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4.
Lipids Health Dis ; 17(1): 145, 2018 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-29925393

RESUMO

BACKGROUND: The beneficial effects in lipid profiles after obesity surgery might be associated with the decrease in cardiovascular risk. However, direct comparison between different surgical techniques has not been extensively performed. METHODS: In the present study we compare 20 obese women submitted to laparoscopic Roux en Y gastric bypass (RYGB) with 20 women submitted to sleeve gastrectomy (SG). Twenty control women matched for age and baseline cardiovascular risk were also included. Both patients and controls were followed up for 1 year after surgery or conventional treatment with diet and exercise, respectively. Lipid profiles were measured at baseline, 6 and 12 months later. Carotid intima-media thickness was measured by ultrasonography at baseline and at the end of the study. RESULTS: Women submitted to bariatric surgery showed a decrease in total cholesterol, triglycerides, oxidized-LDL and ApoB, and an increase in HDL and ApoA concentrations that occurred regardless of the surgical procedure. LDL concentrations, however, decreased only after RYGB whereas Lp(a) showed no changes. We did not observe any correlation between the changes in serum lipid concentrations and those in carotid intima-media thickness. CONCLUSIONS: Sleeve gastrectomy and gastric bypass induce a similar beneficial effect on serum lipids in women with high cardiovascular risk 1 year after surgery.


Assuntos
Gastrectomia/métodos , Derivação Gástrica , Gastroplastia/métodos , Obesidade Mórbida/sangue , Obesidade Mórbida/cirurgia , Adulto , Apolipoproteínas A/sangue , Apolipoproteínas B/sangue , Restrição Calórica , Doenças Cardiovasculares/diagnóstico por imagem , Doenças Cardiovasculares/prevenção & controle , Artérias Carótidas/diagnóstico por imagem , Espessura Intima-Media Carotídea , Estudos de Casos e Controles , Exercício Físico , Feminino , Seguimentos , Humanos , Lipoproteínas HDL/sangue , Lipoproteínas LDL/sangue , Pessoa de Meia-Idade , Obesidade Mórbida/dietoterapia , Obesidade Mórbida/patologia , Risco , Triglicerídeos/sangue , Ultrassonografia
5.
Arch Osteoporos ; 13(1): 56, 2018 05 08.
Artigo em Inglês | MEDLINE | ID: mdl-29736771

RESUMO

The prevalence of asymptomatic vertebral fracture in HIV-infected patients over 50 was 20%, associated with older age, male sex, longer time since HIV diagnosis, and tubular renal alterations. Vertebral fractures were independent of osteoporosis at lumbar spine, and were not predicted by the use of the FRAX equation. PURPOSE: Vertebral fractures (VF) are the hallmark of osteoporotic fractures. Our objective was to determine the prevalence of asymptomatic VF and associated factors in HIV-infected patients over 50 years, and the role of FRAX equation. METHODS: In a cross-sectional study, a diagnosis of VF was established by the semiquantitative method of Genant in thoracic and lumbar radiographs. Simultaneously, a dual X-ray absorptiometry (DXA), bone and kidney-related analytical, calcium intake, physical exercise, HIV-related factors, and FRAX estimation were evaluated. RESULTS: Overall, 128 patients (35 women, 27%) were included. Mean age was 57 years. Hypophosphatemia and tubular renal dysfunction were observed in 13 and 21%. DXA scan showed osteopenia and osteoporosis at hip in 65 and 7% of patients, and in spine in 39 and 34%, respectively. VF were observed in 26 patients (20%), with a trend to be associated with lower serum phosphate, increased alkaline phosphatase, and with lower daily calcium intake. In a multivariate analysis, older age (OR 1.2 per year; 14% of VF at 50-55; 44% at 65-70), male sex (26 vs 6%), longer time since HIV diagnosis, and renal and tubular dysfunction were the associated factors. VF were not related with osteoporosis at lumbar spine, and could not be predicted by the FRAX equation. CONCLUSIONS: The prevalence of asymptomatic vertebral fractures is high in HIV-infected patients older than 50 years, and is not identified by the presence of osteoporosis in spine neither predicted by the FRAX equation. Spine and lumbar X-rays should be routinely performed in this aging population.


Assuntos
Infecções por HIV/complicações , HIV , Vértebras Lombares/lesões , Fraturas por Osteoporose/epidemiologia , Fraturas da Coluna Vertebral/epidemiologia , Absorciometria de Fóton/métodos , Idoso , Doenças Assintomáticas , Densidade Óssea , Estudos Transversais , Feminino , Infecções por HIV/epidemiologia , Humanos , Vértebras Lombares/diagnóstico por imagem , Masculino , Pessoa de Meia-Idade , Fraturas por Osteoporose/complicações , Fraturas por Osteoporose/diagnóstico , Prevalência , Radiografia , Espanha/epidemiologia , Fraturas da Coluna Vertebral/complicações , Fraturas da Coluna Vertebral/diagnóstico
6.
J Nutr Biochem ; 30: 143-52, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27012631

RESUMO

The present study was designed to elucidate the protective effects of dietary apigenin (API) enrichment in a chronic colitis model induced by DSS in mice. Inflammatory mediators and the possible role of canonical and non-canonical NLRP3 inflammasome signaling pathways in the beneficial effects of API under chronic inflammatory conditions were also explored. Six-week-old mice were randomized in four dietary groups: sham and control groups received standard diet (SD), and other two groups were fed with API at 0.1%. After 30days, all groups except sham received 3% DSS in drinking water for 5days followed by a regime of 21days of water. Our results revealed that dietary API supplementation decreased the macroscopic and microscopic damage signs of colitis; also, it was capable to down-regulate mPGES, COX-2 and iNOS enzyme colonic expressions and to decrease serum matrix metalloproteinase (MMP-3) levels. Similarly, API diet reduced IL-1ß and TNF-α proinflammatory cytokine secretions in primary LPS-stimulated splenocytes. Furthermore, we demonstrated that API anti-inflammatory activity was related with an inhibition of both canonical and non-canonical NLRP3 inflammasome pathways by decreasing proinflammatory IL-1ß and IL-18 cytokine levels as a consequence of regulation of cleaved caspase-1 and caspase-11 enzymes. We conclude that API supplement might provide a basis for developing a new dietary strategy for the prevention of chronic ulcerative colitis.


Assuntos
Apigenina/administração & dosagem , Colite/induzido quimicamente , Sulfato de Dextrana/efeitos adversos , Inflamassomos/efeitos dos fármacos , Transdução de Sinais , Animais , Feminino , Camundongos , Camundongos Endogâmicos C57BL
7.
J. physiol. biochem ; 70(1): 163-172, mar. 2014.
Artigo em Inglês | IBECS | ID: ibc-121615

RESUMO

Inflammatory bowel disease (IBD), which includes ulcerative colitis and Crohn’s disease, is a chronic intestinal disorder resultant from a dysfunctional epithelial, innate and adaptive immune response to intestinal microorganisms. Current IBD treatment presents limitations in both efficacy and safety that stimulated for new active drugs. Retama spp. have been traditionally used in the Mediterranean region in treatment of pain and inflammation. In this study, the anti-inflammatory and protective properties of a standardised aqueous extract from Retama monosperma (RmE) was evaluated in vivo, by intra-colonic administration of trinitrobenzene sulfonic acid (TNBS) in rats as a Crohn’s disease model. The qualitative and quantitative analysis of flavonoids from RmE was performed by high-performance liquid chromatography-tandem mass spectrometry. Oral administration of RmE diminished the severity and extension of the intestinal injuries induced by TNBS. In addition, RmE increased mucus production in goblet cells in colon mucosa, decreased neutrophil infiltration and cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) overexpression. Similarly, RmE significantly reduced p38 mitogen-activated protein kinase activation, preventing the inhibitory protein IêB degradation in colonic mucosa. RmE anti-inflammatory effects seem to be related to impairment of neutrophil function and COX-2 and iNOS down-regulation possibly through p38 MAPK and nuclear transcription factor kappa B signalling pathways. These results suggest that RmE might contribute to the development of new pharmaceutical products for inflammatory bowel disease


Assuntos
Animais , Ratos , Anti-Inflamatórios/farmacocinética , Cytisus , Colite Ulcerativa/tratamento farmacológico , Substâncias Protetoras/farmacocinética , Modelos Animais de Doenças , Extratos Vegetais/uso terapêutico
8.
J Physiol Biochem ; 70(1): 163-72, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24057513

RESUMO

Inflammatory bowel disease (IBD), which includes ulcerative colitis and Crohn's disease, is a chronic intestinal disorder resultant from a dysfunctional epithelial, innate and adaptive immune response to intestinal microorganisms. Current IBD treatment presents limitations in both efficacy and safety that stimulated for new active drugs. Retama spp. have been traditionally used in the Mediterranean region in treatment of pain and inflammation. In this study, the anti-inflammatory and protective properties of a standardised aqueous extract from Retama monosperma (RmE) was evaluated in vivo, by intra-colonic administration of trinitrobenzene sulfonic acid (TNBS) in rats as a Crohn's disease model. The qualitative and quantitative analysis of flavonoids from RmE was performed by high-performance liquid chromatography-tandem mass spectrometry. Oral administration of RmE diminished the severity and extension of the intestinal injuries induced by TNBS. In addition, RmE increased mucus production in goblet cells in colon mucosa, decreased neutrophil infiltration and cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) overexpression. Similarly, RmE significantly reduced p38 mitogen-activated protein kinase activation, preventing the inhibitory protein IκB degradation in colonic mucosa. RmE anti-inflammatory effects seem to be related to impairment of neutrophil function and COX-2 and iNOS down-regulation possibly through p38 MAPK and nuclear transcription factor kappa B signalling pathways. These results suggest that RmE might contribute to the development of new pharmaceutical products for inflammatory bowel disease.


Assuntos
Anti-Inflamatórios/farmacologia , Colite Ulcerativa/tratamento farmacológico , Fabaceae/química , Extratos Vegetais/farmacologia , Doença Aguda , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/uso terapêutico , Colite Ulcerativa/induzido quimicamente , Colite Ulcerativa/metabolismo , Colo/efeitos dos fármacos , Colo/enzimologia , Colo/patologia , Ciclo-Oxigenase 2/metabolismo , Avaliação Pré-Clínica de Medicamentos , Etanol , Expressão Gênica , Proteínas I-kappa B/genética , Proteínas I-kappa B/metabolismo , Sistema de Sinalização das MAP Quinases , Masculino , NF-kappa B/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Extratos Vegetais/química , Extratos Vegetais/uso terapêutico , Ratos , Ratos Wistar , Ácido Trinitrobenzenossulfônico
9.
Nutr Cancer ; 65(1): 147-56, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23368925

RESUMO

Oleuropein (OL) is the most prominent phenolic compound in the fruit of olive tree. Although OL has shown powerful anticancer activity the underlying action mechanism remains largely unknown. The present study evaluated the effects of OL on hydroxityrosol (HT)-29 human colon adenocarcinoma cells in comparison to hydroxytyrosol, its hydrolysis product, and to elucidate the underlying anticancer molecular mechanisms involved. Cell proliferation was determined using SRB assay. Cell cycle and apoptosis were assessed by flow cytometry and changes in MAPK cascade protein expression, HIF-1α, p53, PPARγ, and NFKß signaling pathways by Western blot. Although OL showed less potency than HT, in terms of cell growth inhibition, induced significant changes in cell cycle analysis and caused a significant increase in the apoptotic population. Both compounds produced a remarkable decrease in HIF-1α protein and an upregulation of p53 protein expression. However, no significant changes in IkB-α and MAPK cascade protein expressions were observed. HT produced a significant upregulation in peroxisome proliferator-activated receptor gamma (PPARγ) expression whereas OL failed. PPARγ upregulation may be one of the principal mechanisms of the tumor shrinkage by HT. Our novel findings demonstrate that OL limits the growth and induces apoptosis in HT-29 cells via p53 pathway activation adapting the HIF-1α response to hypoxia.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Neoplasias Colorretais/tratamento farmacológico , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Olea/química , Piranos/farmacologia , Apoptose/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Neoplasias Colorretais/metabolismo , Neoplasias Colorretais/patologia , Regulação para Baixo/efeitos dos fármacos , Células HT29/efeitos dos fármacos , Humanos , Proteínas I-kappa B/metabolismo , Glucosídeos Iridoides , Iridoides , Inibidor de NF-kappaB alfa , NF-kappa B/metabolismo , PPAR gama/metabolismo , Álcool Feniletílico/análogos & derivados , Álcool Feniletílico/farmacologia , Proteína Supressora de Tumor p53/metabolismo , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
10.
Eur J Pharm Sci ; 48(3): 572-81, 2013 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-23238173

RESUMO

Extra virgin olive oil (EVOO) has demonstrated immunomodulatory and antiinflammatory properties in murine experimental ulcerative colitis (UC). In addition to its high monounsaturated fatty acid content, evidences have accumulated on the favorable properties of minor, although highly bioactive, components present in the unsaponifiable fraction (UF). The present study was designed to evaluate the effects of dietary EVOO's UF supplementation on acute UC. C57BL/6 mice were fed from weaning with sunflower oil (SD), EVOO diet and UF-enriched SD at 5% oil (SD+UF). After 30 days, mice were exposed to 3% dextran sulfate sodium (DSS) for 5 days developing acute colitis. After 4 days of DSS removal, animals were sacrificed and colons were histological and biochemically processed. Disease activity index and microscopic damage score were significantly improved in EVOO and SD+UF dietary groups versus SD group. In addition, both dietary treatments significantly induced decreases in MCP-1 and TNF-α levels, iNOS and COX-2 overexpression and p38 MAPKs activation in colon mucosa. Moreover, an upregulation of IκB expression was also observed after feeding the animals with both diets. However, no statistically differences between data from mice fed with EVOO or UF+SD diets were observed. Dietary enrichment with EVOO's UF reduces the damage in acute colitis model, alleviating the oxidative events and returning proinflammatory proteins expression to basal levels probably through p38 MAPK and NFκB signalling pathways. EVOO's UF diet might provide a basis for developing a new strategy in dietary supplementation for the prevention of UC.


Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Colite Ulcerativa/prevenção & controle , Colo/imunologia , Suplementos Nutricionais , Mucosa Intestinal/imunologia , Extratos Vegetais/uso terapêutico , Óleos de Plantas/química , Animais , Quimiocina CCL2/antagonistas & inibidores , Quimiocina CCL2/genética , Quimiocina CCL2/metabolismo , Colite Ulcerativa/imunologia , Colite Ulcerativa/metabolismo , Colite Ulcerativa/patologia , Colo/metabolismo , Colo/patologia , Ciclo-Oxigenase 2/química , Ciclo-Oxigenase 2/genética , Ciclo-Oxigenase 2/metabolismo , Ativação Enzimática , Feminino , Regulação da Expressão Gênica , Hidrólise , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Camundongos Endogâmicos C57BL , Óxido Nítrico Sintase Tipo II/antagonistas & inibidores , Óxido Nítrico Sintase Tipo II/genética , Óxido Nítrico Sintase Tipo II/metabolismo , Azeite de Oliva , Óleos de Plantas/normas , Distribuição Aleatória , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/genética , Fator de Necrose Tumoral alfa/metabolismo , Desmame , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
11.
Biochem Pharmacol ; 82(7): 737-45, 2011 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-21763290

RESUMO

Current epidemiological and experimental studies support a beneficial role of dietary polyphenols in several gastrointestinal diseases, including inflammatory bowel disease. The aim of this study was to gain a better understanding of the effects of a naturally occurring polyphenol, ellagic acid, present in some fruits such as pomegranate, raspberries and nuts among others, in an experimental murine model of Crohn's disease by intra-colonic administration of TNBS in rats. Analysis of the lesions were carried out by macroscopic and histological technics. Inflammation response was assessed by histology and myeloperoxidase activity. iNOS and COX-2 are upregulated by MAPKs and NF-κB nuclear transcription factor in intestinal epithelial cells thus, we determined the expression of iNOS, COX-2 and the involvement of the p38, JNK, ERK1/2 MAPKs and NF-κB signalling in the protective effect of EA by western blotting. Oral administration of EA (10-20 mg/kg) diminished the severity and extension of the intestinal injuries induced by TNBS although there was no observed a significant dose-response. In addition, EA increased mucus production in goblet cells in colon mucosa, decreased neutrophil infiltration and pro-inflammatory proteins COX-2 and iNOS overexpression. Also EA was capable of reducing the activation of p38, JNK and ERK1/2 MAPKs, preventing the inhibitory protein IκB-degradation and inducing an inhibition of the nuclear translocation level of p65 in colonic mucosa. In conclusion, EA reduces the damage in a rat model of Crohn's disease, alleviates the oxidative events and returns pro-inflammatory proteins expression to basal levels probably through MAPKs and NF-κB signalling pathways.


Assuntos
Doença de Crohn/prevenção & controle , Ácido Elágico/uso terapêutico , Flavonoides/uso terapêutico , Fenóis/uso terapêutico , Transporte Ativo do Núcleo Celular , Doença Aguda , Animais , Núcleo Celular/metabolismo , Colo/efeitos dos fármacos , Colo/metabolismo , Doença de Crohn/induzido quimicamente , Doença de Crohn/patologia , Ciclo-Oxigenase 2/metabolismo , Quinase I-kappa B/metabolismo , Mucosa Intestinal/efeitos dos fármacos , Mucosa Intestinal/metabolismo , Leucócitos/fisiologia , Masculino , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Peroxidase/metabolismo , Fosforilação , Polifenóis , Ratos , Ratos Wistar , Transdução de Sinais , Fator de Transcrição RelA/metabolismo , Ativação Transcricional , Ácido Trinitrobenzenossulfônico
12.
Inflamm Res ; 57(4): 145-50, 2008 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-18368290

RESUMO

OBJECTIVE: Evaluate the Monocyte Locomotion Inhibitory Factor (MLIF) effect upon the expression of genes encoding human cytokines, receptors and related factors in the human cell line U-937. MLIF (Met-Gln-Cys-Asn-Ser) is an anti-inflammatory pentapeptide produced by Entamoeba histolytica that inhibits many human monocyte functions. MATERIAL AND METHODS: U-937 cell line cultured (24 hrs/RPMI). RNA extracted by Trizol method. 385 genes were analyzed on microarray membranes, complement by real-time RT-PCR and protein expression of some affected genes. RESULTS: MLIF had a preferentially inhibitory effect on gene expression; four genes were over-expressed and 13 underexpressed in MILF vs. simple medium - constitutive expression. Three genes are over-expressed and 19 under-expressed in MLIF/PMA vs. PMA - induced expression. CONCLUSIONS: Many modified genes are products regulated by the Nuclear Factor-kappaB and Mitogen Activated Protein Kinase pathways, suggesting MLIF involvement with these two major pathways for the modulation of the inflammation and immune responses.


Assuntos
Citocinas/metabolismo , Entamoeba histolytica/metabolismo , Expressão Gênica/efeitos dos fármacos , Monócitos/metabolismo , Oligopeptídeos/metabolismo , Oligopeptídeos/farmacologia , Animais , Citocinas/genética , Perfilação da Expressão Gênica , Humanos , Inflamação/metabolismo , Quinases de Proteína Quinase Ativadas por Mitógeno/metabolismo , Modelos Biológicos , Monócitos/citologia , NF-kappa B/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos , Células U937
13.
Mol Genet Genomics ; 266(3): 463-70, 2001 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-11713676

RESUMO

Mucor circinelloides responds to blue light by activating carotene biosynthesis. Wild-type strains grown in darkness contain minimal amounts of beta-carotene because of the low levels of transcription of the structural genes for carotenogenesis. When exposed to a light pulse, the level of transcription of these genes increases strongly, leading to the formation of high concentrations of beta-carotene. The crgA gene is involved in the regulation of light-induced carotenoid biosynthesis. This gene, originally identified as a 3'-truncated ORF which causes carotene over-accumulation in the dark, encodes a protein with a cysteine-rich, zinc-binding, RING-finger motif, as found in diverse groups of regulatory proteins. The expression of the crgA gene is activated by a light pulse, with a time course similar to that of the structural genes for carotenogenesis. To understand the regulatory role of the crgA gene in carotenogenesis, we have used a genetic approach based on the construction of crgA null mutants by gene replacement. Lack of the crgA function provokes the over-accumulation of carotenoids both in the dark and the light. Introduction of the wild-type crgA allele into these mutants restores the wild-type phenotype for carotenogenesis. The high levels of carotenoid accumulation shown by the null crgA mutants are correlated with an increase in the expression of carotenogenic structural genes. These results strongly indicate that crgA acts as a negative regulator of light-inducible carotenogenesis in M. circinelloides.


Assuntos
Carotenoides/biossíntese , Proteínas Fúngicas/genética , Mucor/genética , Primers do DNA/química , DNA Fúngico/genética , Regulação da Expressão Gênica , Genes Fúngicos , Teste de Complementação Genética , Luz , Mucor/metabolismo , Mucor/efeitos da radiação , Mutação , Fenótipo , Plasmídeos/genética , Reação em Cadeia da Polimerase , RNA/metabolismo , Transformação Genética
14.
Clín. salud ; 11(1): 99-134, ene. 2000. tab, ilus, graf
Artigo em Es | IBECS | ID: ibc-15518

RESUMO

Se describe la evaluación y tratamiento de un varón de 24 años que acude consulta por la aparición de vómitos ante situaciones en las que se ve forzado a salir a comer/cenar con personas que no sean de confianza (sobre todo mujeres) o en aquellas situaciones de interacción social con una mujer que pudieran dar lugar a una mayor intimidad heterosexual. El tratamiento se llevó a cabo en 36 sesiones en las que se combinaron varias técnicas orientadas en tres direcciones: 1) reducir la respuesta condicionada de ansiedad somática ante los diversos estímulos utilizando entrenamiento en respiración, desensibilización sistemática y exposición gradual "in vivo"; 2) modificar las ideas irracionales que se presentan ante las situaciones problema específicas y, más en general, las referidas a lo que debe ser una relación de pareja o " la pareja ideal", utilizando terapia racional sistemática y generación de auto-instrucciones positivas; 3) mejorar las habilidades interpersonales del sujeto a través del entrenamiento en negación asertiva y expresión de emociones. Tras la intervención se eliminó la respuesta condicionada de vómito, se redujo notabemente el número de pensamientos negativos antes, durante y después de las situaciones problema, y se produjo un incremento significativo en actividades de ocio e interacciones soiales (bien que pudieran dar lugar a comidas/cenas, bien a situaciones de intimidad con mujeres) y a un mayor disfrute de éstas. Los resultados se mantienen tras seis meses de seguimiento (AU)


Assuntos
Adulto , Masculino , Humanos , Transtornos Fóbicos/terapia , Relações Interpessoais , Heterossexualidade , Vômito/terapia , Transtornos Fóbicos/psicologia , Ansiedade/psicologia , Ansiedade/terapia , Respiração , Seguimentos , Vômito/psicologia
16.
Vet Immunol Immunopathol ; 51(1-2): 147-56, 1996 May.
Artigo em Inglês | MEDLINE | ID: mdl-8797284

RESUMO

Monoclonal antibodies (mAbs) to surface markers of bovine lymphocytes MHC I, MHC II, B-cells, T-cells (CD2, CD4, CD8 and gamma/delta) and interleukin-2 (IL-2) receptor were tested in the goat by flow cytometry and using immunohistochemical methods. Samples from peripheral blood and secondary lymphoid organs (mesenteric lymph nodes, spleen and ileal Peyer's patch) were studied. The percentage of positive cells obtained by flow cytometry and its compartmentalisation in different tissue sections showed that the mAbs against MHC I, MHC II, CD2, CD4, CD8, gamma/delta and IL-2 receptor recognised lymphocyte subpopulations similar to those present in the bovine. However, the mAbs tested on B-cells reacted only partially in the recognition of this subpopulation.


Assuntos
Antígenos de Superfície/análise , Cabras/imunologia , Sistema Linfático/imunologia , Linfócitos/imunologia , Animais , Anticorpos Monoclonais , Linfócitos B/imunologia , Antígenos CD2/imunologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Citometria de Fluxo , Genes MHC Classe I/imunologia , Genes MHC da Classe II/imunologia , Imuno-Histoquímica , Linfonodos/citologia , Linfonodos/imunologia , Nódulos Linfáticos Agregados/citologia , Nódulos Linfáticos Agregados/imunologia , Receptores de Antígenos de Linfócitos T gama-delta/imunologia , Receptores de Interleucina-2/imunologia , Baço/citologia , Baço/imunologia
17.
Eur J Haematol ; 56(5): 301-7, 1996 May.
Artigo em Inglês | MEDLINE | ID: mdl-8641404

RESUMO

We performed a prospective study in 17 consecutive patients following Autologous bone marrow (BM) or rhG-CSF primed peripheral blood item cell (PBSC) transplantation, with the objective of comparing immune recovery between both procedures and to evaluate results in rhG-CSF mobilized peripheral blood stem cell transplantation (PBSCT). Kinetics of immune reconstitution showed differences, with a faster recovery of CD3+ and CD8+ T cells, and a more rapid and sustained recovery of CD8+/-/CD56+ natural killer (NK) cells in the PBCSCT group. Autologous bone marrow transplantation (ABMT) was associated with a improved reconstitution of the CD19+/CD5+/-subpopulation. Moreover, rhG-CSF mobilized PBSCT generated a greater recovery of CD8+/-/CD56+ cells than previous data concerning transplantation with peripheral blood (PB) progenitors collected after myelosuppressive chemotherapy or myelosuppressive therapy plus rhG-CSF. Our results show differences in the rate and pattern of B and T lymphocytes reconstitution after ABMT and PBSCT. Additionally, we state an enhancement of CD56+ cells in patients undergoing PBSCT mobilized solely using rhG-CSF.


Assuntos
Transplante de Medula Óssea/imunologia , Fator Estimulador de Colônias de Granulócitos/uso terapêutico , Transplante de Células-Tronco Hematopoéticas , Células Matadoras Naturais/imunologia , Linfócitos T/imunologia , Adolescente , Adulto , Antígenos CD/análise , Neoplasias da Mama/imunologia , Neoplasias da Mama/terapia , Feminino , Transplante de Células-Tronco Hematopoéticas/métodos , Humanos , Leucemia/imunologia , Leucemia/terapia , Linfoma/imunologia , Linfoma/terapia , Pessoa de Meia-Idade , Neoplasias Ovarianas/imunologia , Neoplasias Ovarianas/terapia , Estudos Prospectivos , Proteínas Recombinantes/uso terapêutico , Células-Tronco/efeitos dos fármacos , Transplante Autólogo/imunologia , Macroglobulinemia de Waldenstrom/imunologia , Macroglobulinemia de Waldenstrom/terapia
18.
Br J Haematol ; 93(2): 464-71, 1996 May.
Artigo em Inglês | MEDLINE | ID: mdl-8639450

RESUMO

The levels of platelet-associated Igs (PAIgs) and plasma circulating antiplatelet antibodies were evaluated by a flow cytometric immunofluorescence assay (FCIFA), an enzyme-linked immunoassay (ELISA), and a platelet radioactive antiglobulin test (PRAT), in a group of 45 human immunodeficiency virus (HIV)-infected intravenous drug users (IVDUs), with or without thrombocytopenia (TCP). Direct tests demonstrated an increased amount of PAIgs in 40% of the patients, irrespective of their platelet count. These PAIgs were mainly of IgG class and could not be eluted with ether. Plasma IgG with antiplatelet activity was found in 70% of the thrombocytopenic individuals, whereas it was detected in only one patient without TCP. The relative frequencies of antibodies against the platelet glycoproteins (GPs) Ib/IX and IIb/IIIa were assessed in plasma from all patients by means of the monoclonal antibody-specific immobilization of platelet antigens assay (MAIPA). Plasmas from all non-thrombocytopenic patients were negative when tested by indirect MAIPA. In contrast, 10/23 plasma from thrombocytopenic patients reacted with either GP IIb/IIIa, GP Ib/IX, or both GPs. Finally, aiming to investigate whether HIV antibodies from these patients are reactive with normal platelets, we performed absorption-elution experiments, followed by evaluation of HIV antibodies in the indirect eluates by ELISA and Western blot. Interestingly, we detected anti-HIV antibodies that bind to normal platelet antigens in 50% of the ether eluates prepared from control platelets sensitized with plasma from patients with TCP, but in only 5% of eluates obtained from platelets sensitized with plasma from non-thrombocytopenic patients. The present study provides direct evidence that specific autoantibodies against platelet membrane GPs Ib/IX and IIb/IIIa are common in HIV positive thrombocytopenic individuals. The finding in these patients of HIV antibodies cross-reactive with normal platelets, suggests that mimicry of human antigens by HIV could play a key role in the pathophysiology of the HIV-related TCP.


Assuntos
Autoanticorpos/análise , Anticorpos Anti-HIV/análise , Transtornos Relacionados ao Uso de Opioides/imunologia , Complexo Glicoproteico GPIIb-IIIa de Plaquetas/imunologia , Complexo Glicoproteico GPIb-IX de Plaquetas/imunologia , Trombocitopenia/imunologia , Adulto , Antígenos de Plaquetas Humanas , Autoanticorpos/imunologia , Western Blotting , Ensaio de Imunoadsorção Enzimática , Feminino , Anticorpos Anti-HIV/imunologia , Humanos , Imunoglobulina G/análise , Masculino , Transtornos Relacionados ao Uso de Opioides/complicações , Abuso de Substâncias por Via Intravenosa/complicações , Abuso de Substâncias por Via Intravenosa/imunologia , Trombocitopenia/virologia
19.
Acta Haematol ; 96(3): 135-9, 1996.
Artigo em Inglês | MEDLINE | ID: mdl-8876609

RESUMO

We have compared three techniques for the detection of plasma circulating antiplatelet antibodies, i.e., the platelet suspension immunofluorescence test (PSIFT), the platelet radioactive antiglobulin test (PRAT), and the monoclonal antibody immobilization of platelet antigens (MAIPA). Frozen plasma samples from patients with idiopathic thrombocytopenic purpura or HIV-associated thrombocytopenia were used in the study. The PSIFT and PRAT showed the appropriate ease of performance necessary for screening purposes. The PSIFT is free of radioactivity hazards, but seemed to be less sensitive than the PRAT. The MAIPA is a useful tool to detect antibodies against glycoproteins (GPs) Ib/IX and IIb/IIIa. However, in comparison to PSIFT and PRAT, MAIPA is more time consuming, requires considerable technical expertise, and the identification of antiplatelet activity is highly dependent on the selection of an appropriate primary anti-GP monoclonal antibody. This could explain the lower prevalence of antiplatelet activity detected by MAIPA, in comparison to the frequency provided by the PSIFT and PRAT.


Assuntos
Autoanticorpos/sangue , Plaquetas/imunologia , Técnicas Imunológicas , Trombocitopenia/diagnóstico , Anticorpos Monoclonais , Antígenos de Superfície/imunologia , Citometria de Fluxo , Técnica Indireta de Fluorescência para Anticorpo/métodos , Humanos , Trombocitopenia/imunologia
20.
Ann Hematol ; 71(3): 105-10, 1995 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-7548327

RESUMO

Several studies have demonstrated that both CD34+/CD38- and CD34+/HLA-DR- human hematopoietic progenitor cells have properties associated with hematopoietic stem cells. However, the kinetics of these two cell populations in human peripheral blood (PB) after priming with granulocyte colony-stimulating factor (rhG-CSF) has not been investigated. By using flow-cytometric analysis we have shown that administration of rhG-CSF to 14 patients eligible for peripheral blood progenitor cell (PBPC) transplantation led to an increment of CD34+/CD38+ and CD34+/HLA-DR+ cells in the PB that paralleled the increase of total CD34+ cells, indicating that such subpopulations are responsible for the major release of CD34+ cells. Furthermore, rhG-CSF priming led to a significant mobilization of fractions of more immature CD34+/CD38- and CD34+/HLA-DR- cells to the PB. In the leukapheresis preparations, the average frequency of CD34+ cells lacking the CD38 or HLA-DR antigens was low (5% and 30%, respectively), with little overlap between the CD38- and HLA-DR- subpopulations. In addition, the yield of each subset of CD34+ cells (CD34+/CD38 +/- and CD34+/HLA-DR +/-) in the PB correlated with the numbers in the collected material. The results of the present study indicate that administration of rhG-CSF causes a significant increase of CD34+/CD38 +/- and CD34+/HLA-DR +/- cells in PB, and that such cells can be then safely harvested by leukapheresis procedures.


Assuntos
Antígenos CD , Antígenos de Diferenciação/biossíntese , Fator Estimulador de Colônias de Granulócitos/administração & dosagem , Antígenos HLA-DR/biossíntese , Células-Tronco Hematopoéticas/efeitos dos fármacos , N-Glicosil Hidrolases/biossíntese , ADP-Ribosil Ciclase , ADP-Ribosil Ciclase 1 , Adolescente , Adulto , Antígenos CD34/biossíntese , Diferenciação Celular/efeitos dos fármacos , Feminino , Citometria de Fluxo , Transplante de Células-Tronco Hematopoéticas , Células-Tronco Hematopoéticas/metabolismo , Células-Tronco Hematopoéticas/patologia , Humanos , Leucaférese , Masculino , Glicoproteínas de Membrana , Pessoa de Meia-Idade , Proteínas Recombinantes/administração & dosagem
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