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1.
BMC Urol ; 21(1): 12, 2021 Jan 28.
Artigo em Inglês | MEDLINE | ID: mdl-33509164

RESUMO

BACKGROUND: Radical prostatectomy is the gold standard treatment for men with localized prostate cancer. This technique is associated with post-operative urinary incontinence. Pelvic floor physiotherapy is a conservative, painless and economical treatment for this specific situation. Kegel exercises and perineal electrostimulation are common techniques to train pelvic floor muscles. The perineal electrostimulation can be applied to the patient with surface electrodes or by an intra-cavitary anal probe. This study proposes that transcutaneous perineal electrostimulation is as effective as intra-cavitary electrostimulation in reducing urinary incontinence secondary to radical prostatectomy. The main objective is to compare the efficacy of the treatment with transcutaneous perineal electrostimulation versus the same intra-cavitary treatment to reduce the magnitude of urinary incontinence after radical prostatectomy, and the impact on the quality of life. METHODS: This single-blind equivalence randomized controlled trial will include 70 man who suffer urinary incontinence post radical prostatectomy. Participants will be randomized into surface electrodes group and intra-anal probe group. The groups will receive treatment for 10 consecutive weeks. Outcomes include changes in the 24-h Pad Test, and ICIQ-SF, SF-12 and I-QoL questionnaires. Clinical data will be collected at baseline, 6 and 10 weeks after the first session, and 6 months after the end of treatment. DISCUSSION: The results will allow us to prescribe the most beneficial perineal electrostimulation technique in the treatment of urinary incontinence derived from radical prostatectomy. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT03587402. 27/06/2018.


Assuntos
Terapia por Estimulação Elétrica/métodos , Complicações Pós-Operatórias/terapia , Prostatectomia , Neoplasias da Próstata/cirurgia , Ensaios Clínicos Controlados Aleatórios como Assunto/métodos , Incontinência Urinária/terapia , Estudos de Equivalência como Asunto , Humanos , Masculino , Períneo , Prostatectomia/métodos , Método Simples-Cego , Estimulação Elétrica Nervosa Transcutânea , Resultado do Tratamento
2.
Trials ; 22(1): 356, 2021 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-34016168

RESUMO

BACKGROUND: Chronic pelvic pain syndrome (CPPS) is a multifactorial disorder that affects 5.7% to 26.6% of women and 2.2% to 9.7% of men, characterized by hypersensitivity of the central and peripheral nervous system affecting bladder and genital function. People with CPPS have much higher rates of psychological disorders (anxiety, depression, and catastrophizing) that increase the severity of chronic pain and worsen quality of life. Myofascial therapy, manual therapy, and treatment of trigger points are proven therapeutic options for this syndrome. This study aims to evaluate the efficacy of capacitive resistive monopolar radiofrequency (CRMRF) at 448 kHz as an adjunct treatment to other physiotherapeutic techniques for reducing pain and improving the quality of life of patients with CPPS. METHODS: This triple-blind (1:1) randomized controlled trial will include 80 women and men with CPPS. Participants will be randomized into a CRMRF activated group or a CRMRF deactivated group and receive physiotherapeutic techniques and pain education. The groups will undergo treatment for 10 consecutive weeks. At the beginning of the trial there will be an evaluation of pain intensity (using VAS), quality of life (using the SF-12), kinesiophobia (using the TSK-11), and catastrophism (using the PCS), as well as at the sixth and tenth sessions. DISCUSSION: The results of this study will show that CRMRF benefits the treatment of patients with CPPS, together with physiotherapeutic techniques and pain education. These results could offer an alternative conservative treatment option for these patients. TRIAL REGISTRATION: ClinicalTrials.gov NCT03797911 . Registered on 8 January 2019.


Assuntos
Dor Crônica , Dor Crônica/diagnóstico , Dor Crônica/terapia , Feminino , Humanos , Masculino , Medição da Dor , Dor Pélvica/diagnóstico , Dor Pélvica/terapia , Qualidade de Vida , Ensaios Clínicos Controlados Aleatórios como Assunto , Síndrome
3.
Int J Pharm ; 494(1): 136-45, 2015 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-26278488

RESUMO

Alternatives to DEHP plasticizers are used in various PVC medical devices (MD) for infusion. As they are able to migrate from these MDs into infused solutions, they may come into contact with patient. Different and specific clinical parameters influence their migration in at-risk situations such as infusion. In contrast to the regulations for Food Contact Materials (MCDA), there is currently no acceptable migration limits for the use of these plasticizers in clinical situations. In order to assess their migration, and thus control the risks linked to these MDs, we developed a migration model for the plasticizers in MDs. To this end, we applied a cross-disciplinary methodological process similar to that used in the food-processing industry, taking into account the MDs' conditions of use in clinical practice. The simulation model is simple and includes the following conditions: MD should be tested with a dynamic method that respects our established clinical assumption (2 L of infused solutions via 13 dm(2) of plasticized PVC), at a temperature of 25 °C and during 24 h of contact, using a 50/50 (v/v) ethanol/water simulant. This model could be proposed as a tool for the safety evaluation of the patients' exposure risk to plasticizers from PVC medical devices for infusions.


Assuntos
Plastificantes/química , Cloreto de Polivinila/química , Dietilexilftalato/química , Humanos , Bombas de Infusão , Modelos Teóricos , Temperatura , Água/química
4.
Mol Plant Microbe Interact ; 8(4): 506-14, 1995.
Artigo em Inglês | MEDLINE | ID: mdl-8589407

RESUMO

Magnaporthe grisea, the fungal pathogen that causes rice blast disease, secretes two endo-beta-1,4-D-xylanases (E. C. 3.2.1.8) when grown on rice cell walls as the only carbon source. One of the xylanases, XYN33, is a 33-kD protein on sodium dodecyl sulfate-polyacrylamide gel and accounts for approximately 70% of the endoxylanase activity in the culture filtrate. The second xylanase, XYN22, is a 22-kD protein and accounts for approximately 30% of the xylanase activity. The two proteins were purified, cloned, and sequenced. XYN33 and XYN22 are both basic proteins with calculated isoelectric points of 9.95 and 9.71, respectively. The amino acid sequences of XYN33 and XYN22 are not homologous, but they are similar, respectively, to family F and family G xylanases from other microorganisms. The genes encoding XYN33 and XYN22, designated XYN33 and XYN22, are single-copy in the haploid genome of M. grisea and are expressed when M. grisea is grown on rice cell walls or on oatspelt xylan, but not when grown on sucrose.


Assuntos
Ascomicetos/enzimologia , Ascomicetos/genética , Xilosidases/genética , Xilosidases/isolamento & purificação , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , Primers do DNA/genética , DNA Fúngico/genética , Endo-1,4-beta-Xilanases , Expressão Gênica , Genes Fúngicos , Ponto Isoelétrico , Dados de Sequência Molecular , Peso Molecular , Oryza/microbiologia , Doenças das Plantas/microbiologia , Reação em Cadeia da Polimerase , Homologia de Sequência de Aminoácidos
5.
FEBS Lett ; 437(3): 319-23, 1998 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-9824316

RESUMO

Two tobacco genes (TOGT) with homology to glucosyltransferase genes known to be induced by salicylic acid (SA) also responded rapidly to a fungal elicitor or to an avirulent pathogen. SA, although an efficient inducer, was shown not to be essential in the signal transduction pathway regulating TOGT gene expression during the resistance response. Recombinant TOGT proteins produced in Escherichia coli exhibited low, but significant, glucosyltransferase activity towards SA, but very high activity towards hydroxycoumarins and hydroxycinnamic acids, with glucose esters being the predominant products. These results point to a possible important function in defense of these glucosyltransferases in conjugating aromatic metabolites prior to their transport and cross-linking to the cell wall.


Assuntos
Benzoatos/metabolismo , Proteínas Fúngicas/farmacologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Glucosiltransferases/genética , Glucosiltransferases/metabolismo , Glicoproteínas de Membrana/farmacologia , Nicotiana/enzimologia , Nicotiana/genética , Doenças das Plantas/genética , Plantas Tóxicas , Ácido Salicílico/metabolismo , Ácido Salicílico/farmacologia , Catálise , Clonagem Molecular , Indução Enzimática/genética , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Glucosiltransferases/biossíntese , Ácido Salicílico/análise , Análise de Sequência de DNA , Nicotiana/virologia , Vírus do Mosaico do Tabaco/metabolismo
6.
Biochimie ; 75(8): 687-706, 1993.
Artigo em Inglês | MEDLINE | ID: mdl-8286442

RESUMO

The hypersensitive reaction to a pathogen is one of the most efficient defense mechanisms in nature and leads to the induction of numerous plant genes encoding defense proteins. These proteins include: 1) structural proteins that are incorporated into the extracellular matrix and participate in the confinement of the pathogen; 2) enzymes of secondary metabolism, for instance those of the biosynthesis of plant antibiotics; 3) pathogenesis-related (PR) proteins which represent major quantitative changes in soluble protein during the defense response. The PRs have typical physicochemical properties that enable them to resist to acidic pH and proteolytic cleavage and thus survive in the harsh environments where they occur: vacuolar compartment or cell wall or intercellular spaces. Since the discovery of the first PRs in tobacco many other similar proteins have been isolated from tobacco but also from other plant species, including dicots and monocots, the widest range being characterized from hypersensitively reacting tobacco. Based first on serological properties and later on sequence data, the tobacco PRs have been classified in five major groups. Group PR-1 contains the first discovered PRs of 15-17 kDa molecular mass, whose biological activity is still unknown, but some members have been shown recently to have antifungal activity. Group PR-2 contains three structurally distinct classes of 1,3-beta-glucanases, with acidic and basic counterparts, with dramatically different specific activity towards linear 1,3-beta-glucans and with different substrate specificity. Group PR-3 consists of various chitinases-lysozymes that belong to three distinct classes, are vacuolar or extracellular, and exhibit differential chitinase and lysozyme activities. Some of them, either alone or in combination with 1,3-beta-glucanases, have been shown to be antifungal in vitro and in vivo (transgenic plants), probably by hydrolysing their substrates as structural components in the fungal cell wall. Group PR-4 is the less studied, and in tobacco contains four members of 13-14.5 kDa of unknown activity and function. Group PR-5 contains acidic-neutral and very basic members with extracellular and vacuolar localization, respectively, and all members show sequence similarity to the sweet-tasting protein thaumatin. Several members of the PR-5 group from tobacco and other plant species were shown to display significant in vitro activity of inhibiting hyphal growth or spore germination of various fungi probably by a membrane permeabilizing mechanism.(ABSTRACT TRUNCATED AT 400 WORDS)


Assuntos
Doenças das Plantas/microbiologia , Proteínas de Plantas/metabolismo , Plantas/imunologia , Quitinases/metabolismo , Glucana 1,3-beta-Glucosidase , Hidrolases/metabolismo , Proteínas de Plantas/biossíntese , Proteínas de Plantas/classificação , Plantas Tóxicas , Nicotiana/metabolismo , beta-Glucosidase/metabolismo
7.
Exp Gerontol ; 35(9-10): 1389-403, 2000 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-11113617

RESUMO

The discovery of two missense mutations (A53T and A30P) in the gene encoding the presynaptic protein alpha-synuclein (alphaSN) that are genetically linked to rare familial forms of Parkinson's disease and its accumulation in Lewy bodies and Lewy neurites has triggered several attempts to generate transgenic mice overexpressing human alphaSN. Analogous to a successful strategy for the production of transgenic animal models for Alzheimer's disease we generated mice expressing wildtype and the A53T mutant of human alphaSN in the nervous system under control of mouse Thy1 regulatory sequences. These animals develop neuronal alpha-synucleinopathy, striking features of Lewy pathology, neuronal degeneration and motor defects. Neurons in brainstem and motor neurons appeared particularly vulnerable. Motor neuron pathology included axonal damage and denervation of neuromuscular junctions, suggesting that alphaSN may interfere with a universal mechanism of synapse maintenance. Thy1-transgene expression of wildtype human alphaSN resulted in comparable pathological changes thus supporting a central role for mutant and wildtype alphaSN in familial and idiopathic forms of diseases with neuronal alpha-synucleinopathy and Lewy pathology. The mouse models provide means to address fundamental aspects of alpha-synucleinopathy and to test therapeutic strategies.


Assuntos
Corpos de Lewy/patologia , Proteínas do Tecido Nervoso/genética , Doença de Parkinson/patologia , Substituição de Aminoácidos , Animais , Encéfalo/metabolismo , Encéfalo/patologia , Modelos Animais de Doenças , Feminino , Expressão Gênica , Genótipo , Humanos , Corpos de Lewy/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , Atividade Motora/genética , Mutação , Doença de Parkinson/genética , Fenótipo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Sinucleínas , Transgenes/genética , alfa-Sinucleína
8.
Ann Fr Anesth Reanim ; 33(5): 304-9, 2014 May.
Artigo em Francês | MEDLINE | ID: mdl-24784917

RESUMO

OBJECTIVES: To assess the stability of cisatracurium besilate solution stored at 5°C and 25°C. MATERIALS AND METHODS: Cisatracurium solutions at 2, 5 and 0.1mg/mL in 0.9 % sodium chloride or 5 % glucose were exposed to 5°C and 25°C under 60 % relative humidity for seven days. The physicochemical stability was assessed at 24, 48hours and seven days with dosage of the active substance, detection of degradation products and a possible racemization, measuring pH, osmolality and turbidity, assessment of coloration, visible particles and invisible particles count. RESULTS: Cisatracurium besilate present good stability for 24hours at 5°C and 25°C for concentrations between 0.1 and 5mg/mL. Beyond 24hours, the solutions at 2 and 5mg/mL remained stable for seven days at 5°C. At 25°C, potentially toxic degradation products appear in solutions of 0.1mg/mL between 24 and 48hours. No racemization was detected, the drug remains in its active form cis. CONCLUSION: Cisatracurium solutions at 2 and 5mg/mL may be stored at 5°C or 25°C for seven days. It's advisable to keep the solutions in a dilution of 0.1mg/mL in 0.9 % sodium chloride or 5 % glucose in the refrigerator. No diluted solution should be stored at room temperature beyond 24hours.


Assuntos
Atracúrio/análogos & derivados , Fármacos Neuromusculares não Despolarizantes/química , Atracúrio/química , Embalagem de Medicamentos , Estabilidade de Medicamentos , Armazenamento de Medicamentos , Glucose , Concentração de Íons de Hidrogênio , Soluções Farmacêuticas , Cloreto de Sódio , Seringas
9.
Ann Fr Anesth Reanim ; 33(4): 227-31, 2014 Apr.
Artigo em Francês | MEDLINE | ID: mdl-24636791

RESUMO

OBJECTIVES: Tracheostomy is a frequent procedure in ICU but patient follow-up and management after ICU has been poorly documented. We conducted a practice survey in French general ICUs and in neurointensive care units concerning tracheostomized ICU patients and their management after ICU. STUDY DESIGN: National observational descriptive transversal study as survey of opinion and practices. MATERIALS AND METHODS: An email, with a link to an automated online questionnaire, was sent to two medical doctors of each French ICU. Demographic data and reported practices concerning indications, technique and post-ICU management were collected. RESULTS: We received 148 intensivists responses from different ICUs, of which 15% from neurointensivists. There was no difference between general intensivists and neurointensivists concerning the reported use of tracheostomy (10±13% vs 20±22%, P=0.05) and concerning the usual timing of the procedure (predominantly between the 10th and the 21th day) (P=0.62). Indications were weaning failure from the ventilator and neurological ventilatory dysfunction. Percutaneous tracheostomy was mainly performed irrespective of the type of unit. Only 48% of doctors declared usually be able to wean patient from the cannula before ICU discharge. Usual difficulties for post-ICU transfer due only to the presence of the cannula were found by 80% of respondants. Eighty-nine per cent of respondents felt that management of tracheostomized patients after the ICU could be improved. CONCLUSION: Tracheostomy is a frequent procedure, mostly percutaneous. Indications and timing for tracheostomy correspond to the recommendations. Reported difficulties in post-ICU management are important and present nationwide.


Assuntos
Unidades de Terapia Intensiva/estatística & dados numéricos , Traqueostomia/estatística & dados numéricos , França , Pesquisas sobre Atenção à Saúde , Humanos , Médicos , Inquéritos e Questionários , Resultado do Tratamento
11.
Ann Fr Anesth Reanim ; 32(1): 12-7, 2013 Jan.
Artigo em Francês | MEDLINE | ID: mdl-23199847

RESUMO

OBJECTIVE: To describe the knowledge of paediatricians regarding the practice of antimicrobial prophylaxis for caesarean section in reference to the Consensus Conference of the French Society of Anesthesia and Intensive Care (SFAR) and assess the feasibility of a change in attitude (injection of the antibiotic prior to incision) among paediatricians Perinatal Health Network of Auvergne (RSPA) working in maternity. STUDY DESIGN: Cross sectional study by survey. METHODS: First questionnaire was sent to 46 RSPA paediatricians working in maternity. Almost one-third of paediatricians who returned the questionnaire said they were not concerned. A second questionnaire was developed with two paediatricians of the CHU and sent to the same 46 paediatricians. The statistical part involved percentages. RESULTS: Response rates were respectively 61% and 67%. For the first questionnaire, only 25% of the paediatricians knew the antibiotic and the time for injection. For the second questionnaire, 87% were in favour of an administration before incision and 42% thought it will not affect the care of the newborn. For 35% of respondents, it could lead to a change in the duration of antibiotic therapy in cases of perinatal infection and for 13% only a delay in the implementation of antibiotic therapy in children. CONCLUSION: The RSPA paediatricians did not know the practices of antibiotic prophylaxis for caesarean section. However, they did not appear opposed to an administration before cord clamping as it would not delay the implementation of any antibiotics in the newborn.


Assuntos
Antibioticoprofilaxia/métodos , Cesárea/métodos , Adulto , Estudos Transversais , Feminino , França , Pesquisas sobre Atenção à Saúde , Conhecimentos, Atitudes e Prática em Saúde , Humanos , Recém-Nascido , Masculino , Pediatria , Assistência Perinatal , Médicos , Gravidez , Inquéritos e Questionários
12.
Fisioterapia (Madr., Ed. impr.) ; 43(5): 264-272, sept.- oct. 2021. tab
Artigo em Espanhol | IBECS (Espanha) | ID: ibc-219247

RESUMO

Introducción y objetivo El dolor miofascial pélvico crónico es una patología con una elevada prevalencia, y de etiología muy diversa, al igual que su tratamiento. No se ha establecido un protocolo de tratamiento, y hasta el momento, se recomienda un abordaje multidisciplinar. El objetivo es evaluar el efecto de un protocolo basado en 10 sesiones de técnicas de inducción miofascial (TIM) en pacientes con dolor miofascial pélvico crónico (DMPC). Material y métodos Se realizó un estudio cuasi experimental (antes-después), con una intervención basada en TIM adaptadas a la localización del dolor de los pacientes. Se evaluó la intensidad del dolor mediante la escala visual analógica (EVA) y la calidad de vida, tanto física como mental, mediante el cuestionario de salud SF-12v2. Para valorar el efecto de las TIM, se compararon los resultados preintervención-postintervención mediante la prueba t de Student para datos apareados y el test no paramétrico de Wilcoxon. Resultados Se incluyeron 50 pacientes en el estudio (27 mujeres y 23 hombres), con una edad media de 44,8 años (desviación estándar [DE] 12,1) y una duración media de los síntomas de 58,3 meses (DE 60,5). El análisis por protocolo mostró que la intensidad del dolor disminuyó en 3,99 puntos al final de la intervención y la calidad de vida mejoró en los sumatorios físico y mental en 5,45 y 5,87 puntos, respectivamente (p < 0,05). El 86,7% de los pacientes completó el protocolo de tratamiento. Conclusiones En un grupo de pacientes con dolor miofascial pélvico crónico, las TIM parecen tener un efecto beneficioso significativo, reduciendo el dolor y mejorando la calidad de vida (AU)


Introduction and objective Myofascial Chronic Pelvic Pain (MCPP) is a pathology with a high prevalence, and an aetiology as diverse is its treatment. A treatment protocol has not yet been established and a multidisciplinary approach is currently recommended. The aim is to evaluate the effect of a Myofascial Induction Techniques (MIT) protocol based on 10 sessions in patients with Myofascial Chronic Pelvic Pain. Material and methods A quasi-experimental study (before-after) was carried out, with an intervention based on MIT adapted to the location of the pain. Pain intensity was evaluated using the Visual Analogue Scale and quality of life, both physical and mental, was assessed using the SF-12v2 Health Survey questionnaire. To assess the effect of MIT, the pre-intervention and post-intervention results were compared using the Student's t test for paired data, and the non-parametric Wilcoxon test. Results 50 patients were included in the study (27 women and 23 men), with a mean age of 44.8 years (SD 12.1), and a mean duration of symptoms of 58.3 months (SD 60.5). The protocol analysis showed that pain intensity decreased by 3.99 points at the end of the intervention and quality of life improved in the physical and mental scores by 5.45 and 5.87 points, respectively (p < 0.05). Of the patients, 86.7% completed the treatment protocol. Conclusions In a group of patients with myofascial chronic pelvic pain, MIT appear to have a significant beneficial effect, reducing pain and improving quality of life (AU)


Assuntos
Humanos , Masculino , Feminino , Adulto , Pessoa de Meia-Idade , Dor Pélvica/terapia , Modalidades de Fisioterapia , Síndromes da Dor Miofascial/terapia , Manipulações Musculoesqueléticas , Resultado do Tratamento , Dor Crônica
15.
Scand J Immunol ; 63(4): 290-8, 2006 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-16623929

RESUMO

The T-cell response to lymphocytic choriomeningitis virus was studied in mice lacking very late antigen-1 (VLA-1). The generation of virus-specific effector T cells was unimpaired in VLA-1(-/-) mice. In the memory phase, VLA-1 deficiency did not influence the number of memory CD8(+) T cells or their distribution between lymphoid and nonlymphoid organs. Regarding a functional role of VLA-1, we found that intracerebral infection of both VLA-1(-/-) and wild-type (wt) mice resulted in lethal T-cell-mediated meningitis, and quantitative and qualitative analyses of the cellular exudate did not reveal any significant differences between the two strains. Expression of VLA-1 was also found to be redundant regarding the ability of effector T cells to eliminate virus from internal organs of i.v. infected mice. Using delayed-type hypersensitivity (DTH) assays to evaluate subdermal CD8(+) T-cell-mediated inflammation, no significant influence of VLA-1 was found either in the primary response or in the memory phase. However, alpha-VLA-4 antibody reduced the DTH-like reaction in VLA-1(-/-) mice to a higher degree than in wt mice, suggesting a synergistic effect of blocking both integrins. Taken together, the current findings indicate that the expression of VLA-1 is not pivotal for T-cell-mediated antiviral immunity to a systemic infection.


Assuntos
Integrina alfa1beta1/imunologia , Vírus da Coriomeningite Linfocítica/imunologia , Linfócitos T/imunologia , Viroses/imunologia , Animais , Linfócitos T CD8-Positivos/imunologia , Hipersensibilidade Tardia/etiologia , Hipersensibilidade Tardia/imunologia , Imunidade Celular , Memória Imunológica , Integrina alfa1beta1/genética , Integrina alfa4beta1/imunologia , Meningite/genética , Meningite/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Knockout , Viroses/patologia
16.
Plant Mol Biol ; 14(3): 381-90, 1990 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-2102821

RESUMO

The purification to homogeneity of pathogenesis-related (PR) proteins R and S from Nicotiana tabacum cv. Samsun NN leaves has been achieved by using a combination of conventional and high-performance chromatographic supports. The same procedure allowed the purification and the characterization of four other proteins which displayed some properties characteristic of tobacco PR proteins and were shown to accumulate in tobacco leaves in response to virus infection. They can be, therefore, considered as new tobacco PR proteins which we designate as PR-s1, -s2, -r1 and -r2. The relative electrophoretic mobilities (Rf) under non-denaturing conditions were estimated to 0.30 for PR-r1 and -r2, 0.25 for Pr-R, 0.20 for PR-s1 and -s2 and 0.15 for PR-S. On SDS gels PR proteins R and S possessed the same apparent molecular weight (Mr 24,000) as did PR-proteins s1 and r1 (Mr 14,500) and PR-s2 and -r2 (Mr 13,000). However, proteins s1, s2, r1 and r2 had identical electrophoretic mobilities on SDS gels when the loading sample buffer contained no reducing agent. Polyclonal antisera were raised against PR proteins R and S and used in immunoblotting experiments. Proteins R and S were shown to be serologically closely related. No cross-reaction was detected with any of the four new tobacco PR proteins r1, r2, s1 and s2 or with the previously described PR proteins, i.e. PR-1a, -1b, -1c, -2, -N, -O, -P and -Q.


Assuntos
Proteínas de Plantas/isolamento & purificação , Plantas/análise , Cromatografia , Reações Cruzadas , Eletroforese em Gel de Poliacrilamida , Imunoquímica , Peso Molecular , Proteínas de Plantas/química , Proteínas de Plantas/imunologia , Plantas/microbiologia , Plantas Tóxicas , Nicotiana/análise , Nicotiana/microbiologia , Vírus do Mosaico do Tabaco/patogenicidade
17.
EMBO J ; 6(11): 3209-12, 1987 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-16453802

RESUMO

Three of the ten acidic ;pathogenesis-related' (PR) proteins known to accumulate in Nicotiana tabacum cv Samsun NN reacting hypersensitively to tobacco mosaic virus, namely -O, -N and -2, have been shown to have 1,3-beta-glucanase (EC 3.2.1.39) activity. By using sera raised against each protein purified to homogeneity close serological relationships have been demonstrated between the three proteins. The same specific sera cross-reacted with a basic protein which is also a 1,3-beta-glucanase induced by virus infection and which can be considered as a new basic pathogenesis-related protein of tobacco. Protein PR-O and the basic 1,3-beta-glucanase display about the same specific enzymatic activity, i.e. 50-fold and 250-fold higher than specific activities of proteins PR-N and -2 respectively.

18.
Proc Natl Acad Sci U S A ; 84(19): 6750-4, 1987 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-16578819

RESUMO

Four endochitinases (poly[1,4-(N-acetyl-beta-D-glucosaminide)] glycanohydrolase, EC 3.2.1.14) have been purified from leaves of Nicotiana tabacum cv. Samsun NN reacting hypersensitively to tobacco mosaic virus. Two of them are acidic proteins of molecular weights 27,500 and 28,500 and have been identified as 2 of the 10 pathogenesis-related proteins that are known to accumulate in tobacco in response to stress or pathogen attack. These two pathogenesis-related proteins, named "P" and "Q" when their biological function was unknown, account for one-third of tobacco mosaic virus-induced chitinase activity of tobacco leaves. They are serologically closely related to the two other chitinases, which can be considered as new basic pathogenesis-related proteins. These two basic chitinases exhibit higher molecular weights (32,000 and 34,000) and higher specific enzyme activity than the two acidic isoforms.

19.
Plant Cell ; 1(3): 285-91, 1989 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-2535503

RESUMO

Samsun NN tobacco cells were transformed with chimeric genes for pathogenesis-related (PR) proteins derived from genomic (PR-1a, GRP) or cDNA (PR-S) clones under the transcriptional control of the cauliflower mosaic virus 35S promoter. Regenerated plants were assayed by RNA and protein gel blotting, and plants showing high specific expression of the inserted genes were selected for self-pollination and seed formation. Inspection of second generation transformants showed that constitutive expression of PR-1a, GRP, and PR-S in tobacco in general does not have an effect on the phenotypic appearance of the plants or the expression of other endogenous PR genes. Furthermore, constitutive expression of the above genes does not affect the susceptibility of the plants to infection with tobacco mosaic virus or alfalfa mosaic virus.


Assuntos
Vírus do Mosaico/fisiologia , Nicotiana/genética , Proteínas de Plantas/genética , Plantas Tóxicas , Sequência de Bases , Células Cultivadas , DNA , Dados de Sequência Molecular , Doenças das Plantas , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Nicotiana/microbiologia , Vírus do Mosaico do Tabaco/fisiologia , Transformação Genética , Replicação Viral
20.
Plant Physiol ; 121(1): 163-72, 1999 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-10482671

RESUMO

H(2)O(2) from the oxidative burst, cell death, and defense responses such as the production of phenylalanine ammonia lyase (PAL), salicylic acid (SA), and scopoletin were analyzed in cultured tobacco (Nicotiana tabacum) cells treated with three proteinaceous elicitors: two elicitins (alpha-megaspermin and beta-megaspermin) and one glycoprotein. These three proteins have been isolated from Phytophthora megasperma H20 and have been previously shown to be equally efficient in inducing a hypersensitive response (HR) upon infiltration into tobacco leaves. However, in cultured tobacco cells these elicitors exhibited strikingly different biological activities. beta-Megaspermin was the only elicitor that caused cell death and induced a strong, biphasic H(2)O(2) burst. Both elicitins stimulated PAL activity similarly and strongly, while the glycoprotein caused only a slight increase. Only elicitins induced SA accumulation and scopoletin consumption, and beta-megaspermin was more efficient. To assess the role of H(2)O(2) in HR cell death and defense response expression in elicitin-treated cells, a gain and loss of function strategy was used. Our results indicated that H(2)O(2) was neither necessary nor sufficient for HR cell death, PAL activation, or SA accumulation, and that extracellular H(2)O(2) was not a direct cause of intracellular scopoletin consumption.


Assuntos
Peróxido de Hidrogênio/metabolismo , Nicotiana/citologia , Fenilalanina Amônia-Liase/metabolismo , Reguladores de Crescimento de Plantas/farmacologia , Plantas Tóxicas , Ácido Salicílico/metabolismo , Escopoletina/metabolismo , Proteínas de Algas/farmacologia , Morte Celular/efeitos dos fármacos , Células Cultivadas , Ativação Enzimática/efeitos dos fármacos , Glicoproteínas/farmacologia , Phytophthora , Doenças das Plantas/induzido quimicamente , Folhas de Planta/citologia , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/enzimologia , Folhas de Planta/metabolismo , Explosão Respiratória/efeitos dos fármacos , Nicotiana/efeitos dos fármacos , Nicotiana/enzimologia , Nicotiana/metabolismo
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