873 research outputs found
The Effectiveness of the Size Matters Handwriting Program
No full textAbstract
Date Presented 3/31/2017
With school-based occupational therapists reporting up to 75% of their caseload related to handwriting, the urgency to identify a proven and efficient instructional program is paramount. Effective, embeddable, measurable, easy, and fast, the Size Matters Handwriting Program promotes collaboration in the natural environment and the Workload model.
Primary Author and Speaker: Beverly Moskowitz
Additional Authors and Speakers: Beth Carswell, Jennifer Kitzmiller, Moira Bushell, Laura Neikrug, Chaya Gottesman
Contributing Authors: Beth Pfeiffer, Gillian Rai, Tammy Murray</jats:p
"Ice Road" by Gillian Slovo. [review - radio script]
No full text"Ice road" is a novel of nineteenth century proportions by prolific British author Gillian
Slovo. With its broad canvas of Russian history and large cast of characters, led by a
young woman called Natasha, it consciously harks back to Tolstoy’s "War and Peace".
The story begins with a cleaner called Irina Davydovna Arbatova, a pragmatic
worker born at the beginning of the twentieth century, who by various chance encounters
becomes involved in the family of Boris Aleksandrovic Ivanov, a party official, one of the new soviet ruling class. The setting is Leningrad, and the year is 1934
Editing Aphra Behn in the Digital Age: An Interview with Gillian Wright and Alan Hogarth
No full textThis interview provides a view of the work in progress for the Cambridge University Press edition of the Complete Works of Aphra Behn. Gillian Wright serves as a general editor (with Elaine Hobby, Claire Bowditch, and Mel Evans) as well as the volume editor for Behn’s poetry. Alan Hogarth is the Postdoctoral Research Associate working with Mel Evans on the computational stylistics and author attribution testing. The discussion focuses on the scope and principles of editing the poetry of Aphra Behn, the role of stylometry in establishing the corpus, the status of work, a few particular poems, and some surprises
Nieznajoma z North Carthage. Dwuznaczne narracje Gillian Flynn w powieści "Zaginiona dziewczyna"
No full textThis article analyzes the use of narration in Gillian Flynn's Gone Girl. The author employs the „missing white woman syndrome” and unreliable narrators to manipulate readers' perceptions and expectations.This article analyzes the use of narration in Gillian Flynn's Gone Girl. The author employs the „missing white woman syndrome” and unreliable narrators to manipulate readers' perceptions and expectations
Green Fluorescent Proteins: Towards Extra-Cellular Applications?
No full textThe Green Fluorescent Protein (GFP) and its numerous variants are applied
extensively in a multitude of in vivo applications and have been studied in this context
at length. In contrast however, the study of GFP’s within the emerging fields of nanoand
micro-technology, which offer broader extracellular applications for GFP and its
derivatives, has only recently begun to gather momentum. This thesis presents the
directed design of a novel series of Enhanced Green and Enhanced Yellow Fluorescent
Proteins (EGFP and EYFP respectively), for implementation in extracellular
applications such as biosensing and fundamental research into fluorescence protein
behaviour. Each parent fluorescent protein (EGFP or EYFP) was altered to display a
single solvent exposed reactive sulfhydryl group with varying degrees of connectivity to
the internal GFP chromophore. These sulfhydryl groups were introduced into the
protein primary structure via point mutation to yield cysteine residues in place of the
targeted native amino acid. Careful examination of the EGFP and EYFP tertiary
structures to identify amino acids within the protein primary sequence that fulfilled
specific criteria, which were defined in our experimental design, resulted in substitution
of amino acids at positions 221, 223, 219, 212 and 97 in EGFP and 221, 223, 212, 95
and 21 in EYFP.
Critical development of supporting methodologies delivered vast improvements
on literature protocols for expression and purification of the GFP variants listed above.
Expression protocol investigation determined that the most prolific E. coli strain for
recombinant fluorescent protein production was BL21, which, coupled with our
methodology, produced up to 13.6 mg of fluorescent protein per gram of wet cell pellet.
The novel purification procedure described in this Thesis delivered highly pure protein
with impressive yields (75-80 %).
Characterisation of the novel proteins that were designed and produced during
this work revealed no change in the proteins’ ability to resist denaturation resulting from
cysteine substitution. Neither was there any change in fluorescence emission or UV
absorption profiles for standard concentrations (< 60 mM) of any of the purified proteins
that were produced. While standard protein solutions returned normal fluorescence
emission profiles, solutions that contained protein concentrations above 60 mM
displayed red shifted emission maximum values. For protein solutions within the mM
concentration range this red shift in fluorescence emission was at times in the order of
30 nm resulting in emission maximums of up to 540 nm for EGFP, and 548 nm for
EYFP and recombinant proteins containing an L221C mutation. Preliminary
investigations into this phenomenon showed that the changes observed in fluorescence
emission were dependent on protein concentration and could be due to dipole-dipole
interactions which may be induced by protein aggregate formation at high protein
concentrations. Manipulations that were performed on fluorescent proteins during this
study included proteolytic digestion with Proteinase K and subsequent testing of the
digested protein product. This work identified an increase in the quantum yield of
proteolytically digested EGFP and EYFP from 0.6 to 0.8 accompanied by no reduction
in the digested proteins resistance to denaturing treatments except when treated with
1 % SDS solution.Thesis (PhD Doctorate)Doctor of Philosophy (PhD)School of Biomolecular and Physical SciencesScience, Environment, Engineering and TechnologyFull Tex
Gillian Dooley interviews Joris Luyendijk, author of 'Fit to Print: Misrepresenting the Middle East'.
No full textInterview with Joris Luyendijk, author of 'Fit to Print: Misrepresenting the Middle East', a book about the problems of foreign journalism in the Middle East
Conjuring our beings: Stacey Gillian Abe and Immy Mali in conversational partnership
No full textThe series of Conversational Partnerships began in 2017 in African Arts vol. 50, no. 2, with a conversation between two artists: Eria Nsubuga SANE from Uganda and Sikhumbuzo Makandula from South Africa. The format of a “conversational partnership” (Rubin and Rubin 2012: 7) emphasizes the cocreation of meaning by the interviewer and interviewee as coauthors. This enables a move away from the art history format of the interviewer (usually a writer) assuming the role of the sole author and the interviewee (often an artist) having no status as an author despite the fact that her or his practice-led creation of knowledge is foundational to the content of the interview. Stacey Gillian Abe and Immy Mali participated in a joint artists' residency as part of the RAW program at Rhodes University in South Africa from November to December 2017. During this time, they engaged with each other's practice-led work, and they created this conversational partnership at a writing breakaway in the Eastern Cape
Terminal Modifications of PNA and Their Use in Diagnostic and Antisense Technologies
No full textPeptide nucleic acids (PNA) are analogues of DNA that bind to DNA and RNA via Watson-Crick base-pairing rules. Due to the lack of a negatively-charged backbone, hybridisation of PNA to DNA or RNA occurs without electrostatic repulsion thus binding is typically stronger and more rapid than when traditional DNA probes are used. This is reflected in the increased melting temperature (Tm) of the conjugates. These properties, as well as the chemical and biological stability of PNA, make these molecules attractive for use in diagnostic and therapeutic applications. Amino acids are routinely conjugated to PNA probes to enhance the synthesis and solubility of the probes or assist with their cellular delivery, however little thought is given to the impact these modifications have on their hybridisation properties. In this work, a series of PNA-peptide chimeric assemblies based around a single PNA sequence were used to investigate the effect different amino acids have on the stability and specificity of PNA/DNA hybridisation. These experiments demonstrated that the positively charged amino acid lysine, which is routinely conjugated to PNA probes, increases the stability of the resultant PNA/DNA duplexes such that at many experimental temperatures, single base mismatched target DNA will also stably hybridise with PNA. In contrast, the negatively charged amino acid glutamic acid decreases the thermal stability of the mismatch duplexes sufficiently so that they are not stable at most experimental temperatures, whilst the fully complementary duplex is. This indicates that glutamic acid should replace lysine as the routine solubility enhancing group used for PNA probes.Thesis (PhD Doctorate)Doctor of Philosophy (PhD)School of Biomolecular and Physical SciencesScience, Environment, Engineering and TechnologyFull Tex
The Adenosine Receptor and Serum Deprivation-Induced Neuronal Differentiation
No full textAdenosine is a multi-functional physiological molecule found abundantly in the body. It is one of the important components of ATP cellular energy metabolism. Adenosine has diverse actions as a ligand on many different types of cells and tissues acting via specific receptors. Currently, four subtypes of adenosine receptors are described, namely, the A1, A2A, A2B and A3 receptors. Neuroblastoma, mostly found in young children, is a malignant tumor derived from peripheral neurons in the body. Several different types of neuroblastoma cell lines of human origin have been established and contributed to the studies of neuroblastoma itself, neuronal differentiation, neurotransmitters, alcoholism, Alzheimer's disease and other neuronal diseases and disorders. In 1987, it was shown by Abbracchio et al. that a human neuroblastoma cell line, IMR32, could be induced to differentiate into cells that have a more neuronal morphology, with long neurites, by an adenosine receptor agonist 5'-N-ethylcarboxamideadenosine (NECA) 2. 'Neuronal differentiation' is expected to be a new alternative to the conventional clinical therapies, such as surgery, chemotherapy and radiotherapy. Unlike IMR32, PC12 cells, a rat adrenal pheochromocytoma cell line, resembling human neuroblastoma cell lines and also expressing the A2 subtype of adenosine receptors, was shown not to differentiate under stimulation of the A2A subtype of adenosine receptors 3. Moreover, adenosine inhibited neuronal differentiation in mouse dorsal root ganglion cells presumably via the A1 subtype 4. The mechanism(s) of these confusing effects of adenosine on neuronal differentiation require examination. First, a detection method for each of the adenosine receptor subtypes was developed using reverse transcriptase polymerase chain reaction (RT-PCR). This provided a sensitive, non-radioactive, analytical tool. Subtype-specific, four pairs of PCR primers, corresponding to the A1, A2A, A2B and A3 receptors, were designed and synthesized. The RT-PCR study revealed the presence of adenosine A1, A2A and A2B receptor mRNAs in untreated SH-SY5Y cells. These PCR primers were also designed so that they would allow multiplex PCR. Optimization of conditions for multiplex PCR was conducted, allowing it to detect several adenosine receptor subtypes simultaneously, and it was proven to be partially successful. In the study of differentiation, the use of the designed PCR primers was not quantitative to measure the levels of adenosine receptors due to variations of the expressions levels of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene, a house-keeping gene commonly used as the internal control in PCR or northern blot analysis. An adequate neuronal differentiation model system was established in order to study the possible role(s) of adenosine in neuronal differentiation. Nerve growth factor (NGF), a well-known inducer of differentiation of rat PC12 cells, did not show any apparent differentiation effects on human neuroblastoma SH-SY5Y cells. All-trans retinoic acid (50 µM) induced distinct neuronal differentiation in SH-SY5Y cells, however ethanol, used as a vehicle for retinoic acid, was also shown to have effects on this cell line causing morphological changes. Adenosine (100 µM) alone also did not induce marked differentiation in this cell line probably due to the presence of adenosine in serum. Adenosine deaminase-resistant, synthetic adenosine analogues were used and demonstrated enhancement of differentiation. A serum deprivation-induced differentiation in SH-SY5Y was found to be a consistent and useful model to evaluate the effects of other factors on differentiation in this cell line. This serum deprivation-induced differentiation was also found to accompany a substantial rise in the expression of neurofilament-H (NF-H), one of the marker proteins for neuronal differentiation, at the protein level. Using this model, the possible involvement of adenosine signaling via its receptors was investigated. Treatment of cells with selective adenosine analogues for the A1 and A2A subtypes, 2-chloro-N6-cyclopentyladenosine (CCPA, 100 nM) and 2-[4-(2-carboxylethyl)phenylamino]-5'-N-ethylcarboxamido (CGS21680, 30 and 100 nM), respectively, enhanced the differentiation induced by serum deprivation at day 7 by approximately 60% and 70%, respectively. These enhancing effects of agonists were blocked by selective antagonists, 8-cyclophenyl-1,3-dipropylxanthine (DPCPX) and 9-chloro-2-(2-furyl)[1,2,4]triazolo[1,5-c]quinalzolin-5-amine (CGS15943), respectively. Simultaneous co-stimulation of the A1 and A2A subtypes with these agonists gave no further effects compared to the enhancing effects exerted by CCPA or CGS21680 alone. Signal transduction pathways were examined using various protein kinase inhibitors. A selective protein kinase A (PKA) inhibitor N-(2-[p-bromocinnamylamino]ethyl)-5-isoquinolinesulfonamide hydrochloride (H-89, 100 nM) alone greatly enhanced the differentiation induced by serum deprivation in this cell line. No additive or synergistic effects of 10 nM H-89 with either the A1 or A2A receptor agonist were seen. A selective mitogen-activated protein kinase kinase (MAPKK) inhibitor 2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one (PD098,059) showed a similar pattern to H-89: 100 nM PD098,059 alone caused enhanced differentiation in serum deprivation-induced SH-SY5Y cells. The combination of PD098,059 and adenosine agonists did not show any further enhancement of differentiation.
On the contrary, a selective protein kinase C (PKC) inhibitor, chelerythrine, suppressed the differentiation (by 51%) by serum deprivation at 1 uM, and at 100 nM, chelerythrine suppressed the enhancement of differentiation caused by CCPA and CGS21680 with no effect on the basic level of differentiation, indicating the possible involvement of PKC both in the differentiation induced by serum deprivation and the adenosine receptor-induced potentiation. Surprisingly, contrary to the assumption that the stimulation of PKA induces or assists neuronal differentiation, H-89 (20 uM) alone exerted a prompt differentiation (44% at day 2) in SH-SY5Y cells in the presence of the normal serum concentration (10%). This data suggests that the previously assumed role of PKA in differentiation must be re-evaluated. This H-89-induced differentiation model was shown to have a different differentiation mechanism to the previous serum deprivation-induced differentiation. Establishment of these new differentiation study models will add further options to explore neuronal differentiation, especially, of human type.Thesis (PhD Doctorate)Doctor of Philosophy (PhD)School of ScienceFull Tex
Analyses of Cytokeratins and p63 Isoforms Expressed by Human Limbal Epithelial Cells
No full textCultivated autologous limbal epithelial grafts have shown promise as a valuable treatment to treat ocular surface injuries. The rationale for this therapy is that the limbus contains progenitor cells of the corneal epithelium and that when expanded and transplanted these cells help to restore a healthy epithelium. Nevertheless, this approach is still experimental and the optimal procedure for producing, grafting and assessing the presence of limbal stem cells in these grafts remains under development.
The first results chapter examines the differences in cytokeratin expression between donor corneoslceral rims and cultivated limbal epithelial cultures derived from these donor rims. The expression of keratins 3 and 14 is more stable between the in situ and in vitro environments, while keratins 6 and 19 appear to be upregulated in limbal epithelial cells in response to their isolation and culture. This highlights the role of keratins in limbal epithelial cell function and provides valuable knowledge for assessing the phenotype of cultivated limbal epithelial grafts. Cytokeratin expression is more important in identifying the transient amplifying population than the progenitor population, indicating the need for more specific markers to identify the progenitor cells.
The second results chapter investigated the potential of p63 in identifying the progenitor population of the limbal epithelium. The use of RT-PCR enabled a more detailed examination of the RNA expression of the p63 isoforms. The experiments performed assessed p63 in situ and in vitro, including conditions designed to expand the progenitor population in culture. This work confirms the value of p63 as a marker for immature limbal epithelium and also demonstrates for the first time the p63 isoforms produced by human limbal epithelial cells in vitro.
The final results chapter describes the application of techniques developed and utilised in the previous 2 chapters upon autologous cell samples and constructed grafts used to treat 5 patients clinically. The phenotypic analyses demonstrate that both the cultivated epithelium and the grafts created from the different donor biopsies exhibit similar properties in the areas examined. In addition, these results allowed comparison between clinically used autologous cultures and the methods employed to cultivate limbal epithelium examined in the previous chapter.
The use of both p63 and the various cytokeratins in assessing the level of differentiation hold merit, however more specific markers for the stem cell population of the limbal epithelium remain elusive. The results present a deeper analysis of the material used to treat limbal stem cell deficiency and give insight into further evolution of these treatments.Thesis (PhD Doctorate)Doctor of Philosophy (PhD)School of Biomolecular and Biomedical SciencesFull Tex
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