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1.
Milbank Q ; 101(3): 881-921, 2023 09.
Article in English | MEDLINE | ID: mdl-37186312

ABSTRACT

Policy Points More rigorous methodologies and systematic approaches should be encouraged in the science of scaling. This will help researchers better determine the effectiveness of scaling, guide stakeholders in the scaling process, and ultimately increase the impacts of health innovations. The practice and the science of scaling need to expand worldwide to address complex health conditions such as noncommunicable and chronic diseases. Although most of the scaling experiences described in the literature are occurring in the Global South, most of the authors publishing on it are based in the Global North. As the science of scaling spreads across the world with the aim of reducing health inequities, it is also essential to address the power imbalance in how we do scaling research globally. CONTEXT: Scaling of effective innovations in health and social care is essential to increase their impact. We aimed to synthesize the evidence base on scaling and identify current knowledge gaps. METHODS: We conducted an umbrella review according to the Joanna Briggs Institute Reviewers' Manual. We included any type of review that 1) focused on scaling, 2) covered health or social care, and 3) presented a methods section. We searched MEDLINE (Ovid), Embase, PsycINFO (Ovid), CINAHL (EBSCO), Web of Science, The Cochrane Library, Sociological Abstracts (ProQuest), Academic Search Premier (EBSCO), and ProQuest Dissertations & Theses Global from their inception to August 6, 2020. We searched the gray literature using, e.g., Google and WHO-ExpandNet. We assessed methodological quality with AMSTAR2. Paired reviewers independently selected and extracted eligible reviews and assessed study quality. A narrative synthesis was performed. FINDINGS: Of 24,269 records, 137 unique reviews were included. The quality of the 58 systematic reviews was critically low (n = 42). The most frequent review type was systematic review (n = 58). Most reported on scaling in low- and middle-income countries (n = 59), whereas most first authors were from high-income countries (n = 114). Most reviews concerned infectious diseases (n = 36) or maternal-child health (n = 28). They mainly focused on interventions (n = 37), barriers and facilitators (n = 29), frameworks (n = 24), scalability (n = 24), and costs (n = 14). The WHO/ExpandNet scaling definition was the definition most frequently used (n = 26). Domains most reported as influencing scaling success were building scaling infrastructure (e.g., creating new service sites) and human resources (e.g., training community health care providers). CONCLUSIONS: The evidence base on scaling is evolving rapidly as reflected by publication trends, the range of focus areas, and diversity of scaling definitions. Our study highlights knowledge gaps around methodology and research infrastructures to facilitate equitable North-South research relationships. Common efforts are needed to ensure scaling expands the impacts of health and social innovations to broader populations.


Subject(s)
Health Personnel , Income , Humans , Social Support , Systematic Reviews as Topic
2.
JMIR Aging ; 6: e43106, 2023 Jan 18.
Article in English | MEDLINE | ID: mdl-36566499

ABSTRACT

BACKGROUND: Older adults with disabilities such as loss of autonomy face the decision of whether to stay at home or move to a health care facility such as a nursing home. Therefore, they may need support for this difficult decision. OBJECTIVE: We assessed the intention of Canadian older adults to use an electronic decision aid (eDA) to make housing decisions and identified the factors that influenced their intention. METHODS: We conducted a cross-sectional study using a web-based survey targeting older adults across 10 Canadian provinces and 3 territories. We included respondents from a web-based panel who were aged ≥65 years, understood English or French, had access to an electronic device with an internet connection, and had made a housing decision over the past few months or were planning to make a decision in the coming year. We based the web-based survey on the Unified Theory of Acceptance and Use of Technology (UTAUT). We adapted 17 UTAUT items to measure respondents' intention to use the eDA for housing decisions, as well as items measuring 4 intention constructs (performance expectancy, effort expectancy, social influence, and facilitating conditions). We also assessed eHealth literacy using both subjective and objective scales. We used descriptive statistics and multivariable linear regression analyses to identify the factors influencing the intention to use the eDA. RESULTS: Of the 11,972 invited panelists, 1176 (9.82%) met the eligibility criteria, and 1000 (85.03%) respondents completed the survey. The mean age was 72.5 (SD 5.59) years. Most respondents were male (548/1000, 54.8%), White (906/1000, 90.6%), English speakers (629/1000, 62.9%), and lived in Ontario or Quebec (628/1000, 62.8%) and in urban areas (850/1000, 85%). The mean scores were 27.8 (SD 5.88) out of 40 for subjective eHealth literacy and 3.00 (SD 0.97) out of 5 for objective eHealth literacy. In our sample, the intention score was 4.74 (SD 1.7) out of 7. The mean scores of intention constructs out of 7 were 5.63 (SD 1.28) for facilitating conditions, 4.94 (SD 1.48) for performance expectancy, 5.61 (SD 1.35) for effort expectancy, and 4.76 (SD 1.59) for social influence. In the final model, the factors associated with intention included mother tongue (ß=.30; P<.001), objective eHealth literacy (ß=-.06; P=.03), performance expectancy (ß=.55; P<.001), social influence (ß=.37; P<.001), and facilitating conditions (ß=.15; P<.001). CONCLUSIONS: Findings from this pan-Canadian web-based survey on Canadian older adults suggest that their intention to use the eDA to make housing decisions is similar to the findings in other studies using UTAUT. The factors identified as influencing intention were mother tongue, objective eHealth literacy, performance expectancy, social influence, and facilitating conditions. These will guide future strategies for the implementation of the eDA.

3.
MDM Policy Pract ; 7(2): 23814683221124090, 2022.
Article in English | MEDLINE | ID: mdl-36132436

ABSTRACT

Background. Older adults receiving home care services often face decisions related to aging, illness, and loss of autonomy. To inform tailored shared decision making interventions, we assessed their decisional needs by asking about the most common difficult decisions, measured associated decisional conflict, and identified factors associated with it. Methods. In March 2020, we conducted a cross-sectional survey with a pan-Canadian Web-based panel of older adults (≥65 y) receiving home care services. For a difficult decision they had faced in the past year, we evaluated clinically significant decisional conflict (CSDC) using the 16-item Decisional Conflict Scale (score 0-100) with a >37.5 cutoff. To identify factors associated with CSDC, we performed descriptive, bivariable, and multivariable analyses using the stepwise selection method with an assumed entry and exit significance level of 0.15 and 0.20, respectively. Final model selection was based on the Bayesian information criterion. Results. Among 460 participants with an average age of 72.5 y, difficult decisions were, in order of frequency, about housing and safety (57.2%), managing health conditions (21.8%), and end-of-life care (8.3%). CSDC was experienced by 14.6% (95% confidence interval [CI]: 11.5%, 18.1%) of respondents on all decision points. Factors associated with CSDC included household size = 1 (OR [95% CI]: 1.81 [0.99, 3.33]; P = 0.27), household size = 3 (2.66 [0.78, 8.98]; P = 0.83), and household size = 4 (6.91 [2.23, 21.39]; P = 0.014); preferred option not matching the decision made (4.05 [2.05, 7.97]; P < 0.001); passive role in decision making (5.13 [1.78, 14.77]; P = 0.002); and lower quality of life (0.70 [0.57, 0.87]; P<0.001). Discussion. Some older adults receiving home care services in Canada experience CSDC when facing difficult decisions. Shared decision-making interventions could mitigate associated factors. Highlights: This is the first study in Canada to assess the decisional needs of older adults receiving care at home and to identify their most common difficult decisions.Difficult decisions most frequently made were about housing and safety. The most significant decisional conflict was experienced by people making decisions about palliative care.When their quality-of-life score was low, older adults experienced clinically significant decision conflict.

4.
BMJ Open ; 12(8): e061215, 2022 08 05.
Article in English | MEDLINE | ID: mdl-36129731

ABSTRACT

OBJECTIVES: Decision aids (DAs) for clients in home and community care can support shared decision-making (SDM) with patients, healthcare teams and informal caregivers. We aimed to identify DAs developed for home and community care, verify their adherence to international DA criteria and explore the involvement of interprofessional teams in their development and use. DESIGN: Systematic review reported following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. DATA SOURCES: Six electronic bibliographic databases (MEDLINE, Embase, CINAHL Plus, Web of Science, PsycINFO and the Cochrane Library) from inception to November 2019, social media and grey literature websites up to January 2021. ELIGIBILITY CRITERIA: DAs designed for home and community care settings or including home care or community services as options. DATA EXTRACTION AND SYNTHESIS: Two reviewers independently reviewed citations. Analysis consisted of a narrative synthesis of outcomes and a thematic analysis. DAs were appraised using the International Patient Decision Aid Standards (IPDAS). We collected information on the involvement of interprofessional teams, including nurses, in their development and use. RESULTS: After reviewing 10 337 database citations and 924 grey literature citations, we extracted characteristics of 33 included DAs. DAs addressed a variety of decision points. Nearly half (42%) were relevant to older adults. Several DAs did not meet IPDAS criteria. Involvement of nurses and interprofessional teams in the development and use of DAs was minimal (33.3% of DAs). CONCLUSION: DAs concerned a variety of decisions, especially those related to older people. This reflects the complexity of decisions and need for better support in this sector. There is little evidence about the involvement of interprofessional teams in the development and use of DAs in home and community care settings. An interprofessional approach to designing DAs for home care could facilitate SDM with people being cared for by teams. PROSPERO REGISTRATION NUMBER: CRD42020169450.


Subject(s)
Home Care Services , Patient Participation , Aged , Decision Making , Decision Making, Shared , Decision Support Techniques , Humans , Patient Care Team
5.
MDM Policy Pract ; 7(2): 23814683221116304, 2022.
Article in English | MEDLINE | ID: mdl-35983319

ABSTRACT

Background. In Canada, caregivers of older adults receiving home care face difficult decisions that may lead to decision regret. We assessed difficult decisions and decision regret among caregivers of older adults receiving home care services and factors associated with decision regret. Methods. From March 13 to 30, 2020, at the outbreak of the COVID-19 pandemic, we conducted an online survey with caregivers of older adults receiving home care in the 10 Canadian provinces. We distributed a self-administered questionnaire through Canada's largest and most representative private online panel. We identified types of difficult health-related decisions faced in the past year and their frequency and evaluated decision regret using the Decision Regret Scale (DRS), scored from 0 to 100. We performed descriptive statistics as well as bivariable and multivariable linear regression to identify factors predicting decision regret. Results. Among 932 participants, the mean age was 42.2 y (SD = 15.6 y), and 58.4% were male. The most frequently reported difficult decisions were regarding housing and safety (75.1%). The mean DRS score was 28.8/100 (SD = 8.6). Factors associated with less decision regret included higher caregiver age, involvement of other family members in the decision-making process, wanting to receive information about the options, and considering organizations interested in the decision topic and health care professionals as trustworthy sources of information (all P < 0.001). Factors associated with more decision regret included mismatch between the caregiver's preferred option and the decision made, the involvement of spouses in the decision-making process, higher decisional conflict, and higher burden of care (all P < 0.001). Discussion. Decisions about housing and safety were the difficult decisions most frequently encountered by caregivers of older adults in this survey. Our results will inform future decision support interventions. Highlights: This is one of the first studies to assess decision regret among caregivers of older adults receiving home and community care services and to identify their most frequent difficult decisions.Difficult decisions were most frequently about housing and safety. Most caregivers of older adults in all 10 provinces of Canada experienced decision regret.Factors associated with less decision regret included higher caregiver age, the involvement of other family members in the decision-making process, wanting to receive information about the options, considering organizations interested in the decision topic, and health care professionals as trustworthy sources of information. Factors associated with more decision regret included mismatch between the caregiver's preferred option and the decision made, the involvement of spouses in the decision-making process, higher decisional conflict, and higher burden of care.

6.
Nurs Health Sci ; 24(2): 487-498, 2022 Jun.
Article in English | MEDLINE | ID: mdl-35460164

ABSTRACT

Interprofessional care teams can play a key role in supporting older adults (and caregivers) in making informed health decisions, yet shared decision making is not widely practiced in home care. Based on an earlier needs assessment with older adults (and caregivers) with home care experience, we aimed to explore the perceptions of home care teams on the decisions facing their clients and their perceived involvement in shared decision making. A cross-sectional study was conducted with 614 home care providers (nurses, personal support workers, rehabilitation professionals) in three Canadian provinces (Quebec, Ontario, and Alberta). Home care providers considered the decision "to stay at home or move" as the most difficult for older adults. Those most frequently involved in decision making with older adults were family members and least involved were physicians. Although all home care providers reported high levels of shared decision-making, we detected an effect of respondent's discipline on self-perceived shared decision-making; nurses and rehabilitation professionals reported significantly higher levels of shared decision making than personal support workers. A more tailored approach is required to support shared decision making in interprofessional care teams.


Subject(s)
Caregivers , Home Care Services , Aged , Canada , Cross-Sectional Studies , Decision Making , Decision Making, Shared , Humans
7.
JMIR Med Educ ; 8(2): e36948, 2022 May 02.
Article in English | MEDLINE | ID: mdl-35318188

ABSTRACT

BACKGROUND: Continuing professional development (CPD) is essential for physicians to maintain and enhance their knowledge, competence, skills, and performance. Web-based CPD plays an essential role. However, validated theory-informed measures of their impact are lacking. The CPD-REACTION questionnaire is a validated theory-informed tool that evaluates the impact of CPD activities on clinicians' behavioral intentions. OBJECTIVE: We aimed to review the use of the CPD-REACTION questionnaire, which measures the impact of CPD activities on health professionals' intentions to change clinical behavior. We examined CPD activity characteristics, ranges of intention, mean scores, score distributions, and psychometric properties. METHODS: We conducted a systematic review informed by the Cochrane review methodology. We searched 8 databases from January 1, 2014, to April 20, 2021. Gray literature was identified using Google Scholar and Research Gate. Eligibility criteria included all health care professionals, any study design, and participants' completion of the CPD-REACTION questionnaire either before, after, or before and after a CPD activity. Study selection, data extraction, and study quality evaluation were independently performed by 2 reviewers. We extracted data on characteristics of studies, the CPD activity (eg, targeted clinical behavior and format), and CPD-REACTION use. We used the Mixed Methods Appraisal Tool to evaluate the methodological quality of the studies. Data extracted were analyzed using descriptive statistics and the Student t test (2-tailed) for bivariate analysis. The results are presented as a narrative synthesis reported according to the PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) guidelines. RESULTS: Overall, 65 citations were eligible and referred to 52 primary studies. The number of primary studies reporting the use of CPD-REACTION has increased continuously since 2014 from 1 to 16 publications per year (2021). It is available in English, French, Spanish, and Dutch. Most of the studies were conducted in Canada (30/52, 58%). Furthermore, 40 different clinical behaviors were identified. The most common CPD format was e-learning (34/52, 65%). The original version of the CPD-REACTION questionnaire was used in 31 of 52 studies, and an adapted version in 18 of 52 studies. In addition, 31% (16/52) of the studies measured both the pre- and postintervention scores. In 22 studies, CPD providers were university-based. Most studies targeted interprofessional groups of health professionals (31/52, 60%). CONCLUSIONS: The use of CPD-REACTION has increased rapidly and across a wide range of clinical behaviors and formats, including a web-based format. Further research should investigate the most effective way to adapt the CPD-REACTION questionnaire to a variety of clinical behaviors and contexts. TRIAL REGISTRATION: PROSPERO CRD42018116492; https://www.crd.york.ac.uk/prospero/display_record.php?RecordID=116492.

8.
Syst Rev ; 10(1): 261, 2021 09 30.
Article in English | MEDLINE | ID: mdl-34593027

ABSTRACT

BACKGROUND: There is a growing interest in scaling effective health innovations to promote equitable access to high-quality health services worldwide. However, multiple challenges persist in scaling innovations. In this study, we aim to summarize the scaling evidence in the health and social care literature and identify current knowledge gaps. METHODS: We will conduct a living umbrella review according to the Joanna Briggs Institute Reviewers' Manual. We will consider all knowledge syntheses addressing scaling in health or social care (e.g., any setting, any clinical area) and conducted in a systematic way. We will search the following electronic databases: MEDLINE (Ovid), Embase, PsychINFO (Ovid), CINAHL (EBSCO), Web of Science, The Cochrane Library, Sociological Abstract (Proquest), Academic Search Premier (EBSCO), and Proquest Dissertations & Theses Global, from inception. Furthermore, we will conduct searches of the grey literature. No restriction regarding date or language will be applied. Each phase of the review will be processed by two independent reviewers. We will develop a data extraction form on Covidence. We will assess the methodological quality of the included reviews using AMSTAR2 and the risk of bias using ROBIS. Results will be presented in tabular form and accompanied by a narrative synthesis covering the traditional themes of scaling science that emerge from the analysis, such as coverage, range, and sustainability, as well as themes less covered in the literature, including reporting guidance, models, tools, barriers, and/or facilitators to scaling innovations, evidence regarding application in high-income or low-income countries, and end-user engagement. We will disseminate the findings via publications and through relevant networks. DISCUSSION: The findings of the umbrella review will facilitate access to scaling evidence in the literature and help strengthen the science of scaling for researchers, policy makers, and program managers. Finally, this work will highlight important knowledge gaps and help prioritize future research questions. SYSTEMATIC REVIEW REGISTRATION: This protocol was registered with the International Prospective Register of Systematic Reviews (PROSPERO) on November 11, 2020 (registration number: CRD42020183774 ).


Subject(s)
Health Services , Research Design , Humans , Research Report , Review Literature as Topic , Social Support , Systematic Reviews as Topic
9.
Syst Rev ; 10(1): 55, 2021 02 11.
Article in English | MEDLINE | ID: mdl-33573701

ABSTRACT

BACKGROUND: The scale-up of evidence-based innovations is required to reduce waste and inequities in health and social services (HSS). However, it often tends to be a top-down process initiated by policy makers, and the values of the intended beneficiaries are forgotten. Involving multiple stakeholders including patients and the public in the scaling-up process is thus essential but highly complex. We propose to identify relevant strategies for meaningfully and equitably involving patients and the public in the science and practice of scaling up in HSS. METHODS: We will adapt our overall method from the RAND/UCLA Appropriateness Method. Following this, we will perform a two-prong study design (knowledge synthesis and Delphi study) grounded in an integrated knowledge translation approach. This approach involves extensive participation of a network of stakeholders interested in patient and public involvement (PPI) in scaling up and a multidisciplinary steering committee. We will conduct a systematic scoping review following the methodology recommended in the Joanna Briggs Institute Reviewers Manual. We will use the following eligibility criteria: (1) participants-any stakeholder involved in creating or testing a strategy for PPI; (2) intervention-any PPI strategy proposed for scaling-up initiatives; (3) comparator-no restriction; (4) outcomes: any process or outcome metrics related to PPI; and (5) setting-HSS. We will search electronic databases (e.g., Medline, Web of Science, Sociological Abstract) from inception onwards, hand search relevant websites, screen the reference lists of included records, and consult experts in the field. Two reviewers will independently select and extract eligible studies. We will summarize data quantitatively and qualitatively and report results using the PRISMA extension for Scoping Reviews (PRISMA-ScR) checklist. We will conduct an online Delphi survey to achieve consensus on the relevant strategies for PPI in scaling-up initiatives in HSS. Participants will include stakeholders from low-, middle-, and high-income countries. We anticipate that three rounds will allow an acceptable degree of agreement on research priorities. DISCUSSION: Our findings will advance understanding of how to meaningfully and equitably involve patients and the public in scaling-up initiatives for sustainable HSS. SYSTEMATIC REVIEW REGISTRATION: We registered this protocol with the Open Science Framework on August 19, 2020 ( https://osf.io/zqpx7/ ).


Subject(s)
Research Design , Research Report , Humans , Knowledge , Patient Participation , Review Literature as Topic , Social Work , Systematic Reviews as Topic
10.
BMJ Open ; 10(11): e039102, 2020 11 09.
Article in English | MEDLINE | ID: mdl-33168556

ABSTRACT

INTRODUCTION: Shared decision making is an interpersonal process whereby healthcare providers collaborate with and support patients in decision-making. Older adults receiving home care need support with decision-making. We will explore what older adults receiving home care and their caregivers need for making better health-related decisions. METHODS AND ANALYSIS: This two-phase sequential exploratory mixed methods study will be conducted in a pan-Canadian healthcare organisation, SE Health. First, we will create a participant advisory group to advise us throughout the research process. In phase 1 (qualitative), we will recruit a convenience sample of 15-30 older adults and caregivers receiving home care to participate in open-ended semi-structured interviews. Phase 1 participants will be invited to share what health-related decisions they face at home and what they need for making better decisions. In phase 2 (quantitative), interdisciplinary health and social care providers will be invited to answer a web-based survey to share their views on the decisional needs of older adults and their caregivers. The survey will include questions informed by findings from qualitative interviews in phase 1, and a workbook for assessing decisional needs based on the Ottawa Decision Support Framework. Finally, qualitative and quantitative results will be triangulated (by methods, investigator, theory and source) to develop a comprehensive understanding of decision-making needs from the perspective of older adults, caregivers and health and social care providers. We will use the quality of mixed methods studies in health services research guidelines and the Checklist for Reporting the Results of Internet E-Surveys checklist. ETHICS AND DISSEMINATION: Ethics approval was obtained from the research ethics boards at Southlake Regional Health Centre and Université Laval. This study will inform the design of decision support interventions. Further dissemination plans include summary briefs for study participants, tailored reports for home care decision makers and policy makers, and peer-reviewed publications. TRIAL REGISTRATION NUMBER: NCT04327830.


Subject(s)
Caregivers , Decision Making , Aged , Humans , Reproducibility of Results
11.
Epigenomics ; 8(9): 1209-26, 2016 09.
Article in English | MEDLINE | ID: mdl-27586997

ABSTRACT

AIM: The goal of this study is to characterize the specific methylation profile triggered by DNMT3B protein isoforms expressed at different levels in breast cell lines. MATERIALS & METHODS: Microarray DNA methylation data were analyzed and associated with functional genome annotation data. RESULTS: A large spectrum of DNMT3B3/DNMT3B2 expression ratio values was observed in parental breast cell lines. According to their methylation profiles, hierarchical clustering of untransfected cell lines revealed clustering based on their ER/PR status. Overexpression of DNMT3B3 triggered methylation changes of thousands of CpG sites in breast cells. Based on the trend of methylation changes, the results suggest an antiproliferative action of the DNMT3B3 isoform through a dominant negative effect on its wild-type counterpart DNMT3B2. CONCLUSION: This study revealed specific pathways modulated by DNMT3B isoforms, which could regulate cell proliferation and other biological mechanisms. This illustrates the importance of multiple interactions between isoforms in the complexity of methylation processes.


Subject(s)
DNA (Cytosine-5-)-Methyltransferases/genetics , DNA Methylation , Cell Proliferation , CpG Islands , DNA (Cytosine-5-)-Methyltransferases/metabolism , Epigenesis, Genetic , Gene Expression Regulation, Neoplastic , HEK293 Cells , Humans , MCF-7 Cells , Protein Isoforms/genetics , Protein Isoforms/metabolism , DNA Methyltransferase 3B
12.
J Hum Genet ; 58(2): 59-66, 2013 Feb.
Article in English | MEDLINE | ID: mdl-23151675

ABSTRACT

ZNF350/ZBRK1 is a transcription factor, which associates with BRCA1 to co-repress GADD45A to regulate DNA damage repair, and the expression of ZNF350 is altered in different human carcinomas. In a previous study, we identified ZNF350 genomic variants potentially involved in breast cancer susceptibility in high-risk non-BRCA1/2 breast cancer individuals, which pointed toward a potential association for variants in the 5'-UTR and promoter regions. Therefore, direct sequencing was undertaken and identified 12 promoter variants, whereas haplotype analyses put in evidence four common haplotypes with a frequency>2%. However, based on their frequency observed in breast cancer and unrelated healthy individuals, these are not statistically associated with breast cancer risk. Luciferase promoter assays in two breast cancer cell lines identified two haplotypes (H11 and H12) stimulating significantly the expression of ZNF350 transcript compared with the common haplotype H8. The high expression of the H11 allele was associated with the variant c.-874A. Using MatInspector and Transcription Element Search softwares, in silico analyses predicted that the variant c.-874A created a binding site for the factors c-Myc and myogenin. This study represents the first characterization step of the ZNF350 promoter. Additional studies in larger cohorts and other populations will be needed to further evaluate whether common and/or rare ZNF350 promoter variants and haplotypes could be associated with a modest risk of breast cancer.


Subject(s)
Breast Neoplasms/genetics , Genes, BRCA1 , Genes, BRCA2 , Genetic Predisposition to Disease , Promoter Regions, Genetic , Repressor Proteins/genetics , Base Sequence , Canada , DNA Primers , Female , Haplotypes , Humans , Linkage Disequilibrium , Polymerase Chain Reaction
13.
Can J Microbiol ; 54(9): 797-802, 2008 Sep.
Article in English | MEDLINE | ID: mdl-18772944

ABSTRACT

We used insertional mutagenesis to produce genetically tagged mutants of the Dutch elm disease fungus Ophiostoma novo-ulmi subsp. novo-ulmi. We first optimized transformation of O. novo-ulmi protoplasts by the restriction enzyme mediated integration method. A concentration of 80 U of HindIII with 108 fungal protoplasts and 5 microg of plasmid DNA was the most efficient for generating a high number of O. novo-ulmi mutants carrying a single insertion in their genome. Mycelium- and yeast-like growth kinetics of 24 O. novo-ulmi mutants were evaluated in vitro. Flanking sequences were successfully recovered in 8% of the transformants analyzed. Some mutant phenotypes appeared to result from gene disruption events, whereas others likely involved modifications of noncoding regions. Several nuclear loci that control vegetative growth and could potentially impact parasitic fitness were successfully tagged.


Subject(s)
Ascomycota/genetics , Mutagenesis, Insertional , Plant Diseases/microbiology , Ulmus/microbiology , Ascomycota/growth & development , Ascomycota/metabolism , Fungal Proteins/genetics , Fungal Proteins/metabolism , Molecular Sequence Data , Phenotype , Transformation, Genetic
14.
Fungal Genet Biol ; 45(4): 565-78, 2008 Apr.
Article in English | MEDLINE | ID: mdl-18255325

ABSTRACT

The mobility of transposable elements (TEs) can contribute to genome plasticity, under- or over-expression of genes and ectopic recombination. The data collected in this study provide evidence of stress-induced mobility of OPHIO1 and OPHIO2 transposons, recently detected in Ophiostoma ulmi and O. novo-ulmi, the causal agents of Dutch elm disease (DED). The analyses of OPHIO UTRs and TIRs indicated the presence of two potential binding site motifs and a heat shock protein (hsp) promoter which could be involved in the mobility of OPHIO1 following a heat shock stress. The exact position of the hsp promoter was determined by 5' RACE PCR. After confirmation of the expression by RT-PCR of both OPHIO1 and OPHIO2 transposases in the absence of stress factors, we tested two experimental procedures to induce mobility of OPHIO TEs: (1) an exogenous (cloned) copy of OPHIO1 was introduced into the O. novo-ulmi subsp. americana strain W2 (OPHIO1 free strain) to give mutant strain W2:OPHIO1. After exposure of W2:OPHIO1 to a 55 degrees C heat shock treatment, some of the survivors showed signs of incomplete transposition (excision without reinsertion) of OPHIO1. (2) The O. novo-ulmi subsp. novo-ulmi strain AST27, introgressed from O. ulmi and carrying a distinct endogenous copy of OPHIO2 (OPHIO2-int.), was subjected to a series of abiotic stress treatments. Although a promoter sequence could not be identified, both exposures to UV light and to a 4 degrees C cold treatment caused perfect excision of OPHIO2-int. In contrast to OPHIO1, heat shock stress did not induce OPHIO2-int. mobility. Taken together, these results allow us to hypothesize a potential interspecific invasion of OPHIO transposons due to their mobility in Ophiostoma spp.


Subject(s)
Ascomycota/genetics , DNA Transposable Elements , Recombination, Genetic , 5' Untranslated Regions , Ascomycota/radiation effects , Base Sequence , Binding Sites , Cold Temperature , Hot Temperature , Molecular Sequence Data , Promoter Regions, Genetic , RNA, Fungal/genetics , RNA, Messenger/genetics , Transposases/biosynthesis , Ultraviolet Rays
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