305 research outputs found
Picturing Distance: Ed Ruschaâs Los Angeles Photobooks
Ed Ruscha's photobooks create a deliberate type of low-key provocation the artist has referred to as a type of âHuh?â â a presentation of the American landscape in a book format that makes it very difficult to locate unambiguous cues for meaning or clear affect in the photographs. This essay lays out an extended reading of the notion of âdistanceâ as uniting these photobooks in their physicality, in the ways in which the postwar industrial landscape of Los Angeles is presented to the viewer obliquely, and in the cooled manner of affect that was so influential for later artists
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Exploring Non-transgenic CRISPR-Cas9 gRNA Delivery Using Transactivation in Nicotiana benthamiana
Gene editing via CRISPR-Cas9 (Clustered, Regularly Interspaced, Short Palindromic Repeats â CRISPR-associated protein 9) is a powerful tool in biotechnology. The method involves an endonuclease Cas9 forming a complex with a guide RNA (gRNA) that matches a distinct DNA target sequence, causing double-stranded breaks in the DNA site. The break induces imperfect DNA repair resulting in insertions or deletions that render the target gene non-functional. It is also possible to mutate the Cas9 protein so that it loses its cutting function, but can still sit on the target DNA when in a complex with gRNA. This deactivated Cas9 (dCas9) can be fused with transcriptional activators or repressors to adjust the expression of a target gene.
While CRISPR-Cas9 is already widely used in plants, there are limitations to the systemâs delivery and efficiency. The aims of the research in this thesis were to generate a dCas9-activator-reporter system and to use the system to explore the possibility of two different non-transgenic methods of delivering gRNA.
The system consists of two plant expression plasmids. One encodes a constitutively expressed dCas9 fused to a transcriptional activator and a reporter gene driven by a minimal promoter. The second contains the gRNA, designed to target the minimal promoter. When the constructs are co-expressed, the gRNA-bound dCas9-activator sits on the minimal promoter and drives increased reporter expression.
Transient assays in N. benthamiana established the functionality of the system, showing increased reporter levels when the two constructs were co-expressed compared to the activator- reporter construct alone. Stable transgenic N. benthamiana lines of the separate constructs were generated and taken to the T2 generation.
To explore non-transgenic gRNA introduction methods, I tested viral delivery and grafting. Tobacco Rattle Virus (TRV) engineered to contain gRNA for the system was able to activate the reporter. However, the results suggest that viral recombination of the gRNA insert may cause the effect to be lost over time.
The positive readout transactivation system developed in this thesis will be a valuable tool for future CRISPR development in plants. In addition, the data reported present opportunities for further exploration on potential delivery methods, spatial specificity of CRISPR, and the mobility characteristics of synthetic gRNA compared to various endogenous RNAs
Sewing Lives: Mary Shelleyâs \u3cem\u3eFrankenstein\u3c/em\u3e and the Global Garment Industry
This paper takes Mary Shelleyâs Frankenstein and uses it as an extended metaphor to investigate the points of destructive alienation and disassociation within the globalized consumption of clothing. The promise of new clothing is a set of garments that function like Victorâs dream of creation; materials are stitched together to give objects that match our closest-held ideals. And yet, because of our quick Victor-Frankenstein-like alienation from these âfast fashionâ objects when they no longer please us, clothing becomes, like the monster, an abjected figure for waste and shame, moving around the globe destructively, created from the bodies of the poor and having lost the care of its creator or consumer. Solving the problem of sustainability in the fashion industry involves, this paper argues, taking account of that difficult relationship with the global monstrous, and of the unique ways in which clothes galvanize some of our deepest emotions
Sewing Lives: Mary Shelleyâs Frankenstein and the Global Garment Industry
This paper takes Mary Shelleyâs Frankenstein and uses it as an extended metaphor to investigate the points of destructive alienation and disassociation within the globalised consumption of clothing. The promise of new clothing is a set of garments that function like Victorâs dream of creation; materials are stitched together to give objects that match our closest-held ideals. And yet, because of our quick Victor-Frankenstein-like alienation from these âfast fashionâ objects when they no longer please us, clothing becomes, like the monster, an abjected figure for waste and shame, moving around the globe destructively, created from the bodies of the poor and having lost the care of its creator or consumer. Solving the problem of sustainability in the fashion industry involves, this paper argues, taking account of that difficult relationship with the global monstrous, and of the unique ways in which clothes galvanise some of our deepest emotions
An explorative study of the perceptions of social workers who are exposed to work-related, secondary traumatic experiences through their clientele, of the psychosocial support Equine Assisted Therapy (E.A.T) could provide
Includes abstract.Includes bibliographical references.This study emanates from the fact that trauma is an increasing reality in our society and it has a particular focus on social workers who work with clientele that have been exposed to and who have suffered trauma. As a result and by using clinical judgment of social workers working in such environments, the social workers may have developed Secondary Traumatic Stress (S.T.S.) symptoms. Psychosocial support functions as a preventative and supportive tool for social workers and emotion-focussed programmes have beneficial effects on the psychosocial wellbeing of social workers by reducing their distress. This study will be exploring a psychosocial intervention called Equine Assisted Therapy (E.A.T.), which makes use of horses in a therapeutic model and enables clients to interact with horses on an emotional and metaphorical level that empowers clients to experience emotional and behavioural change and growth. The significance of this study is to enable the researcher to explore E.A.T. as a possible psychosocial intervention that can provide support for social workers and health care professionals alike who work in environments where they are exposed to high levels of trauma. The researcher made use of a qualitative paradigm that was explorative in nature to gain perspective of the chosen topic. The population for the study comprised of 16 social workers that work in the family violence, child welfare and sexual assault field and who attended a once-off Equine Assisted Therapy (E.A.T.) session to expose them to the therapy model being explored. The participants then volunteered for a semi-structured interview with the researcher which was digitally recorded, thereby allowing the researcher to collect the data and then analyse the data
Novel Determinants That Influence Azole Susceptibility in Candida glabrata and Candida albicans
Despite the scientific and medical communitiesâ best efforts, the incidence of fungal infections in susceptible populations continues to rise. The most common cause of these opportunistic fungal infections is Candida. In fact, Candida is the fourth most common pathogen associated with nosocomial blood stream infections. Reported mortality rates for patients with candidemia vary, but have not decreased in the past fifteen years and are reported to be as high as 50%. Candida glabrata, second only to Candida albicans among Candida infections, expresses high rates of resistance to treatment with arguably the best class of currently available antifungals - the azoles.
Other available antifungals have associated toxicities, are not available as oral dosage forms or are cost prohibitive. As multidrug resistant C. glabrata have been reported, the need to find ways to overcome resistance to azoles is more pressing than ever. The work described here highlights our efforts to develop a better understanding of azole resistance in Candida, with a focus on C. glabrata, which can then be utilized to inform better strategies for decreasing or preventing resistance.
In C. glabrata clinical azole resistance is mediated almost exclusively by activating mutations in the zinc cluster transcription factor Pdr1, which controls the genes encoding the multidrug resistance transporters Cdr1, Pdh1, and Snq2. However, the specific relative contribution of these transporters to resistance is not known. In order to determine this, the SAT1 flipper method was used to delete CDR1, PDH1, and SNQ2 in a strain of C. glabrata engineered to carry a clinically relevant activating mutation in PDR1. Susceptibility testing was performed according to the CLSI guidelines with minor modifications and confirmed with Etest strips. Of the single transporter deletion strains, only CDR1 deletion resulted in decreased azole MIC. Deletion of PDH1 in combination with CDR1 resulted in a moderate decrease in MIC from that observed with deletion of CDR1 alone. SNQ2 deletion only decreased the MIC in the triple deletion strain in the absence of both CDR1 and PDH1. Deletion of all three transporters in combination decreased the MIC to the level observed in the PDR1 deletion strains for some, but not all of the azoles tested, which indicates additional Pdr1 targets likely play a minor role in this process. These results demonstrate that Cdr1 is the most important Pdr1-mediated multidrug resistance transporter for azole resistance in C. glabrata, suggesting that targeting this transporter alone might be sufficient to overcome this clinical problem.
Upc2 and Ecm22 in S. cerevisiae and Upc2 in C. albicans are the transcriptional regulators of ERG11, the gene encoding the target of azoles in the ergosterol biosynthesis pathway. Recently two homologs for these transcription factors, UPC2A and UPC2B, were identified in C. glabrata. One of these, UPC2A, was shown to influence azole susceptibility. We hypothesized that due to the global role for Upc2 in sterol biosynthesis in S. cerevisiae and C. albicans, disruption of UPC2A would enhance the activity of fluconazole in both azole-susceptible-dose dependent (SDD) and -resistant C. glabrata clinical isolates. To test this hypothesis, we constructed mutants disrupted for UPC2A and UPC2B alone and in combination in a matched pair of clinical azole-SDD and - resistant isolates. Disruption of UPC2A in both the SDD and resistant isolates resulted in increased susceptibility to sterol biosynthesis inhibitors, including a reduction in fluconazole minimum inhibitory concentration and minimum fungicidal concentration, enhanced azole activity by time-kill analysis, a decrease in ergosterol content, and downregulation of baseline and inducible expression of several sterol biosynthesis genes. Our results indicate that Upc2A is a key regulator of ergosterol biosynthesis and is essential for resistance to sterol biosynthesis inhibitors in C. glabrata. As such, the UPC2A pathway may represent a potential co-therapeutic target for enhancing azole activity against this organism.
The importance of Pdr1 in azole resistance in C. glabrata is well established. Our understanding of how Pdr1 is being regulated, however, is predominantly informed by regulation of similar systems in other organisms. In order to identify genes that interact with the Pdr1 transcriptional pathway, and influence the susceptibility of C. glabrata to fluconazole, we screened a collection of deletion mutants for those exhibiting increased resistance to fluconazole. Deletion of the gene coding for a protein homologous to the S. cerevisiae J protein Jjj1 resulted in decreased fluconazole susceptibility. We used the SAT1 flipper method to generate independent deletion mutants for JJJ1 in a SDD clinical isolate. Expression of both CDR1 and PDR1 was increased in the absence of JJJ1. In the absence of CDR1 or PDR1, deletion of JJJ1 had only a modest effect on fluconazole susceptibility. Transcriptional profiling using RNA-Seq revealed up-regulation of genes of the Pdr1 regulon in the absence of JJJ1. Jjj1 appears to be a negative regulator of fluconazole resistance in C. glabrata and acts primarily through up-regulation of the ABC transporter gene CDR1 via activation of the Pdr1 transcriptional pathway.
Unlike C. glabrata which has essentially one mechanism of resistance, in C. albicans clinical azole resistance can be attributed to multiple mechanisms, often in combination. The RTA3 gene, coding for a member of the Rta1p-like lipid-translocating exporter family, is coordinately upregulated with the ABC transporter genes CDR1 and CDR2 in azole-resistant clinical isolates of C. albicans that carry activating mutations in the transcription factor Tac1p. We show here that deleting RTA3 in an azole-resistant clinical isolate carrying a Tac1p activating mutation lowered fluconazole resistance by two-fold, while overexpressing RTA3 in an azole-susceptible clinical isolate resulted in enhanced fluconazole tolerance associated with trailing growth in a liquid microtiter plate assay. We also demonstrate that an Rta3p-GFP fusion protein localizes predominantly to the plasma membrane, consistent with a putative function for Rta3p as a lipid translocase
Fighting the Drop-Out Crisis
Describes data-driven efforts to raise high school graduation rates in New York, Philadelphia, and Portland, Oregon by replacing low-performing schools with smaller schools or offering alternative schools and programs, as well as their outcomes to date
An Integrated Mechanistic Model of Mindfulness-Oriented Recovery Enhancement for Opioid-Exposed MotherâInfant Dyads
A growing body of neurobiological and psychological research sheds light on the mechanisms underlying the development and maintenance of opioid use disorder and its relation to parenting behavior. Perinatal opioid use is associated with risks for women and children, including increased risk of child maltreatment. Drawing from extant data, here we provide an integrated mechanistic model of perinatal opioid use, parenting behavior, infant attachment, and child well-being to inform the development and adaptation of behavioral interventions for opioid-exposed motherâinfant dyads. The model posits that recurrent perinatal opioid use may lead to increased stress sensitivity and reward dysregulation for some mothers, resulting in decreased perceived salience of infant cues, disengaged parenting behavior, disrupted infant attachment, and decreased child well-being. We conclude with a discussion of Mindfulness-Oriented Recovery Enhancement as a means of addressing mechanisms undergirding perinatal opioid use, parenting, and attachment, presenting evidence on the efficacy and therapeutic mechanisms of mindfulness. As perinatal opioid use increases in the United States, empirically informed models can be used to guide treatment development research and address this growing concern
Elder Abuse in the European Union
The rapid rise in persons over age 60 has created a platform for the rise of elder abuse all over the world. Increasing strains on caregivers and the realization that elder abuse is a serious issue have only recently produced research on the topic. In light of this, this presentation evaluates the state of the aging population in the European Union where more research has been done on elder abuse than in other areas of the world. This presentation focuses specifically on elder abuse in Germany and in Greece, as these countries represent the cultural and geographical extremes in Europe today. First, the presentation evaluates the risk factors and forms that elder abuse is taking today in the European Union. The presentation then describes the state of elder abuse in both Germany and Greece, taking into consideration demographics, culture, and current responses. Finally, this presentation evaluates possible steps to implement for effective change in the area of elder abuse
Non-expert practice application of an AI vision systems in design engineering projects
Design projects units for BSc (Hons) Design Engineering students at Bournemouth University integrate and apply knowledge from a range of taught units together with self-directed learning and towards solving design problems. Recently, level 6 (FHEQ) project students have proposed and designed solutions that require AI vision-systems. These projects presented a problem for supervision, with limited, or no expertise in the technology or available equipment; students therefore treated these subsystems as a âblack-boxâ exercise. To address these issues a set of technical requirements were compiled, a range of AI technology solutions were identified before selecting the Nvidia Jetson Nano. From the literature, a stream-lined practical program was developed to introduce the technology to level 5 and level 6 project students as part of their design education. This provided hands on experience through familiarization with the interface and the use of pretrained models before students re-trained networks with their own datasets. Level 5 students utilised the technology to develop a scratch detection machine for sorting damaged components. Level 6 students were provided with the opportunity to integrate the technology into projects where appropriate and two students did so; one developed a device to identify people trapped in buildings after an earthquake, the second developed a device for monitoring chili-plants when grown under polytunnels. Developing and delivering the introductory programme as a non-expert learning pathway has enhanced the student experience within design education, provided a simple workflow that students can utilise and build upon, and led to successful student outcomes
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