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1.
Front Rehabil Sci ; 4: 1281680, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-38078068

RESUMO

Background: Living with chronic pain (CP) often implies major lifestyle changes, including modifications of daily routines and work. Surprisingly, few validated and effective interventions specifically target functional outcomes in this population. Redesign your Everyday Activities and Lifestyle with Occupational Therapy [REVEAL(OT)] is a lifestyle-oriented intervention led by occupational therapists that directly targets the daily functional challenges of living with CP. The intervention was initially developed and studied as an add-on to standard treatment delivered by Danish multidisciplinary specialized pain clinics. Adapting, implementing, and evaluating REVEAL(OT) within the Canadian healthcare system will contribute to broadening the scope of treatments offered in specialized pain clinics that do not yet include occupational therapy. Objective: The proposed study aims to define and refine REVEAL(OT)/CA with partners (authors of original intervention, people with lived experience, clinicians, managers). Methods: This participatory action research will use a multi-method design and follow the ORBIT model for developing behavioral treatments for chronic diseases. A process of co-construction with partners and an advisory committee will take place in two Montreal specialized pain clinics. It consists of two related work packages (WPs). In WP1, a first series of focus groups with partners (n = 86) and workshops with the advisory committee will be conducted to co-develop the hypothetical pathway describing intervention components and their potential mechanisms of action on targeted outcomes, as well as the first version of the adapted intervention manual. WP2 will co-refine REVEAL(OT)/CA by exploring its acceptability, feasibility and mechanisms of action through intervention deliveries (at least twice in each of two specialized pain clinics; n ≥ 60 patients) and focus groups and/or individual interviews with participating patients and partners. At the end of this study, the intervention manual will be generated both in French and English. Discussion: This study will set the stage for subsequent implementation and effectiveness assessment projects and be an important step towards the deployment of interventions aiming to improve engagement in meaningful daily activities among adults living with CP. Registration: OSF Registries, osf.io/8gksa. Registered 3 August 2023, https://osf.io/8gksa.

2.
J Prev Alzheimers Dis ; 8(2): 142-150, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33569560

RESUMO

Importance/Objective: To describe the feasibility and acceptability of a 6-month web-based multidomain lifestyle training intervention for community-dwelling older people and to test the effects of the intervention on both function- and lifestyle-related outcomes. DESIGN: 6-month, parallel-group, randomized controlled trial (RCT). SETTING: Toulouse area, South-West, France. PARTICIPANTS: Community-dwelling men and women, ≥ 65 years-old, presenting subjective memory complaint, without dementia. INTERVENTION: The web-based multidomain intervention group (MIG) received a tablet to access the multidomain platform and a wrist-worn accelerometer measuring step counts; the control group (CG) received only the wrist-worn accelerometer. The multidomain platform was composed of nutritional advices, personalized exercise training, and cognitive training. Main outcomes and measures: Feasibility, defined as the proportion of people connecting to ≥75% of the prescribed sessions, and acceptability, investigated through content analysis from recorded semi-structured interviews. Secondary outcomes included clinical (eg, cognitive function, mobility, health-related quality of life (HRQOL)) and lifestyle (eg, step count, food intake) measurements. RESULTS: Among the 120 subjects (74.2 ±5.6 years-old; 57.5% women), 109 completed the study (n=54, MIG; n=55, CG). 58 MIG subjects connected to the multidomain platform at least once; among them, adherers of ≥75% of sessions varied across multidomain components: 37 people (63.8% of 58 participants) for cognitive training, 35 (60.3%) for nutrition, and three (5.2%) for exercise; these three persons adhered to all multidomain components. Participants considered study procedures and multidomain content in a positive way; the most cited weaknesses were related to exercise: too easy, repetitive, and slow progression. Compared to controls, the intervention had a positive effect on HRQOL; no significant effects were observed across the other clinical and lifestyle outcomes. CONCLUSIONS AND RELEVANCE: Providing multidomain lifestyle training through a web-platform is feasible and well-accepted, but the training should be challenging enough and adequately progress according to participants' capabilities to increase adherence. Recommendations for a larger on-line multidomain lifestyle training RCT are provided.


Assuntos
Envelhecimento , Cognição/fisiologia , Exercício Físico/fisiologia , Estilo de Vida , Qualidade de Vida , Idoso , Idoso de 80 Anos ou mais , Transtornos Cognitivos/fisiopatologia , Feminino , Humanos , Masculino
3.
Sci Rep ; 10(1): 21785, 2020 12 11.
Artigo em Inglês | MEDLINE | ID: mdl-33311585

RESUMO

The main function of pain is to automatically draw attention towards sources of potential injury. However, pain sometimes needs to be inhibited in order to address or pursue more relevant tasks. Elucidating the factors that influence how people manage this relationship between pain and task performance is essential to understanding the disruptive nature of pain and its variability between individuals. Here, 41 healthy adults completed a challenging working memory task (2-back task) while receiving painful thermal stimulations. Examining the trial-by-trial relationship between pain perception and task performance revealed that pain's disruptive effects on performance were mediated by self-reported pain intensity, and that the analgesic effects of a competing task were influenced by task performance. We found that higher pain catastrophizing, higher trait anxiety, and lower trait mindfulness were associated with larger trade-offs between pain perception and task performance, suggesting that these psychological factors can predict increased fluctuations between disruption by pain and analgesia from a competing task. Altogether these findings provide an important and novel perspective on our understanding of individual differences in the interplay between pain and ongoing task performance.


Assuntos
Analgesia , Ansiedade , Catastrofização , Cognição , Memória de Curto Prazo , Percepção da Dor , Adulto , Ansiedade/tratamento farmacológico , Ansiedade/fisiopatologia , Ansiedade/psicologia , Catastrofização/tratamento farmacológico , Catastrofização/fisiopatologia , Catastrofização/psicologia , Feminino , Humanos , Masculino
4.
Implement Sci ; 14(1): 55, 2019 06 06.
Artigo em Inglês | MEDLINE | ID: mdl-31171011

RESUMO

BACKGROUND: Elders living with polypharmacy may be taking medications that do not benefit them. Polypharmacy can be associated with elevated risks of poor health, reduced quality of life, high care costs, and persistently complex care needs. While many medications could be problematic, this project targets medications that should be deprescribed for most elders and for which guidelines and evidence-based deprescribing tools are available. These are termed potentially inappropriate prescriptions (PIPs) and are as follows: proton pump inhibitors, benzodiazepines, antipsychotics, and sulfonylureas. Implementation strategies for deprescribing PIPs in complex older patient populations are needed. METHODS: This will be a pragmatic cluster randomized controlled trial in community-based primary care practices across Canada. Eligible practices provide comprehensive primary care and have at least one physician that consents to participate. Community-dwelling patients aged 65 years and older with ten or more unique medication prescriptions in the past year will be included. The objective is to assess whether the intervention reduces targeted PIPs for these patients compared with usual care. The intervention, Structured Process Informed by Data, Evidence and Research (SPIDER), is a collaboration between quality improvement (QI) and research programs. Primary care teams will form interprofessional Learning Collaboratives and work with QI coaches to review electronic medical record data provided by their regional Practice Based Research Networks (PBRNs), identify areas of improvement, and develop and implement changes. The study will be tested for feasibility in three PBRNs (Toronto, Montreal, and Edmonton) using prospective single-arm mixed methods. Findings will then guide a pragmatic cluster randomized controlled trial in five PBRNs (Calgary, Winnipeg, Ottawa, Montreal, and Halifax). Seven practices per PBRN will be recruited for each arm. The analysis will be by intention to treat. Ten percent of patients who have at least one PIP at baseline will be randomly selected to participate in the assessment of patient experience and self-reported outcomes. Qualitative methods will be used to explore patient and physician experience and evaluate SPIDER's processes. CONCLUSION: We are testing SPIDER in a primary care population with complex care needs. This could provide a widely applicable model for care improvement. TRIAL REGISTRATION: Clinicaltrials.gov NCT03689049 ; registered September 28, 2018.


Assuntos
Polimedicação , Atenção Primária à Saúde/normas , Melhoria de Qualidade , Idoso , Idoso de 80 Anos ou mais , Canadá , Humanos , Prescrição Inadequada , Masculino , Qualidade de Vida , Projetos de Pesquisa
5.
Aging Clin Exp Res ; 30(9): 1127-1135, 2018 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-29368298

RESUMO

BACKGROUND: Multidomain interventions composed of nutritional counseling, exercise and cognitive trainings have shown encouraging results as effective preventive strategies delaying age-related declines. However, these interventions are time- and resource-consuming. The use of Information and Communication Technologies (ICT) might facilitate the translation from research into real-world practice and reach a massive number of people. AIM: This article describes the protocol of the eMIND study, a randomized controlled trial (RCT) using a web-based multidomain intervention for older adults. METHODS: One hundred and twenty older adults (≥ 65 years), with a spontaneous memory complaint, will be randomly assigned to a six-month web-based multidomain (nutritional counseling, physical and cognitive trainings) intervention group with a connected accelerometer (number of steps, energy expenditure), or to a control group with access to general information on healthy aging plus the accelerometer, but no access to the multidomain intervention. The main outcome is the feasibility/acceptability of the web-based intervention. Secondary clinical outcomes include: cognitive functions, physical performance, nutritional status and cost-effectiveness. RESULTS: We expect a high amount of adherers (ie, > 75% compliance to the protocol) to reflect the feasibility. Acceptability, assessed through interviews, should allow us to understand motivators and barriers to this ICT intervention. We also expect to provide data on its effects on various clinical outcomes and efficiency. CONCLUSION AND DISCUSSION: The eMIND study will provide crucial information to help developing a future and larger web-based multidomain lifestyle RCT, which should facilitate the translation of this ICT intervention from the research world into real-life clinical practice for the healthcare of older adults.


Assuntos
Cognição , Internet , Estilo de Vida , Memória , Idoso , Exercício Físico , Terapia por Exercício/métodos , Humanos , Projetos Piloto , Projetos de Pesquisa
6.
J Perinatol ; 36(9): 758-62, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27228507

RESUMO

OBJECTIVE: Evaluate the impact of a non-neonatal intensive care unit (NICU)-specific peer counseling (PC) program on the cessation of human milk receipt at and post-NICU discharge. STUDY DESIGN: A multivariable logistic regression model used data from 400 mother-infant dyads from a level IV NICU to compare cessation of human milk receipt at NICU discharge by PC program status. Kaplan-Meier distributions and a multivariable Cox proportional hazards model assessed the relationship between participants/non-participants and cessation of human milk post-NICU discharge. RESULTS: No statistically significant differences between groups in cessation of human milk either by or post-discharge were observed. Identified variables associated with the outcome(s) of interest included maternal and infant age, length of stay, presence of a breastfeeding duration goal and frequency of NICU lactation consultant contact. CONCLUSION: Exposure to a non NICU-specific PC program was not associated with human milk receipt either by or post-NICU discharge.


Assuntos
Aconselhamento/métodos , Leite Humano , Mães , Grupo Associado , Connecticut , Feminino , Humanos , Lactente , Recém-Nascido , Unidades de Terapia Intensiva Neonatal , Estimativa de Kaplan-Meier , Modelos Logísticos , Masculino , Análise Multivariada , Alta do Paciente , Avaliação de Programas e Projetos de Saúde
7.
Scand J Med Sci Sports ; 24(1): 234-42, 2014 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22537000

RESUMO

The purpose of this study was to characterize the effect of a 2-week overload period immediately followed by a 1-week taper period on different cognitive processes including executive and nonexecutive functions, and related heart rate variability. Eleven male endurance athletes increased their usual training volume by 100% for 2 weeks, and decreased it by 50% for 1 week. A maximal graded test, a constant speed test at 85% of peak treadmill speed, and a Stroop task with the measurement of heart rate variability were performed at each period. All participants were considered as overreached. We found a moderate increase in the overall reaction time to the three conditions of the Stroop task after the overload period (816 ± 83 vs 892 ± 117 ms, P = 0.03) followed by a return to baseline after the taper period (820 ± 119 ms, P = 0.013). We found no association between cognitive performance and cardiac parasympathetic control at baseline, and no association between changes in these measures. Our findings clearly underscore the relevance of cognitive performance in the monitoring of overreaching in endurance athletes. However, contrary to our hypothesis, we did not find any relationship between executive performance and cardiac parasympathetic control.


Assuntos
Sistema Nervoso Autônomo/fisiologia , Cognição/fisiologia , Função Executiva/fisiologia , Exercício Físico/fisiologia , Frequência Cardíaca/fisiologia , Desempenho Psicomotor/fisiologia , Adulto , Teste de Esforço , Humanos , Masculino , Consumo de Oxigênio/fisiologia , Resistência Física/fisiologia , Tempo de Reação/fisiologia , Teste de Stroop , Adulto Jovem
8.
J Gerontol A Biol Sci Med Sci ; 65(12): 1344-52, 2010 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-20837662

RESUMO

BACKGROUND: There is growing evidence of the involvement of executive control in the maintenance of balance in old age. We examined whether healthy older adults who completed five sessions of nonmotor cognitive dual-task training would show significant improvements on measures of dual-task standing balance and mobility, compared with an untrained control group. METHODS: Twenty healthy older adults were assigned to either training or control groups. In the pre- and post-training sessions, all participants performed tests of cognition, balance, and mobility (single-support balance, dynamic posturography, sit-to-stand, 40-foot walk) under single- and dual-task conditions. The training group completed five sessions of cognitive dual-task training spaced at least 2 days apart. The two tasks involved making two-choice decisions to visually presented stimuli. Participants completed multiple blocks of single-task (task A or B, blockwise) and mixed (A, B, or A + B) trials in each training session. RESULTS: The training group showed significant improvements in body sway during single-support balance and center of gravity alignment during double-support dynamic balance. The control group showed no appreciable improvements. CONCLUSIONS: This study is the first to demonstrate training-related benefits to gross motor performance stemming from cognitive dual-task training. The results support the view that motor control in aging is influenced by executive control and have implications for theories of cognitive training and transfer.


Assuntos
Atenção/fisiologia , Cognição/fisiologia , Educação , Movimento/fisiologia , Equilíbrio Postural/fisiologia , Análise e Desempenho de Tarefas , Idoso , Idoso de 80 Anos ou mais , Retroalimentação Sensorial , Feminino , Gravitação , Humanos , Masculino , Propriocepção/fisiologia , Valores de Referência , Fatores de Tempo , Caminhada/fisiologia
9.
Rev Neurol (Paris) ; 162(5): 628-36, 2006 May.
Artigo em Francês | MEDLINE | ID: mdl-16710129

RESUMO

INTRODUCTION: Misidentification syndromes are the main symptoms in Alzheimer's disease. Underlain by complex cognitive, agnosic, and amnesic disturbances of degenerative etiology, they can be expressed by misidentification delusions, as in psychoses. To date, research has focused on identification disturbances of persons selected according to various definitions. OBJECTIVE: Our main objective was to evaluate the frequency of identification disturbances among patients suffering from Alzheimer's disease within the current conditions of diagnosis and treatment. The secondary objective aimed to establish a detailed analysis of symptoms and clinical correlations, and evaluate the effects of the troubles on the caregiver. METHODS: We conducted a regional survey using a questionnaire designed for the caregiver, proposed to the 60 geriatric doctors and neurologists in the Poitou-Charentes region of France, for all patients suffering from Alzheimer's disease (defined according to DSM IV criteria) seen between June 1st and August 31st of 2003. Statview software was used for statistical analysis. RESULTS: The survey was completed for 104 patients: 69.5 percent women and 30.5 percent men, with a mean age of 79 years. The majority of the patients were seen by a neurologist and presented an average cognitive deficiency (MMS ranging from 11 to 20). An identification disturbance, whether or not it was delusional, all domains included, was found in 81.6 percent of the patients and was related to a more severe cognitive deficiency and greater hardship on the part of the caregiver. DISCUSSION: We observed a high prevalence of identification disturbance, which can be explained by the systematic and exhaustive search for identification disturbance. The most frequent disturbance concerned the identification of places, whereas self-identification was less often affected; the authentic Capgras delusion was found less often. CONCLUSION: The study of identification disturbances in Alzheimer's disease can contribute to a better understanding of the cognitive, psychopathological, and physiopathological aspects of the disease as well as to a better knowledge and better care for the patient.


Assuntos
Doença de Alzheimer/epidemiologia , Síndrome de Capgras/epidemiologia , Delusões/epidemiologia , Idoso , Idoso de 80 Anos ou mais , Doença de Alzheimer/diagnóstico , Doença de Alzheimer/psicologia , Síndrome de Capgras/diagnóstico , Síndrome de Capgras/psicologia , Cuidadores/psicologia , Transtornos Cognitivos/diagnóstico , Transtornos Cognitivos/epidemiologia , Transtornos Cognitivos/psicologia , Comorbidade , Efeitos Psicossociais da Doença , Estudos Transversais , Delusões/diagnóstico , Delusões/psicologia , Feminino , França , Inquéritos Epidemiológicos , Humanos , Masculino , Estatística como Assunto
10.
J Nutr Health Aging ; 6(5): 301-5, 2002.
Artigo em Inglês | MEDLINE | ID: mdl-12474018

RESUMO

BACKGROUND: No previous studies have demonstrated either a nutritional improvement, or a survival benefit from tube placement in an institutionalized population. OBJECTIVE: The aim of this study was to determine current indications for tube feeding in French geriatric centers and to evaluate clinical outcome and mortality rates in these frail very old patients. DESIGN: Between November 1, 2000 and April 31, 2001, we prospectively recruited all hospitalized or institutionalized patients who received enteral nutrition (EN) in 7 Departments of Geriatric Medicine in France. Nutritional parameters and main indications of EN were recorded at the time of feeding tube placement. Pneumonia and mortality rates were observed over a period of one year. RESULTS: 57 patients of mean age 81.6 7.8 yrs underwent placement of a feeding tube. Mean BMI value was 20.7 4.8 and mean serum albumin level 26.1 6.1 g/L. The most frequent indications for EN included stoke (39%) and other neurologic diseases (42%). Fourteen patients (25%) died within 30 days, and 27 (47%) died over the 12-month follow-up period. During the first month, an episode of pneumonia was noted in 26 cases (55%). CONCLUSION: The similarity between rates of early mortality reported in our study and those reported in several previous studies involving younger, ambulatory subjects is surprising because we might expect poorer survival in our frail elderly patients. We can think that French geriatric teams have changed their attitudes toward EN in recent years, EN being less frequently used in patients with advanced dementia and at the end-stage of life.

11.
J Appl Physiol (1985) ; 90(5): 1685-90, 2001 May.
Artigo em Inglês | MEDLINE | ID: mdl-11299256

RESUMO

The respective oxidation of glycerol and glucose (0.36 g/kg each) ingested simultaneously immediately before exercise (120 min at 68 +/- 2% maximal oxygen uptake) was measured in six subjects using (13)C labeling. Indirect respiratory calorimetry corrected for protein and glycerol oxidation was used to evaluate the effect of glucose + glycerol ingestion on the oxidation of glucose and fat. Over the last 80 min of exercise, 10.0 +/- 0.8 g of exogenous glycerol were oxidized (43% of the load), while exogenous glucose oxidation was 21% higher (12.1 +/- 0.7 g or 52% of the load). However, because the energy potential of glycerol is 18% higher than that of glucose (4.57 vs. 3.87 kcal/g), the contribution of both exogenous substrates to the energy yield was similar (4.0-4.1%). Total glucose and fat oxidation were similar in the placebo (144.4 +/- 13.0 and 60.5 +/- 4.2 g, respectively) and the glucose + glycerol (135.2 +/- 12.0 and 59.4 +/- 6.5 g, respectively) trials, whereas endogenous glucose oxidation was significantly lower than in the placebo trial (123.7 +/- 11.7 vs. 144.4 +/- 13.0 g). These results indicate that exogenous glycerol can be oxidized during prolonged exercise, presumably following conversion into glucose in the liver, although direct oxidation in peripheral tissues cannot be ruled out.


Assuntos
Exercício Físico/fisiologia , Glucose/metabolismo , Glicerol/metabolismo , Consumo de Oxigênio , Esforço Físico/fisiologia , Administração Oral , Adulto , Testes Respiratórios , Calorimetria Indireta/métodos , Dióxido de Carbono/análise , Isótopos de Carbono , Glucose/administração & dosagem , Glicerol/administração & dosagem , Humanos , Masculino , Oxirredução
12.
Patient Educ Couns ; 45(4): 275-84, 2001 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-11755773

RESUMO

The aim of this study was to describe the frequency and content of discussions on lifestyle issues during an annual examination and to identify characteristics of patients, physicians, and visits associated with lifestyle discussions. Audio-recordings of the visits of 35 family physicians with 148 of their patients were analyzed. Bivariate descriptive and multivariate analyses were used. On average, the visits contained discussions of 3.6 different issues for a total time of 2.9 min. Of the 11 topics of interest, weight, diet and nutrition, physical activity, and tobacco use were the most frequently discussed. Consultation with a female physician and perception by the physician of a poorer patient mental health status were associated with the number of themes discussed and the duration of exchanges. Results suggest that although, discussions on lifestyle issues are frequently observed during these visits, they remain limited in scope. Physicians appear to concentrate their energy on targeted patients, and female doctors are more active in this domain.


Assuntos
Medicina de Família e Comunidade/normas , Estilo de Vida , Aceitação pelo Paciente de Cuidados de Saúde/psicologia , Educação de Pacientes como Assunto/normas , Relações Médico-Paciente , Adulto , Idoso , Análise de Variância , Comunicação , Feminino , Promoção da Saúde/normas , Pesquisa sobre Serviços de Saúde , Humanos , Masculino , Pessoa de Meia-Idade , Motivação , Aceitação pelo Paciente de Cuidados de Saúde/estatística & dados numéricos , Quebeque , Fatores Sexuais , Inquéritos e Questionários , Gravação em Fita
13.
Int J Psychoanal ; 81 ( Pt 4): 667-86, 2000 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-11028233

RESUMO

The author investigates the origins of 'Mourning and melancholia', which has been the standard work of psychoanalytic reference on mourning since its publication. She notes that the existence of this paper has always tended to be taken for granted and that it is therefore important to identify the foundations on which Freud developed his conception of mourning, which seem to have remained almost unexamined in the literature. The internal and external sources are discussed. The former are defined as biographical data, involving a correlation between the theme of mourning and Freud's own bereavements, as well as a possible causal link between these instances and his creative activity, while the latter comprise Freud's reading and his scientific exchanges. The external sources may be subdivided into anthropological, religious (mainly Jewish), psychiatric and psychoanalytic sources. The author points out that in his essay Freud has far less to say about mourning than about melancholia and concludes that the new view of this founding text accruing from the consideration of all the above factors shows that Freud was relatively uninterested in the normal model of mourning, especially when compared with all that flowed from his work on dreams.


Assuntos
Luto , Transtorno Depressivo Maior/psicologia , Transtorno Depressivo Maior/terapia , Teoria Freudiana , Humanos , Terapia Psicanalítica
15.
Glycobiology ; 9(10): 1045-51, 1999 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-10521541

RESUMO

The genome of Saccharomyces cerevisiae contains five genes that encode type II transmembrane proteins with significant amino acid similarity to the alpha-1,3-mannosyltransferase Mnn1p. The roles of the three genes most closely related to MNN1 were examined in mutants carrying single and multiple combinations of the disrupted genes. Paper chromatographic analysis of [2-3H]mannose-labeled O-linked oligosaccharides released by beta-elimination showed that the MNT2 (YGL257c) and MNT3 (YIL014w) genes in combination with MNN1 have overlapping roles in the addition of the fourth and fifth alpha-1,3-linked mannose residues to form Man4 and Man5 oligosaccharides whereas MNT4 (YNR059w) does not appear to be required for O-glycan synthesis.


Assuntos
Manosiltransferases/metabolismo , Glicoproteínas de Membrana/metabolismo , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae/enzimologia , Sequência de Aminoácidos , Genes Fúngicos , Manose/metabolismo , Manosiltransferases/genética , Glicoproteínas de Membrana/genética , Dados de Sequência Molecular , Família Multigênica , Mutação , Oligossacarídeos/biossíntese , Oligossacarídeos/química , Saccharomyces cerevisiae/genética , Homologia de Sequência de Aminoácidos
16.
Science ; 285(5429): 901-6, 1999 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-10436161

RESUMO

The functions of many open reading frames (ORFs) identified in genome-sequencing projects are unknown. New, whole-genome approaches are required to systematically determine their function. A total of 6925 Saccharomyces cerevisiae strains were constructed, by a high-throughput strategy, each with a precise deletion of one of 2026 ORFs (more than one-third of the ORFs in the genome). Of the deleted ORFs, 17 percent were essential for viability in rich medium. The phenotypes of more than 500 deletion strains were assayed in parallel. Of the deletion strains, 40 percent showed quantitative growth defects in either rich or minimal medium.


Assuntos
Deleção de Genes , Genes Essenciais , Genoma Fúngico , Fases de Leitura Aberta , Saccharomyces cerevisiae/genética , Meios de Cultura , Regulação Fúngica da Expressão Gênica , Marcação de Genes , Genes Fúngicos , Fenótipo , Reação em Cadeia da Polimerase , Recombinação Genética , Saccharomyces cerevisiae/crescimento & desenvolvimento
17.
Yeast ; 15(10B): 1001-8, 1999 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10407279

RESUMO

The complete yeast genome contains a large number of genes of unknown biological function. Simple, rapid and reliable specific screens are valuable tools in exploring gene function via systematic phenotypic analysis of large mutant collections. This report provides a new approach for monitoring changes in cell wall strength, based on the deleterious effects caused by ultrasound on the yeast cell surface. Sonication can thus be used for the screening of mutants affected in the architecture or stability of the cell wall, since such mutants are expected to have an altered sensitivity to this treatment compared to that of a wild-type. The experimental procedure, consisting in the quantification of damaged cells after a mild sonication treatment, by means of flow cytometry, can be applied on a large scale. The usefulness of the sonication assay as a primary screen for cell wall-related mutants is evaluated on the collection of calcofluor white-hypersensitive and -resistant mutants obtained by Lussier et al. (1997). A further phenotypic characterization of the sonication-hypersensitive mutants within the calcofluor white collection is also presented.


Assuntos
Parede Celular/fisiologia , Genes Fúngicos , Mutação , Leveduras/genética , Benzenossulfonatos/farmacologia , Cafeína/farmacologia , Cálcio/farmacologia , Parede Celular/química , Citometria de Fluxo , Fenótipo , Sonicação , Leveduras/efeitos dos fármacos , Leveduras/ultraestrutura
18.
Proc Natl Acad Sci U S A ; 96(3): 1065-70, 1999 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-9927694

RESUMO

Tissue-type plasminogen activator (tPA) catalyzes the rate-limiting initial step in the fibrinolytic cascade. Systemic infusion of tPA has become the standard of care for acute myocardial infarction. However, even the relatively short-duration protocols currently employed have encountered significant hemorrhagic complications, as well as complications from rebound thrombosis. Gene therapy offers a method of local high-level tPA expression over a prolonged time period to avoid both systemic hemorrhage and local rebound thrombosis. To examine the impact of local tPA overexpression, an adenoviral vector expressing tPA was created. The construct was characterized functionally in vitro, and the function of the vector was confirmed in vivo by delivery to the rabbit common femoral artery. Systemic coagulation parameters were not perturbed at any of the doses examined. The impact of local overexpression of tPA on in vivo thrombus formation was examined subsequently in a stasis/injury model of arterial thrombosis. The construct effectively prevented arterial thrombosis in treated animals, whereas viral and nonviral controls typically developed occluding thrombi. This construct thus offers a viable technique for promoting a locally thromboresistant small-caliber artery.


Assuntos
Terapia Genética , Trombose/prevenção & controle , Ativador de Plasminogênio Tecidual/metabolismo , Adenoviridae , Animais , Células Cultivadas , Endotélio Vascular/citologia , Endotélio Vascular/fisiologia , Artéria Femoral , Vetores Genéticos , Humanos , Coelhos , Proteínas Recombinantes/biossíntese , Trombose/patologia , Ativador de Plasminogênio Tecidual/biossíntese , Ativador de Plasminogênio Tecidual/genética , Veias Umbilicais
19.
Biochim Biophys Acta ; 1426(2): 323-34, 1999 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-9878809

RESUMO

Glycosylation constitutes one of the most important of all the post-translational modifications and may have numerous effects on the function, structure, physical properties and targeting of particular proteins. Eukaryotic glycan structures are progressively elaborated in the secretory pathway. Following the addition of a core N-linked carbohydrate in the endoplasmic reticulum, glycoproteins move to the Golgi complex where the elongation of O-linked sugar chains and processing of complex N-linked oligosaccharide structures take place. In order to better define how such post-translational modifications occur, we have been studying the yeast KTR and MNN1 mannosyltransferase gene families. The KTR family contains nine members: KRE2, YUR1, KTR1, KTR2, KTR3, KTR4, KTR5, KTR6 and KTR7. The MNN1 family contains six members: MNN1, TTP1, YGL257c, YNR059w, YIL014w and YJL86w. In this review, we address protein structure, sequence similarities and enzymatic activity in the context of each gene family. In addition, a description of the known function of many family members in O- and N-linked glycosylation is included. Finally, the genetic interactions and functional redundancies within a gene family are also discussed.


Assuntos
Manosiltransferases/genética , Glicoproteínas de Membrana/genética , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae/enzimologia , Sequência de Aminoácidos , Proteínas Fúngicas/biossíntese , Proteínas Fúngicas/genética , Glicoproteínas/biossíntese , Glicosilação , Dados de Sequência Molecular , Nitrogênio/química , Oxigênio/química , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/ultraestrutura
20.
J Bacteriol ; 180(19): 5020-9, 1998 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9748432

RESUMO

The Candida glabrata KRE9 (CgKRE9) and KNH1 (CgKNH1) genes have been isolated as multicopy suppressors of the tetracycline-sensitive growth of a Saccharomyces cerevisiae mutant with the disrupted KNH1 locus and the KRE9 gene placed under the control of a tetracycline-responsive promoter. Although a cgknh1Delta mutant showed no phenotype beyond slightly increased sensitivity to the K1 killer toxin, disruption of CgKRE9 resulted in several phenotypes similar to those of the S. cerevisiae kre9Delta null mutant: a severe growth defect on glucose medium, resistance to the K1 killer toxin, a 50% reduction of beta-1,6-glucan, and the presence of aggregates of cells with abnormal morphology on glucose medium. Replacement in C. glabrata of the cognate CgKRE9 promoter with the tetracycline-responsive promoter in a cgknh1Delta background rendered cell growth tetracycline sensitive on media containing glucose or galactose. cgkre9Delta cells were shown to be sensitive to calcofluor white specifically on glucose medium. In cgkre9 mutants grown on glucose medium, cellular chitin levels were massively increased.


Assuntos
Candida/genética , Proteínas Fúngicas/genética , Glucanos/análise , Glicoproteínas/genética , Proteínas de Saccharomyces cerevisiae , Saccharomyces cerevisiae/genética , Homologia de Sequência de Aminoácidos , beta-Glucanas , Sequência de Aminoácidos , Sequência de Bases , Benzenossulfonatos/farmacologia , Candida/citologia , Candida/efeitos dos fármacos , Parede Celular/química , Quitina/análise , Clonagem Molecular , Resistência Microbiana a Medicamentos , Genes Fúngicos/genética , Teste de Complementação Genética , Fatores Matadores de Levedura , Dados de Sequência Molecular , Mutação , Micotoxinas/farmacologia , Regiões Promotoras Genéticas/genética , Proteínas Recombinantes de Fusão , Mapeamento por Restrição , Análise de Sequência de DNA
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