequences to the human genome (UCSC hg17) identified 9595 distinct genomic loci; 64% were defined by a single clone while the remaining 36% were represented by multiple, redundant clones. Approximately 68% of the loci were located near a transcription start site. The distribution of these loci covered all 23 chromosomes, with 63% overlapping a bioinformatically identified CGI. The high representation of genomic CGI in this rich collection of clones supports the utilization of microarrays produced with this library for the study of global epigenetic mechanisms and protein–chromatin interactions. A browsable database is available on-line to facilitate exploration of the CGIs in this library and their association with annotated genes or promoter elements

Feed-forward: Future questions, future maps

"Feed-forward" is a technique that encourages families to imagine the pattern of their relationships at some future point in time. Questions about the future, in conjunction with positive connotation, put families in a metaposition to their own dilemmas and thus facilitate change by opening up new solutions for old problems. What does e.e. cummings say? "Always the most beautiful answer, who asks the more difficult question." You see I am not asking another question each time. I am making the same question bigger. Bateson, Gregory 1980 This paper describes a technique I call "feed-forward," which is based on the family's consideration of how the pattern of their relationships will continue in the future. Since the maps for the future are not yet set, the family are free to construct or imagine a different set of alternatives to their dilemma. The consideration of these future maps places the family in a metaposition to their own dilemma, and the system increases its view of its own evolutionary potential. Pragmatically, future questions, in combination with positive connotation, promote the rehearsal of new solutions, suggest alternative actions, foster learning, discard ideas of predetermination, and address the system's specific change model. In particular, future questions are useful in working with families with a chronic illness, whose concept of future time is often frozen. The questions contrast and separate the system's ongoing pattern from the illness by comparing the present relationships with the relationships that predated the illness and the relationships the family anticipate on recovery or stabilization. Two years ago I wrote a paper, "Circular Questioning" 9, in which I analyzed the Milan associates' technique of circular questioning. One of the questions asks the family to describe to the therapists the changes in their relationships before and after an onset event. I used the image of an arc suspended between the "before" and "after" of the event, connecting the onset of the problem to the present dilemma, and vice versa. I would now like to move that arc and connect the present and the future, asking questions of now and when, now and if, or now and suppose, etc. 1 For example, "If your parents were divorced, who would you live with?" Or, "When you go to college, which parent will miss you most?" In this way, new information about the future is introduced into the system. One can easily recognize that these questions have a loose hypnotic structure; if one accepts the first proposition of the question"When you go to college"the second part, containing the question"which parent will miss you most?"may be entertained as new information based on the understanding of the present plus the consideration of that particular future. These questions elicit relationship information and introduce ideas into the system about the stability, endurance, or change in its patterns over time. 2 According to the Milan associates, future questions break the pervasive rules that govern communication in the familyi.e., the rules for who is allowed to say what. Since the future is often indicated but not "set," no one is bound by formal contextual rules, and a different pattern may be imagined. For example, if you ask a family member a hypothetical question regarding future events, because the event is only now being considered, the system is free to create a new map. Then the communication of these new ideas about the future becomes important information introduced back into the present "time" of the system. They include fantasies, wishes, opinions, hopes, etc., all a part of the ongoing system and now unexpectedly called into play as part of the family's expressed interactions. In fact, repeated hypothetical questioning of an outcomeif this or that event obtainedgives the family a sense of their own potential to imagine new solutions. At that moment I would say the family are in the process of feed-forward. In considering how things could turn out if, you are addressing a basic descriptor of the system: its capacity to evolve. It is that much harder for the system to restabilize when its evolutionary potential is evoked. The question is how (through what therapeutic mechanism) can one leave context-bound experiences and move ahead to new organizations. 3,4 Positive Connotation One of the most important propositions of Milan systemic therapy is the use of the positive connotation, a technique that describes positively the current organization of the system (11). Blaming in any form is omitted, and instead a perception is offered that defines positively the family's dilemma, regards it as context-bound, and implies that contexts themselves are relative and changeable. I emphasize the importance of using positive connotation in constructing future maps, for

The CBC Newsworld holodeck

For the past 73 years, the CBC has disseminated a unique Canadian perspective across the world, producing a phenomenally rich multimedia record of the country and our social, political and cultural heritage and news. This project utilizes visualization and sonification of portions of an enormous historical CBC Newsworld data corpus to enable an "on this day" experience for viewers. The digitized collection of 24-hour news videos spans a 24-year period (1989-2013) within an immersive multiscreen environment, to enable gesture-driven context-aware browsing, information seeking, and segment review. Employing natural language processing technologies, the interface displays keywords and key phrases identified in the transcripts, enabling serendipitous video search and display and offering a unique browsing opportunity within this rich "big data" corpus

Analysis of Myc bound loci identified by CpG island arrays shows that Max is essential for Myc-dependent repression

1Ontario Cancer Institute/Princess Margaret upon Myc [2, 3]. To evaluate if Myc bound to the regulatory regions of these genes, chromatin immunoprecipitation (ChIP) assays were conducted by using an antic-Myc antibody as described previously [4]. Myc binding to the 5 � regulatory region of a known activated gene (cad) was detected in human HL60 cells, while no bind-Hospital ing was evident at the negative control region, a chromo-2Department of Medical Biophysics some 22 E box (chrm22 E box) (Figure 1, compare lane

CpG Island microarray probe sequences derived from a physical library are representative of CpG Islands annotated on the human genome. Nucleic Acids Res 33:2952–2961

ABSTRACT An effective tool for the global analysis of both DNA methylation status and protein-chromatin interactions is a microarray constructed with sequences containing regulatory elements. One type of array suited for this purpose takes advantage of the strong association between CpG Islands (CGIs) and gene regulatory regions. We have obtained 20 736 clones from a CGI Library and used these to construct CGI arrays. The utility of this library requires proper annotation and assessment of the clones, including CpG content, genomic origin and proximity to neighboring genes. Alignment of clone sequences to the human genome (UCSC hg17) identified 9595 distinct genomic loci; 64% were defined by a single clone while the remaining 36% were represented by multiple, redundant clones. Approximately 68% of the loci were located near a transcription start site. The distribution of these loci covered all 23 chromosomes, with 63% overlapping a bioinformatically identified CGI. The high representation of genomic CGI in this rich collection of clones supports the utilization of microarrays produced with this library for the study of global epigenetic mechanisms and proteinchromatin interactions. A browsable database is available on-line to facilitate exploration of the CGIs in this library and their association with annotated genes or promoter elements

CpG Island microarray probe sequences derived from a physical library are representative of CpG Islands annotated on the human genome.

An effective tool for the global analysis of both DNA methylation status and protein-chromatin interactions is a microarray constructed with sequences containing regulatory elements. One type of array suited for this purpose takes advantage of the strong association between CpG Islands (CGIs) and gene regulatory regions. We have obtained 20,736 clones from a CGI Library and used these to construct CGI arrays. The utility of this library requires proper annotation and assessment of the clones, including CpG content, genomic origin and proximity to neighboring genes. Alignment of clone sequences to the human genome (UCSC hg17) identified 9595 distinct genomic loci; 64% were defined by a single clone while the remaining 36% were represented by multiple, redundant clones. Approximately 68% of the loci were located near a transcription start site. The distribution of these loci covered all 23 chromosomes, with 63% overlapping a bioinformatically identified CGI. The high representation of genomic CGI in this rich collection of clones supports the utilization of microarrays produced with this library for the study of global epigenetic mechanisms and protein-chromatin interactions. A browsable database is available on-line to facilitate exploration of the CGIs in this library and their association with annotated genes or promoter elements