A study of CIS-acting elements required for dosage compensation in Drosophila Melanogaster : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Genetics at Massey University, Palmerston North, New Zealand

Dosage compensation (the equalisation of X-linked gene products) occurs in Drosophila melanogaster by a two fold transcriptional up-regulation of X-linked gene expression in males. This involves the binding of five proteins, MSL-1, MSL-2, MSL-3, MLE, MOF, and potentially an RNA (roXl or roX2), to hundreds of sites along the male X chromosome. The cis-acting X-linked DNA sequences required for dosage compensation (called dosage compensation regulatory elements or DCREs) remain elusive, despite numerous attempts of identify them. An insulated reporter gene assay system has been developed to minimise problems previously encountered with identification of these elements. The reporter system consists of the constitutive armadillo promoter fused to the lacZ reporter gene (called arm-lacZ). This reporter construct is flanked by SCS/SCS' insulator elements to block potential repressive effects of an autosomal chromatin environment. The role of the roX genes during dosage compensation was investigated. Initially both the roXl and roX2 RNAs were expressed from within the arm-lacZ insulated system. Expression of either RNA lead to a significant increase in lacZ expression in males, although consistently less than two-fold. These results suggested that either the MSL complex was binding to the roX genes or the expression of the roX RNAs in cis lead to male-specific hypertranscription of lacZ. To test these possibilities roX1 and roX2 cDNAs were inserted into the arm-lacZ reporter. Insertion of either cDNA lead to a significant increase in lacZ expression in males, suggesting that the transcribed regions of the roX genes contain binding site(s) for the MSL complex. Interestingly the level of lacZ hypertranscription in males was significantly higher in homozygous roX1 cDNA lines than homozygous roX1 gene lines. This may indicate that too high a local concentration of roX1 RNA has a dampening effect on the level of hypertranscription meditated by the MSL complex. In a set of experiments designed to identify the MSL binding site(s) in roX1, two regions of the cDNA sequence were amplified and inserted into the arm-lacZ system. One of these fragments, containing a proposed DNAseI hypersensitivity site and possible GAGA binding sites, increased lacZ expression in males, but to levels lower than the entire cDNA. This suggests there may be more than one MSL biding site in roX1. A second method of dosage compensation is thought to occur in Drosophila, independently of the MSL proteins. The arm-lacZ insulated reporter system was used to investigate the hypothesis that some genes may be dosage compensated due to repression by Sex-lethal (Sxl) in females. Several genes have been found to contain three or more Sxl binding sites in their 3' UTRs. with some also carrying Sxl binding sites in the 5' UTR. Fragments from the Sxl, Cut and Small Forked genes, containing numerous Sxl binding sites from the 3' UTR, were inserted into the 3' UTR region of arm-lacZ. Males carrying autosomal insertions of the construct had on average 1.07 - 1.50 times the level of β-galactosidase in females. This suggests that some genes could be partially compensated through Sxl repression in females. In addition to inserting 3' UTR fragments into arm-lacZ, a synthetic oligonucleotide containing a long Sxl binding site was inserted into the 5' region of an arm-lacZ construct already carrying the Runt 3' UTR fragment. Males carrying autosomal insertions of the construct had levels of β-galactosidase activity similar to those lines carrying autosomal insertions of the 3' UTR fragments alone. This suggests that other factors such as RNA binding proteins or RNA secondary structure may be required in order to obtain efficient translation repression by Sxl. Finally three X-linked DNA fragments, from the 1C region, were inserted individually between the SCS' element and the armadillo promoter. If the X-linked fragment contained a DCRE then males carrying autosomal insertions of the construct would produce twice the β-galactosidase activity of females. However, males and females expressed the same levels of lacZ

The Evaluation of Capacity Building Grants: Key Learnings for a Successful Program

Whether prompted by venture philanthropy, high engagement grantmaking, or a growing interest in nonprofit management, many foundations now have programs aimed at capacity building for their grantees. Grants from these programs are often highly targeted to meet the specific organizational needs of individual grantees. But how can a foundation get from the apples and oranges of individual grantee results to a succinct way of reporting overall program achievements? This was the question that the Maine Community Foundation (MCF) brought to FSG. Our analysis not only helped them evaluate their program, it also highlighted three basic lessons that can increase the likelihood of success for any capacity building initiative

\u3ci\u3eConcerto for Violin and Orchestra\u3c/i\u3e

The Concerto for Violin and Orchestra is a three-movement composition for solo violin accompanied by a medium-sized symphonic ensemble. This piece is composed in partial fulfillment of the requirements of the degree Master of Music with a concentration in Composition from the University of Tennessee, Knoxville. The Concerto was composed during the 2012-2013 academic year. This paper provides a narrative analysis of the Concerto in terms of the parameters of its musical content and their relationships. Furthermore, references are made to related aspects of influential compositions of the last century. Compositions by Samuel Barber, Béla Bartók, Alban Berg, Paul Hindemith, Dmitry Kabalevsky, Sergey Prokofiev, Jean Sibelius, and Igor Stravinsky are included in this study. Chapter topics include genre, form, melody, harmony, rhythm, and orchestration

Art ConText: On This Ground: An Installation by Rebecca Belmore

Exhibition Notes, Number 11, Summer 2000. On getting acquainted with a community and making art in strange places: An Interview. Between March and June 2000, Rebecca Belmore was the Art ConText artist-in-residence at the Providence Public Library’s Smith Hill Branch. She participated in the transformation of an empty lot behind the library; worked with a group of immigrants in a library literacy program; and engaged students at the nearby Camden Avenue Elementary School; all while creating the artwork that now dominates the Museum’s Farago Entry Gallery. Following are her thoughts on questions posed by David Henry, Head of Education, The RISD Museum.https://digitalcommons.risd.edu/risdmuseum_journals/1008/thumbnail.jp

Systems and methods for altering visual acuity

Provided are systems operable to effect a temporary change in a modulation transfer function (MTF) of a target imaging system. The systems include a light source operable to produce light for transient propagation onto at least a portion of the target imaging system, a power source in operative communication with the light source and configured to effect the production of light from the light source, and a transmission unit in operative communication with the light source and configured to propagate the produced light onto at least a portion of the target imaging system. The propagated light is configured for absorbance by the portion of the target imaging system; the absorbance causes an increase in temperature and a change in a refractive index profile of at least the portion of the imaging system. The change in refractive index profile produces a temporary change in the MTF of the imaging system.Board of Regents, University of Texas Syste

Being uninformed on informed consent: a pilot survey of medical education faculty

BACKGROUND: This paper describes a pilot survey of faculty involved in medical education. The questionnaire focuses on their understanding of IRB policies at their institution, specifically in relation to the use of student assessment and curriculum evaluation information for scholarship. METHODS: An anonymous survey was distributed to medical educators in a variety of venues. Two brief scenarios of typical student assessment or curriculum evaluation activities were presented and respondents were asked to indicate their likely course of action related to IRB approval. The questionnaire also asked respondents about their knowledge of institutional policies related to IRB approval. RESULTS: A total of 121 completed surveys were obtained; 59 (50%) respondents identified themselves as from community-based medical schools. For the first scenario, 78 respondents (66%) would have contact with the IRB; this increased to 97 respondents (82%) for the second scenario. For both scenarios, contact with the IRB was less likely among respondents from research-intensive institutions. Sixty respondents (55%) were unsure if their institutions had policies addressing evaluation data used for scholarship. Fifty respondents (41%) indicated no prior discussions at their institutions regarding IRB requirements. CONCLUSION: Many faculty members are unaware of IRB policies at their medical schools related to the use of medical student information. To the extent that policies are in place, they are highly variable across schools suggesting little standardization in faculty understanding and/or institutional implementation. Principles to guide faculty decision-making are provided

Ability of γδ T cells to modulate the Foxp3 T cell response is dependent on adenosine.

Whether γδ T cells inhibit or enhance the Foxp3 T cell response depends upon their activation status. The critical enhancing effector in the supernatant is adenosine. Activated γδ T cells express adenosine receptors at high levels, which enables them to deprive Foxp3+ T cells of adenosine, and to inhibit their expansion. Meanwhile, cell-free supernatants of γδ T cell cultures enhance Foxp3 T cell expansion. Thus, inhibition and enhancement by γδ T cells of Foxp3 T cell response are a reflection of the balance between adenosine production and absorption by γδ T cells. Non-activated γδ T cells produce adenosine but bind little, and thus enhance the Foxp3 T cell response. Activated γδ T cells express high density of adenosine receptors and have a greatly increased ability to bind adenosine. Extracellular adenosine metabolism and expression of adenosine receptor A2ARs by γδ T cells played a major role in the outcome of γδ and Foxp3 T cell interactions. A better understanding of the functional conversion of γδ T cells could lead to γδ T cell-targeted immunotherapies for related diseases

Role of Spore-Associated Inosine-Uridine Nucleoside Hydrolase IunA in Bacillus anthracis Spores

The abstract for this presentation can be downloaded by clicking on the blue download button