44 resultados para Reading frames

em Brock University, Canada


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ABSTRACT Recombinant adenoviruses are currently under intense investigation as potential gene delivery and gene expression vectors with applications in human and veterinary medicine. As part of our efforts to develop a bovine adenovirus type 2 (BAV2) based vector system, the nucleotide sequence of BAV2 was determined. Sixty-six open reading frames (ORFs) were found with the potential to encode polypeptides that were at least 50 amino acid (aa) residue long. Thirty-one of the BAV2 polypeptide sequences were found to share homology to already identified adenovirus proteins. The arrangement of the genes revealed that the BAV2 genomic organization closely resembles that of well-characterized human adenoviruses. In the course of this study, continuous propagation of BAV2 over many generations in cell culture resulted in the isolation of a BAV2 spontaneous mutant in which the E3 region was deleted. Restriction enzyme, sequencing and PCR analyses produced concordant results that precisely located the deletion and revealed that its size was exactly 1299 bp. The E3-deleted virus was plaque-purified and further propagated in cell culture. It appeared that the replication of such a virus lacking a portion of the E3 region was not affected, at least in cell culture. Attempts to rescue a recombinant BAV2 virus with the bacterial kanamycin resistance gene in the E3 region yielded a candidate as verified with extensive Southern blotting and PCR analyses. Attempts to purify the recombinant virus were not successful, suggesting that such recombinant BAV2 was helper-dependent. Ten clones containing full-length BAV2 genomes in a pWE15 cosmid vector were constructed. The infectivity of these constructs was tested by using different transfection methods. The BAV2 genomic clones did appear to be infectious only after extended incubation period. This may be due to limitations of various transfection methods tested, or biological differences between virus- and E. co//-derived BAV2 DNA.

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Fire blight is an economically important disease of apples and pears that is caused by the bacterium Erwinia amylovora. Control of the disease depends on limiting primaly blosson1 infection in the spring, and rapidly removing infected tissue. The possibility of using phages to control E.amylovora populations has been suggested, but previous studies have. failed to show high treatment efficacies. This work describes the development of a phage-based biopesticide that controls E. amylovora populations under field conditions, and significantly reduces the incidence of fire blight. This work reports the first use ofPantoea agglomerans, a non-pathogenic relative ofE. amylovora, as a carrier for E. amylovora.phages. Its role is to support a replicating population of these phages on blossom surfaces during the period when the flowers are most susceptible to infection. Seven phages and one carrier isolate were selected for field trials from existing collections of 56 E. amylovora phages and 249 epiphytic orchard bacteria. Selection of the . /' phages and carrier was based on characteristics relevant to the production and field perfonnance of a biopesticide: host range, genetic diversity, growth under the conditions of large-scale production, and the ability to prevent E. amylovora from infecting pear blossoms. In planta assays showed that both the phages and the carrier make significant contributions to reducirig the development of fire blight symptoms in pear blossoms. Field-scale phage production and purification methods were developed based on the growth characteristics of the phages and bacteria in liquid culture, and on the survival of phages in various liquid media. Six of twelve phage-carrier biopesticide treatments caused statistically signiflcant reductions in disease incidence during orchard trials. Multiplex real-time PCR was used to simultaneously monitor the phage, carrier, and pathogen populations over the course of selected treatments. In all cases. the observed population dynamics of the biocontrol agents and the pathogen were consistent with the success or failure of each treatment to control disease incidence. In treatments exhibiting a significantly reduced incidel1ce of fire blight, the average blossom population ofE.amylovora had been reduced to pre-experiment epiphytic levels. In successful treatments the phages grew on the P. agglomerans carrier for 2 to 3 d after treatment application. The phages then grew preferentially on the pathogen, once it was introduced into this blossom ecosystem. The efficacy of the successful phage-based treatnlents was statistically similar to that of streptomycin, which is the most effective bactericide currently available for fire blight prevention. The in planta behaviour ofE. amylovora was compared to that ofErwinia pyrifoliae, a closely related species that causes fire blight-like synlptoms on pears in southeast Asia. Duplex real-time PCR was used to monitor the population dynamics of both species on single blossonls. E. amylovora exhibited a greater competitive fitness on Bartlett pear blossoms than E. pyrifoliae. The genome ofErwinia phage Ea21-4 was sequenced and annotated. Most of the 8-4.7 kB genome is substantially different from previously described sequences, though some regions are notably similar to Salmonella phage Felix 01 . Putative functions were assigned to approximately 30% of the predicted open reading frames based on amino acid sequence comparisons and N-terminal sequencing of structural proteins.

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Adenoviruses are nonenveloped icosahedral shaped particles. The double stranded DNA viral genome is divided into 5 major early transcription units, designated E1 A, E1 B, and E2 to E4, which are expressed in a regulated manner soon after infection. The gene products of the early region 3 (E3), shown to be nonessential for viral replication in vitro, are believed to be involved in counteracting host immunosurveillance. In order to sequence the E3 region of Bovine adenovirus type 2 (BAV2) it was necessary to determine the restriction map for the plasmid pEA48. A physical restriction endonuclease map for BamHl, Clal, Eco RI, Hindlll, Kpnl, Pstt, Sail, and Xbal was constructed. The DNA insert in pEA48 was determined to be viral in origin using Southern hybridization. A human adenovirus type 5 recombinant plasmid, containing partial DNA fragments of the two transcription units L4 and L5 that lie just outside the E3, was used to localize this region. The recombinant plasmid pEA was subcloned to facilitate sequencing. The DNA sequences between 74.8 and 90.5 map units containing the E3, the hexon associated protein (pVIII), and the fibre gene were determined. Homology comparison revealed that the genes for the hexon associated pV11I and the fibre protein are conserved. The last 70 amino acids of the BAV2 pV11I were the most conserved, showing a similarity of 87 percent with Ad2 pV1I1. A comparison between the predicted amino acid sequences of BAV2 and Ad40, Ad41 , Ad2 and AdS, revealed that they have an identical secondary structure consisting of a tail, a shaft and a knob. The shaft is composed of 22, 15 amino acid motifs, with periodic glycines and hydrophobic residues. The E3 region was found to consist of about 2.3 Kbp and to encode four proteins that were greater than 60 amino acids. However, these four open reading frames did not show significant homology to any other known adenovirus DNA or protein sequence.

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Adenoviruses are non-enveloped icosahedral-shaped particles which possess a double-stranded DNA genome. Currently, nearly 100 serotypes of adenoviruses have been identified, 48 of which are of human origin. Bovine adenoviruses (BAVs), causing both mild respiratory and/or enteral diseases in cattle, have been reported in many countries all over the world. Currently, nine serotypes of SAVs have been isolated which have been placed into two subgroups based on a number of characteristics which include complement fixation tests as well as the ability to replicate in various cell lines. Bovine adenovirus type 2 (BAV2), belonging to subgroup I, is able to cause pneumonia as well as pneumonic-like symptoms in calves. In this study, the genome of BAV2 (strain No. 19) was subcloned into the plasmid vector pUC19. In total, 16 plasmids were constructed; three carry internal San fragments (spanning 3.1 to 65.2% ), and 10 carry internal Pstl fragments (spanning 4.9 to 97.4%), of the viral genome. Each of these plasmids was analyzed using twelve restriction endonucleases; BamHI, CiaI, EcoRl, HiOOlll, Kpnl, Noll, NS(N, Ps~, Pvul, Saj, Xbal, and Xhol. Terminal end fragments were also cloned and analyzed, sUbsequent to the removal of the 5' terminal protein, in the form of 2 BamHI B fragments, cloned in opposite orientations (spanning 0 to 18.1°k), and one Pstll fragment (spanning 97.4 to 1000/0). These cloned fragments, along with two other plasmids previously constructed carrying internal EcoRI fragments (spanning 20.6 to 90.5%), were then used to construct a detailed physical restriction map using the twelve restriction endonucleases, as well as to estimate the size of the genome for BAV2(32.5 Kbp). The DNA sequences of the early region 1 (E1) and hexon-associated gene (protein IX) have also been determined. The amino acid sequences of four open reading frames (ORFs) have been compared to those of the E1 proteins and protein IX from other Ads.

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Recombinant Adenoviruses (Ads) have been shown to have potential applications in three areas: gene therapy, high level protein expression and recombinant vaccines.' At least three different locations within the Ad genome can be deleted and subsequently used for the insertion of foreign sequences. These include the Early 3 (E3), Early 1 (E1) and Early 4 (E4) regions. Viral vectors of this type have been well studied in Human Ads 2 and 5, however one has not yet been constructed for Bovine Adenovirus Type 2 (BAV2). The E3 region is located between 76.6 and 86 m.u. on the r-strand and is transcribed in a rightward direction. The gene products of the Early 3 region (E3) have been shown to be non-essential for viral replication, in vitro, but are required for host immunosurveillance. This study represents the cloning and reconstitution of a BAV2 E3 deletion mutant. A deletion of 1800bp was made within the E3 region of BAV2 and the thymidine kinase gene was subsequently inserted in the deleted area . . The plasmid pdlE3-4tk1 (23.4Kbp) was constructed and used to to facilitate homologous recombination with the wild type BAV2 to produce a mutant. Southern Blotting and Hybridization results suggest the presence of a BAV2 E3 deletion mutant with thymidine kinase sequences present. The E4 region of Human Adenovirus types 2 and 5 is located at the extreme right end of the genome (91.3 map units - 99.1 map units) and is transcribed in a leftward direction giving rise to a complicated set of differentially spliced mRNAs. Essentially there are 7 open reading frames (ORFs) encoding for at least 7 polypeptides. The gene products encoded by the E4 region have been shown to be essential for the expression of late viral genes, host cell shutoff and normal viral growth. We have cloned and sequenced the right end segment between 90.5 map units and 100 map units of the BAV2 genome. The results show several open reading frames which encode polypeptides exhibiting homology to three polypeptides encoded by the E4 region of human adenovirus type 2. These include the 14kDa protein encoded by ORF1, the 34kDa protein encoded by ORF6 and the 13kDa protein encoded by ORF3. The nucleotide sequence, restriction enzyme map, and ORF map of the E4 region could be very useful in future molecular manipulation of this region and could possibly explain the slow growth rate of BAV2 in MDBK cells.

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Dr. Gibson reading a book while standing in front of a collection of shelfed science texts.

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This study examined the effect of expHcitly instructing students to use a repertoire of reading comprehension strategies. Specifically, this study examined whether providing students with a "predictive story-frame" which combined the use of prediction and summarization strategies improved their reading comprehension relative to providing students with generic instruction on prediction and summarization. Results were examined in terms of instructional condition and reading ability. Students from 2 grade 4 classes participated in this study. The reading component of the Canadian Achievement Tests, Second Edition (CAT/2) was used to identify students as either "average or above average" or "below average" readers. Students received either strategic predication and summarization instruction (story-frame) or generic prediction and summarization instruction (notepad). Students were provided with new but comparable stories for each session. For both groups, the researcher modelled the strategic tools and provided guided practice, independent practice, and independent reading sessions. Comprehension was measured with an immediate and 1-week delayed comprehension test for each of the 4 stories, hi addition, students participated in a 1- week delayed interview, where they were asked to retell the story and to answer questions about the central elements (character, setting, problem, solution, beginning, middle, and ending events) of each story. There were significant differences, with medium to large effect sizes, in comprehension and recall scores as a fimction of both instructional condition and reading ability. Students in the story-frame condition outperformed students in the notepad condition, and average to above average readers performed better than below average readers. Students in the story-frame condition outperformed students in the notepad condition on the comprehension tests and on the oral retellings when teacher modelling and guidance were present. In the cued recall sessions, students in the story-frame instructional condition recalled more correct information and generated fewer errors than students in the notepad condition. Average to above average readers performed better than below average readers across comprehension and retelling measures. The majority of students in both instructional conditions reported that they would use their strategic tool again.

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The main thrust of this thesis is the re-exploration of Friedrich Nietzsche's "critique of nihilism" through the lenses of Gilles Deleuze. A Deleuzian reading of Nietzsche is motivated by a post-deconstrnctive style of interpretation, inasmuch as Deleuze goes beyond, or in between, henneneutics and deconstrnction. Deleuze's post-deconstrnctive reading of Nietzsche is, however, only secondary to the main aim of this thesis. The primary thrust of this study is the critique of a way of thinking characterized by Nietzsche as nihilistic. Therefore, it should be noted that this study is not about Deleuze's reading per se; rather, it is an appraisal of Nietzsche's "critique of nihilism" using Deleuze's experimental reading. We will accrue Nietzsche's critique and Deleuze's post-deconstrnctive reading in order to appraise Nietzsche's critique itself. Insofar as we have underscored Deleuze's purported experimentation of Nietzschean themes, this study is also an experiment in itself. Through this experimentation, we will find out whether it is possible to partly gloss Nietzsche's critique of nihilism through Deleuzian phraseology. Far from presenting a mere exposition of Nietzsche's text, we are, rather, re-reading, that is, re-evaluating Nietzsche's critique of nihilism through Deleuze's experimentation. This is our way of thinking with Nietzsche. Nihilism is the central problem upon which Nietzsche's philosophical musings are directed; he deems nihilism as a cultural experience and, as such, a phenomenon to be reckoned with. In our reconstruction of Nietzsche's critique of nihilism, we locate two related elements which constitute the structure of the prescription of a cure, Le., the ethics of affirmation and the ontology of becoming. Appraising Nietzsche's ethics and ontology amounts to clarifying what Deleuze thinks as the movement from the "dogmatic image of thought" to the "new image of thought." Through this new image of thought, Deleuze makes sense of a Nietzschean counterculture which is a perspective that resists traditional or representational metaphysics. Deleuze takes the reversal of Platonism or the transmutation of values to be the point of departure. We have to, according to Deleuze, abandon our old image of the world in order to free ourselves from the obscurantism of foundationalist or essentialist thinking. It is only through the transmutation of values that we can make sense of Nietzsche's ethics of affirmation and ontology of becoming. We have to think of Nietzsche's ethics as an "ethics" and not a moral philosophy, and we have to think of his ontology as 1/ ontology" and not as metaphysics. Through Deleuze, we are able to avoid reading Nietzsche as a moral philosopher and metaphysician. Rather, we are able to read Nietzsche as one espousing an ethical imperative through the thought of the eternal return and one advocating a theory of existence based on an immanent, as opposed to transcendent, image of the world.

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It has bee1l said that feminism is dead, but in fact feminism is alive in popular cultural fonlls that offer pleasure, style, fUll and advice, as well as political messages that are internalized alld continuously enacted in the lives of North American female youth. This thesis discusses popular feminism with respect to mainstream girls' cultural discourses in music alld magazine reading. Specifically this thesis examines the importance of Madonna, Gwen Stefani, and the Spice Girls, in addition to the numerous girl magazines available on the market today, such as Seventeen and YM. Focusing on the issue of the feminine versus feminist polarity and its importance to girls' culture, this thesis attempts to demonstrate how popular feminism can be used as a mode of empowerment and illustrates the mode of consumption of popular feminist texts that frames female selfimage, attitude, behaviour and speech. Through the employment of popular feminist theories and a discourse alld semiotic analysis of musical lyrics, performance and style, in addition to magazine reading and advertisements, this thesis highlights the use of active media reading and being by girls to gaill an understanding with regards to social positioning and postmodern political identity. More fundamentally, this thesis questions how popular feminism disables, questions and critiques popular ideologies ill a patriarchal society.

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This study examined the effects of providing students with explicit instruction in how to use a repertoire of reading comprehension strategies and test taking skills when reading and responding to three types of questions (direct, inferential, critical). Specifically, the study examined whether providing students with a "model" of how to read and respond to the text and to the comprehension questions improved their reading comprehension relative to providing them with implicit instruction on reading comprehension strategies and test taking skills. Students' reading comprehension and test taking performance scores were compared as a function of instructional condition. Students from 2 grade 8 classes participated in this study. The reading component of the Canadian Achievement Tests, Third Edition (CAT/3) was used to identify students' level of reading comprehension prior to the formal instructional sessions. Students received either explicit instruction, which involved modelling, or implicit instruction, which consisted of review and discussion of the strategies to be used. Comprehension was measured through the administration of formative tests after each instructional session. The formative tests consisted of reading comprehension questions pertaining to a specific form of text (narrative, informational, graphic). In addition, students completed 3 summative tests and a delayed comprehension test which consisted of the alternative version of the CAT/3 standardized reading assessment. These data served as a posttest measure to determine whether students had shown an improvement in their reading comprehension skills as a result of the program delivery. There were significant differences in students' Canadian Achievement Test performance scores prior to the onset of the study. Students in the implicit group attained significantly higher comprehension scores than did students in the explicit group. The results from the program sessions indicated no significant differences in reading comprehension between the implicit and explicit conditions, with the exception of the 6th session involving the reading and interpreting of graphic text. Students in the explicit group performed significantly better when reading and interpreting graphic text than those in the implicit group. No significant differences were evident between the two study conditions across the three summative tests. Upon completion of the study, the results from the Canadian Achievement Test indicated no significant differences in performance between the two study conditions. The findings from this study reveal the effectiveness of providing students with explicit strategy instruction when reading and responding to various forms of text. Modelling the appropriate reading comprehension strategies and test taking skills enabled students to apply the same thought processes to their own independent work. This form of instruction enabled students in the explicit group to improve in their abilities to comprehend and respond to text and therefore should be incorporated as an effective form of classroom teaching.

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In this thesis, three main questions were addressed using event-related potentials (ERPs): (1) the timing of lexical semantic access, (2) the influence of "top-down" processes on visual word processing, and (3) the influence of "bottom-up" factors on visual word processing. The timing of lexical semantic access was investigated in two studies using different designs. In Study 1,14 participants completed two tasks: a standard lexical decision (LD) task which required a word/nonword decision to each target stimulus, and a semantically primed version (LS) of it using the same category of words (e.g., animal) within each block following which participants made a category judgment. In Study 2, another 12 participants performed a standard semantic priming task, where target stimulus words (e.g., nurse) could be either semantically related or unrelated to their primes (e.g., doctor, tree) but the order of presentation was randomized. We found evidence in both ERP studies that lexical semantic access might occur early within the first 200 ms (at about 170 ms for Study 1 and at about 160 ms for Study 2). Our results were consistent with more recent ERP and eye-tracking studies and are in contrast with the traditional research focus on the N400 component. "Top-down" processes, such as a person's expectation and strategic decisions, were possible in Study 1 because of the blocked design, but they were not for Study 2 with a randomized design. Comparing results from two studies, we found that visual word processing could be affected by a person's expectation and the effect occurred early at a sensory/perceptual stage: a semantic task effect in the PI component at about 100 ms in the ERP was found in Study 1 , but not in Study 2. Furthermore, we found that such "top-down" influence on visual word processing might be mediated through separate mechanisms depending on whether the stimulus was a word or a nonword. "Bottom-up" factors involve inherent characteristics of particular words, such as bigram frequency (the total frequency of two-letter combinations of a word), word frequency (the frequency of the written form of a word), and neighborhood density (the number of words that can be generated by changing one letter of an original word or nonword). A bigram frequency effect was found when comparing the results from Studies 1 and 2, but it was examined more closely in Study 3. Fourteen participants performed a similar standard lexical decision task but the words and nonwords were selected systematically to provide a greater range in the aforementioned factors. As a result, a total of 18 word conditions were created with 18 nonword conditions matched on neighborhood density and neighborhood frequency. Using multiple regression analyses, we foimd that the PI amplitude was significantly related to bigram frequency for both words and nonwords, consistent with results from Studies 1 and 2. In addition, word frequency and neighborhood frequency were also able to influence the PI amplitude separately for words and for nonwords and there appeared to be a spatial dissociation between the two effects: for words, the word frequency effect in PI was found at the left electrode site; for nonwords, the neighborhood frequency effect in PI was fovind at the right elecfrode site. The implications of otir findings are discussed.

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In this thesis, I critically examine the discourses that inform how we conceptualise HIV/AIDS in Sub-Saharan Africa as they are produced in a sample of Canadian news articles, two nonfiction texts - Stephanie Nolen's 28 Stories of AIDS in Africa and Jonathan Morgan and the Bambanani Women's Group's Long Life ... Positive HIV Stories - as well as two literary texts - John Le Carre's popular fiction novel The Constant Gardener and an anthology of stories and poems from Southern Africa titled Nobody Ever Said AIDS, compiled and edited by Nobantu Rasebotsa, Meg Samuelson and Kylie Thomas. Paying particular attention to the role of metaphor in discursive formation, I have found that military metaphors, usually used in conjunction with biomedical discourses, continue to dominate what is said about HIV/AIDS. However, the use of military metaphors to conceptualise HIV/AIDS contributes to stigma and limits the effectiveness of responses to the pandemic. I argue that accessing alternative metaphors and discourses, such as biopsychosocial discourse, can lead to a more layered - and more beneficial - conceptualisation of HIV/AIDS, encouraging a more active response to the pandemic.

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This study compared the relative effectiveness of two computerized remedial reading programs in improving the reading word recognition, rate, and comprehension of adolescent readers demonstrating significant and longstanding reading difficulties. One of the programs involved was Autoskill Component Reading Subskills Program, which provides instruction in isolated letters, syllables, and words, to a point of rapid automatic responding. This program also incorporates reading disability subtypes in its approach. The second program, Read It Again. Sam, delivers a repeated reading strategy. The study also examined the feasibility of using peer tutors in association with these two programs. Grade 9 students at a secondary vocational school who satisfied specific criteria with respect to cognitive and reading ability participated. Eighteen students were randomly assigned to three matched groups, based on prior screening on a battery of reading achievement tests. Two I I groups received training with one of the computer programs; the third group acted as a control and received the remedial reading program offered within the regular classroom. The groups met daily with a trained tutor for approximately 35 minutes, and were required to accumulate twenty hours of instruction. At the conclusion of the program, the pretest battery was repeated. No significant differences were found in the treatment effects of the two computer groups. Each of the two treatment groups was able to effect significantly improved reading word recognition and rate, relative to the control group. Comprehension gains were modest. The treatment groups demonstrated a significant gain, relative to the control group, on one of the three comprehension measures; only trends toward a gain were noted on the remaining two measures. The tutoring partnership appeared to be a viable alternative for the teacher seeking to provide individualized computerized remedial programs for adolescent unskilled readers. Both programs took advantage of computer technology in providing individualized drill and practice, instant feedback, and ongoing recordkeeping. With limited cautions, each of these programs was considered effective and practical for use with adolescent unskilled readers.

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The gene,nil purpose (If this thesis was to examine certain variables considered to be associated with reading readiness in kindergarten children Twenty-four children from a parent-funded parochial school i.n 5t Catharines. Ontaj-io were exanlined in this study, The children.'s reading readiness level, measured by the Test of Ea1'1y Reading Ability (JERA:) and by teacher"s ratings was correlated with various cognitive variables, These variables consisted (If a rough index of intelligence as measured by the Peabody Picture Vocabulary Test - Revised CPPVT-Rl Form M, mean len.gth of utterance (MUn, the sum of relative,. subordinate and coordinate clauses, the !wmber of core statements children make when telling stories and a memory test. All but the memory test related to at least one measure of reading readiness, Although the memory test did not correlate significantly with reading readiness, the children appeaj~ed to be sensitive to the stimulus set size in terms of their study time, In addition. interi'uption in the interval between studying the stimuli and the ,reca1i test had a negathre effect on pe.rfornlance and set size had a substantial effect 011 recognition performance, The educational implications of these correlates of reading readiness are discussed as weH as the implications for future research.

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The major hypothesis of this paper is that any deviance in syntax present in oral language will be evident in oral r eading behaviour. Using Lee and Canter's Developmental i 1 Sentence Scoring technique (1971) and Y. Goodman and Burke's Reading Miscue Inventory (1972) linguistic competence was established in t hree male children. ages 10 to 11. patterns of strengths and weaknesses in reading were determined. and the relationships t hat were established, were examined. Results of the study i ndicate that oral language behaviour is closely tied to oral r eading behaviour. This type of approach can be used as a basis for a diagnosis of a reading difficulty and then a prescription for language and reading skills.