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2.
Am J Obstet Gynecol ; 2024 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-38367755

RESUMO

BACKGROUND: The potential association between mode of obstetrical delivery and subsequent sexual outcomes of the birthing parent remains uncertain and has not been well investigated from the perspective of positive sexual life satisfaction. OBJECTIVE: This study aimed to investigate if there was any association between mode of delivery and subsequent sexual life satisfaction of the birthing parent. A secondary aim was to assess the extent to which this association changed when stratified by time elapsed since delivery. STUDY DESIGN: The study matched participants in the Stockholm Public Health Cohort with deliveries recorded in the Swedish Medical Birth Register. Any deliveries recorded in the registry before the participation in the Stockholm Public Health Cohort were included (n=46,078). The length of time from delivery to outcome assessment varied from 1 month to 41 years (mean, 18 years [±10.8]). Mode of delivery was retrieved from the same registry, whereas self-perceived sexual life satisfaction was retrieved from the Stockholm Public Health Cohort Questionnaires where participants had assessed their sexual life satisfaction as 1 out of 5 mutually exclusive options. Multinomial logistic regression was used to test for any association between mode of delivery (cesarean, instrumental, and spontaneous vaginal delivery) and sexual life satisfaction, both overall and stratified by time elapsed since delivery. RESULTS: After adjusting for covariates, no statistically significant (P < .05) difference in subsequent sexual life satisfaction of the birthing parent between modes of delivery was identified. Adjusted odds ratios for assessing sexual life satisfaction as the lowest level ("very unsatisfactory") were 1.11 (95% confidence interval, 0.98-1.25) for cesarean delivery and 1.16 (95% confidence interval, 0.99-1.35) for instrumental delivery, compared with spontaneous vaginal delivery. The difference in covariate-adjusted prevalence of the lowest level of sexual life satisfaction among the different groups categorized by time since delivery was small: 4.0% (95% confidence interval, 2.4%-5.6%) for cesarean delivery as opposed to 2.8% (95% confidence interval, 2.1%-3.6%) for spontaneous vaginal delivery within 2 years since delivery. CONCLUSION: These findings do not support any impact of mode of delivery on the subsequent self-perceived sexual life satisfaction among birthing people, either overall or across different time periods since delivery.

3.
BJOG ; 131(1): 15-25, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37340193

RESUMO

OBJECTIVE: To examine antiseizure medication (ASM) prescription during pregnancy. DESIGN: Population-based drug utilisation study. SETTING: UK primary and secondary care data, 1995-2018, from the Clinical Practice Research Datalink GOLD version. POPULATION OR SAMPLE: 752 112 completed pregnancies among women registered for a minimum of 12 months with an 'up to standard' general practice prior to the estimated start of pregnancy and for the duration of their pregnancy. METHODS: We described ASM prescription across the study period, overall and by ASM indication, examined patterns of prescription during pregnancy including continuous prescription and discontinuation, and used logistic regression to investigate factors associated with those ASM prescription patterns. MAIN OUTCOME MEASURES: Prescription of ASMs during pregnancy and discontinuation of ASMs before and during pregnancy. RESULTS: ASM prescription during pregnancy increased from 0.6% of pregnancies in 1995 to 1.6% in 2018, driven largely by an increase in women with indications other than epilepsy. Epilepsy was an indication for 62.5% of pregnancies with an ASM prescription and non-epilepsy indications were present for 66.6%. Continuous prescription of ASMs during pregnancy was more common in women with epilepsy (64.3%) than in women with other indications (25.3%). Switching ASMs was infrequent (0.8% of ASM users). Factors associated with discontinuation included age ≥35, higher social deprivation, more frequent contact with the GP and being prescribed antidepressants or antipsychotics. CONCLUSIONS: ASM prescription during pregnancy increased between 1995 and 2018 in the UK. Patterns of prescription around the pregnancy period vary by indication and are associated with several maternal characteristics.


Assuntos
Prescrições de Medicamentos , Epilepsia , Gravidez , Feminino , Humanos , Estudos de Coortes , Reino Unido , Família , Epilepsia/tratamento farmacológico , Anticonvulsivantes/uso terapêutico
4.
TechTrends ; 67(3): 561-571, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36249671

RESUMO

Elementary school children are spending more time using digital technologies. Teachers and schools are concerned about the student's digital safety. We interviewed ten elementary school teachers virtually to understand their concerns and understand the actions they take regarding elementary school children's digital safety. Using thematic analysis, we identified themes of concerns and actions of teachers and schools on elementary school children's digital safety. Some digital safety concerns discussed by the teachers included content-related concerns such as accessing inappropriate content, contact-related concerns, inappropriate contact with others online, sharing personal information, lack of understanding of danger, conduct-related concerns regarding cyberbullying and digital footprint, contract-related concerns such as digital security and privacy, and home-related concerns. Teachers and schools have taken several actions to address these concerns. Some of the digital safety actions included security measures and limits, monitoring student activities, providing education on digital safety, and support from guidance counselors. This study has implications for elementary school educators, administrators, parents, and students on the safe use of digital technologies. Supplementary Information: The online version contains supplementary material available at 10.1007/s11528-022-00803-z.

5.
J Am Heart Assoc ; 11(19): e025102, 2022 10 04.
Artigo em Inglês | MEDLINE | ID: mdl-36172928

RESUMO

Background Alcohol intake increases blood pressure yet estimates of associations between maternal intake and hypertensive disorders of pregnancy (HDP) are sparse and range from null to a protective effect. Here we estimated the association of maternal drinking during pregnancy with preeclampsia and gestational hypertension (separately and jointly, as HDP). We used partner's alcohol intake as a negative control exposure, beverage type-specific models, and a range of sensitivity analyses to strengthen causal inference and reduce the influence of bias. Methods and Results We performed a longitudinal analysis of prospectively collected data on self-reported alcohol intake and presence of HDP from the UK ALSPAC (Avon Longitudinal Study of Parents and Children) cohort. Multivariable multinomial regression models were adjusted for confounders and mutually adjusted for partner's or maternal alcohol intake in the negative control analysis. We also performed a beverage type analysis of the effect of beer and wine separately on HDP risk, owing to different social patterning associated with different drinks. Sensitivity analyses assessed the robustness of results to assumptions of no recall bias, no residual confounding, and no selection bias. Of the 8999 women eligible for inclusion, 1490 fulfilled the criteria for HDP (17%). Both maternal and partner's drinking were associated with decreased HDP odds (mutually adjusted odds ratio [OR], 0.86; [95% CI, 0.77-0.96], P=0.008 and OR, 0.82; [95% CI, 0.70-0.97], P=0.018, respectively). We demonstrate the validity of the negative control analyses using the same approach for smoking as the exposure. This confirmed an inverse association for maternal but not partner's smoking, as expected. Estimates were more extreme for increasing levels of wine intake compared with increasing levels of beer. Multiple sensitivity analyses did not alter our conclusions. Conclusions We observed an inverse relationship between alcohol intake during pregnancy and risk of HDP for both maternal and, more surprisingly, partner's drinking. We speculate that this is more likely to be due to common environmental exposures shared between pregnant women and their partners rather than a true causal effect. This warrants further investigation using different study designs, including Mendelian randomization.


Assuntos
Hipertensão Induzida pela Gravidez , Pré-Eclâmpsia , Consumo de Bebidas Alcoólicas/efeitos adversos , Consumo de Bebidas Alcoólicas/epidemiologia , Criança , Estudos de Coortes , Feminino , Humanos , Hipertensão Induzida pela Gravidez/epidemiologia , Estudos Longitudinais , Gravidez
6.
BJOG ; 129(12): 2010-2018, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-35856885

RESUMO

OBJECTIVES: To investigate the association between mode of delivery and subsequent maternal sexual wellbeing. DESIGN: Prospective birth cohort study. SETTING: Avon (in Bristol area), UK. POPULATION: Participants in the Avon Longitudinal Study of Parents and Children (ALSPAC). METHODS: Mode of delivery was abstracted from obstetric records and sexual wellbeing measures were collected via a self-report questionnaire. Missing data were imputed using multiple imputation, and ordinal logistic regression models for ordered categorical outcomes were adjusted for the covariates maternal age at delivery, pre-pregnancy body mass index, diabetes during pregnancy, socio-economic position, parity, depression and anxiety. MAIN OUTCOME MEASURES: Sexual enjoyment and frequency at four time points postpartum (between 33 months and 18 years) and two types of sex-related pain (pain in the vagina during sex and elsewhere after sex) at 11 years postpartum. RESULTS: We found no association between mode of delivery and sexual enjoyment (e.g. adjusted odds ratio [OR] 1.11, 95% confidence interval [95% CI] 0.97-1.27 at 33 months) or sexual frequency (OR 0.99, 95% CI 0.88-1.12 at 33 months). Caesarean section was associated with an increased odds of pain in the vagina during sex at 11 years postpartum as compared with vaginal delivery in the adjusted model (OR 1.74, 95% CI 1.46-2.08). CONCLUSIONS: These findings provide no evidence supporting associations between caesarean section and sexual enjoyment or frequency. However, mode of delivery was shown to be associated with dyspareunia, which may not be limited to abdominal scarring.


Assuntos
Cesárea , Parto Obstétrico , Criança , Estudos de Coortes , Parto Obstétrico/efeitos adversos , Feminino , Humanos , Estudos Longitudinais , Dor , Gravidez , Estudos Prospectivos
7.
NPJ Genom Med ; 7(1): 36, 2022 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-35672413

RESUMO

Despite the growing accessibility of clinical sequencing, functional interpretation of variants remains a major hurdle to molecular diagnostics of Mendelian diseases. We aimed to describe a new adult-onset myopathy with muscle weakness and hyperCKemia caused by a nonsense variant in muscular LMNA-interacting protein (MLIP). Following RNA-sequencing, differential expression analysis uncovered a significant downregulation of this gene, which had a surprisingly mild effect on MLIP protein expression. RT-PCR and long-read sequencing (LRS) both support an important transcriptome shift in the patient, where decreased MLIP levels are seemingly due to nonsense-mediated decay of transcripts containing the exon 5 mutation. Moreover, a compensatory mechanism upregulates the functionally lacking isoforms and generates novel transcripts. These results support the recently discovered clinical implications of MLIP variants in myopathies, highlighting for the first time its relevance in adult-onset cases. These results also underline the power of LRS as a tool for the functional assessment of variants of unknown significance (VUS), as well as the definition of accurate isoform profile annotations in a tissue-specific manner.

8.
TechTrends ; 66(3): 483-494, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35434728

RESUMO

Current research surrounding online computer science education emphasizes the need for high-quality professional development opportunities. However, there is a gap in research in the inclusion of online computer science educators to identify needs and strategies that make the online computer science courses effective. Through a Research-to-Practice Partnership (RPP), this paper examines the instructional strategies and recommendations from online Computer Science teachers. This study seeks to better understand (1) What design, facilitation, and assessment strategies do teachers use to teach programming online? and (2) What recommendations do teachers have for those interested in teaching programming online? The feedback teachers provided during the study assisted in identifying the current needs in online AP Computer Science. The participants suggested additional ways the RPP could support teachers in strengthening their practice, which has assisted in the production of high-quality professional development to support novice teachers entering the field of Computer Science.

9.
J Comput High Educ ; 34(2): 297-320, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-34305360

RESUMO

Learning and Development (L&D) roles are important to organizations for improving employee's knowledge and skills. This study examined various roles and competency domains required of learning and development professionals in higher education. Ten different roles of L&D professionals were examined for 20 competency domains through a qualitative coding process. We extracted and analyzed 294 unique postings from the job board, higheredjobs.com. Results indicated that designers and directors are the most advertised L&D postings. The top five competency domains required of L&D professionals were collaboration, communication, content development, project management, and assessment and evaluation. Communication and collaboration skills were required for most of the roles. Leadership and people management were ranked highest for executives and directors. In addition, competency domains aggregated by roles are provided. Implications are provided for employees, L&D graduate programs and professionals, and researchers.

10.
Epileptic Disord ; 23(4): 649-654, 2021 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-34240707

RESUMO

Musicogenic seizures and ictal asystole are two rare features mainly encountered in temporal lobe epilepsy. We present the first case of musicogenic epilepsy with ictal asystole in a patient with right temporal lobe epilepsy, confirmed by video-EEG. The pathophysiological mechanism of both conditions remains uncertain. These two features appeared after many years of epilepsy in our patient, and although this association might be coincidental, common mechanisms could be involved, including remodelling of neural connections or seizure propagation to a singular zone. This case also highlights the need for precise electroclinical assessment of seizures that change clinically over time, particularly when falls occur after a certain period of latency, suggesting cardiac consequences of the seizures.


Assuntos
Epilepsia Reflexa , Eletroencefalografia , Epilepsia do Lobo Temporal/complicações , Epilepsia do Lobo Temporal/diagnóstico , Parada Cardíaca/complicações , Humanos , Convulsões/diagnóstico , Convulsões/etiologia
11.
J Exp Bot ; 72(10): 3881-3901, 2021 05 04.
Artigo em Inglês | MEDLINE | ID: mdl-33758916

RESUMO

Plants need to cope with strong variations of nitrogen availability in the soil. Although many molecular players are being discovered concerning how plants perceive NO3- provision, it is less clear how plants recognize a lack of nitrogen. Following nitrogen removal, plants activate their nitrogen starvation response (NSR), which is characterized by the activation of very high-affinity nitrate transport systems (NRT2.4 and NRT2.5) and other sentinel genes involved in N remobilization such as GDH3. Using a combination of functional genomics via transcription factor perturbation and molecular physiology studies, we show that the transcription factors belonging to the HHO subfamily are important regulators of NSR through two potential mechanisms. First, HHOs directly repress the high-affinity nitrate transporters, NRT2.4 and NRT2.5. hho mutants display increased high-affinity nitrate transport activity, opening up promising perspectives for biotechnological applications. Second, we show that reactive oxygen species (ROS) are important to control NSR in wild-type plants and that HRS1 and HHO1 overexpressors and mutants are affected in their ROS content, defining a potential feed-forward branch of the signaling pathway. Taken together, our results define the relationships of two types of molecular players controlling the NSR, namely ROS and the HHO transcription factors. This work (i) up opens perspectives on a poorly understood nutrient-related signaling pathway and (ii) defines targets for molecular breeding of plants with enhanced NO3- uptake.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Proteínas de Transporte de Ânions/genética , Proteínas de Transporte de Ânions/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas , Nitratos/metabolismo , Nitrogênio/metabolismo , Raízes de Plantas/metabolismo , Espécies Reativas de Oxigênio , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
12.
Comput Educ ; 159: 104009, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32921895

RESUMO

Systematic reviews were conducted in the nineties and early 2000's on online learning research. However, there is no review examining the broader aspect of research themes in online learning in the last decade. This systematic review addresses this gap by examining 619 research articles on online learning published in twelve journals in the last decade. These studies were examined for publication trends and patterns, research themes, research methods, and research settings and compared with the research themes from the previous decades. While there has been a slight decrease in the number of studies on online learning in 2015 and 2016, it has then continued to increase in 2017 and 2018. The majority of the studies were quantitative in nature and were examined in higher education. Online learning research was categorized into twelve themes and a framework across learner, course and instructor, and organizational levels was developed. Online learner characteristics and online engagement were examined in a high number of studies and were consistent with three of the prior systematic reviews. However, there is still a need for more research on organization level topics such as leadership, policy, and management and access, culture, equity, inclusion, and ethics and also on online instructor characteristics.

13.
Educ Technol Res Dev ; 68(4): 1613-1633, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32837122

RESUMO

In this introduction to the special issue on systematic reviews on emerging learning environments and technologies, we introduce best practices for conducting systematic reviews and meta-analysis and discuss the need for a systematic review on emerging learning environments and technologies. We synthesize research on seven primary areas of emerging learning environments and technologies that include: (1) social media, (2) massive open online courses, (3) special education technology, (4) mobile learning, (5) game-based learning and gamification, (6) adaptive learning, and (7) learning analytics and introduce the thirteen articles that were included in this special issue. This article also provides implications for the readers on using and conducting systematic reviews.

14.
Health Qual Life Outcomes ; 17(1): 86, 2019 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-31118046

RESUMO

BACKGROUND AND AIMS: Patients with age -related hearing loss (ARHL) and their natural caregivers have to confront a disability that produces progressive lifestyle changes. There is an interest in studying the ability of patients and their caregivers to cope with the difficulties that affect quality of life (QoL). In a sample of patient-caregiver dyads in the specific context of ARHL, we examine whether the QoL of patients and caregivers is influenced by the coping processes they use from a specific actor-partner interdependence model (APIM). METHODS: This cross-sectional study involved dyads with patients having a diagnosis of ARHL. The self-reported data included QoL (WHOQoL-BREF) and coping strategies (BriefCope). The APIM was used to test the dyadic effects of coping strategies on QoL. RESULTS: A total of 448 dyads were included; the patients and caregivers were love partners for 59% of the dyads. Coping strategies, such as social support, avoidance, problem solving, and positive thinking, exhibited evidence of actor effects (degree to which the individual's coping strategies are associated with their own QoL). Effects on the partner (degree to which the individual's coping strategies are associated with the QoL of the other member of the dyad) were found, i.e., when the patients mobilized their coping strategy based on social support and problem-solving, their caregivers reported higher environmental QoL. CONCLUSION: This study emphasizes that the QoL for patients and their caregivers was directly related to the coping strategies they used. This finding suggests that targeted interventions should be offered to help patients and their relatives to implement more effective coping strategies.


Assuntos
Adaptação Psicológica , Cuidadores/psicologia , Perda Auditiva/psicologia , Qualidade de Vida/psicologia , Adulto , Idoso , Envelhecimento/fisiologia , Estudos Transversais , Feminino , Perda Auditiva/etiologia , Humanos , Masculino , Pessoa de Meia-Idade , Autorrelato , Apoio Social
15.
Plant Biotechnol J ; 16(1): 322-336, 2018 01.
Artigo em Inglês | MEDLINE | ID: mdl-28626940

RESUMO

Ethylene response factor 1 (ERF1) is an essential integrator of the jasmonate and ethylene signalling pathways coordinating a large number of genes involved in plant defences. Its orthologue in Hevea brasiliensis, HbERF-IXc5, has been assumed to play a major role in laticifer metabolism and tolerance to harvesting stress for better latex production. This study sets out to establish and characterize rubber transgenic lines overexpressing HbERF-IXc5. Overexpression of HbERF-IXc5 dramatically enhanced plant growth and enabled plants to maintain some ecophysiological parameters in response to abiotic stress such as water deficit, cold and salt treatments. This study revealed that HbERF-IXc5 has rubber-specific functions compared to Arabidopsis ERF1 as transgenic plants overexpressing HbERF-IXc5 accumulated more starch and differentiated more latex cells at the histological level. The role of HbERF-IXc5 in driving the expression of some target genes involved in laticifer differentiation is discussed.


Assuntos
Hevea/metabolismo , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Hevea/genética , Reguladores de Crescimento de Plantas/metabolismo , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
16.
Comput Med Imaging Graph ; 52: 58-71, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27025904

RESUMO

Studying morphological changes of subcortical structures often predicate neurodevelopmental and neurodegenerative diseases, such as Alzheimer's disease and schizophrenia. Hence, methods for quantifying morphological variations in the brain anatomy, including groupwise shape analyses, are becoming increasingly important for studying neurological disorders. In this paper, a novel groupwise shape analysis approach is proposed to detect regional morphological alterations in subcortical structures between two study groups, e.g., healthy and pathological subjects. The proposed scheme extracts smoothed triangulated surface meshes from segmented binary maps, and establishes reliable point-to-point correspondences among the population of surfaces using a spectral matching method. Mean curvature features are incorporated in the matching process, in order to increase the accuracy of the established surface correspondence. The mean shapes are created as the geometric mean of all surfaces in each group, and a distance map between these shapes is used to characterize the morphological changes between the two study groups. The resulting distance map is further analyzed to check for statistically significant differences between two populations. The performance of the proposed framework is evaluated on two separate subcortical structures (hippocampus and putamen). Furthermore, the proposed methodology is validated in a clinical application for detecting abnormal subcortical shape variations in Alzheimer's disease. Experimental results show that the proposed method is comparable to state-of-the-art algorithms, has less computational cost, and is more sensitive to small morphological variations in patients with neuropathologies.


Assuntos
Algoritmos , Doença de Alzheimer/diagnóstico por imagem , Adulto , Doença de Alzheimer/patologia , Estudos de Casos e Controles , Criança , Hipocampo/diagnóstico por imagem , Hipocampo/patologia , Humanos , Imageamento por Ressonância Magnética , Masculino , Putamen/diagnóstico por imagem , Putamen/patologia , Esquizofrenia/diagnóstico por imagem , Esquizofrenia/patologia
17.
Eur J Paediatr Neurol ; 20(2): 252-260, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26778232

RESUMO

BACKGROUND: The mechanisms of childhood and perinatal arterial ischemic stroke (AIS) are poorly understood. Multiple risk factors include cerebral arteriopathy, congenital cardiac disease, infection, sickle cell disease, and maternal-fetal conditions in neonates. For infections and parainfectious conditions being the most important a possible inflammatory pathophysiology has long been suspected. This pilot study aims to detect, whether there are any abnormalities of inflammatory markers associated with childhood and neonatal stroke. METHODS: The concentration of 23 different metalloproteinases (MMPs), tissue inhibitors of MMPs (TIMPs), endothelial factors, vascular cell adhesion proteins, and cytokines in plasma were measured in 12 children with AIS, 7 healthy age matched controls and 6 full term neonates with perinatal AIS. RESULTS: At the time of the acute event children with AIS had significantly elevated levels of MMP-9, TIMP4, IL-6, IL-8 and CRP compared to controls (p < 0.05). Except for lower IL-6 and CRP levels the pattern of children with a history of varizella-zoster virus (VZV) and other viral infections did not differ to the non-infectious group. Median levels of MMP-1, MMP-2, TIMP-1, TIMP-2, sE-selectin, sICAM-1, sVCAM-1, IL-8, IL-10, TNF-alpha, VEGF, Fetuin A were found to be higher in the neonatal group when compared with older children. CONCLUSION: This pilot study supports the assumption of an inflammatory process and up-regulation of metalloproteinases and their inhibitors, and altered pattern of circulating pro-inflammatory cytokines, CRP and vWF levels in pediatric and neonatal AIS. It highlights the feasibility but also difficulties for similar larger future studies that should aim to clarify childhood stroke etiopathogenesis and consecutive further therapeutic options.


Assuntos
Biomarcadores/sangue , Acidente Vascular Cerebral/sangue , Acidente Vascular Cerebral/fisiopatologia , Criança , Pré-Escolar , Feminino , Humanos , Lactente , Inflamação/complicações , Masculino , Projetos Piloto , Fatores de Risco
18.
Plasmid ; 81: 50-4, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26210260

RESUMO

pCAMBIA vectors have become popular for their easy handling, stability and the existence of a range of selection and reporter genes. However, these vectors have yet to integrate the Gateway® cloning system, which has enabled site-specific recombination without the need for restriction enzymes and ligases. This paper sets out to convert the pCambia2300 binary vector into a destination vector with the Gateway® cassette driven by the CaMV35S promoter. The destination vector, pCamway35S, was then evaluated using the uidA reporter gene. Transient and stable transformation experiments were successfully assayed, either by particle bombardment or by Agrobacterium tumefaciens in Allium cepa and Hevea embryogenic calli. After counting the transformation units, the statistical analysis performed on the data showed that the pCamway 35S::uidA vector was as efficient as pCambia2301, a pCAMBIA2300 containing the uidA reporter gene under the CaMV 35S promoter.


Assuntos
Clonagem Molecular , Vetores Genéticos/genética , Plasmídeos/genética , Agrobacterium/genética , Clonagem Molecular/métodos , Expressão Gênica , Ordem dos Genes , Genes Reporter , Reprodutibilidade dos Testes , Transformação Bacteriana
19.
Appl Environ Microbiol ; 80(21): 6591-600, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25128340

RESUMO

Ring-hydroxylating dioxygenases (RHDs) play a crucial role in the biodegradation of a range of aromatic hydrocarbons found on polluted sites, including polycyclic aromatic hydrocarbons (PAHs). Current knowledge on RHDs comes essentially from studies on culturable bacterial strains, while compelling evidence indicates that pollutant removal is mostly achieved by uncultured species. In this study, a combination of DNA-SIP labeling and metagenomic sequence analysis was implemented to investigate the metabolic potential of main PAH degraders on a polluted site. Following in situ labeling using [(13)C]phenanthrene, the labeled metagenomic DNA was isolated from soil and subjected to shotgun sequencing. Most annotated sequences were predicted to belong to Betaproteobacteria, especially Rhodocyclaceae and Burkholderiales, which is consistent with previous findings showing that main PAH degraders on this site were affiliated to these taxa. Based on metagenomic data, four RHD gene sets were amplified and cloned from soil DNA. For each set, PCR yielded multiple amplicons with sequences differing by up to 321 nucleotides (17%), reflecting the great genetic diversity prevailing in soil. RHDs were successfully overexpressed in Escherichia coli, but full activity required the coexpression of two electron carrier genes, also cloned from soil DNA. Remarkably, two RHDs exhibited much higher activity when associated with electron carriers from a sphingomonad. The four RHDs showed markedly different preferences for two- and three-ring PAHs but were poorly active on four-ring PAHs. Three RHDs preferentially hydroxylated phenanthrene on the C-1 and C-2 positions rather than on the C-3 and C-4 positions, suggesting that degradation occurred through an alternate pathway.


Assuntos
Betaproteobacteria/genética , Dioxigenases/genética , Dioxigenases/metabolismo , Poluição Ambiental , Metagenoma , Hidrocarbonetos Policíclicos Aromáticos/metabolismo , Microbiologia do Solo , Betaproteobacteria/classificação , Betaproteobacteria/enzimologia , Biotransformação , Clonagem Molecular , DNA Bacteriano/química , DNA Bacteriano/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Variação Genética , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Análise de Sequência de DNA
20.
Eur J Pediatr ; 173(12): 1691-4, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-24706162

RESUMO

UNLABELLED: We describe a 5-year-old girl with marked hypotonia, poor feeding and reduced facial expression since birth. Congenital myopathy was suspected; muscle biopsy showed unspecific type 1 fibre predominance. The possibility of a ryanodine receptor 1 gene (RYR1)-associated myopathy was considered, but not further investigated. At the age of 2 years, she presented with exophthalmos. Brain MRI revealed optic pathway glioma. On clinical examination, she had six café-au-lait spots, thus fulfilling the diagnostic criteria for neurofibromatosis type 1 (NF1). The hypotonia was then attributed to NF1. At the age of 3 years, she developed scoliosis and had an unusually severe motor delay for NF1, as she was not able to walk independently. Dual pathology was suspected, and muscle MRI showed the typical pattern for RYR1-related myopathy. This was genetically confirmed with the discovery of two heterozygous mutations. CONCLUSION: NF1 is one of the most frequent genetic diseases in children. RYR1-related myopathy is one of the most frequent causes of congenital myopathy. The combination of these two pathologies has not yet been described. In cases of unusual presentations or clinical course, the possibility of genetic "double trouble" should be considered.


Assuntos
Anormalidades Múltiplas , DNA/genética , Genes da Neurofibromatose 1 , Mutação de Sentido Incorreto , Miopatia da Parte Central/genética , Neurofibromatose 1/genética , Canal de Liberação de Cálcio do Receptor de Rianodina/genética , Biópsia , Pré-Escolar , Análise Mutacional de DNA , Diagnóstico Diferencial , Feminino , Humanos , Imageamento por Ressonância Magnética , Miopatia da Parte Central/diagnóstico , Miopatia da Parte Central/metabolismo , Neurofibromatose 1/diagnóstico , Neurofibromatose 1/metabolismo , Fenótipo , Músculo Quadríceps/metabolismo , Músculo Quadríceps/patologia , Canal de Liberação de Cálcio do Receptor de Rianodina/metabolismo
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