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1.
Arch Oral Biol ; 56(7): 634-42, 2011 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-21429473

RESUMO

OBJECTIVE: The objective of this study was to describe the changes in salivary protein profiles in infants between the ages of 3 and 6 months, and to evaluate the impact of teeth eruption and introduction of solid foods on such profiles. DESIGN: 73 infants were followed longitudinally at 3 and 6 months of age. Their whole saliva proteins were separated by SDS-PAGE electrophoresis and semi-quantified by image analysis. Amylase activity was also measured on a sub-sample of the population (n=42 infants). Bands which abundance was significantly different between the two ages according to paired comparisons were identified by mass spectrometry techniques. RESULTS: Out of 21 bands, 13 were significantly different between 3 and 6 months of age. Two short variants of amylase increased in abundance with age, as did amylase activity. Other changes possibly translated developmental physiological events, for example maturation of the adaptive immune system. The balance between S-type cystatins and cystatins A and B was modified, in favour of S-type cystatins at 6 months of age. Teeth eruption resulted in an increase in albumin abundance, whilst introduction of solid foods was associated with higher levels of ß-2 microglobulin and S-type cystatins. CONCLUSIONS: Salivary profiles were modified substantially between the ages of 3 and 6 months. Both teeth eruption and diet had an impact on abundance changes for some proteins, revealing dynamic interactions between saliva proteome, oral physiology and diet.


Assuntos
Dieta , Eletroforese em Gel de Poliacrilamida , Proteínas e Peptídeos Salivares/análise , Erupção Dentária/fisiologia , Amilases/análise , Cromatografia Líquida , Cistatina A/análise , Cistatina B/análise , Feminino , Seguimentos , Humanos , Lactente , Alimentos Infantis , Fórmulas Infantis , Estudos Longitudinais , Masculino , Leite Humano , Estudos Prospectivos , Inibidores de Proteases/análise , Cistatinas Salivares/análise , Componente Secretório/análise , Albumina Sérica/análise , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Espectrometria de Massas em Tandem , Desmame , Microglobulina beta-2/análise
2.
Am J Obstet Gynecol ; 204(3): 254.e1-7, 2011 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-21167469

RESUMO

OBJECTIVE: The purpose of this study was to investigate the temporal changes in immunoreactive cystatin A and the enzymatic activity of cathepsins B, H, L, and S in human cervicovaginal fluid (CVF) in late pregnancy and spontaneous labor. STUDY DESIGN: CVF was collected weekly (n = 95 women) from 36 weeks gestation until spontaneous term labor. Cystatin A was quantified using enzyme-linked immunosorbent assay. The enzyme activity of cathepsins B, H, L, and S was measured with fluorometric enzyme assay kits. RESULTS: Cystatin A significantly decreased towards (P = .016, 2-way analysis of variance) and during labor (P < .001, 2-way analysis of variance). Enzymatic activity of cathepsins B, H, and S did not change with labor onset (P = .452, P = .703, P = .411, respectively, 2-way analysis of variance). CONCLUSION: In late gestation, CVF-decreased expression of the cysteine protease inhibitor, cystatin A, is associated with labor. Although the role and contribution of cystatin A to increased extracellular matrix remodeling has yet to be elucidated, the data that were obtained are consistent with this hypothesis.


Assuntos
Líquidos Corporais/enzimologia , Catepsinas/análise , Cistatina A/análise , Cisteína Proteases/análise , Adulto , Feminino , Humanos , Trabalho de Parto , Gravidez , Inibidores de Proteases/análise , Nascimento a Termo
3.
Radiother Oncol ; 93(3): 575-80, 2009 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19850367

RESUMO

PURPOSE: To assess radiosensitivity of neck metastases of squamous cell carcinoma of the head and neck (SCCHN) by immunocytochemical profiling of fine-needle aspiration biopsy (FNAB) cell specimens. PATIENTS AND METHODS: Immunocytochemical reactions to p53, cyclin D1, stefin A and Ki-67 were determined in FNAB cell samples of neck metastases from 21 patients treated with concomitant chemoradiotherapy and correlated to clinical characteristics and response to therapy. RESULTS: Six (28.6%), eight (38.1%), 15 (71.4%) and nine (42.9%) FNAB cell samples were classified as p53, cyclin D1, stefin A and Ki-67 positive, respectively. Statistically significant predictors of favorable nodal response to chemoradiation were p53 (P=0.025) and cyclin D1 (cytoplasmic fraction, P=0.048) negativity and Ki-67 positivity (P=0.045). Regional recurrence correlated with low Ki-67 immunoreactivity. A favorable profile of cyclin D1 and Ki-67 (one or both of the two) further improved the predictive strength of these markers. CONCLUSIONS: FNAB is a non-invasive, simple and cheap procedure, which could serve simultaneously for diagnostic purposes and for radiosensitivity testing. Immunocytochemical determination of cyclin D1 and Ki-67 in FNAB cell samples from neck metastases of SCCHN seems to be a valuable marker for predicting regional response to radiotherapy and might assist when deciding on appropriate primary therapy.


Assuntos
Biópsia por Agulha Fina , Carcinoma de Células Escamosas/radioterapia , Neoplasias de Cabeça e Pescoço/radioterapia , Linfonodos/metabolismo , Linfonodos/patologia , Metástase Linfática/radioterapia , Pescoço , Tolerância a Radiação , Adulto , Idoso , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patologia , Ciclina D1/análise , Cistatina A/análise , Feminino , Neoplasias de Cabeça e Pescoço/metabolismo , Neoplasias de Cabeça e Pescoço/patologia , Humanos , Imuno-Histoquímica , Antígeno Ki-67/análise , Masculino , Pessoa de Meia-Idade , Proteína Supressora de Tumor p53/análise
4.
Arch Oral Biol ; 54(5): 437-44, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-19268279

RESUMO

OBJECTIVE: The aim of this study was to investigate the type and the nature of peptides present in the in vivo formed human acquired enamel pellicle. DESIGN: Pellicle material was collected from 10 volunteers and subjected to sample preparations consisting of centrifugal filtration using a 10 kDa molecular weight cut-off membrane and high-resolution gel filtration chromatography. The fractions containing peptides <10 kDa obtained by both methods were analyzed by LC-ESI-MS/MS. RESULTS: 78 natural pellicle peptides with molecular weights ranging from 766.9 Da to 3981.4 Da were identified originating from 29 different proteins. CONCLUSIONS: The number of peptides present in acquired enamel pellicle appears to be large and this is likely to enhance the functional spectrum of this protein film. The presence of small peptides in pellicle may be functionally important since structure/function studies of many salivary proteins have shown that specific domains within these native proteins retain or even exhibit enhanced biological activities. The data present the basis for determining the precise function of these pellicle peptides and for gaining insights into the role pellicle plays in the oral cavity.


Assuntos
Película Dentária/química , Proteínas e Peptídeos Salivares/análise , Adulto , Anexina A1/análise , Proteínas de Ligação ao Cálcio/análise , Cromatografia em Gel , Cromatografia Líquida , Cistatina A/análise , Feminino , Humanos , Ponto Isoelétrico , Masculino , Filtros Microporos , Peso Molecular , Fosfoproteínas/análise , Inibidores de Proteases/análise , Proteoma/análise , Cistatinas Salivares/análise , Proteínas Salivares Ricas em Prolina/análise , alfa-Amilases Salivares/análise , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em Tandem , Adulto Jovem
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