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1.
Artigo em Inglês | MEDLINE | ID: mdl-37539856

RESUMO

BACKGROUND AND OBJECTIVES: Cypress and olive pollen are the most prevalent sensitizer trees in the Mediterranean area. Some patients exhibit a dual sensitization which has not been well documented yet. To identify the allergens involved in the dual cypress and olive allergy (C+O) and study the relationship between phenotype and allergen sensitization. METHODS: C+O patients were selected. Monosensitized subjects to olive or cypress were used as reference. Specific IgE to whole extracts and purified allergens from olive and cypress were performed. Immunoblotting was done to analyze IgG and IgE-binding using olive polyclonal antibodies and patients' sera, respectively. Mutual immunoblotting inhibition of olive and cypress extracts, and inhibition of cypress extract immunoblotting with olive allergens were performed. Multiple correspondence analysis and hierarchical cluster classifications were conducted to analyze the relationships between C+O clinical presentation (symptoms, seasonality) and allergen profile. RESULTS: C+O patients were clustered in 4 phenotypes. The most frequent one (58.4%) was rhinoconjunctivitis in winter (February) and spring (May), with asthma in 38% of subjects. Ole e 1 and Cup s 1 were the major allergens. Homologous proteins to Ole e 1, Ole e 9 and Ole e 11 in cypress pollen were identified and these olive allergens inhibit IgE-binding to cypress extract. CONCLUSIONS: The exclusive C+O allergy results from co-sensitization to Cup s 1 and Ole e 1, and to cross-reactivity due to Ole e 1-like, Ole e 9-like and Ole e 11-like allergens not described previously, and translates into 4 clinical phenotypes of winter and/or spring or perennial rhinoconjunctivitis with and without asthma.

4.
J Investig Allergol Clin Immunol ; 32(5): 375-382, 2022 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-34085936

RESUMO

BACKGROUND: Given the increased popularity of flaxseed in meals, several cases of allergy to these seeds have been reported. Little is known about the allergens implicated in hypersensitivity reactions to flaxseed. The present study aimed to identify the allergens involved in IgE-mediated reactions in 5 patients with a clinical history of severe systemic symptoms after flaxseed consumption. METHODS: Proteins that were potential allergens with IgE-binding capacity were purified from flaxseed extract using chromatography and identified via MALDI-TOF mass spectrometry. Immunoassays were performed using the 5 allergic patients' sera tested individually and as a pool. RESULTS: Immunoblotting of the flaxseed extract revealed a low-molecular-mass protein (around 13 kDa) in 4 of the 5 patients, while a protein of approximately 55 kDa was detected in 2 patients. The proteins were identified by mass spectrometry as flaxseed 2S albumin, which is included in the WHO/IUIS allergen nomenclature as Lin u 1, and 11S globulin. Inhibition assays revealed in vitro IgE-mediated cross-reactivity between Lin u 1 and peanut and cashew nut proteins, while IgE-mediated recognition of 11S globulin by patients' sera was partially inhibited by several plant-derived sources. CONCLUSIONS: Seed storage proteins from flaxseed were involved in the development of severe symptoms in the 5 patients studied and exhibited cross-reactivity with other allergenic sources. Besides the severity of flaxseed allergy in patients sensitized to 2S albumin, this is the first time that 11S globulin has been identified as a potential allergen. Taking these data into account should ensure a more accurate diagnosis.


Assuntos
Linho , Globulinas , Hipersensibilidade a Noz , Proteínas de Nozes , Albuminas , Alérgenos/efeitos adversos , Sequência de Aminoácidos , Antígenos de Plantas , Reações Cruzadas , Linho/efeitos adversos , Humanos , Imunoglobulina E/metabolismo , Extratos Vegetais , Proteínas de Plantas/efeitos adversos , Proteínas de Armazenamento de Sementes/metabolismo
7.
Obes Rev ; 18 Suppl 2: 7-18, 2017 07.
Artigo em Inglês | MEDLINE | ID: mdl-28741907

RESUMO

The prevalence of overweight and obesity is rapidly increasing among Latin American children, posing challenges for current healthcare systems and increasing the risk for a wide range of diseases. To understand the factors contributing to childhood obesity in Latin America, this paper reviews the current nutrition status and physical activity situation, the disparities between and within countries and the potential challenges for ensuring adequate nutrition and physical activity. Across the region, children face a dual burden of undernutrition and excess weight. While efforts to address undernutrition have made marked improvements, childhood obesity is on the rise as a result of diets that favour energy-dense, nutrient-poor foods and the adoption of a sedentary lifestyle. Over the last decade, changes in socioeconomic conditions, urbanization, retail foods and public transportation have all contributed to childhood obesity in the region. Additional research and research capacity are needed to address this growing epidemic, particularly with respect to designing, implementing and evaluating the impact of evidence-based obesity prevention interventions.


Assuntos
Dieta , Exercício Físico , Estado Nutricional , Sobrepeso/epidemiologia , Obesidade Infantil/epidemiologia , Magreza/epidemiologia , Adolescente , Criança , Humanos , América Latina/epidemiologia , Sobrepeso/etiologia , Obesidade Infantil/etiologia , Prevalência , Fatores Socioeconômicos , Magreza/etiologia
8.
Acta neurol. colomb ; 30(1): 22-31, ene.-mar. 2014. ilus, tab
Artigo em Espanhol | LILACS | ID: lil-724885

RESUMO

Introducción. Diferentes pruebas neuropsicológicas permiten explorar las funciones cognitivas del adulto mayor, en un tiempo corto. En Colombia se dispone de pocos estudios sobre puntuaciones y puntos de corte para el MMSE y para el MoCA en relación al diagnóstico de deterioro cognitivo. Objetivo. Describir la distribución de las puntuaciones del MMSE y el MoCA y los puntos de corte con mejor discriminación, para el diagnóstico de deterioro cognitivo leve y demencia, en una muestra de pacientes de Bogotá. Material y métodos. Se evaluaron 248 pacientes por un equipo multidisciplinario, que consultaron a la Clínica de Memoria del HIUSJ entre 2009-2012, siguiendo un protocolo establecido. Se identificaron las puntuaciones del MoCA y MMSE, que permitieron obtener el mayor porcentaje de pacientes correctamente clasificados. Resultados. En el 70% de los pacientes con DCL y en el 69 % de los sujetos normales, se encontraron puntuaciones del MMSE inferiores o iguales a 28. En 91% de pacientes con DCL y 84% de los sujetos normales, se presentaron puntuaciones del MoCA inferiores o iguales a 25. Los pacientes con cualquier tipo de demencia, presentaron puntuaciones del MMSE inferiores o iguales a 27 e inferiores o iguales a 24 en el MoCA. Conclusión. Según el presente estudio, el tamizaje de funciones cognitivas, utilizando el MoCA, clasifica de manera más acertada que el MMSE, a los sujetos con deterioro cognitivo. Creemos que en atención primaria, estos puntos de corte del MoCA, pueden ser considerados por ahora, cuando se trate especialmente de sujetos con alta escolaridad.


Introduction. Some cognitive tests allow the evaluation of cognitive functions on the elderly in a short period of time. There are few studies in Colombia about cut-off point for the MMSE and the MoCA test. Objectives. To describe the distribution on scores on MMSE and MoCA test and the cut-off point with a better discrimination criteria for the diagnosis of mild cognitive impairment and dementia, in a sample of patients from Bogotá. Materials and methods. Two hundred forty eight patients were included in this study, being evaluated by a multidisciplinary team that followed an established protocol, on patients who attended to the Memory Clinic of HIUSJ between 2009-2012. MoCA test and MMSE scores that allow higher percentages of correctly classified patients were identified. Results. Seventy percent of patients with mild cognitive impairment and 69% of normal individuals had scores on MMSE below or equal to 28. Ninety-one percent of patients with MCI and 89% of normal patients, had scores below or equal to 25. Patients with any type of dementia had scores on MMSE below or equal to 27 and below or equal to 24 in MoCA test. Conclusion. According to the study, the screening of cognitive functions, using MoCA test, is more accurate than MMSE in patients with cognitive decline. The cut-off points, identified in our study, can be considered useful until now in primary attention, in patients with a high level of education.

9.
Arch. alerg. inmunol. clin ; 44(2): 55-61, 2013. tab
Artigo em Espanhol | LILACS | ID: biblio-947749

RESUMO

Antecedentes. Los estados de inmunodeficiencia impactan en el funcionamiento familiar. Objetivo. Comparar el funcionamiento familiar y aspectos clínico-nutricionales entre niños con alto nivel de sospecha de inmunodeficiencias primarias y aquellos con menor nivel de sospecha. Lugar de aplicación. Clínica de atención primaria. Diseño. Estudio transversal con grupos comparados. Población. 400 niños entre 1 año y 7 años 11 meses de edad, asignados a dos grupos, uno integrado por aquellos con puntaje ≥25 en la escala PRIDE (primary immunodeficiency disease evaluation), n=9 y otro integrado por aquellos que alcanzaron un puntaje ≤25 en la evaluación con la escala PRIDE, n=391. Material y métodos. Los grupos se compararon con respecto a funcionamiento familiar, estado nutricional y criterios clínicos de riesgo para inmunodeficiencias primarias. Resultados. Se encontraron asociaciones estadísticamente significativas de puntaje PRIDE con infecciones respiratorias altas >8, episodios de otitis media aguda ≥4, episodios de sinusitis aguda ≥2, infecciones de vías respiratorias bajas ≥2, no respuesta a ≥3 esquemas de antibióticos para un mismo cuadro de infección, infecciones urinarias >6, infecciones del sistema nervioso central, enfermedades alérgicas o autoinmunes concomitantes, caída del cordón umbilical > 30 días y funcionamiento familiar. Las áreas más dañadas del funcionamiento familiar fueron roles y armonía. Conclusiones. Hubo diferencias en el funcionamiento familiar y factores de riesgo relacionados con infección y predisposición a ella entre los niños con puntaje PRIDE ≥25 y aquellos con puntaje <25. (AU)


Background. Immunodeficiency states impact on family functioning. Objective. To compare family functioning, clinical and nutritional status among children with high suspicion of primary immunodeficiencies versus those in which the level of suspicion is lower. Design. Cross-sectional study with compared groups. Population. 400 children between 1 year and 7 years 11 months of age, assigned to two groups according to PRIDE (primary immunodeficiency disease evaluation) ³25 average, n=9 and the other composed of those who scored below 25 on the assessment with that scale, n = 391. Material and methods. The groups were compared regarding: family functioning, nutritional status and clinical risk criteria for primary immunodeficiencies. Results. Statistically significant associations were found among PRIDE scoring with following variables: upper respiratory infections >8, episodes of acute media otitis ³4, two o more episodes of lower respiratory infections, sinusitis ³ 2, three or more schedules of antibiotics to treat the same infection, more than six urinary tract infections in the past year, central nervous system infections, allergic or autoimmune concomitant diseases, drop of umbilical cord > 30 days and family functioning. The most damaged areas of family functioning were roles and harmony. Conclusions. There were differences in family functioning and related risk factors for both infection and predisposition to that between children receiving 25 or more points after PRIDE evaluation and those with PRIDE score below 25(AU)


Assuntos
Humanos , Masculino , Feminino , Pré-Escolar , Criança , Família , Estado Nutricional , Fatores de Risco , Síndromes de Imunodeficiência , Cordão Umbilical , Infecções
10.
Neurol Res ; 31(3): 228-33, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19040797

RESUMO

BACKGROUND: Gonyautoxin are phycotoxins, whose molecular mechanism of action is a reversible block of the voltage-gated sodium channels at axonal level, impeding nerve impulse propagation. OBJECTIVE: To evaluate clinical efficacy of gonyautoxin in the treatment of patients with chronic tensional-type headache. METHODS: Open trial from September 2004 to 2005 in Hospital Clínico Universidad de Chile. Twenty-seven patients with chronic tension-type headache were locally infiltrated with gonyautoxins (50 micrograms) in ten sites considered as pain trigger points in a fixed infiltration protocol. In each site, a volume of 200 microlitres was injected. EMG recording was performed before and immediately after infiltrations. Main outcome measures are where a significantly drop-off in acute headache pain score occurs and number of days without headache pain. RESULTS: No side effects were detected in the follow-up period. From base line of 2 weeks, 19 patients of 27 (70%) are the successfully responders to the treatment. They showed the remarkable immediate effect after infiltration demonstrated by trapezium EMG recording. Patients reported a fall in pain score 5 minutes post-injection from 5.0 +/- 2.8 to 1.6 +/- 1.6 (mean +/- SD). The responder showed an average of 8.1 +/- 9.9 weeks of headache pain-free, all of them without a second infiltration or use of any additional analgesic medication. DISCUSSION: The therapeutic properties of gonyautoxin local infiltration in chronic tension-type headache patients are shown to be safe and effective. This report describes a new therapy for chronic tension-type headache involving local infiltrations of gonyautoxins. The immediate headache pain relief effect shown only minutes after toxin infiltrations were the most remarkable feature of this protocol. This is the first gonyautoxins testing report in the treatment of chronic tension-type headache.


Assuntos
Saxitoxina/análogos & derivados , Cefaleia do Tipo Tensional/tratamento farmacológico , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Doença Crônica , Eletromiografia/efeitos dos fármacos , Feminino , Humanos , Injeções Intramusculares/métodos , Masculino , Toxinas Marinhas/administração & dosagem , Toxinas Marinhas/efeitos adversos , Pessoa de Meia-Idade , Medição da Dor/efeitos dos fármacos , Saxitoxina/administração & dosagem , Saxitoxina/efeitos adversos
11.
P. R. health sci. j ; 24(2): 137-140, Jun. 2005.
Artigo em Inglês | LILACS | ID: lil-472969

RESUMO

BACKGROUND: Respiratory syncytial virus (RSV) is an important cause of respiratory tract disease in infants and young children. Immunoprophylaxis is available for high-risk infants. This study reviews infants with bronchiolitis at three primary care hospitals to describe the epidemiology of RSV infection in Puerto Rico. METHODS: We analyzed data from 2 hospitals by obtaining the number of infants diagnosed with bronchiolitis and estimating the percentage of cases due to RSV. A third hospital included patients with confirmed RSV infection. RESULTS: There were 4,557 patients in the study. RSV infection occurred throughout the year. Data shows a progressive decrease in RSV-positive infections. CONCLUSIONS: Data confirms year-round incidence of RSV in Puerto Rico. A standardized surveillance system in Puerto Rico is imperative to determine RSV epidemiology. The decrease in positive RSV infections may be due to the introduction of immunoprophylaxis to high-risk patients.


Assuntos
Humanos , Lactente , Recém-Nascido , Bronquiolite/epidemiologia , Infecções por Vírus Respiratório Sincicial/epidemiologia , Vírus Sincicial Respiratório Humano/isolamento & purificação , Bronquiolite/diagnóstico , Bronquiolite/virologia , Estudos Transversais , Infecções por Vírus Respiratório Sincicial/diagnóstico , Porto Rico/epidemiologia , Estudos Retrospectivos
13.
Biochem Soc Trans ; 31(Pt 3): 584-6, 2003 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-12773160

RESUMO

We have identified three groups of growth-constraint genes using mosaic genetic screens in Drosophila melanogaster, including PTEN (phosphatase and tensin homologue deleted on chromosome 10), and the tuberous sclerosis complex (TSC) genes, Tsc1 and Tsc2. Our studies show that all three groups of genes participate in mechanisms that regulate organ and organism size in animals. We propose that mechanisms of organ size control are critical targets for diseases, such as tumorigenesis, which require an increase in tissue size and total mass, and for evolutionary events that alter the size of organisms. Using genetic and biochemical methods, we have shown that Tsc1 and Tsc2 function in the insulin/phosphoinositide 3-kinase (PI3K)/Akt pathway. We have shown that Akt regulates the Tsc1-Tsc2 complex by directly phosphorylating Tsc2. We have shown further that S6 kinase (S6K) is a downstream component of the PI3K/Akt/TSC pathway and reduction of S6K activity can block TSC defects. Recent studies from many laboratories have now confirmed our findings in mice, rats and human patients, and have shown that drugs that antagonize S6K activities, such as rapamycin, diminish tumours in TSC-deficient mice and rats. Clinical trials based on these findings have begun. Given that other components of the pathway, such as PTEN, are also mutated in a large number of cancer patients and that these components regulate intracellular insulin signalling, therapeutics based on the knowledge of the pathway could have effects beyond the TSC patient population.


Assuntos
Drosophila melanogaster/genética , Genes Supressores de Tumor , Proteínas/genética , Proteínas Repressoras/genética , Animais , Constituição Corporal , Drosophila melanogaster/anatomia & histologia , Mamíferos , Camundongos , Camundongos Knockout , Fosforilação , Proteínas/metabolismo , Proteínas Repressoras/metabolismo , Proteína 1 do Complexo Esclerose Tuberosa , Proteína 2 do Complexo Esclerose Tuberosa , Proteínas Supressoras de Tumor
14.
Oral Oncol ; 38(5): 460-7, 2002 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-12110340

RESUMO

In this double-blind, randomized trial, we compared the clinical efficacy of intralesional vinblastine (VNB) and 3% sodium tetradecyl sulfate (STS) in the treatment of oral Kaposi's sarcoma (OKS). Subjects with OKS were randomly assigned to receive a single intralesional injection of either VNB or STS, at a standard dose (0.2 mg/cm(2)). Differences were evaluated by the Mann-Whitney U and Fisher's exact tests. Sixteen HIV-infected patients were included, eight received VNB and eight received STS; clinical response was evaluated at days 7, 14, and 28 following treatment. Tumor size reduction was 0.68 and 0.61 cm in the VNB and STS groups, respectively (P=0.80). Two VNB patients had complete or partial response whereas four STS subjects had partial responses (P=0.61). Patients in both groups experienced minimal toxicity. We conclude that intralesional vinblastine or STS are adequate for the management of OKS. The benefits of STS are its low cost and ease of use.


Assuntos
Antineoplásicos Fitogênicos/uso terapêutico , Neoplasias Bucais/tratamento farmacológico , Sarcoma de Kaposi/tratamento farmacológico , Soluções Esclerosantes/uso terapêutico , Tetradecilsulfato de Sódio/uso terapêutico , Vimblastina/uso terapêutico , Adulto , Método Duplo-Cego , Seguimentos , Infecções por HIV/complicações , Humanos , Injeções Intralesionais , Masculino , Pessoa de Meia-Idade , Neoplasias Bucais/complicações , Sarcoma de Kaposi/complicações , Resultado do Tratamento
16.
Neuron ; 30(2): 335-44, 2001 May.
Artigo em Inglês | MEDLINE | ID: mdl-11394997

RESUMO

On axonal surfaces that flank the node of Ranvier and in overlying glial paranodal loops, proteins are arranged within circumscribed microdomains that defy explanation by conventional biosynthetic mechanisms. We postulate that the constraint of proteins to these loci is accomplished in part by discriminative membrane-embedded molecular sieves and diffusion barriers, which serve to organize and redistribute proteins after delivery by vesicular transport to neural cell plasma membranes. One sieve likely comprises a moveable, macromolecular scaffold of axonal and glial cell-derived transmembrane adhesion molecules and their associated cytoplasmic binding partners, located at the ends of each elongating myelin internode; this sieve contributes to restricting the sodium channel complexes to the node. We also anticipate the existence of a passive paranodal diffusion barrier at the myelin/noncompact membrane border, which prohibits protein diffusion out of contiguous paranodal membranes.


Assuntos
Axônios/fisiologia , Proteínas do Tecido Nervoso/fisiologia , Neuroglia/fisiologia , Nós Neurofibrosos/fisiologia , Animais , Membrana Celular/fisiologia , Humanos , Bainha de Mielina/fisiologia , Neurônios/fisiologia , Nós Neurofibrosos/ultraestrutura
17.
Proc AMIA Symp ; : 731-5, 2001.
Artigo em Inglês | MEDLINE | ID: mdl-11825282

RESUMO

Critical parts of the software development life cycle are concerned with eliciting, understanding, and managing requirements. Though the literature on this subject dates back for several decades, practicing effective requirements development remains a current and challenging area. Some projects flourish with a requirements development process (RDP) that is implicit and informal, but this approach may be overly risky, particularly for large projects that involve multiple individuals, groups, and systems over time. At Partners HealthCare System in Boston, Massachusetts, we have applied a more formal approach for requirements development to the Patient Computing Project. The goal of the project is to create web-based software that connects patients electronically with their physician's offices and has the potential to improve care efficiency and quality. It is a large project, with over 500 function points. Like most technological innovation, the successful introduction of this system requires as much attention to understanding the business needs and workflow details as it does to technical design and implementation. This paper describes our RDP approach, and key business requirements discovered through this process. We believe that a formal RDP is essential, and that informatics as a field must include proficiencies in this area.


Assuntos
Internet , Validação de Programas de Computador , Software/normas , Telemedicina/normas , Boston , Prestação Integrada de Cuidados de Saúde , Humanos , Pacientes , Relações Médico-Paciente
18.
J Cell Biol ; 150(3): 657-66, 2000 Aug 07.
Artigo em Inglês | MEDLINE | ID: mdl-10931875

RESUMO

Two major isoforms of the cell adhesion molecule neurofascin NF186 and NF155 are expressed in the central nervous system (CNS). We have investigated their roles in the assembly of the node of Ranvier and show that they are targeted to distinct domains at the node. At the onset of myelination, NF186 is restricted to neurons, whereas NF155 localizes to oligodendrocytes, the myelin-forming glia of the CNS. Coincident with axon ensheathment, NF155 clusters at the paranodal regions of the myelin sheath where it localizes in apposition to the axonal adhesion molecule paranodin/contactin-associated protein (Caspr1), which is a constituent of the septate junction-like axo-glial adhesion zone. Immunoelectron microscopy confirmed that neurofascin is a glial component of the paranodal axo-glial junction. Concentration of NF155 with Caspr1 at the paranodal junctions of peripheral nerves is also a feature of Schwann cells. In Shiverer mutant mice, which assemble neither compact CNS myelin nor normal paranodes, NF155 (though largely retained at the cell body) is also distributed at ectopic sites along axons, where it colocalizes with Caspr1. Hence, NF155 is the first glial cell adhesion molecule to be identified in the paranodal axo-glial junction, where it likely interacts with axonal proteins in close association with Caspr1.


Assuntos
Moléculas de Adesão Celular/isolamento & purificação , Junções Intercelulares/fisiologia , Bainha de Mielina/fisiologia , Fatores de Crescimento Neural/isolamento & purificação , Neuroglia/fisiologia , Oligodendroglia/fisiologia , Animais , Técnicas de Cocultura , Imunofluorescência , Gânglios Espinais/citologia , Glicoproteínas de Membrana/isolamento & purificação , Camundongos , Camundongos Mutantes , Neuropeptídeos/isolamento & purificação , Nervo Óptico/citologia , Epitélio Pigmentado Ocular/ultraestrutura , Isoformas de Proteínas/isolamento & purificação , Nós Neurofibrosos/fisiologia , Ratos , Ratos Sprague-Dawley , Células de Schwann/citologia , Nervo Isquiático/citologia , Canais de Sódio/genética
19.
Int J Dermatol ; 39(7): 501-5, 2000 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-10940113

RESUMO

BACKGROUND: Oral mucosal manifestations may be the initial feature, the most florid clinical feature, or the only sign of mucocutaneous diseases. METHODS: Sixty adult patients (48 women, 12 men) with oral mucous complaints were referred to and evaluated at a dermatology clinic in a tertiary care hospital in Mexico City between November 1996 and September 1998. RESULTS: The most frequent oral conditions observed were pemphigus vulgaris (18.3%), lichen planus (8.3%), candidiasis (8.3%), recurrent aphthous ulcers (6.7%), herpetic lesions (6.7%), xerostomia (6.7%), and traumatic lesions (6.7%). Oral affection in mucocutaneous conditions was observed in 21 (35%) patients; the diagnosis was based on oral signs in 10 (48%) of these patients. A large number of oral conditions had previously been misdiagnosed. CONCLUSIONS: The importance of the diagnosis of oral conditions in dermatology has been underlined in this study due to the frequency and diversity of oral lesions. The benefits of an interdisciplinary approach in the management of patients has been highlighted.


Assuntos
Doenças da Boca/epidemiologia , Adolescente , Adulto , Idoso , Dermatologia , Diagnóstico Diferencial , Feminino , Humanos , Masculino , México/epidemiologia , Pessoa de Meia-Idade , Doenças da Boca/diagnóstico , Mucosa Bucal/patologia , Ambulatório Hospitalar
20.
J Neurosci Res ; 60(6): 697-703, 2000 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-10861781

RESUMO

We present here a new approach which permits us to follow myelin proteins within living, actively myelinating cells. We have developed probes to study the spatial and temporal incorporation of proteins into the myelin sheath by expressing myelin proteins fused to the green fluorescent protein (GFP). GFP from the jellyfish Aequorea victoria and its derivatives, e.g., blue fluorescent protein (BFP) were used as molecular reporters to monitor the intracellular distribution of myelin proteins. Fusion proteins (14 kD myelin basic protein [MBP]-GFP, 21 kD MBP-GFP) were expressed in primary Schwann cells (SCs) and their distribution was monitored by confocal microscopy. The autofluorescent chimeric proteins were readily visualized and their subcellular localization was unaffected by the GFP reporter. However, because of the length of culturing time necessary to establish permanent cell lines, we found that it was not possible to obtain MBP-GFP stable SCs that also were capable of myelinating neuronal axons. We therefore devised a way of introducing vectors under conditions where cells are dividing in response to endogenous stimuli, and therefore are still capable of myelinating. We designed a protocol in which SCs cocultured with dorsal root ganglion (DRG) neurons are transfected while they are actively dividing. SCs transfected in this way exhibit a good level of protein expression and retain their myelinating phenotype. The fusion protein expression lasts long enough to observe "green myelin. " These fluorescently tagged myelin proteins will allow high-resolution examination of the protein and membrane traffic in normal myelinating cells.


Assuntos
Indicadores e Reagentes , Proteínas Luminescentes/genética , Proteínas da Mielina/genética , Proteínas da Mielina/fisiologia , Bainha de Mielina/fisiologia , Proteínas Recombinantes de Fusão , Axônios/metabolismo , Membrana Celular/metabolismo , Núcleo Celular/metabolismo , Técnicas de Cocultura , Gânglios Espinais/citologia , Gânglios Espinais/fisiologia , Proteínas de Fluorescência Verde , Células HeLa , Humanos , Indicadores e Reagentes/metabolismo , Proteínas Luminescentes/química , Proteínas Luminescentes/metabolismo , Peso Molecular , Proteínas da Mielina/química , Proteínas da Mielina/metabolismo , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/metabolismo , Células de Schwann/metabolismo , Células de Schwann/fisiologia
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