882 resultados para Transcripts


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Most breast cancer risk factors are associated with prolonged exposure of the mammary gland to high levels of estrogens. The actions of estrogens are predominantly mediated by two receptors, ERα and ERβ, which act as transcription factors binding with high affinity to estrogen response elements in the promoter region of target genes. However, most target genes do not contain the consensus estrogen response elements, but rather degenerated palindromic sequences showing one or more mutations and other ER-binding sites such as AP-1 and SP-1. Using the differential display reverse transcription-polymerase chain reaction technique, our group identified several genes differentially expressed in normal tissue and in ER-positive and ER-negative primary breast tumors. One of the genes shown to be down-regulated in breast tumors compared to normal breast tissue was the PHLDA1 (Pleckstrin homology-like domain, family A, member 1). In the present study, we investigated the potential of PHLDA1 to be regulated by estrogen via ER in MCF-7 breast cancer cells. The promoter region of PHLDA1 shows an imperfect palindrome, an AP-1- and three SP-1-binding sites potentially regulated by estrogens. We also assessed the effects of 17β-estradiol on PHLDA1 mRNA expression in MCF-7 breast cancer cells. MCF-7 cells exposed to 10 nM 17β-estradiol showed more than 2-fold increased expression of the PHLDA1 transcripts compared to control cells (P = 0.05). The anti-estrogen ICI 182,780 (1 µM) inhibited PHLDA1 mRNA expression and completely abolished the effect of 10 nM 17β-estradiol on PHLDA1 expression (P < 0.05), suggesting that PHLDA1 is regulated by estrogen via ER.

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Insertional mutagenesis is an important tool for functional genomics in Drosophila melanogaster. The insertion site in the KG00562 mutant fly line has been mapped to the CG8709 (herein named DmLpin) locus and to the 3’ of kermit (also called dGIPC). This mutant line presents a high lethality rate resulting from a gain of function. To obtain some insight into the biological role of the mutated locus, we have characterized the mutation and its relation to the high mortality of the KG00562 fly line. In this mutant, we did not detect one of the DmLpin transcripts, namely DmLpinK, but we did detect an unusual 2.3-kb mRNA (LpinK-w). Further investigation revealed that the LpinK-w transcript results from an aberrant splicing between the untranslated first exon of DmLpinK and the mini-white marker gene. Lack of DmLpinK or LpinK-w expression does not contribute to lethality, since heterozygous KG00562/Def7860 animals presented lethality rates comparable to those of the wild type. In contrast, the overexpression of kermit was associated with lethality of the KG00562 fly line. Significantly higher levels of kermit were detected in the Malpighian tubules of KG00562/+ flies that presented higher lethality rates than wild-type or KG00562/Def7860 animals, in which the lethality was rescued. In agreement with a recently reported study, our data support the hypothesis that misexpression of kermit/dGIPC could interfere with Drosophila development, with further investigations being needed in this direction.

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The actions of thyroid hormone (TH) on pancreatic beta cells have not been thoroughly explored, with current knowledge being limited to the modulation of insulin secretion in response to glucose, and beta cell viability by regulation of pro-mitotic and pro-apoptotic factors. Therefore, the effects of TH on proinsulin gene expression are not known. This led us to measure: a) proinsulin mRNA expression, b) proinsulin transcripts and eEF1A protein binding to the actin cytoskeleton, c) actin cytoskeleton arrangement, and d) proinsulin mRNA poly(A) tail length modulation in INS-1E cells cultured in different media containing: i) normal fetal bovine serum - FBS (control); ii) normal FBS plus 1 µM or 10 nM T3, for 12 h, and iii) FBS depleted of TH for 24 h (Tx). A decrease in proinsulin mRNA content and attachment to the cytoskeleton were observed in hypothyroid (Tx) beta cells. The amount of eEF1A protein anchored to the cytoskeleton was also reduced in hypothyroidism, and it is worth mentioning that eEF1A is essential to attach transcripts to the cytoskeleton, which might modulate their stability and rate of translation. Proinsulin poly(A) tail length and cytoskeleton arrangement remained unchanged in hypothyroidism. T3 treatment of control cells for 12 h did not induce any changes in the parameters studied. The data indicate that TH is important for proinsulin mRNA expression and translation, since its total amount and attachment to the cytoskeleton are decreased in hypothyroid beta cells, providing evidence that effects of TH on carbohydrate metabolism also include the control of proinsulin gene expression.

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Anastomotic dehiscence is the most severe complication of colorectal surgery. Metalloproteinases (MMPs) and interleukins (ILs) can be used to analyze the healing process of anastomosis. To evaluate the effects of bromopride on MMP and cytokine gene expression in left colonic anastomoses in rats with or without induced abdominal sepsis, 80 rats were divided into two groups for euthanasia on the third or seventh postoperative day (POD). They were then divided into subgroups of 20 rats for sepsis induction or not, and then into subgroups of 10 rats for administration of bromopride or saline. Left colonic anastomosis was performed and abdominal sepsis was induced by cecal ligation and puncture. A colonic segment containing the anastomosis was removed for analysis of gene expression of MMP-1α, MMP-8, MMP-13, IL-β, IL-6, IL-10, tumor necrosis factor-α (TNF-α), and interferon-γ (IFN-γ). On the third POD, bromopride was associated with increased MMP-1α, MMP-13, IL-6, IFN-γ, and IL-10 gene expression. On the seventh POD, all MMP transcripts became negatively modulated and all IL transcripts became positively modulated. In the presence of sepsis, bromopride administration increased MMP-8 and IFN-γ gene expression and decreased MMP-1, TNF-α, IL-6, and IL-10 gene expression on the third POD. On the seventh POD, we observed increased expression of MMP-13 and all cytokines, except for TNF-α. In conclusion, bromopride interferes with MMP and IL gene expression during anastomotic healing. Further studies are needed to correlate these changes with the healing process.

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Nuclear receptor subfamily 1, group I, member 3 (NR1I3) is reported to be a possible novel therapeutic target for some cancers, including lung, brain and hematopoietic tumors. Here, we characterized expression of NR1I3 in a mouse model of lung carcinogenesis induced by 4-(methylnitrosamino)-4-(3-pyridyl)-1-butanone (NNK), the most potent tobacco carcinogen. Lung tumors were collected from mice treated with NNK (400 mg/kg) and euthanized after 52 weeks. Benign and malignant lesions were formalin-fixed and paraffin-embedded for histology and immunohistochemistry, with samples snap-frozen for mRNA analysis. Immunohistochemically, we found that most macrophages and type I and II pneumocytes expressed NR1I3, whereas fibroblasts and endothelial cells were NR1I3−. Compared with benign lesions, malignant lesions had less NR1I3+ tumor cells. Gene expression analysis also showed an inverse correlation between NR1I3 mRNA expression and tumor size (P=0.0061), suggesting that bigger tumors expressed less NR1I3 transcripts, in accordance with our immunohistochemical NR1I3 tests. Our results indicate that NR1I3 expression decreased during progression of malignant lung tumors induced by NNK in mice.

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The aim of this dissertation was to examine the skills and knowledge that pre-service teachers and teachers have and need about working with multilingual and multicultural students from immigrant backgrounds. The specific goals were to identify pre-service teachers’ and practising teachers’ current knowledge and awareness of culturally and linguistically responsive teaching, identify a profile of their strengths and needs, and devise appropriate professional development support and ways to prepare teachers to become equitable culturally responsive practitioners. To investigate these issues, the dissertation reports on six original empirical studies within two groups of teachers: international pre-service teacher education students from over 25 different countries as well as pre-service and practising Finnish teachers. The international pre-service teacher sample consisted of (n = 38, study I; and n = 45, studies II-IV) and the pre-service and practising Finnish teachers sample encompassed (n = 89, study V; and n = 380, study VI). The data used were multi-source including both qualitative (students’ written work from the course including journals, final reflections, pre- and post-definition of key terms, as well as course evaluation and focus group transcripts) and quantitative (multi-item questionnaires with open-ended options), which enhanced the credibility of the findings resulting in the triangulation of data. Cluster analytic procedures, multivariate analysis of variance (MANOVA), and qualitative analyses mostly Constant Comparative Approach were used to understand pre-service teachers’ and practising teachers’ developing cultural understandings. The results revealed that the mainly white / mainstream teacher candidates in teacher education programmes bring limited background experiences, prior socialisation, and skills about diversity. Taking a multicultural education course where identity development was a focus, positively influenced teacher candidates’ knowledge and attitudes toward diversity. The results revealed approaches and strategies that matter most in preparing teachers for culturally responsive teaching, including but not exclusively, small group activities and discussions, critical reflection, and field immersion. This suggests that there are already some tools to address the need for the support needed to teach successfully a diversity of pupils and provide in-service training for those already practising the teaching profession. The results provide insight into aspects of teachers’ knowledge about both the linguistic and cultural needs of their students, as well as what constitutes a repertoire of approaches and strategies to assure students’ academic success. Teachers’ knowledge of diversity can be categorised into sound awareness, average awareness, and low awareness. Knowledge of diversity was important in teachers’ abilities to use students’ language and culture to enhance acquisition of academic content, work effectively with multilingual learners’ parents/guardians, learn about the cultural backgrounds of multilingual learners, link multilingual learners’ prior knowledge and experience to instruction, and modify classroom instruction for multilingual learners. These findings support the development of a competency based model and can be used to frame the studies of pre-service teachers, as well as the professional development of practising teachers in increasingly diverse contexts. The present set of studies take on new significance in the current context of increasing waves of migration to Europe in general and Finland in particular. They suggest that teacher education programmes can equip teachers with the necessary attitudes, skills, and knowledge to enable them work effectively with students from different ethnic and language backgrounds as they enter the teaching profession. The findings also help to refine the tools and approaches to measuring the competencies of teachers teaching in mainstream classrooms and candidates in preparation.

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Lysinuric protein intolerance (LPI) is a recessively inherited disorder characterised by reduced plasma and increased urinary levels of cationic amino acids (CAAs), protein malnutrition, growth failure and hyperlipidemia. Some patients develop severe immunological, renal and pulmonary complications. All Finnish patients share the same LPIFin mutation in the SLC7A7 gene that encodes CAA transporter y+LAT1. The aim of this study was to examine molecular factors contributing to the various symptoms, systemic metabolic and lipid profiles, and innate immune responses in LPI. The transcriptomes, metabolomes and lipidomes were analysed in whole-blood cells and plasma using RNA microarrays and gas or liquid chromatography-mass spectrometry techniques, respectively. Toll-like receptor (TLR) signalling in monocyte-derived macrophages exposed to pathogens was scrutinised using qRT-PCR and the Luminex technology. Altered levels of transcripts participating in amino acid transport, immune responses, apoptosis and pathways of hepatic and renal metabolism were identified in the LPI whole-blood cells. The patients had increased non-essential amino acid, triacylglycerol and fatty acid levels, and decreased plasma levels of phosphatidylcholines and practically all essential amino acids. In addition, elevated plasma levels of eight metabolites, long-chain triacylglycerols, two chemoattractant chemokines and nitric oxide correlated with the reduced glomerular function in the patients with kidney disease. Accordingly, it can be hypothesised that the patients have increased autophagy, inflammation, oxidative stress and apoptosis, leading to hepatic steatosis, uremic toxicity and altered intestinal microbe metabolism. Furthermore, the LPI macrophages showed disruption in the TLR2/1, TLR4 and TLR9 pathways, suggesting innate immune dysfunctions with an excessive response to bacterial infections but a deficient viral DNA response.

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This study focuses on teacher practices in publicly funded music schools in Finland. As views on the aims of music education change and broaden, music schools across Europe share the challenge of developing their activities in response. In public and scholarly debate, there have been calls for increased diversity of contents and concepts of teaching. In Finland, the official national curriculum for state-funded music schools builds on the ideal that teaching and learning should create conditions which promote ‘a good relationship to music’. The meaning of this concept has been deliberately left open in order to leave room for dialogue, flexibility, and teacher autonomy. Since what is meant by ‘good’ is not defined in advance, the notion of ‘improving’ practices is also open to discussion. The purpose of the study is to examine these issues from teachers’ point of view by asking what music school teachers aim to accomplish as they develop their practices. Methodologically, the study introduces a suggestion for building empirical research on Alperson’s ‘robust’ praxial approach to music education, a philosophical theory which is strongly committed to practitioner perspectives and musical diversity. A systematic method for analysing music education practices, interpretive practice analysis, is elaborated with support from interpretive research methods originally used in policy analysis. In addition, the research design shows how reflecting conversations (a collaborative approach well-known in Nordic social work) can be fruitfully applied in interpretive research and combined with teacher inquiry. Data have been generated in a collaborative project involving five experienced music school teachers and the researcher. The empirical material includes transcripts from group conversations, data from teacher inquiry conducted within the project, and transcripts from follow-up interviews. The teachers’ aspirations can be understood as strivings to reinforce the connection between musical practices and various forms of human flourishing such that music and flourishing can sustain each other. Examples from their practices show how the word ‘good’ receives its meaning in context. Central among the teachers’ concerns is their hope that students develop a free and sustainable interest in music, often described as inspiration. I propose that ‘good relationships to music’ and ‘inspiration’ can be understood as philosophical mediators which support the transition from an indeterminate ‘interest in music’ towards specific ways in which music can become a (co-)constitutive part of living well in each person’s particular circumstances. Different musical practices emphasise different aspects of what is considered important in music and in human life. Music school teachers consciously balance between a variety of such values. They also make efforts to resist pressure which might threaten the goods they think are most important. Such goods include joy, participation, perseverance, solid musical skills related to specific practices, and a strong sense of vitality. The insights from this study suggest that when teachers are able to create inspiration, they seem to do so by performing complex work which combines musical and educational aims and makes general positive contributions to their students’ lives. Ensuring that teaching and learning in music schools remain as constructive and meaningful as possible for both students and teachers is a demanding task. The study indicates that collaborative, reflective and interdisciplinary work may be helpful as support for development processes on both individual and collective levels of music school teacher practices.

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This study examined the influence of training on Asian learners' beliefs, interaction, and attitudes during collaborative learning (CL) and explored the processes of their CL in pairs. The literature contains few studies on the effect of collaborative training in language learning. In addition, it shows gaps between SLA theory and practice resulting from learners' cultural differences. Although second/subsequent language acquisition (SLA) theory assumes that CL contributes to language learning, implementing CL in a multicultural classroom is often considered to be unsuccessful by teachers. The research questions designed to address this gap explore: (a) the extent to which tra~ng affects Asian learners' attitudes towards and interaction during CL; (b) how Asian learners accomplish collaborative tasks in pairs. In the quasi-experimental research design, the learners in the treatment group received special training in CL for 5 weeks while the learners in the comparison group did not receive similar training. Data were collected from 45 McMaster University students through pre- and posttests, pre- and postintervention questionnaires, student information, and informal classroom observations. To detennine the influence of training, the frequency of communication units (c-units), Language Related Episodes (LREs), Collaborative Dialogue (CD) from audio-taped data, and the fmal draft scores were compared between pre- and posttests. The learners' pre- and postintervention questionnaires were also compared. Transcripts from audio-taped data, students' information, their responses and comments from questionnaires, and informal observations served to investigate the processes of Asian learners' CL. Overall, this study found that training had significant influence on the frequency of c-units and CD, and considerable impact on the draft scores, although little influence on the frequency of LREs was observed. The results from the questionnaires in the treatment group showed positive changes in the learners' beliefs on pair work after training. On the other hand, analyses of the transcription data showed that the learners did not conduct enough discussion for a resolution of problems with peers. In conclusion, results suggested the need for teacher intervention, a longer period of collaborative training, and an implementation of self-evaluation into the course grade to encourage the learners to succeed in collaborative learning.

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There are a considerable number of programs and agencies that count on the existence of a unique relationship between nature and human development. In addition, there are significant bodies of literature dedicated to understanding developmentally focused nature-based experiences. This research project was designed to flirther the understanding of this phenomenon. Consequently, the purpose of this research endeavour was to discover the essence ofthe intersection ofpersonal transformation and nature-based leisure, culminating in a rich and detailed account of this otherwise tacit phenomenon. As such, this research built on the assumption of this beneficial intersection of nature and personal transformation and contributes to the understanding ofhow this context is supporting or generating of selfactualization and positive development. Heuristic methods were employed because heuristics is concerned with the quality and essence of an experience, not causal relationships (Moustakas, 1990). Heuristic inquiry begins with the primary researcher and her personal experience and knowledge of the phenomenon. This study also involved four other coresearchers who had also experienced this phenomenon intensely. Co-researchers were found through purposeful and snowball sampling. Rich narrative descriptions of their experiences were gathered through in-depth, semi-structured interviews, and artifact elicitation was employed as a means to get at co-researchers' tacit knowledge. Each coresearcher was interviewed twice (the first interview focused on personal transformation, the second on nature) for approximately four and a half hours in total. Transcripts were read repeatedly to discern patterns that emerged from the study of the narratives and were coded accordingly. Individual narratives were consolidated to create a composite narrative of the experience. Finally, a creative synthesis was developed to represent the essence of this tacit experience. In conclusion the essence of the intersection of nature-based leisure and personal transformation was found to lie in the convergence of the lived experience of authenticity. The physical environment of nature was perceived and experienced to be a space and context of authenticity, leisure experiences were experienced as an engagement of authenticity, and individuals themselves encountered a true or authentic self that emanated from within. The implications of these findings are many, offering suggestions, considerations and implications from reconsidered approaches to environmental education to support for selfdirected human development.

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This thesis explored 5 public elementary school teachers ' perceptions of spirituality and its implications for classroom practice. A generic qualitative study was conducted where each teacher shared her experiences and perceptions in one audiotaped semistructured interview. Transcripts were generated and coded for themes which emerged, resulting in the findings of the study. Following this process, the participants verified the accuracy of the transcripts and findings through a member-checking system. The research found that each teacher has her own definition of spirituality. Furthermore, one's personal connection with spirituality can involve a relationship with religion, the self, a higher being, others, and nature. These spiritual relationships were nurtured through a variety of methods outlined by the teachers. This resulted in the creation of a personal spiritual profile for each teacher which contained each teacher's spiritual connections or facets and the methods used to develop these facets. The teachers identified spiritual needs in their students warranting the need for and importance of spiritual education. Given this, a number of classroom practices were identified with the intention of meeting the spiritual needs of students. Among these practices, the teacher as role model was identified as a significant practice for students' spiritual development in the classroom. The teachers further outlined a number of professional development initiatives with the intention to promote greater awareness for spiritual education and to provide resources for educators.

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The purpose of this qualitative study was to understand the relationships between creativity and the working artist/teacher employed by an art college. The topic emerged from my job as an instructor at The Ontario College of Art which was used as the primary data resource and provided the highest caliber of professionals to chose from. Existent data were used to facilitate the study generated by the research of Cawelti, Rappaport, and Wood (1992). The data were generated by a group of 5 faculty members from The University of Northern Iowa, recognized for their expertise in the arts (a painter, a poet, a sculptor, a novelist, and a photographer). They were asked to respond to the following statement: "In as much detail as you like, list the things that you did, thought, or felt the last time you created an artistic product. II Cawelti, Rappaport, and Wood (1992) produced three models of the creative process, each building on the previous, with the resultant third,being in my opinion, an excellent illustration (text/visual) of the creative process. Model three (Appendix D) presented a "multi-dimensional view of the creative process: time, space, observatility, and consciousnessll (p. 90). Model three utilized a visual mapping device along the bottom of the page linked to text segments above. Both the visual and the text were interrelated so that they harmonized into a comprehensive "picture." The parti'cipants of this qualitative study were asked to consider model three from their professional perspective as artist/teachers. The interpretive sciences directed the methodology. The hermeneutic circle of continuous reflection from the whole to the part and back to the whole was an important aspect of the data analyses. Four members of the Foundation Department at The Ontario College of Art were the key participants. A series of conversational interviews was the primary source of data collection, this was augmented by observation, fie,ldnotes, and follow up telephone interviews. Transcripts of interviews were returned to participants for reflection and the telephone was used to discuss any additional -points raised. Analysis consisted of coding and organizing data according to emerging themes. These themes formed the basis for the narrative stories. The text of the narrative stories were given back to each participant for further comment. Revisions were made until both the researcher and the participants felt that the stories reflected reality. The resultant whole was critiqued from the researcher's perspective. The significance of this study was discussed as it pertains to the working artist/teacher and areas in need of further study are pointed out.

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The concept of self-directed learning was examined from the nurse educators' perspective. One structured interview, lasting between one and one-and-a half hours, was conducted with each of 14 nurse educators in two community college continuing education programs in nursing. One community college program encourages selfdirected learning; the other encourages self-study and active participation in the teaching/learning process. All 14 interviews were tape-recorded and transcribed verbatim. Verbatim transcripts were analyzed for themes, patterns, and relationships utilizing analyst-constructed typologies. six prerequisites or necessary conditions for facilitating self-directed learning in a community college continuing education program in nursing were identified. ~he crucial issue in facilitating self-directed learning was found to be the issue of teacher-control.

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Human adenoviruses (Ads), members of the family adenoviridae, are medium-sized DNA viruses which have been used as valuable research tools for the study of RNA processing, oncogenic transformation, and for the development of viral vectors for use in gene delivery and immunization technology. The left 12% of the linear Ad genollle codes for products which are necessary for the efficient replication of the virus, as well as being responsible for the forlllation of tumors in animallllodels. The establishlllent of the 293 cell line, by immortalization of human embryonic kidney cells with th~ E1 region of Ad type S (AdS), has facilitated extensive manipulation of the Ads and the development of recombinant Ad vectors. The study of bovine adenoviruses (BAVs), which cause mild respiratory and gastrointestinal infections in cattle has, on the other hand, been limited primarily to that of infectivity, immunology and clinicallllanifestations. As a result, any potential as gene delivery vehicles has not yet been realized. Continued research into the molecular biolo~gy of BAVs and the development of recolllbinant vectors would benefit from the development of a cell line analogous to that of the 293 cells. In an attelllpt to establish such a cell line, the recombinant plaslllid pKC-neo was constructed, containing the left 0-19.7% of the BAV type 3 (BAV3) genome, and the selectable marker for resistance to the aminoglycoside G418, a neomycin derivative. The plasmid construct was then used to transfect both the Madin-Darby bovine kidney (MDBK) -iicell line and primary bovine lung cells, after which G418-resistant foci were selected for analysis. Two cell lines, E61 (MDBK) and E24 (primary lung), were subsequently selected and analysed for DNA content, revealing the presence of the pKC-neo sequences in their respective genomes. In addition, BAV3 RNA transcripts were detected in the E61 cells. Although the presence of E1 products has yet to be confirmed in both cell lines, the E24 cells exhibit a phenotype characteristic of partial transformation by E1. The apparent immortalization of the primary lung cells will permit exploitation of their ability to take up exogenous DNA at high efficiency.

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This qualitative study examines teachers' experiences implementing new standardized curricula in Ontario schools. This new curricula contained several policy changes and an expectations based format which directed what knowledge and skills students were to demonstrate in each subject. This level of specificity of subject-content served to control teachers in relation to curricula; however, data suggested that at the same time, teachers had enormous flexibility in terms of pedagogy. Four secondary teachers who were implementing a Grade 10 course in the 2000-2001 school year participated in the study. The qualitative framework supported the researcher's emphasis on examining the participants' perspectives on the implementation of expectation-based curricula. Data collected included transcripts from interviews conducted with teacher participants and a representative of the Ontario Ministry of Education and Training, field notes, and a research journal. Many of the factors often cited in the literature as influencing implementation practices were found to have affected the participants' experiences of curriculum implementation: time, professional development, and teachers' beliefs, particularly concerning students. In addition, the format of the policy documents proved to both control and free teachers during the implementation process. Participants believed that the number of specific expectations did not provide them an opportunity to add content to the curriculum; at the same time, teachers also noted that the general format of the policy document allowed them to direct instruction to match students' needs and their own teaching preferences. Alignment between teachers' beliefs about education and their understanding of the new curriculum affected the ways in which many participants adapted during the implementation process.