RESUMO
INTRODUCTION: Lung transplantation (LT) is associated with an increased risk of infection, cancer, chronic renal failure, cardiovascular disease and osteoporosis. Some risk factors precede transplantation and could benefit for early diagnosis and optimised care. METHODS: The incidence of comorbidities and their treatment before referral were assessed in 157 consecutive lung transplant candidates between 2008 and 2011. RESULTS: The median age was 37years [25; 51]. Fifty-six percent had a body mass index below 19kg/m(2). In the COPD group, only 50 % had undergone a pulmonary rehabilitation program in the preceding 2 years. Osteoporosis was present in 42 %, of whom 36 % were on bisphophonate therapy. Vitamin D deficiency was present in 65 %. Previously undiagnosed cardiovascular risk factors were discovered during LT assessment: hypertension in one patient, hypercholesterolemia in 6 % and diabetes in 4 %. Poor dental condition necessitating extractions were found in 41 % of patients. Protective anti-HBs antibodies levels were present in 50 % of the patients at the time of referral. CONCLUSION: The assessment and early treatment of nutritional disorders, osteoporosis and risk factors for infection as well as addressing associated cardiovascular risk factors should be optimised in the care of patients with chronic respiratory insufficiency. The potential for becoming a lung transplant candidate in the future should be kept in mind early in the global management of those patients.
Assuntos
Transplante de Pulmão , Cuidados Pré-Operatórios/métodos , Insuficiência Respiratória/cirurgia , Adulto , Doenças Cardiovasculares/epidemiologia , Comorbidade , Complicações do Diabetes/epidemiologia , Feminino , Humanos , Hipercolesterolemia/tratamento farmacológico , Hipercolesterolemia/epidemiologia , Masculino , Desnutrição/epidemiologia , Desnutrição/terapia , Pessoa de Meia-Idade , Osteoporose/tratamento farmacológico , Osteoporose/epidemiologia , Insuficiência Respiratória/complicações , Estudos Retrospectivos , Doenças Dentárias/epidemiologia , Extração DentáriaRESUMO
Numerous compounds acting on the venous system have been developed over the last 50 years, including and often associating flavonoids, pycnogenols, saponosides, nucleosides and bilobatides. Some products have proved highly successful such as Cyclo 3 Fort (Fabroven), Phlebodril, made of ruscus extract (flavonoids, saponins), hesperidin-methyl-chalone and ascorbic acid. This presentation is an attempt to classify the scientifically proven mechanisms of such substances. All selected substances have been tested in clinical trials for their action on the symptoms of chronic venous insufficiency (CVI), leg pain and edema. As the distensibility of the leg veins is the main pathological factor determined by plethysmogaphy, it was at first thought that leg pain would be related to it. But observations have shown that this factor is often unrelated to the enlargement of the veins but rather to a whole chain of factors. The effects of different drugs on venous distensibility, rheological disorders, the prevention of wall dystrophy and action on the microcirculation are examined.
Assuntos
Adenosina/farmacologia , Perna (Membro)/irrigação sanguínea , Extratos Vegetais/farmacologia , Insuficiência Venosa/tratamento farmacológico , Animais , Hipóxia Celular/efeitos dos fármacos , Doença Crônica , Humanos , Sistema Linfático/efeitos dos fármacosRESUMO
The aim was to compare, in terms of cost-effectiveness, two diagnostic strategies for finding out the primary site of tumors revealed by metastasis, adopting the hospital's perspective. The observed strategy reflected the usual practices of doctors at the Regional Cancer Center in Toulouse (France), and was based on a sample of 202 patients of this Center. The standardized strategy, which reflected limited diagnostic investigation, was simulated using the same sample of patients to whom we applied the recommendations of local experts. In the low assumption regarding the effectiveness of the standardized strategy, the observed strategy compared to the standardized one raised the life expectancy from 407 to 418 days at an incremental cost of $US 1,236 per patient (1996 values). In this case, one day of additional life induced a cost of $US 112 per patient. In the high assumption, the incremental effectiveness was null and the incremental cost was $US 1,236 per patient. In conclusion, the effectiveness of the observed strategy as compared to the standardized strategy was highly questionable, given that the patients' quality of life was not taken into account.
Assuntos
Neoplasias Primárias Desconhecidas/diagnóstico , Adulto , Idoso , Idoso de 80 Anos ou mais , Algoritmos , Análise Custo-Benefício , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Primárias Desconhecidas/economia , Neoplasias Primárias Desconhecidas/mortalidade , Neoplasias Primárias Desconhecidas/terapia , Análise de Sobrevida , Fatores de TempoRESUMO
Treatment of venous and lymphatic insufficiency of the lower limbs is based on 3 components: elastic support, venotonic drugs and radical treatments (surgery or sclerotherapy) of insufficient veins. Venotonic drugs have specific indications limited to functional impairment: heavy feeling in the legs, pain and impatience in the evening. There are different categories of venolymphatic drugs. Flavonoids have various pharmacological actions, most notably an increase in venous tone, reduction of capillary permeability and increase of capillary resistance. Choice of a venotonic drug is funded on knowledge of pharmacodynamics and pharmacokinetics of the molecule, critical evaluation of clinical studies, physician's personal experience and drug cost. Venotonic drugs are useful when venous insufficiency leads to functional manifestations. They are especially the treatment of heavy leg syndromes during warm seasons when elastic support is uncomfortable.
Assuntos
Vasodilatadores/uso terapêutico , Insuficiência Venosa/tratamento farmacológico , Difosfato de Adenosina/uso terapêutico , Ácido Ascórbico/uso terapêutico , Permeabilidade Capilar/efeitos dos fármacos , Resistência Capilar/efeitos dos fármacos , Custos de Medicamentos , Flavonoides/uso terapêutico , Ginkgo biloba/uso terapêutico , Humanos , Seleção de Pacientes , Fitoterapia , Plantas Medicinais , Vasodilatadores/classificação , Vasodilatadores/economia , Vasodilatadores/farmacologia , Insuficiência Venosa/etiologia , Insuficiência Venosa/fisiopatologiaRESUMO
BACKGROUND: Some forms of chemoresistance in leukemia may start from failure of tumour cells to successfully undergo apoptosis and Bcl-2 may play a role in this defect. Therefore, we evaluated the Bcl-2 content and synthesis in relation with the apoptotic potential in leukemic cell lines after anthracycline treatment. METHODS: U937, HL60, and K562 cells and their drug resistant (DR) variants were treated with varying concentrations of Idarubicin (IDA). Apoptosis was evaluated by fluorescence microscopy after acridine orange staining. Bcl-2 and Bax content were evaluated either by flow cytometry after indirect immunolabelling or by Western blot. RESULTS: High Bcl-2 contents were not related to a poor ability to undergo apoptosis in U937, HL60, K562 and their DR variants. IDA induced a concentration-dependent increase in Bcl-2 content in all cell lines as long as they do not perform apoptosis. Enhanced Bcl-2 expression was inhibited by cycloheximide, actinomycin D, or antisense oligonucleotide directed against bcl-2 mRNA. Bcl-2 expression was also increased in the resistant U937 variant after serum deprivation or C2-ceramide treatment. The synthesis of Bcl-2 led to an increased Bcl-2/Bax ratio solely in the cells with an apoptosis-resistance phenotype. CONCLUSIONS: These data suggest that exposure to IDA induces Bcl-2 expression in leukemic cell lines, and that this mechanism could contribute to apoptosis resistance and participate in the acquisition of chemoresistance. They also confirm that the evolution of the Bcl-2/Bax ratio reflects apoptotic ability better than the steady state level of Bcl-2 expression.
Assuntos
Antibióticos Antineoplásicos/uso terapêutico , Apoptose/efeitos dos fármacos , Idarubicina/uso terapêutico , Leucemia Mieloide/tratamento farmacológico , Leucemia Mieloide/patologia , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , Proteínas Proto-Oncogênicas c-bcl-2/fisiologia , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/genética , Antibióticos Antineoplásicos/farmacologia , Apoptose/genética , Divisão Celular/efeitos dos fármacos , Ceramidas/farmacologia , Relação Dose-Resposta a Droga , Resistencia a Medicamentos Antineoplásicos/genética , Citometria de Fluxo , Regulação da Expressão Gênica , Células HL-60 , Humanos , Idarubicina/efeitos adversos , Idarubicina/farmacologia , Células K562 , Leucemia Mieloide/genética , Fenótipo , Proteínas Proto-Oncogênicas/biossíntese , Proteínas Proto-Oncogênicas c-bcl-2/genética , Células U937 , Proteína X Associada a bcl-2RESUMO
We evaluated reticulocyte counting and measurement of immature reticulocyte fraction (IRF) with the ABX PENTRA 120 Retic blood analyzer on 300 blood samples. Reticulocyte counts were compared with those obtained by visual counting of 2,000 RBCs, by the TOA (Kobe, Japan) Sysmex R-2000 and a flow cytometry method. The parameters analyzed were the percentages of reticulocytes on all analyzers and the IRF with different modalities. The Retic Count kit (Becton Dickinson, San Jose, CA) was used with the Coulter (Hialech, FL) XL, and a mean channel of fluorescence (MCF) was calculated to fit the reticulocyte maturation. Reticulocyte counting with the ABX (Montpellier, France) PENTRA 120 Retic showed excellent precision and linearity with no significant carryover. Reticulocyte counts were stable after blood storage for 72 hours at 4 degrees C but not at room temperature (RT). IRF parameters values were stable for only 8 hours at 4 degrees C and 6 hours at RT. Comparisons of the methods showed good intraclass correlation (RI) for reticulocyte percentages between ABX PENTRA 120 Retic and Sysmex R-2000, ABX PENTRA 120 Retic and flow cytometry, Sysmex R-2000 and flow cytometry, and ABX PENTRA 120 Retic and manual counting. IRF values were correlated between fluorescence rates and RNA content, but in each case, low RI values were found, showing that Sysmex and ABX IRF values were not concordant. We obtained a significant correlation between mean fluorescence index and the MCF measured by flow cytometry, but the 2 methods were not concordant using the RI. The ABX PENTRA 120 Retic is a good instrument for analyzing reticulocyte count and percentage and allows a good analysis of IRF with several modalities.
Assuntos
Contagem de Reticulócitos/métodos , Reticulócitos/citologia , Doença Aguda , Automação , Doença Crônica , Estudos de Avaliação como Assunto , Citometria de Fluxo/métodos , Corantes Fluorescentes , Humanos , Leucemia/sangue , Reprodutibilidade dos Testes , Contagem de Reticulócitos/instrumentação , Sensibilidade e EspecificidadeRESUMO
Patients infected with HIV are at increased risk of atherosclerosis, and have evidence of endothelium dysfunction. The hypothesis was tested that HIV-related endothelium dysfunction is related to loss of antioxidants. This was done by the supplementation of the antioxidants selenium and beta-carotene. We supplemented the diet of 10 HIV-seropositive subjects with 100 microg selenium daily, 11 subjects with 30 mg beta-carotene twice daily while 15 subjects were not supplemented. Plasma was obtained at outset and after a year, and tested by ELISA for endothelial cell, platelet and inflammatory markers. The non-supplemented patients experienced increases in von Willebrand factor and soluble thrombomodulin (both p <0.01). There were no changes in any of the indices in the patients taking selenium or beta-carotene. Increased von Willebrand factor and soluble thrombomodulin in the non-supplemented patients imply increased damage to the endothelium over the year of the study. Therefore we interpret the lack of increase in the patients taking antioxidants as evidence of the protection of the endothelium by these agents.
Assuntos
Antioxidantes/farmacologia , Endotélio Vascular/efeitos dos fármacos , Infecções por HIV/patologia , Selenometionina/farmacologia , beta Caroteno/farmacologia , Arteriosclerose/epidemiologia , Biomarcadores , Dieta , Suscetibilidade a Doenças , Selectina E/análise , Endotélio Vascular/fisiopatologia , Feminino , Humanos , Molécula 1 de Adesão Intercelular/análise , Masculino , Estresse Oxidativo , Projetos Piloto , Testes de Função Plaquetária , Fatores de Risco , Trombomodulina/análise , Molécula 1 de Adesão de Célula Vascular/análise , Fator de von Willebrand/análiseRESUMO
Expression of the multidrug resistance (MDR) phenotype is responsible for chemotherapy failure in numerous cancers. This phenotype is generally due to the expression of the mdr1 gene-encoded P-gp. Modulation of P-gp activity by chemotherapy has limited possibilities because of toxicity and poor specificity. In contrast, specific transcription blockage of the mdr1 gene can be obtained by oligonucleotides forming a triple helix structure at the DNA level. We used here immunofluorescence and both flow cytometry and image analysis to evaluate surface and total P-gp content in K562 MDR cells. The mdr1 mRNA content was measured by RT-PCR. We confirm the capacity of a 27-mer oligodeoxynucleotide, targeted to an mdr1 DNA fragment, to cause a 10-fold decrease in mdr1 mRNA level. However, this specific genetic inhibition was functionally limited because cellular growth was not modified in a cytotoxic environment. We found that total P-gp content was reduced in resistant cells treated with the mdr1-targeted oligonucleotide, while it remained in high levels on the cell surface, suggesting the existence of a large cytoplasmic pool of P-gp (approximately 50% of the total cellular P-gp). Moreover, when cycloheximide was used for 72 h to suppress protein synthesis, surface P-gp expression showed no decrease, whereas total P-gp was considerably lowered. A rapid 35% decrease in surface P-gp level was reached when resistant cells were treated for 24 h with brefeldin A, an inhibitor of intracellular protein trafficking. Simultaneously, the total P-gp level remained stable, thus indicating a probable accumulation of cytoplasmic P-gp, in agreement with the interruption of protein migration. We propose that the cytoplasmic P-gp pool could be a storage pool consumed for maintaining a steady-state level of surface P-gp. Cytometry could be a useful tool to study such a mechanism of P-gp trafficking and cellular distribution, which could explain the difficulties encountered in achieving stable and rapid effects of MDR reversal with oligonucleotides.
Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/biossíntese , Resistência a Múltiplos Medicamentos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Antibacterianos/farmacologia , Sequência de Bases , Brefeldina A , Divisão Celular/efeitos dos fármacos , Cicloeximida/farmacologia , Ciclopentanos/farmacologia , Citoplasma/metabolismo , Primers do DNA , Citometria de Fluxo/métodos , Imunofluorescência , Células HL-60 , Humanos , Cinética , Leucemia , Macrolídeos , Oligodesoxirribonucleotídeos/farmacologia , Reação em Cadeia da Polimerase , Inibidores da Síntese de Proteínas/farmacologia , Proteínas Proto-Oncogênicas c-abl/biossíntese , RNA Mensageiro/metabolismo , Transfecção , Células Tumorais CultivadasRESUMO
An increasing number of methods are being described to detect apoptotic cells. However, attempts to detect apoptotic cells in clinical samples are rarely successful. A hypothesis is that apoptotic cells are cleared from the circulation by phagocytosis before they become detectable by conventional morphological or cytometric methods. Using LR73 adhering cells as phagocytes in a model of in vitro phagocytosis, we found that phagocytosis of daunorubicin (DNR)-treated U937, HL60, or K562 leukemia cell lines occurred prior to phosphatidylserine externalization, DNA hydrolysis, chromatin condensation, nuclear fragmentation, or mitochondrial potential alteration. Moreover DNR-treated K562 cells were eliminated by phagocytes while apoptosis was never observed by any of the above methods. By contrast, using a fluorometric batch analysis assay to detect caspase activity in ceramide- or DNR-treated cells (fluorogenic substrate for caspase), we found that caspase activity increased in apoptosis-committed cells before they were detected by flow cytometry or recognized by phagocytes. Similarly a caspase activity increase was detected in circulating mononuclear cells of luekemic patients 15 h after the beginning of anthracyclin treatment. We suggest that recent findings on enzymatic events (caspase activation) occurring in the early events of apoptosis must now allow the development of new markers for apoptosis, irrespective of the morphological features or internucleosomal fragmentation which are late events in apoptosis.
Assuntos
Antibióticos Antineoplásicos/farmacologia , Apoptose , Caspases , Cisteína Endopeptidases/metabolismo , Daunorrubicina/farmacologia , Fagócitos/fisiologia , Animais , Anexina A5 , Antígenos CD36/metabolismo , Células CHO , Caspase 3 , Cumarínicos/metabolismo , Cricetinae , Ativação Enzimática , Inibidores Enzimáticos/farmacologia , Corantes Fluorescentes/metabolismo , Células HL-60 , Humanos , Oligopeptídeos/metabolismo , Fagocitose , Esfingosina/análogos & derivados , Esfingosina/farmacologia , Especificidade por Substrato , Células Tumorais CultivadasRESUMO
Vitronectin (VN) is a plasma glycoprotein that promotes cell attachment and induces migration of human smooth muscle cells (SMCs) in culture. VN has been observed to accumulate in human atherosclerotic plaques, although its origin and role in atherosclerosis are not yet established. In the present experiments, synthesis of VN by intimal cells and its colocalization with receptors, alphavbeta3 and alphavbeta5, were studied by in situ hybridization and immunohistochemistry on 15 human atherosclerotic plaques from carotid arteries obtained after surgery. Strong VN protein and mRNA expression was observed in the intima and in the media. In the intima, VN mRNA expression was colocalized with SMCs, indicating that these cells produce VN, which may account for its accumulation in atherosclerotic plaques. In SMCs in culture, immunoprecipitation after metabolic labeling demonstrated that human SMCs do synthesize vitronectin. Confocal microscopic examination showed that VN colocalized with its receptors, alphavbeta3 and alphavbeta5, in the atherosclerotic intima. However, the distribution of the VN receptors on SMCs in culture in contact with VN was different. These observations suggest that VN plays various parts in atherogenesis via different SMC membrane receptors.
Assuntos
Arteriosclerose/metabolismo , Músculo Liso Vascular/metabolismo , Receptores de Vitronectina/metabolismo , Vitronectina/metabolismo , Arteriosclerose/patologia , Doenças das Artérias Carótidas/metabolismo , Doenças das Artérias Carótidas/patologia , Artéria Carótida Interna/metabolismo , Artéria Carótida Interna/patologia , Células Cultivadas , Humanos , Integrinas/metabolismo , Músculo Liso Vascular/patologia , RNA Mensageiro/metabolismo , Vitronectina/genéticaRESUMO
A flow cytometry method has been introduced into the routine investigation of whole bone marrow samples following red blood cell lysis on the basis of a primary CD45/side scatter (SSC) gating procedure. Blast cells were first identified by CD45/SSC gating in 74 cases of acute myeloid leukemia (AML) and the results were compared to a conventional FSC/SSC gating procedure and to MGG-staining smears. The percentages of blast cells in these samples as defined by the morphological analysis of MGG smears correlated better with the values determined by CD45/SSC gating (r = 0.94) than with the blast cell counts recorded with FSC/SSC gating (r = 0.76). These findings were not surprising because while CD45 expression was regularly lower on leukemic blasts than on normal lymphoid and monocytic cells, the FCS/SSC characteristics of these populations were overlapping. In 53 samples, the blast cell populations were also analyzed with a panel of FITC-conjugated monoclonal antibodies that were utilized in double labeling with CD45-PE. We show that the CD45/SSC gating procedure improved phenotypic determination of the blast cells in three ways: (1) by discriminating between leukemic blast cells and residual normal cells; (2) by excluding normal cells from the phenotypic analysis of leukemic blast cells; and (3) by identifying blast cell heterogeneity in many cases of leukemia on the basis of different CD45 display. Moreover, this immunophenotyping procedure on whole bone marrow samples also allowed an efficient discrimination between the various cell lineages and facilitated the analysis of leukemic blasts present in low proportions.
Assuntos
Citometria de Fluxo/métodos , Imunofenotipagem/métodos , Leucemia Mieloide/diagnóstico , Antígenos Comuns de Leucócito , Doença Aguda , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígenos CD/genética , Antígenos CD/imunologia , Contagem de Células , Feminino , Técnica Direta de Fluorescência para Anticorpo , Humanos , Leucemia Mieloide/classificação , Leucemia Mieloide/imunologia , Masculino , Pessoa de Meia-IdadeRESUMO
Endothelial damage is present in HIV infection but our understanding of markers and mechanisms is incomplete. We found increased levels of markers of endothelial cell damage such as von Willebrand factor (vWf), soluble thrombomodulin (sTM) and adhesion molecule E-selectin in 90 subjects seropositive for HIV relative to healthy controls. sTM was strongly raised in those patients with the lowest CD4+ cell count (p < 0.001), but levels of vWf increased at each incremental fall in CD4+ cell count and the two indices correlated significantly (r = -0.485, p < 0.001). vWf correlated strongly with levels of the inflammatory cytokines tumor necrosis factor (TNF-alpha) and alpha interferon (IFN-alpha) but sTM correlated only weakly with IFN-alpha. We suggest that increased vWf is largely the result of inflammatory stimulus of the endothelium but that sTM is found only in those patients with more severe disease, and so truly represents endothelial damage.
Assuntos
Selectina E/sangue , Endotélio Vascular/patologia , Infecções por HIV/sangue , Trombomodulina/metabolismo , Fator de von Willebrand/metabolismo , Adolescente , Adulto , Idoso , Biomarcadores/sangue , Estudos Transversais , Feminino , Infecções por HIV/patologia , Humanos , Masculino , Pessoa de Meia-Idade , Valores de Referência , SolubilidadeRESUMO
A flow cytometric method for simultaneous apoptotic cell detection and cell cycle analysis was applied on the U937 cell line. Four antitumoral drugs currently used in the treatment of acute myeloid leukaemia were studied in vitro: DNR, IDR, MITO and Ara-C. Our results show a dissociation between the cytostatic effect (the block in the cell cycle observed for low drug concentrations) and the cytotoxic effect (the induction of apoptosis induced by higher concentrations) for all the tested molecules. Low concentrations of Ara-C induced a block in the S phase while higher concentrations (>10(-7) M) induced apoptosis at the G1-S boundary. Low concentrations of anthracyclines (<40 nM DNR and <20nM IDR) induced a block in G2 without apoptosis. Apoptosis was induced in G1 and/or early S phases by higher concentrations of anthracyclines. The concentration inducing 50% apoptosis (IC50) was found to be, respectively, 200 and 40 nM for DNR and IDR. Analysis of MITO-treated cells showed a parallel increase in the percentages of S phase and apoptotic cells. However, the bivariate analysis showed that apoptosis did occur in a population with G1 DNA content. For two other drugs (CAM and COLC), apoptosis occurred for the same concentrations and in the same phase as the block (in S and G2M, respectively). The IC50 of MITO was found to be 100 nM. Cotreatment of the cells with colchicin and either Ara-C or IDR showed that the passage through mitosis was not necessary for the completion of apoptosis at the G1-S boundary. Short incubations of U937 cells with high concentrations of anthracyclines were found to be efficient in inducing further apoptosis. We conclude that, for all the assayed molecules, the cytotoxic and/or cytostatic effects of the antitumoral drugs tested greatly depend on the concentrations used and that, depending on their in vivo pharmacokinetics, the induction of apoptosis could be an important mechanism of action for some of these drugs.
Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Leucemia/tratamento farmacológico , Doença Aguda , Antibióticos Antineoplásicos/farmacologia , Antimetabólitos Antineoplásicos/farmacologia , Ciclo Celular/efeitos dos fármacos , Citarabina/farmacologia , Daunorrubicina/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais , Citometria de Fluxo , Humanos , Idarubicina/farmacologia , Cinética , Leucemia/patologia , Mitoxantrona/farmacologiaRESUMO
Apoptosis was studied in parental and mdr-1 expressing U937, HL60 and K562 myeloid leukemic cell lines using mdr unrelated inducers of apoptosis such as Ara-C, cycloheximide, serum deprivation, ceramide, monensin and UV irradiation. Apoptosis was efficiently induced by all these treatments in U937 and HL60 cells while K562 cells exhibited an apoptosis-resistant phenotype except with UV and monensin. The pattern of apoptosis resistance in mdr-1 expressing U937 (U937-DR) and HL60 (HL60-DR100) was similar to that presented by K562. This apoptosis-resistant phenotype of mdr cells was not overcome by concentrations of verapamil inhibiting the P-gp 170 pump. The acquisition of this phenotype was posterior to the mdr-1 expressing phenotype since a HL60-DR5 variant, selected at the beginning of the induction of resistance, presented a low level of mdr-1 expression without resistance to apoptosis. The variations observed in the Fas (CD95) expression between sensitive and resistant cells were not sufficient to account for apoptosis resistance. However, a high expression in Abl antigen was found in all the apoptosis-resistant cells. RT-PCR and Western blot analysis showed that this increase in Abl antigen content was accompanied by the expression in U937-DR and HL60-DR100 cells of a hybrid bcr/abl mRNA and a 210 kD Bcr/Abl protein which was constitutive in K562. This expression was due to the translocation of abl and the amplification of the bcr-abl translocated gene. These results are in agreement with the role of Bcr/Abl tyrosine protein kinase as an inhibitor of apoptosis independently of the mdr-1 expression. They also suggest that translocation of the abl gene in the bcr region is a highly probable rearrangement in the mdr-1 expressing myeloid cells and that Bcr/Abl tyrosine kinase effect on apoptosis needs the regulation of intracellular pH and is inactive against UV-induced apoptosis.
RESUMO
Changes in endothelial cell activity are likely to play a role in the thrombotic complications of bone marrow transplantation (BMT) such as the development of veno-occlusive disease. Accordingly, we measured established plasma endothelial cell markers von Willebrand factor (vWf), soluble thrombomodulin (sTM), soluble ICAM-1 (sICAM-1), and possible inducers of these molecules, TNF alpha and elastase, in the plasma of 25 patients, 1 week before as well as 1 and 3 weeks after BMT. Compared to healthy age and sex-matched controls, patients exhibited increased vWf and sTM. One week after transplantation, there were significant increases in vWf and sICAM-1, with a significant fall in elastases. Three weeks after the transplantation, sICAM-1 and, to a lesser extent, vWf increased still further, whereas elastases were unchanged. There were no significant changes in sTM and in TNF alpha through the serial study. Our data suggest that before conditioning, vascular endothelium is damaged by both injury and activation, as seen by the variations of vWf and sTM. After transplantation, the enhancement of this damage seems to be more specifically related to activation, since we observed a strong subsequent increase in vWf and markedly in sICAM-1.
Assuntos
Transplante de Medula Óssea/efeitos adversos , Endotélio Vascular/patologia , Molécula 1 de Adesão Intercelular/análise , Trombomodulina/análise , Fator de von Willebrand/análise , Adolescente , Adulto , Biomarcadores , Criança , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Elastase Pancreática/análise , Fator de Necrose Tumoral alfa/análiseRESUMO
Circulating levels of the adhesion molecule P-selectin (CD62P) are increased in the plasma of patients with atherosclerosis, but its relationship to the anatomical location of symptomatic disease or extent of symptomatic disease is unknown. The influence of the risk factors for atherosclerosis on soluble P-selectin is also unclear. To clarify these questions we analysed plasma samples from 170 patients with symptomatic peripheral vascular disease and 119 asymptomatic controls who were, as a group, age- and sex-matched. Soluble P-selectin (ELISA) was increased in 83 patients with symptomatic disease of the iliac and/or femoral arteries alone (P < 0.05, ANOVA) but not in 37 patients with symptomatic carotid artery disease alone compared with controls. Soluble P-selectin was equally raised in 120 patients with disease at one arterial site and in 50 patients with disease at two or more arterial sites (both P < 0.05) compared with controls. Smoking and atherosclerosis were both independent predictors of raised soluble P-selectin. We conclude that increased soluble P-selectin may have value as a marker of peripheral vascular disease of the iliac and/or femoral arteries in group comparisons only, as the poor discrimination and wide variation of data make comparisons at the individual level difficult.
Assuntos
Selectina-P/sangue , Doenças Vasculares Periféricas/sangue , Idoso , Arteriosclerose/sangue , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fatores de Risco , FumarRESUMO
In hypoxia/ischaemia and ischaemia/reperfusion, human neutrophils are likely to play an important role in the development of endothelial cell damage in the microcirculation. Buflomedil hypochloride improves the capillary perfusion in such related situations, evoking a possible effect upon neutrophils. Using in vitro models of cell adhesion, buflomedil decreased 100% of histamine related neutrophil adhesion (flow system) and partially inhibited adhesion after IL-1-4 hours (flow and stable systems). Hypoxia induced neutrophil adhesion (4 hours) was also reduced by buflomedil, which decreased the expression of P-selectin at the surface of endothelial cells. As adenosine (NECA) exhibited the same results in hypoxia and theophylline inhibited them, such results support an action of buflomedil presumably via the A2 receptor.
Assuntos
Antagonistas de Receptores Adrenérgicos alfa 2 , Antagonistas Adrenérgicos alfa/farmacologia , Hipóxia Celular , Endotélio Vascular/efeitos dos fármacos , Inflamação/patologia , Neutrófilos/efeitos dos fármacos , Pirrolidinas/farmacologia , Adenosina/análogos & derivados , Adenosina/farmacologia , Adenosina-5'-(N-etilcarboxamida) , Adesão Celular/efeitos dos fármacos , Células Cultivadas , Citocinas/metabolismo , Endotélio Vascular/citologia , Humanos , Molécula 1 de Adesão Intercelular/biossíntese , Interleucina-1/farmacologia , Selectina-P/biossíntese , Teofilina/farmacologia , Veias UmbilicaisRESUMO
Twenty-three patients with acute myelogenous leukemia (AML) in first relapse were treated with high-dose cytosine-arabinoside (Ara-C) and amsacrine or idarubicin. To prime the cells, the patients were given rhGM-CSF. We studied the influence of 48-h infusion of rhGM-CSF on proliferation and Ara-C sensitivity of leukemic cells both ex vivo and in vitro. We found that a 48-h infusion of rhGM-CSF increased both white blood cell counts and peripheral blood blast cell percentages. Using a Bromodeoxyuridine/DNA (BrdUrd/DNA) staining in flow cytometry, we found an non-constant increase in cells in the S-phase. Ex vivo 48-h culture of leukemic cells with or without rhGM-CSF, with or without other hematopoietic growth factors (HGFs), showed a greater increase of the cells in the S-phase with GF but no correlation with the ex vivo results. We used a method of quantitation of the DNA synthesis previously described (Lacombe F., et al. (1992) Cytometry 13, 730) to monitor the Ara-C sensitivity of the cells in S-phase before and after 48-h infusion with rhGM-CSF. We observed a great variation in the Ara-C sensitivity of the leukemic cells before and after infusion with rhGM-CSF from one patient to another. The BrdUrd/DNA method seems a convenient method to study the influence of HGFs on Ara-C sensitivity of the patients.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Leucemia Mieloide/tratamento farmacológico , Doença Aguda , Adolescente , Adulto , Idoso , Bromodesoxiuridina/análise , Ciclo Celular , Citarabina/administração & dosagem , DNA de Neoplasias/análise , Relação Dose-Resposta a Droga , Interações Medicamentosas , Resistencia a Medicamentos Antineoplásicos , Ensaios de Seleção de Medicamentos Antitumorais , Feminino , Citometria de Fluxo , Fator Estimulador de Colônias de Granulócitos e Macrófagos/administração & dosagem , Humanos , Leucemia Mieloide/patologia , Masculino , Pessoa de Meia-Idade , Prognóstico , Proteínas Recombinantes/administração & dosagem , Fase S/efeitos dos fármacosRESUMO
Preoperative normovolaemic acute haemodilution (PNAH) is used to reduce major blood loss during elective surgery. Considerable attention has been paid to colloid osmotic pressure, index of diffusibility and intravascular half-life of the currently available substitutes, but there is little information on their rheological properties from in vivo studies. Forty patients undergoing elective aortic reconstruction were given 4% human albumin (HA), 3.5% dextran 40 (Dxt 40), 6% dextran 60 (Dxt 60), 6% hydroxyethylstarch 200 (HES) or modified fluid gelatin (Gel) during PNAH to produce a packed cell volume (PCV) of approximately 30%. Mean volumes of more than 1000 ml were infused. Blood samples were obtained before infusion, immediately after, and 1.5 h after the end of haemodilution. The following variables were measured: PCV, plasma viscosity, whole blood viscosity at measured and corrected PCV (0.45), and erythrocyte aggregation. Haemodynamic and metabolic variables were determined at the same time. The five substitutes had very different effects on red blood cell aggregation and low shear rate viscosity at corrected PCV. Red blood cell aggregation was reduced in the presence of HA, Dxt 40, but was increased moderately to markedly in the presence of the other substitutes in the following order: HES < Dxt 60 < Gel. The influence of the rheological conditions on tissue oxygenation was assessed by measuring the concentration of lactic acid; this was unchanged after PNAH with HA or Dxt 40, but was increased in the presence of HES, Dxt 60 or Gel.