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1.
Braz. j. med. biol. res ; 41(7): 571-578, July 2008. ilus, tab, graf
Article Dans Anglais | LILACS | ID: lil-489516

Résumé

Apoptotic protease activating factor 1 (APAF-1) has a critical role in the regulation of apoptosis. In the present study, the mRNA expression analysis of different APAF-1 transcripts (APAF-1S, APAF-1LC, APAF-1LN, and APAF-1XL) was analyzed in bone marrow samples from 37 patients with acute myeloid leukemia (newly diagnosed, with no previous treatment). APAF-1XL and APAF-1LN transcripts (with and without an extra WD-40 repeat region, respectively) were detected in all samples, although the major form expressed was APAF-1XL in 65 percent of the samples (group 1), while 35 percent of the samples expressed primarily APAF-1LN (group 2). Only 46 percent of the patients presented complete remission in response to remission induction therapy (represented by less than 5 percent marrow blasts and hematological recovery), all but 2 cases being from group 1, 21.6 percent did not attain complete remission (only 1 case from group 1), and 32.4 percent of the patients died early. Lower expression of APAF-1XL (APAF-1XL/APAF-1LN ratio <1.2) was associated with a poor response to therapy (P = 0.0005, Fisher exact test). Both groups showed similar characteristics regarding white blood cell counts, cytogenetic data or presence of gene rearrangements associated with good prognosis as AML1-ETO, CBFB-MYH11 and PML/RARA. Since it has been shown that only the isoforms with the extra WD-40 repeat region activate procaspase-9, we suggest that low procaspase-9 activation may also be involved in the deregulation of apoptosis and chemotherapy resistance in acute myeloid leukemia.


Sujets)
Adulte , Sujet âgé , Sujet âgé de 80 ans ou plus , Femelle , Humains , Mâle , Adulte d'âge moyen , Jeune adulte , Facteur-1 activateur des protéases apoptotiques/génétique , Leucémie aigüe myéloïde/génétique , Protocoles de polychimiothérapie antinéoplasique/usage thérapeutique , Cellules de la moelle osseuse/composition chimique , Études cas-témoins , Densitométrie , ADN complémentaire/génétique , Régulation de l'expression des gènes tumoraux , Leucémie aigüe myéloïde/traitement médicamenteux , RT-PCR , ARN messager/génétique , Facteurs de transcription , Échec thérapeutique , Transcription génétique/génétique , Marqueurs biologiques tumoraux/génétique , Jeune adulte
2.
Experimental & Molecular Medicine ; : 273-283, 2006.
Article Dans Anglais | WPRIM | ID: wpr-96563

Résumé

To increase the biocompatibility and durability of glutaraldehyde (GA)-fixed valves, a biological coating with viable endothelial cells (ECs) has been proposed. However, stable EC layers have not been formed successfully on GA-fixed valves due to their inability to repopulate. In this study, to improve cellular adhesion and proliferation, the GA-fixed prostheses were detoxified by treatment with citric acid to remove free aldehyde groups. Canine bone marrow mononuclear cells (MNCs) were differentiated into EC-like cells and myofibroblast-like cells in vitro. Detoxified prostheses were seeded and recellularized with differentiated bone marrow-derived cells (BMCs) for seven days. Untreated GA-fixed prostheses were used as controls. Cell attachment, proliferation, metabolic activity, and viability were investigated and cell-seeded leaflets were histologically analyzed. On detoxified GA-fixed prostheses, BMC seeding resulted in uninhibited cell proliferation after seven days. In contrast, on untreated GA-fixed prostheses, cell attachment was poor and no viable cells were observed. Positive staining for smooth muscle a-actin, CD31, and proliferating cell nuclear antigen was observed on the luminal side of the detoxified valve leaflets, indicating differentiation and proliferation of the seeded BMCs. These results demonstrate that the treatment of GA-fixed valves with citric acid established a surface more suitable for cellular attachment and proliferation. Engineering heart valves by seeding detoxified GA-fixed biological valve prostheses with BMCs may increase biocompatibility and durability of the prostheses. This method could be utilized as a new approach for the restoration of heart valve structure and function in the treatment of end-stage heart valve disease.


Sujets)
Chiens , Animaux , Fixation tissulaire , Ingénierie tissulaire/méthodes , Suidae , Antigène nucléaire de prolifération cellulaire/analyse , Muscles lisses/composition chimique , Microscopie électronique à balayage , Immunohistochimie , Valves cardiaques/cytologie , Prothèse valvulaire cardiaque , Glutaraldéhyde/composition chimique , Cellules endothéliales/cytologie , Survie cellulaire/physiologie , Prolifération cellulaire , Différenciation cellulaire/physiologie , Techniques de culture cellulaire/méthodes , Adhérence cellulaire/physiologie , Cellules de la moelle osseuse/composition chimique , Antigènes CD31/analyse , Actines/analyse
3.
Braz. j. med. biol. res ; 34(11): 1457-1463, Nov. 2001. tab
Article Dans Anglais | LILACS | ID: lil-303323

Résumé

In a previous study we demonstrated that the incidence of fibroblast colony-forming units (CFU-F) was very low in bone marrow primary cultures from the majority of untreated advanced non-small lung cancer patients (LCP) compared to normal controls (NC). For this reason, we studied the ability of bone marrow stromal cells to achieve confluence in primary cultures and their proliferative capacity following four continuous subcultures in consecutive untreated LCP and NC. We also evaluated the production of interleukin-1ß (IL-1ß) and prostaglandin E2 (PGE2) by pure fibroblasts. Bone marrow was obtained from 20 LCP and 20 NC. A CFU-F assay was used to investigate the proliferative and confluence capacity. Levels of IL-1ß and PGE2 in conditioned medium (CM) of pure fibroblast cultures were measured with an ELISA kit and RIA kit, respectively. Only fibroblasts from 6/13 (46 percent) LCP confluent primary cultures had the capacity to proliferate following four subcultures (NC = 100 percent). Levels of spontaneously released IL-1ß were below 10 pg/ml in the CM of LCP, while NC had a mean value of 1,217 + or - 74 pg/ml. In contrast, levels of PGE2 in these CM of LCP were higher (77.5 + or - 23.6 pg/ml) compared to NC (18.5 + or - 0.9 pg/ml). In conclusion, bone marrow fibroblasts from LCP presented a defective proliferative and confluence capacity, and this deficiency may be associated with the alteration of IL-1ß and PGE2 production


Sujets)
Humains , Mâle , Femelle , Adulte , Adulte d'âge moyen , Carcinome pulmonaire non à petites cellules , Cellules de la moelle osseuse/anatomopathologie , Fibroblastes , Tumeurs du poumon , Études cas-témoins , Cellules de la moelle osseuse/composition chimique , Test clonogénique , Milieux de culture conditionnés , Dinoprostone , Test ELISA
4.
Braz. j. med. biol. res ; 33(8): 889-95, Aug. 2000. tab, ilus
Article Dans Anglais | LILACS | ID: lil-265845

Résumé

The objective of the present study was to determine the effect of protein malnutrition on the glycoprotein content of bone marrow extracellular matrix (ECM). Two-month-old male Swiss mice were submitted to protein malnutrition with a low-protein diet containing 4 percent casein as compared to 20 percent casein in the control diet. When the experimental group had attained a 20 percent loss of their original body weight, we extracted the ECM proteins from bone marrow with PBS buffer, and analyzed ECM samples by SDS-PAGE (7.5 percent) and ECL Western blotting. Quantitative differences were observed between control and experimental groups. Bone marrow ECM from undernourished mice had greater amounts of extractable fibronectin (1.6-fold increase) and laminin (4.8-fold increase) when compared to the control group. These results suggest an association between fluctuations in the composition of the hematopoietic microenvironment and altered hematopoiesis observed in undernourished mice


Sujets)
Animaux , Mâle , Souris , Cellules de la moelle osseuse/composition chimique , Matrice extracellulaire/composition chimique , Fibronectines/métabolisme , Laminine/métabolisme , Malnutrition protéinocalorique/métabolisme , Technique de Western , Études cas-témoins , Électrophorèse sur gel de polyacrylamide , Fibronectines/analyse , Glycoprotéines/analyse , Hématopoïèse extramédullaire/physiologie , Laminine/analyse
5.
Journal of Korean Medical Science ; : 217-223, 2000.
Article Dans Anglais | WPRIM | ID: wpr-18567

Résumé

Due to their high immunostimulatory ability as well as the critical role they play in the maintenance of self-tolerance, dendritic cells have been implicated in the pathogenesis of autoimmune diseases. The non-obese diabetic (NOD) mouse is an animal model of autoimmune type 1 diabetes, in which pancreatic beta cells are selectively destroyed mainly by T cell-mediated immune responses. To elucidate initiation mechanisms of beta cell-specific autoimmunity, we attempted to generate bone marrow-derived dendritic cells from NOD mice. However, our results showed low proliferative response of NOD bone marrow cells and some defects in the differentiation into the myeloid dendritic cells. NOD dendritic cells showed lower expressions of MHC class II, B7-1, B7-2 and CD40, compared with C57BL/6 dendritic cells. In mixed lymphocyte reactions, stimulatory activities of NOD dendritic cells were also weak. Treatment with LPS, INF-gamma and anti-CD40 stimulated NOD dendritic cells to produce IL-12p70. The amount of IL-12, however, appeared to be lower than that of C57BL/6. Results of the present study indicated that there may be some defects in the development of NOD dendritic cells in the bone marrow, which might have an impact on the breakdown of self tolerance.


Sujets)
Souris , Animaux , Maladies auto-immunes/anatomopathologie , Maladies auto-immunes/immunologie , Cellules de la moelle osseuse/anatomopathologie , Cellules de la moelle osseuse/immunologie , Cellules de la moelle osseuse/composition chimique , Différenciation cellulaire/immunologie , Différenciation cellulaire/effets des médicaments et des substances chimiques , Cellules dendritiques/anatomopathologie , Cellules dendritiques/immunologie , Cellules dendritiques/composition chimique , Diabète de type 1/anatomopathologie , Diabète de type 1/immunologie , Test ELISA , Facteur de stimulation des colonies de granulocytes et de macrophages/pharmacologie , Interleukine-12/analyse , Interleukine-4/pharmacologie , Lipopolysaccharides/pharmacologie , Souris de lignée BALB C , Souris de lignée C57BL , Souris de lignée NOD , Obésité
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