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1.
Proc Natl Acad Sci U S A ; 117(42): 26389-26397, 2020 10 20.
Artigo em Inglês | MEDLINE | ID: mdl-33020260

RESUMO

Agrobacterium tumefaciens is the causal agent of crown gall disease. The bacterium is capable of transferring a segment of single-stranded DNA (ssDNA) into recipient cells during the transformation process, and it has been widely used as a genetic modification tool for plants and nonplant organisms. Transferred DNA (T-DNA) has been proposed to be escorted by two virulence proteins, VirD2 and VirE2, as a nucleoprotein complex (T-complex) that targets the host nucleus. However, it is not clear how such a proposed large DNA-protein complex is delivered through the host nuclear pore in a natural setting. Here, we studied the natural nuclear import of the Agrobacterium-delivered ssDNA-binding protein VirE2 inside plant cells by using a split-GFP approach with a newly constructed T-DNA-free strain. Our results demonstrate that VirE2 is targeted into the host nucleus in a VirD2- and T-DNA-dependent manner. In contrast with VirD2 that binds to plant importin α for nuclear import, VirE2 directly interacts with the host nuclear pore complex component nucleoporin CG1 to facilitate its nuclear uptake and the transformation process. Our data suggest a cooperative nuclear import model in which T-DNA is guided to the host nuclear pore by VirD2 and passes through the pore with the assistance of interactions between VirE2 and host nucleoporin CG1. We hypothesize that this large linear nucleoprotein complex (T-complex) is targeted to the nucleus by a "head" guide from the VirD2-importin interaction and into the nucleus by a lateral assistance from the VirE2-nucleoporin interaction.


Assuntos
Proteínas de Bactérias/metabolismo , Proteínas de Ligação a DNA/metabolismo , Canais Iônicos/metabolismo , Complexo de Proteínas Formadoras de Poros Nucleares/metabolismo , Transporte Ativo do Núcleo Celular/fisiologia , Agrobacterium tumefaciens/genética , Núcleo Celular/metabolismo , DNA Bacteriano/genética , DNA de Cadeia Simples/metabolismo , Células Vegetais/metabolismo , Rhizobium/genética , Nicotiana/genética , Transformação Genética/genética , Virulência , Fatores de Virulência/metabolismo
2.
Int J Mol Sci ; 24(9)2023 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-37175754

RESUMO

Nicotinamide adenine dinucleotide (NAD+) is a critical cofactor essential for various cellular processes. Abnormalities in NAD+ metabolism have also been associated with a number of metabolic disorders. The regulation and interconnection of NAD+ metabolic pathways are not yet completely understood. By employing an NAD+ intermediate-specific genetic system established in the model organism S. cerevisiae, we show that histone deacetylases (HDACs) Hst1 and Rpd3 link the regulation of the de novo NAD+ metabolism-mediating BNA genes with certain aspects of the phosphate (Pi)-sensing PHO pathway. Our genetic and gene expression studies suggest that the Bas1-Pho2 and Pho2-Pho4 transcription activator complexes play a role in this co-regulation. Our results suggest a model in which competition for Pho2 usage between the BNA-activating Bas1-Pho2 complex and the PHO-activating Pho2-Pho4 complex helps balance de novo activity with PHO activity in response to NAD+ or phosphate depletion. Interestingly, both the Bas1-Pho2 and Pho2-Pho4 complexes appear to also regulate the expression of the salvage-mediating PNC1 gene negatively. These results suggest a mechanism for the inverse regulation between the NAD+ salvage pathways and the de novo pathway observed in our genetic models. Our findings help provide a molecular basis for the complex interplay of two different aspects of cellular metabolism.


Assuntos
Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae , Saccharomyces cerevisiae/metabolismo , NAD/metabolismo , Histona Desacetilases/genética , Histona Desacetilases/metabolismo , Proteínas de Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/metabolismo , Fosfatos/metabolismo , Sirtuína 2/genética , Sirtuína 2/metabolismo , Transativadores/metabolismo , Proteínas de Homeodomínio/metabolismo
3.
Comput Educ ; 203: 104849, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37287756

RESUMO

The COVID-19 pandemic has forced universities and schools around the world to adopt online learning. Teachers may wonder if their students can attain satisfactory learning performance in an online learning environment without teachers' on-the-spot attention. In order to develop students' skills in programming, promote their enjoyment of learning and intention to learn to program, the researchers integrated two innovative teaching approaches, using online peer-facilitated learning and distributed pair programming, and investigated the effects of these on students' online learning performance. This study conducted an experiment that included 128 undergraduates from four class sections of Department of Finance. Thus, the experimental design in this research was a 2 (Peer-facilitated learning vs. non-peer-facilitated learning) × 2 (Distributed pair programming vs. non-distributed pair programming) factorial pretest/post-test design. The participants in this research mainly consisted of four classes of students from a non-computer or information department who took a compulsory course on programming design.' Both quantitative and qualitative data were collected in this study. According to the results, the peer-facilitated learning group exhibited significantly better development of programming skills, enjoyment of learning, and intention to learn, than the non-peer-facilitated learning group. However, expected effects of enhancing the learning of the students in this study who received the distributed pair programming were not found. The design of online pedagogy can be a reference for online educators. The implications of applying online peer-facilitated learning and distributed pair programming to support students' learning and the design of online programming courses are discussed.

4.
BMC Public Health ; 22(1): 469, 2022 03 09.
Artigo em Inglês | MEDLINE | ID: mdl-35264136

RESUMO

BACKGROUND: Combating viral outbreaks extends beyond biomedical and clinical approaches; thus, public health prevention measures are equally important. Public engagement in preventive efforts can be viewed as the social responsibility of individuals in controlling an infectious disease and are subjected to change due to human behaviour. Understanding individuals' perception of social responsibility is crucial and is not yet explored extensively in the academic literature. We adopted the grounded theory method to develop an explanatory substantive theory to illustrate the process of how individual responded to the outbreak from a social responsibility perspective. METHODS: In-depth interviews were conducted among 23 Malaysians either through telephone or face-to-face depending on the participant's preference. Both purposive and theoretical sampling were used. Participants were invited to share their understanding, perceptions and activities during the COVID-19 pandemic. They were further probed about their perceptions on complying with the public health interventions imposed by the authorities. The interviews were audio-recorded and transcribed verbatim. Data was analysed via open coding, focus coding and theoretical coding, facilitated by memoing, sketching and modelling. RESULTS: Study findings showed that, social responsibility is perceived within its role, the perceived societal role responsibility. In a particular context, an individual assumed only one of the many expected social roles with their perceived circle of responsibility. Individuals negotiated their actions from this perspective, after considering the perceived risk during the outbreak. The four types of behaviour depicted in the matrix diagram facilitate the understanding of the abstract concept of negotiation in the human decision-making process, and provide the spectrum of different behaviour in relation to public response to the COVID-19 pandemic. CONCLUSIONS: Our study adopted the grounded theory approach to develop a theoretical model that illustrates how individual response to COVID-19 preventive measures is determined by the negotiation between perceived societal role responsibility and perceived infection risk. This substantive theoretical model is abstract, thus has relevance for adoption within similar context of an outbreak.


Assuntos
COVID-19 , Teoria Fundamentada , Humanos , Pandemias/prevenção & controle , SARS-CoV-2 , Responsabilidade Social
5.
BMC Health Serv Res ; 22(1): 141, 2022 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-35115006

RESUMO

INTRODUCTION: Evidence shows physical distancing of one metre or more is important to reduce person-to-person SARS-CoV-2 transmission. This puts the Malaysian public healthcare system to a test when overcrowding has always been an issue. A new clinical appointment structure was proposed in the Malaysian public healthcare system amidst the pandemic to reduce the transmission risk. We aim to explore the general public's view on the proposed clinic appointment structure. METHODS: A cross-sectional anonymous web-based survey was conducted between 10th September 2020 and 30th November 2020. The survey was open to Malaysian aged 18 years and older via various social media platforms. The questionnaire consists of sociodemographic, experience of utilising healthcare facilities, and views on clinic appointment structure. RESULTS: A total of 1,144 complete responses were received. The mean age was 41.4 ± 12.4 years and more than half of the respondents had a preference for public healthcare. Among them, 77.1% reported to have a clinical appointment scheduled in the past. Less than a quarter experienced off-office hour appointments, mostly given by private healthcare. 70.2% answered they would arrive earlier if they were given a specific appointment slot at a public healthcare facility, as parking availability was the utmost concern. Majority hold positive views for after office hour clinical appointments, with 68.9% and 63.2% agreed for weekend and weekday evening appointment, respectively. The top reason of agreement was working commitment during office hours, while family commitment and personal resting time were the main reasons for disagreeing with off-office hour appointments. CONCLUSION: We found that majority of our respondents chose to come early instead of arriving on time which disrupts the staggered appointment system and causes over crowdedness. Our findings also show that the majority of our respondents accept off-office hour appointments. This positive response suggests that off-office hour appointments may have a high uptake amongst the public and thus be a possible solution to distribute the patient load. Therefore, this information may help policy makers to initiate future plans to resolve congestions within public health care facilities which in turn eases physical distancing during the pandemic.


Assuntos
COVID-19 , Pandemias , Adulto , Instituições de Assistência Ambulatorial , Estudos Transversais , Atenção à Saúde , Humanos , Pessoa de Meia-Idade , Pandemias/prevenção & controle , SARS-CoV-2
6.
Univers Access Inf Soc ; : 1-16, 2022 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-36530862

RESUMO

With the development of technology and demand for online courses, there have been considerable quantities of online, blended, or flipped courses designed and provided. However, in the technology-enhanced learning environments, which are also full of social networking websites, shopping websites, and free online games, it is challenging to focus students' attention and help them achieve satisfactory learning performance. In addition, the instruction of programming courses constantly challenges both teachers and students, particularly in online learning environments. To overcome and solve these problems and to facilitate students' learning, the researchers in this study integrated two teaching approaches, using meta-cognitive learning strategy (MCLS) and team regulation (TR), to develop students' regular learning habits and further contribute to their programming skills, academic motivation, and refusal self-efficacy of Internet use, in a cloud classroom. In this research, a quasi-experiment was conducted to investigate the effects of MCLS and TR adopting the experimental design of a 2 (MCLS vs. non-MCLS) × 2 (TR vs. non-TR) factorial pre-test/post-test. In this research, the participants consisted of four classes of university students from non-information or computer departments enrolled in programming design, a required course. The experimental groups comprised three of the classes, labelled as G1, G2, and G3. G1 concurrently received both the online MCLS and TR intervention, while G2 only received the online MCLS intervention, and G3 only received the online TR intervention. Serving as the control group, the fourth class (G4) received traditional teaching. This study investigated the effects of MCLS, TR, and their combination, on improving students' programming skills, academic motivation, and refusal self-efficacy of Internet use in an online computing course. According to the results, students who received online TR significantly enhanced their programming design skills and their refusal self-efficacy of Internet use a cloud classroom. However, the expected effects of MCLS on developing students' programming skills, academic motivation, and refusal self-efficacy of Internet use were not found in this study. The teaching strategy of integrating MCLS and TR in an online programming course in this study can serve as a reference for educators when conducting online, blended, or flipped courses during the COVID-19 pandemic.

7.
New Phytol ; 229(5): 2859-2872, 2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33105034

RESUMO

Integration of Agrobacterium tumefaciens transferred DNA (T-DNA) into the plant genome is the last step required for stable plant genetic transformation. The mechanism of T-DNA integration remains controversial, although scientists have proposed the participation of various nonhomologous end-joining (NHEJ) pathways. Recent evidence suggests that in Arabidopsis, DNA polymerase θ (PolQ) may be a crucial enzyme involved in T-DNA integration. We conducted quantitative transformation assays of wild-type and polQ mutant Arabidopsis and rice, analyzed T-DNA/plant DNA junction sequences, and (for Arabidopsis) measured the amount of integrated T-DNA in mutant and wild-type tissue. Unexpectedly, we were able to generate stable transformants of all tested lines, although the transformation frequency of polQ mutants was c. 20% that of wild-type plants. T-DNA/plant DNA junctions from these transformed rice and Arabidopsis polQ mutants closely resembled those from wild-type plants, indicating that loss of PolQ activity does not alter the characteristics of T-DNA integration events. polQ mutant plants show growth and developmental defects, perhaps explaining previous unsuccessful attempts at their stable transformation. We suggest that either multiple redundant pathways function in T-DNA integration, and/or that integration requires some yet unknown pathway.


Assuntos
Arabidopsis , Agrobacterium tumefaciens/genética , Arabidopsis/genética , DNA Bacteriano/genética , DNA Polimerase Dirigida por DNA/genética , Plantas Geneticamente Modificadas , Transformação Genética , DNA Polimerase teta
8.
J Biomed Sci ; 28(1): 85, 2021 Dec 09.
Artigo em Inglês | MEDLINE | ID: mdl-34886870

RESUMO

Parkinson's disease (PD), or paralysis agitans, is a common neurodegenerative disease characterized by dopaminergic deprivation in the basal ganglia because of neuronal loss in the substantia nigra pars compacta. Clinically, PD apparently involves both hypokinetic (e.g. akinetic rigidity) and hyperkinetic (e.g. tremor/propulsion) symptoms. The symptomatic pathogenesis, however, has remained elusive. The recent success of deep brain stimulation (DBS) therapy applied to the subthalamic nucleus (STN) or the globus pallidus pars internus indicates that there are essential electrophysiological abnormalities in PD. Consistently, dopamine-deprived STN shows excessive burst discharges. This proves to be a central pathophysiological element causally linked to the locomotor deficits in PD, as maneuvers (such as DBS of different polarities) decreasing and increasing STN burst discharges would decrease and increase the locomotor deficits, respectively. STN bursts are not so autonomous but show a "relay" feature, requiring glutamatergic synaptic inputs from the motor cortex (MC) to develop. In PD, there is an increase in overall MC activities and the corticosubthalamic input is enhanced and contributory to excessive burst discharges in STN. The increase in MC activities may be relevant to the enhanced beta power in local field potentials (LFP) as well as the deranged motor programming at the cortical level in PD. Moreover, MC could not only drive erroneous STN bursts, but also be driven by STN discharges at specific LFP frequencies (~ 4 to 6 Hz) to produce coherent tremulous muscle contractions. In essence, PD may be viewed as a disorder with deranged rhythms in the cortico-subcortical re-entrant loops, manifestly including STN, the major component of the oscillating core, and MC, the origin of the final common descending motor pathways. The configurations of the deranged rhythms may play a determinant role in the symptomatic pathogenesis of PD, and provide insight into the mechanism underlying normal motor control. Therapeutic brain stimulation for PD and relevant disorders should be adaptively exercised with in-depth pathophysiological considerations for each individual patient, and aim at a final normalization of cortical discharge patterns for the best ameliorating effect on the locomotor and even non-motor symptoms.


Assuntos
Córtex Motor/fisiopatologia , Neurônios/fisiologia , Doença de Parkinson/fisiopatologia , Doença de Parkinson/terapia , Núcleo Subtalâmico/fisiopatologia , Fenômenos Eletrofisiológicos , Humanos
9.
BMC Public Health ; 20(1): 1639, 2020 Nov 02.
Artigo em Inglês | MEDLINE | ID: mdl-33138800

RESUMO

BACKGROUND: Extracurricular sport participation and exercise (ESPE) refers to regular exercise/sport participation in addition to the physical education in school among a school-aged population. Rather than general physical activity, ESPE is typically deliberately initiated and presents an efficient target for interventions. However, compared to physical activity, relatively few studies have investigated sex differences in the development of and factors associated with ESPE using a person-centered approach. This study aimed to examine the latent trajectories of ESPE from childhood to emerging adulthood across sexes, and to identify the associated sex-specific individual (i.e., body mass index, body dissatisfaction, stress, and screen behavior) and parental (i.e., parental exercise and parental screen behavior) factors. METHODS: This study used data from part of the Child and Adolescent Behavior in Long-term Evolution (CABLE) project, which comprised 2072 fourth graders (aged 9 years) in Northern Taiwan followed annually from 2001 to 2013 (13 waves). Repeated-measures latent class analysis was used to identify the trajectories of ESPE for males and females, respectively. Multinomial logistic regression was further used to identify sex-specific factors related to ESPE. RESULTS: Four trajectories of ESPE were identified for males and females. For males, these trajectories were Rarely-to-Never (20%), Often-to-Rarely (32%), Always-to-Never (21%), and Always (27%). For females, these trajectories were Rarely-to-Never (34%), Rarely (23%), Always-to-Rarely (33%), and Always (10%). We observed that the developmental patterns of ESPE varied by sex such that there was an earlier decline in the trajectories of ESPE in females than in males and that, compared with males, fewer females maintained exercise habits in young adulthood. Furthermore, we found several sex-specific factors related to ESPE, namely, stress, BMI, and parental exercise. Body dissatisfaction and individual screen behavior were associated with trajectories of ESPE for both sexes. CONCLUSIONS: We found distinct trajectories of ESPE from childhood to emerging adulthood for both sexes. The trajectories of ESPE for males and females, however, differ in terms of patterns and associated factors. Our findings suggest that efforts to increase ESPE should be initiated early, and may be made more effective by considering sex differences.


Assuntos
Caracteres Sexuais , Esportes , Adolescente , Adulto , Criança , Estudos de Coortes , Exercício Físico , Feminino , Humanos , Estudos Longitudinais , Masculino , Taiwan/epidemiologia , Adulto Jovem
10.
Appl Psychophysiol Biofeedback ; 45(4): 275-282, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-32997269

RESUMO

Abnormal autonomic nervous system (ANS) function may result in poor outcomes in patients with schizophrenia. Altered cardio-respiratory coupling, which indicates suppression of vagal activity, was identified as an important trait in patients with schizophrenia and their unaffected relatives. Heart rate variability (HRV) in standardized bedside reflex tests has been studied, mostly in medicated patients with schizophrenia whose ANS function could be influenced by medication. Our study aimed to explore the autonomic function differences between drug-naïve patients with schizophrenia and healthy individuals during challenge tests combining respiration and HRV analysis. Forty-two drug-naïve patients with schizophrenia were matched with 42 healthy controls in terms of age and gender. Their beat-to-beat blood pressure and heart rate were monitored in the supine position as a survey of ANS function, and the mean heart rate range (MHRR) was measured under deep-breathing challenge. A decreased MHRR, a sensitive sign indicating an impaired parasympathetic response, during the deep-breathing challenge among the drug-naïve patients with schizophrenia was found. Drug-naïve patients with schizophrenia may have a parasympathetic dysfunction in the early stages of schizophrenia before medication is introduced, which could be considered a neurobiological marker in the pathophysiology of schizophrenia.


Assuntos
Sistema Nervoso Autônomo/fisiopatologia , Frequência Cardíaca/fisiologia , Respiração , Esquizofrenia/fisiopatologia , Adulto , Pressão Sanguínea/fisiologia , Feminino , Humanos , Masculino , Sistema Nervoso Parassimpático/fisiologia
11.
Plant Mol Biol ; 101(4-5): 355-371, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31401729

RESUMO

KEY MESSAGE: Protoplasts can be used for genome editing using several different CRISPR systems, either separately or simultaneously, and that the resulting mutations can be recovered in regenerated non-chimaeric plants. Protoplast transfection and regeneration systems are useful platforms for CRISPR/Cas mutagenesis and genome editing. In this study, we demonstrate the use of Cpf1 (Cas12a) and nCas9-activation-induced cytidine deaminase (nCas9-Target-AID) systems to mutagenize Nicotiana tabacum protoplasts and to regenerate plants harboring the resulting mutations. We analyzed 20 progeny plants of Cas12a-mediated phytoene desaturase (PDS) mutagenized regenerants, as well as regenerants from wild-type protoplasts, and confirmed that their genotypes were inherited in a Mendelian manner. We used a Cas9 nickase (nCas9)-cytidine deaminase to conduct C to T editing of the Ethylene receptor 1 (ETR1) gene in tobacco protoplasts and obtained edited regenerates. It is difficult to obtain homozygous edits of polyploid genomes when the editing efficiency is low. A second round of mutagenesis of partially edited regenerants (a two-step transfection protocol) allowed us to derive ETR1 fully edited regenerants without the need for sexual reproduction. We applied three different Cas systems (SaCas9, Cas12a, and nCas9-Traget AID) using either a one-step or a two-step transfection platform to obtain triply mutated and/or edited tobacco regenerants. Our results indicate that these three Cas systems can function simultaneously within a single cell.


Assuntos
Edição de Genes/métodos , Nicotiana/genética , Proteínas de Bactérias/genética , Sistemas CRISPR-Cas , Francisella/genética , Homozigoto , Proteínas de Plantas/genética , Tetraploidia
12.
BMC Health Serv Res ; 19(1): 248, 2019 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-31018843

RESUMO

BACKGROUND: Health Research Priority Setting (HRPS) in the Ministry of Health (MOH) Malaysia was initiated more than a decade ago to drive effort toward research for informed decision and policy-making. This study assessed the impact of funded prioritised research and identified research gaps to inform future priority setting initiatives for universal access and quality healthcare in Malaysia. METHODS: Research impact of universal access and quality healthcare projects funded by the National Institutes of Health Malaysia were assessed based on the modified Payback Framework, addressing categories of informing policy, knowledge production, and benefits to health and health sector. For the HRPS process, the Child Health and Nutrition Research Initiative methodology was adapted and adopted, with the incorporation of stakeholder values using weights and monetary allocation survey. Workshop discussions and interviews with stakeholders and research groups were conducted to identify research gaps, with the use of conceptual frameworks to guide the search. RESULTS: Seventeen ongoing and 50 completed projects were identified for research funding impact analysis. Overall, research fund allocation differed from stakeholders' expectation. For research impact, 48 out of 50 completed projects (96.0%) contributed to some form of policy-making efforts. Almost all completed projects resulted in outputs that contributed to knowledge production and were expected to lead to health and health sector benefits. The HRPS process led to the identification of research priority areas that stemmed from ongoing and new issues identified for universal access and quality healthcare. CONCLUSION: The concerted efforts of evaluation of research funding impact, prioritisation, dissemination and policy-maker involvement were valuable for optimal health research resource utilisation in a resource constrained developing country. Embedding impact evaluation into a priority setting process and funding research based on national needs could facilitate health research investment to reach its potential.


Assuntos
Prioridades em Saúde , Pesquisa sobre Serviços de Saúde/economia , Qualidade da Assistência à Saúde , Apoio à Pesquisa como Assunto , Pessoal Administrativo , Tomada de Decisões , Países em Desenvolvimento , Humanos , Malásia , Formulação de Políticas , Projetos de Pesquisa , Alocação de Recursos
13.
BMC Health Serv Res ; 19(1): 497, 2019 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-31311538

RESUMO

BACKGROUND: Amid the current burden of non-communicable (NCD) diseases in Malaysia, there is a growing demand for more efficient service delivery of primary healthcare. A complex intervention is proposed to improve NCD management in Malaysia. This exploratory study aimed to assess primary healthcare providers' receptiveness towards change prior to implementation of the proposed complex intervention. METHOD: This study was conducted using an exploratory qualitative approach on purposely selected healthcare providers at primary healthcare clinics. Twenty focus group discussions and three in-depth interviews were conducted using a semi-structured interview guide. Consent was obtained prior to interviews and for audio-recordings. Interviews were transcribed verbatim and thematically analysed, guided by the Consolidated Framework for Implementation Research (CFIR), a framework comprised of five major domains promoting implementation theory development and verification across multiple contexts. RESULTS: The study revealed via CFIR that most primary healthcare providers were receptive towards any proposed changes or intervention for the betterment of NCD care management. However, many challenges were outlined across four CFIR domains-intervention characteristics, outer setting, inner setting, and individual characteristics-that included perceived barriers to implementation. Perception of issues that triggered proposed changes reflected the current situation, including existing facilitating aspects that can support the implementation of any future intervention. The importance of strengthening the primary healthcare delivery system was also expressed. CONCLUSION: Understanding existing situations faced at the primary healthcare setting is imperative prior to implementation of any intervention. Healthcare providers' receptiveness to change was explored, and using CFIR framework, challenges or perceived barriers among healthcare providers were identified. CFIR was able to outline the clinics' setting, individual behaviour and external agency factors that have direct impact to the organisation. These are important indicators in ensuring feasibility, effectiveness and sustainability of any intervention, as well as future scalability considerations.


Assuntos
Atitude do Pessoal de Saúde , Pessoal de Saúde/psicologia , Atenção Primária à Saúde/organização & administração , Grupos Focais , Pesquisa sobre Serviços de Saúde , Humanos , Malásia/epidemiologia , Doenças não Transmissíveis/epidemiologia , Doenças não Transmissíveis/prevenção & controle , Pesquisa Qualitativa
14.
Plant Mol Biol ; 97(4-5): 407-420, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29948659

RESUMO

KEY MESSAGE: Arabidopsis and human ARM protein interact with telomerase. Deregulated mRNA levels of DNA repair and ribosomal protein genes in an Arabidopsis arm mutant suggest non-telomeric ARM function. The human homolog ARMC6 interacts with hTRF2. Telomerase maintains telomeres and has proposed non-telomeric functions. We previously identified interaction of the C-terminal domain of Arabidopsis telomerase reverse transcriptase (AtTERT) with an armadillo/ß-catenin-like repeat (ARM) containing protein. Here we explore protein-protein interactions of the ARM protein, AtTERT domains, POT1a, TRF-like family and SMH family proteins, and the chromatin remodeling protein CHR19 using bimolecular fluorescence complementation (BiFC), yeast two-hybrid (Y2H) analysis, and co-immunoprecipitation. The ARM protein interacts with both the N- and C-terminal domains of AtTERT in different cellular compartments. ARM interacts with CHR19 and TRF-like I family proteins that also bind AtTERT directly or through interaction with POT1a. The putative human ARM homolog co-precipitates telomerase activity and interacts with hTRF2 protein in vitro. Analysis of Arabidopsis arm mutants shows no obvious changes in telomere length or telomerase activity, suggesting that ARM is not essential for telomere maintenance. The observed interactions with telomerase and Myb-like domain proteins (TRF-like family I) may therefore reflect possible non-telomeric functions. Transcript levels of several DNA repair and ribosomal genes are affected in arm mutants, and ARM, likely in association with other proteins, suppressed expression of XRCC3 and RPSAA promoter constructs in luciferase reporter assays. In conclusion, ARM can participate in non-telomeric functions of telomerase, and can also perform its own telomerase-independent functions.


Assuntos
Arabidopsis/enzimologia , Proteínas do Domínio Armadillo/metabolismo , Telomerase/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas do Domínio Armadillo/genética , Genes Reporter , Holoenzimas , Humanos , Telomerase/genética , Técnicas do Sistema de Duplo-Híbrido
15.
Plant Biotechnol J ; 16(7): 1295-1310, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29230929

RESUMO

Plant protoplasts are useful for assessing the efficiency of clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) mutagenesis. We improved the process of protoplast isolation and transfection of several plant species. We also developed a method to isolate and regenerate single mutagenized Nicotianna tabacum protoplasts into mature plants. Following transfection of protoplasts with constructs encoding Cas9 and sgRNAs, target gene DNA could be amplified for further analysis to determine mutagenesis efficiency. We investigated N. tabacum protoplasts and derived regenerated plants for targeted mutagenesis of the phytoene desaturase (NtPDS) gene. Genotyping of albino regenerants indicated that all four NtPDS alleles were mutated in amphidiploid tobacco, and no Cas9 DNA could be detected in most regenerated plants.


Assuntos
Sistemas CRISPR-Cas , Mutagênese Sítio-Dirigida/métodos , Protoplastos , Arabidopsis/genética , Brassica/genética , Sistemas CRISPR-Cas/genética , Genes de Plantas/genética , Milhetes/genética , Mutação/genética , Oryza/genética , Oxirredutases/genética , Sasa/genética , Nicotiana/genética , Zea mays/genética
16.
Int J Eat Disord ; 51(11): 1277-1284, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30488563

RESUMO

OBJECTIVE: Nationwide data on eating disorders (EDs) among East Asian populations are scarce. This study aimed to estimate the prevalence, incidence, and psychiatric comorbidities of EDs in Taiwan. METHOD: Drawing from the National Health Research Institute database, we identified the study subjects aged 11-34 years who received an ED diagnosis during ambulatory visits or hospitalization from 2001 to 2012. We analyzed the subjects by age and gender. Diagnoses for comorbid psychiatric disorders were entered in the analysis only if they occurred within 1 year before or after the ED diagnosis date. Rates were age- and sex-adjusted to the average population within the study period. RESULTS: The estimated mean annual incidence was approximately 11.11 (95% CI 10.51-11.72) per 100,000 residents, and the 1 year prevalence was 20.87 (95% CI 20.05-21.70) per 100,000 residents, with evidence of female predominance (female/male ratio = 10.95, 95% CI = 8.97-13.36). Incidence and prevalence peaked at ages 20-24 in females and at ages 25-29 in males. Both rates nearly doubled within a decade. Specifically, the anorexia nervosa trend recently stabilized in females and decreased in males, while that for bulimia nervosa increased in both females and males. The most common comorbid diagnoses were anxiety (53%) and major depressive disorders (22%). DISCUSSION: The incidence and prevalence of EDs were lower in Taiwan than in Western countries. However, these rates were rapidly rising, notably in males. The majority of Taiwanese ED patients also received a diagnosis for various psychiatric comorbidities, a finding worthy of clinical attention.


Assuntos
Transtornos da Alimentação e da Ingestão de Alimentos/psicologia , Adolescente , Adulto , Criança , Comorbidade , Feminino , Humanos , Incidência , Masculino , Prevalência , Taiwan , Adulto Jovem
17.
Eur Addict Res ; 24(2): 71-78, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29902803

RESUMO

BACKGROUND: Animal studies have demonstrated that oxytocin can influence addiction behaviors and might interact with the dopaminergic system, which is a key component of addiction behaviors. However, related evidence from clinical studies is scarce. The aim of our study was to explore the relationship between plasma oxytocin level and heroin craving among patients receiving methadone maintenance treatment, and to ascertain whether this relationship is moderated by novelty-seeking. METHODS: The study was conducted in a methadone maintenance therapy clinic of a medical center in Taiwan. Seventy-seven patients with heroin addiction were enrolled. Plasma oxytocin was measured using an ELISA kit. Craving was assessed using an established instrument, the Chinese Craving Scale. RESULTS: A significant negative association was found between the plasma oxytocin level and craving score, which remained robust after controlling the effects of social support and low-density lipoprotein cholesterol. An interaction between oxytocin and novelty-seeking indicated that this relationship was stronger among patients with a lower level of novelty-seeking. CONCLUSION: This finding may be taken into account in future studies and may provide a basis for the development of potential treatment for addiction. The effect of oxytocin for the treatment of opioid dependence might be modulated by some psychological factors.


Assuntos
Fissura/efeitos dos fármacos , Dependência de Heroína/tratamento farmacológico , Metadona/uso terapêutico , Ocitocina/sangue , Adulto , Comportamento Aditivo/tratamento farmacológico , Feminino , Dependência de Heroína/reabilitação , Humanos , Masculino , Tratamento de Substituição de Opiáceos , Síndrome de Abstinência a Substâncias/sangue
18.
Aggress Behav ; 44(1): 69-82, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-28857191

RESUMO

Adolescence is a developmental period with high vulnerability to sleep problems. However, research identifying distinct patterns and underlying determinants of sleep problems is scarce. This study investigated discrete subgroups of, changes in, and stability of sleep problems. We also examined whether peer victimization influenced sleep problem subgroups and transitions in patterns of sleep problems from late adolescence to young adulthood. Sex differences in the effects of peer victimization were also explored. In total, 1,455 male and 1,399 female adolescents from northern Taiwan participated in this longitudinal study. Latent transition analysis was used to examine changes in patterns of sleep problems and the effects of peer victimization on these changes. We identified three subgroups of sleep problems in males and two in females, and found that there was a certain level of instability in patterns of sleep problems during the study period. For both sexes, those with greater increases in peer victimization over time were more likely to change from being a good sleeper to a poor sleeper. The effects of peer victimization on baseline status of sleep problems, however, was only significant for males, with those exposed to higher levels of peer victimization more likely to be poor sleepers at baseline. Our findings reveal an important role of peer victimization in predicting transitions in patterns of sleep problems. Intervention programs aimed at decreasing peer victimization may help reduce the development and escalation of sleep problems among adolescents, especially in males.


Assuntos
Bullying , Vítimas de Crime/psicologia , Grupo Associado , Transtornos do Sono-Vigília/psicologia , Adolescente , Feminino , Humanos , Estudos Longitudinais , Masculino , Caracteres Sexuais , Taiwan
19.
Plant J ; 85(5): 648-59, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26833589

RESUMO

Rice (Oryza sativa) is one of the world's most important crops. Rice researchers make extensive use of insertional mutants for the study of gene function. Approximately half a million flanking sequence tags from rice insertional mutant libraries are publicly available. However, the relationship between genotype and phenotype is very weak. Transgenic plant assays have been used frequently for complementation, overexpression or antisense analysis, but sequence changes caused by callus growth, Agrobacterium incubation medium, virulence genes, transformation and selection conditions are unknown. We used high-throughput sequencing of DNA from rice lines derived from Tainung 67 to analyze non-transformed and transgenic rice plants for mutations caused by these parameters. For comparison, we also analyzed sequence changes for two additional rice varieties and four T-DNA tagged transformants from the Taiwan Rice Insertional Mutant resource. We identified single-nucleotide polymorphisms, small indels, large deletions, chromosome doubling and chromosome translocations in these lines. Using standard rice regeneration/transformation procedures, the mutation rates of regenerants and transformants were relatively low, with no significant differences among eight tested treatments in the Tainung 67 background and in the cultivars Taikeng 9 and IR64. Thus, we could not conclusively detect sequence changes resulting from Agrobacterium-mediated transformation in addition to those caused by tissue culture-induced somaclonal variation. However, the mutation frequencies within the two publically available tagged mutant populations, including TRIM transformants or Tos17 lines, were about 10-fold higher than the frequency of standard transformants, probably because mass production of embryogenic calli and longer callus growth periods were required to generate these large libraries.


Assuntos
Estudos de Associação Genética/métodos , Variação Genética , Oryza/genética , Transformação Genética/genética , Agrobacterium/genética , Células Clonais/metabolismo , Produtos Agrícolas/genética , DNA Bacteriano/genética , DNA de Plantas/química , DNA de Plantas/genética , Genótipo , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Mutação INDEL , Mutagênese Insercional , Oryza/classificação , Fenótipo , Plantas Geneticamente Modificadas , Ploidias , Polimorfismo de Nucleotídeo Único , Especificidade da Espécie , Taiwan , Técnicas de Cultura de Tecidos/métodos
20.
Fam Process ; 56(3): 734-751, 2017 09.
Artigo em Inglês | MEDLINE | ID: mdl-27660107

RESUMO

The study examined how child and parent characteristics, and contextual sources of stress, such as marital conflict predict initial status and trajectories of parent involvement, support, and harsh control, over a 4-year period in families in Taiwan (n = 4,754). Based on Belsky's (1984) ecological model of parenting, three domains predicting parenting were tested, child characteristics (age cohort and gender), father and mother characteristics (education and depressive symptoms), and contextual sources of stress (marital conflict). The study followed two cohorts of children; the younger cohort was followed from first to fourth grade and the older cohort from fourth to seventh grade. Initially, fourth graders reported more parental involvement, support, and harsh control than first graders. However, involvement, support, and harsh control decreased across the 4 years for the older cohort as they transitioned to early adolescence. In the first year, girls reported more parental involvement and support and less harsh control than boys. Across the 4 years, involvement and support increased, and harsh control decreased for boys; whereas involvement stayed the same, support slightly decreased, and harsh control slightly increased for girls. Children whose parents were more educated reported more parent involvement, support, and harsh control in the first year. Children whose fathers were chronically depressed and whose parents were experiencing marital conflict reported decreasing parent involvement and support over the years.


Assuntos
Depressão/fisiopatologia , Escolaridade , Conflito Familiar , Poder Familiar/psicologia , Poder Familiar/tendências , Pais/psicologia , Adolescente , Criança , Estudos de Coortes , Depressão/epidemiologia , Feminino , Humanos , Masculino , Inquéritos e Questionários , Taiwan/epidemiologia
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