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1.
Arch Pediatr ; 12(11): 1613-6, 2005 Nov.
Artigo em Francês | MEDLINE | ID: mdl-16084073

RESUMO

An ileal perforation occurred shortly after birth in 4 very premature newborns. Diagnosis was made on an abdominal distension with a pneumoperitoneum on X-ray. There were no biological, radiological nor histological signs of necrotizing enterocolitis. There were no digestive short- or long-term complications. According to the few authors who described this syndrome, there are some risk factors, but they were not clearly involved in our cases. Ileal perforation in the absence of signs of necrotizing enterocolitis is rarely reported but should be well known because of its good prognosis.


Assuntos
Doenças do Íleo/patologia , Recém-Nascido Prematuro , Perfuração Intestinal/patologia , Diagnóstico Diferencial , Humanos , Doenças do Íleo/diagnóstico , Recém-Nascido , Perfuração Intestinal/diagnóstico , Prognóstico , Fatores de Risco
2.
Exp Hematol ; 20(8): 962-8, 1992 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-1354620

RESUMO

High-dose chemotherapy followed by autologous bone marrow transplantation (ABMT) in the treatment of malignancies is often associated with immune deficiency following transplantation, possibly contributing to tumor relapse or fatal infection. Interleukin 2 (IL-2), which enhances major histocompatibility complex (MHC)-unrestricted cytotoxicity in vitro, can be applied in vivo as immunotherapy to reduce these potential complications. Recombinant human IL-2 (rIL-2) was administered by continuous i.v. infusion for four courses in 19 patients following ABMT for lymphomas and solid tumors. The patients were assigned to five groups of escalating doses of rIL-2 ranging from 3 to 30 x 10(6) IU/m2/day. The immunological effects and toxicity were monitored. After a transient reduction of lymphocytes in the peripheral blood, a significant lymphocytosis was observed during the rIL-2 infusion with an augmentation of CD2+, CD25+, and CD8(+)-Ia+ T cells and a dose-related increase of CD56+ lymphocytes. Natural killer (NK) activity appeared enhanced in patients treated with as little as 6 x 10(6) IU/m2/day. No statistically significant increase in lymphokine-activated killer (LAK) activity was seen after rIL-2, when compared to LAK activity following ABMT prior to rIL-2 administration. Administration of exogenous rIL-2 to patients who have undergone ablative chemotherapy and ABMT has a role in restoring defective T-cell function. Further trials defining those patients most likely to benefit from rIL-2 integrated with ablative chemotherapy and ABMT are now warranted.


Assuntos
Transplante de Medula Óssea/patologia , Carcinoma de Células Pequenas/cirurgia , Coriocarcinoma/cirurgia , Interleucina-2/uso terapêutico , Neoplasias Pulmonares/cirurgia , Linfoma/cirurgia , Neoplasias Ovarianas/cirurgia , Proteínas Recombinantes/uso terapêutico , Linfócitos T/imunologia , Adulto , Antígenos de Diferenciação de Linfócitos T/análise , Contagem de Células Sanguíneas/efeitos dos fármacos , Antígenos CD2 , Antígenos CD8/análise , Morte Celular/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Feminino , Humanos , Síndromes de Imunodeficiência/tratamento farmacológico , Infusões Intravenosas , Interleucina-2/administração & dosagem , Interleucina-2/efeitos adversos , Células Matadoras Naturais/efeitos dos fármacos , Células Matadoras Naturais/fisiologia , Masculino , Pessoa de Meia-Idade , Fenótipo , Receptores Imunológicos/análise , Receptores de Interleucina-2/análise , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/efeitos adversos , Linfócitos T/ultraestrutura , Transplante Autólogo
3.
Exp Hematol ; 15(10): 1048-54, 1987 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-3311785

RESUMO

Immune reconstitution after autologous bone marrow transplantation (ABMT) was studied in peripheral blood by phytohemagglutinin stimulated T-cell colony formation (CFU-TL) and by surface phenotype analysis of T-lymphocytes with monoclonal antibodies. Twenty-six patients (15 small-cell lung cancer, 5 lymphoma, 3 acute myeloid leukemia [AML], 2 germ cell cancer, and 1 melanoma) were conditioned with high-dose multiple drug combinations (plus total body irradiation in AML patients). No maintenance chemotherapy was given following treatment. Despite a rapid return to normal values of peripheral T3+, T11+ lymphocytes, the T4/T8 ratio remained below 1.0 up to 24 months after transplant, regardless of infection by cytomegalovirus (CMV). A high percentage (26% +/- 3%) of lymphocyte cells with immature phenotype (T8+, Ia+) was found during the first 6 months after transplant. Out of 84 cultures, performed in 26 patients, no growth was observed in 47 instances (22 patients) up to 28 months after grafting. Growth occurred in 37 cultures (11 patients, from 1 to 51 months after transplant) although it never reached the colony numbers of normal controls. Recombinant human interleukin-2 (rIL-2) added to lymphocyte culture induced proliferation in 8 (4 CMV-positive and 4 CMV-negative patients) out of 12 instances of no growth. In cases of depressed CFU-TL (20 cultures), rIL-2 induced a 48% and 92% increase in six CMV-positive patients and nine CMV-negative patients, respectively. These observations show that after ABMT and regardless of CMV status, defects in CFU-TL can be partially corrected by rIL-2.


Assuntos
Transplante de Medula Óssea , Interleucina-2/farmacologia , Leucócitos Mononucleares/citologia , Proteínas Recombinantes/farmacologia , Células-Tronco/citologia , Linfócitos T/citologia , Células Sanguíneas/citologia , Células Sanguíneas/efeitos dos fármacos , Humanos , Leucócitos Mononucleares/efeitos dos fármacos , Fenótipo , Células-Tronco/efeitos dos fármacos , Linfócitos T/classificação , Linfócitos T/efeitos dos fármacos
4.
Bone Marrow Transplant ; 13(3): 261-4, 1994 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-7515299

RESUMO

A pilot study was devised to assess tolerance of combined administration of interleukin-3 (IL-3) and granulocyte-colony stimulating factor (G-CSF) given after chemotherapy to mobilize peripheral blood progenitors cells (PBPC). Eight patients with advanced malignancies received 1 week courses of both IL-3 and G-CSF in one of three schedules: simultaneous 7 days administration (3 patients), sequential administration (3 patients) or partial (3 days) overlap of the two growth factors (2 patients). IL-3 (7.5 micrograms/kg/day) and G-CSF (5 micrograms/kg/day for the simultaneous schedule and 12 micrograms/kg/day for the partial overlapping and sequential schedules) were administered subcutaneously. Side-effects during cytokine administration included WHO grade I-II fever in 6 of 8 patients, flu-like symptoms (including myalgias and arthralgias) in 4 of 8, WHO grade I-II headache in 2 of 8 and WHO grade II nausea and vomiting in 1 of 8. Overall, side-effects appeared similar during combined administration of IL-3 and G-CSF to those observed during administration of IL-3 alone. No fever was observed when G-CSF was administered alone. Two leukaphereses were performed following the treatment with cytokines. Only the seven patients who received cytokines following chemotherapy were analyzed for PBPC mobilization. The median collection of CFU-GM/kg per patient in the seven analyzed patients was 1.3 x 10(5) (range 5.7 x 10(2)-3.6 x 10(5)). In two patients, a second cycle of mobilization with either granulocyte macrophage-colony stimulating factor (GM-CSF) or G-CSF was administered to allow safe engraftment.(ABSTRACT TRUNCATED AT 250 WORDS)


Assuntos
Células Sanguíneas/citologia , Transplante de Medula Óssea/métodos , Fator Estimulador de Colônias de Granulócitos/efeitos adversos , Células-Tronco Hematopoéticas/citologia , Interleucina-3/efeitos adversos , Adulto , Relação Dose-Resposta a Droga , Quimioterapia Combinada , Feminino , Fator Estimulador de Colônias de Granulócitos/administração & dosagem , Fator Estimulador de Colônias de Granulócitos/uso terapêutico , Humanos , Interleucina-3/administração & dosagem , Interleucina-3/uso terapêutico , Leucaférese , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Proteínas Recombinantes/administração & dosagem , Proteínas Recombinantes/efeitos adversos , Proteínas Recombinantes/uso terapêutico , Fatores de Tempo
5.
Anticancer Res ; 18(5A): 3347-53, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-9858907

RESUMO

Platelet production is a regulated phenomenon. Indeed, megakaryocyte volume is inversely correlated to the platelet count not only in normal individuals and immune thrombocytopenic purpura patients, but also, and surprisingly, in chronic myeloid leukemia patients. Patients with polycythemia vera and essential thrombocythemia are located outside this regression line. Herein, we describe morphometrical data confirmed by the flow cytometric measurement of the megakaryocyte endomitotic index (EI). The EI is a value which reflects the mean ploidy of megakaryocytes and corresponds to the mean of (¿log2 DNA content expressed in N¿-1). In this study, the megakaryocyte endomitotic index of 14 normal individuals was compared to those of chronic myeloid leukemia (CML) patients (n = 16), immune thrombocytopenic purpura (ITP) patients (n = 11), essential thrombocythemia (ET) patients (n = 10) and polycythemia vera (PV) patients (n = 12). The megakaryocyte EI was significantly lower in CML patients than in normal individuals. In contrast, in ET, PV and ITP patients, megakaryocyte EI was higher than in normal individuals. An inverse relationship between the endomitotic index estimated by flow cytometry and the mean megakaryocyte volume performed by morphometry was observed in normal individuals, CML and ITP patients. In conclusion, the endomitotic index is higher in ITP, ET and PV patients and lower in CML patients when compared to normal individuals and is an interesting tool which can help to diagnose rapidly hematological disorders with abnormal platelet counts.


Assuntos
Doenças Hematológicas/diagnóstico , Megacariócitos , Índice Mitótico , Ploidias , Adolescente , Adulto , Idoso , Feminino , Citometria de Fluxo , Doenças Hematológicas/sangue , Doenças Hematológicas/patologia , Humanos , Leucemia Mielogênica Crônica BCR-ABL Positiva/sangue , Leucemia Mielogênica Crônica BCR-ABL Positiva/diagnóstico , Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , Masculino , Megacariócitos/patologia , Pessoa de Meia-Idade , Contagem de Plaquetas , Policitemia Vera/sangue , Policitemia Vera/diagnóstico , Policitemia Vera/patologia , Púrpura Trombocitopênica Idiopática/sangue , Púrpura Trombocitopênica Idiopática/diagnóstico , Púrpura Trombocitopênica Idiopática/patologia , Trombocitemia Essencial/sangue , Trombocitemia Essencial/diagnóstico , Trombocitemia Essencial/patologia
6.
Anticancer Res ; 18(1A): 459-64, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-9568120

RESUMO

Megakaryocyte polyploidization is an advantageous and regulated mechanism which leads to an increase in platelet production. In megakaryocyte cell lines, polyploidization can be obtained by using cytochalasin B, an inhibitor of the actin polymerization. The Nucleolar Organizer Regions (AgNORs) are parts of nucleolar DNA transcribed into ribosomal RNA. They are detected by silver staining technique and their number is proportional to protein synthesis. In order to estimate protein synthesis in polyploidizing megakaryocytes, AgNORs were measured in three cell lines with megakaryocyte properties (DAMI, HEL and K562) after a 4-day culture in the presence or absence of cytochalasin B, an inhibitor of the actin polymerization. The mean number of AgNORs per cell was 16.4 +/- 4.3 (m +/- SEM); 24.4 +/- 2.5 and 13.6 +/- 3.1 for DAMI, HEL and K-562 cell lines, respectively. The addition of cytochalasin B (2 micrograms/ml) increased significantly the number of AgNORs per cell (DAMI: 437%, HEL: 384% and K-562: 345% of controls, p < 0.05 by t-test). Moreover, the numbers of nucleoles per cell after addition of cytochalasin B were augmented significantly (DAMI: 258%, HEL: 271% and K-562: 264% of controls, p < 0.05 by t-test). The total protein content estimated by Bradford's method increased significantly to 938%, 326% and 388% of controls in DAMI, HEL and K562, respectively (p < 0.05 by t-test) in cells where actin was inhibited by cytochalasin B. In the presence of cytochalasin B, the endomitotic index (EI) [mean of (log2 DNA content expressed in N) 1] measured by flow cytometry increased to 368%, 207% and 538%, for DAMI, HEL and K-562 cell lines, respectively (p < 0.05 by t-test) after treatment with cytochalasin B. In contrast, the number of AgNORs per unit of DNA (EI) and the total protein content per unit of DNA did not change for DAMI, HEL and K-562 cell lines (p < 0.05 by t-test) after treatment with cytochalasin B. In conclusion, the increase in the number of the Nucleolar Organizer Regions by an agent known to stimulate polyploidization of megakaryocytic cell lines suggests that polyploidization occurs by enhanced protein production proportionally to DNA synthesis.


Assuntos
Actinas/efeitos dos fármacos , Citocalasina B/farmacologia , Megacariócitos/efeitos dos fármacos , Região Organizadora do Nucléolo/efeitos dos fármacos , Poliploidia , Divisão Celular/efeitos dos fármacos , Linhagem Celular , Núcleo Celular/efeitos dos fármacos , Humanos , Megacariócitos/ultraestrutura
7.
Anticancer Res ; 18(3A): 1553-61, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-9673370

RESUMO

Megakaryocyte polyploidization responds to platelet demand and results from the lack of cytoplasmic separation while the nucleus keeps dividing. In normal telophase, the plane of the actin constriction ring is determined by the tubulin spindle. In order to investigate the role of tubulin in the megakaryocyte polyploidization, two cell lines with megakaryocyte properties (DAMI and HEL) were incubated for 4 days in the presence or absence of colchicine (10 ng/ml), an inhibitor of the tubulin spindle. As compared to control conditions, cell cultured in the presence of colchicine reveal an augmentation of cell size, the apparition of multilobed nuclei and an increase in the cytoplasm basophilia, suggesting a megakaryocyte morphology. Furthermore, when cells are cultured in the presence of colchicine, diameters measured by morphometry augment from 17.4 microns +/- 1.7 to 34.5 microns +/- 2.0 and from 27.3 microns +/- 0.3 to 40.2 microns +/- 0.6 for DAMI and HEL cell lines, respectively (p < 0.05 by t-test). After four days of culture in the presence of colchicine, cells undergo arrest proliferation. Ploidy measured by flow cytometry, shows that control cells predominantly diploid (2N) become polyploid with the appearance of 8N, 16N and 32N cells after addition of colchicine. Moreover, the endomitotic index ¿mean of (log2 DNA content expressed in N)-l¿ increases significantly from 0.5 +/- 0.1 to 1.2 +/- 0.1 and from 0.6 +/- 0.1 to 1.4 +/- 0.0 after treatment with colchicine for the DAMI and HEL cell lines, respectively. To identify the nature of the molecules involved in this phenomenon, both forms of actin (monomeric, G- and polymerized, F-) were evaluated by a DNase I inhibition assay. G-actin contents in pg per 10(6) cells are 13.0 pg +/- 2.8 (m +/- SEM) and 1.0 pg +/- 0.1 for unstimulated DAMI and HEL cells. F-actin contents per 10(6) cells are 5.8 pg +/- 1.5 and 0.1 pg +/- 0.0 for DAMI and HEL cells. The addition of colchicine for four days of culture significantly increased the G-actin content (251% and 475% of controls) and F-actin content (170% and 619% of controls) for DAMI and HEL cell lines, respectively. In contrast, the G/F-actin ratio was not affected by colchicine. DAMI cells from each ploidy class were then sorted on an ELITE Coulter and assayed for actin content. While total actin, G-actin and F-actin per cell were augmented in polyploid cells cultured with colchicine, there was a reduction in G-, F- and total actin contents per diploid equivalent when cells become polyploid. In conclusion, these data suggest that inhibition of the tubulin spindle by colchicine induces polyploidization of megakaryocytes by a reduction of both forms of actin, possibly by preventing the actin constriction ring in the telophase.


Assuntos
Actinas/metabolismo , Colchicina/farmacologia , Megacariócitos/metabolismo , Poliploidia , Tubulina (Proteína)/metabolismo , Linhagem Celular , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/ultraestrutura , Citoplasma/efeitos dos fármacos , Citoplasma/ultraestrutura , Humanos , Leucemia , Leucemia Megacarioblástica Aguda , Megacariócitos/citologia , Megacariócitos/efeitos dos fármacos , Ploidias , Tubulina (Proteína)/química , Tubulina (Proteína)/efeitos dos fármacos , Células Tumorais Cultivadas
8.
Anticancer Res ; 19(6B): 5187-9, 1999.
Artigo em Inglês | MEDLINE | ID: mdl-10697532

RESUMO

Megakaryocytes are platelet forming cells and are characterized by polyploidization, a phenomenon by which nuclear division occurs without corresponding cytoplasmic separation. Among the markers allowing to identify megakaryocytes, glycoprotein (GP) IIIa with GPIb and GPIIb are the most important. Using GPIIIa as a marker to recognize megakaryocytes in the bone marrow, we have estimated GPIIIa expression by flow cytometry in megakaryocyte populations from normal individuals and from patients with chronic myelogenous leukemia, immune thrombocytopenic purpura or polycythemia vera. We showed that the expression of GPIIIa is decreasing during megakaryocyte polyploidization in normal and pathological situations.


Assuntos
Antígenos CD/metabolismo , Megacariócitos/metabolismo , Glicoproteínas da Membrana de Plaquetas/metabolismo , Poliploidia , Citometria de Fluxo , Humanos , Integrina beta3 , Leucemia Mielogênica Crônica BCR-ABL Positiva/metabolismo , Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , Megacariócitos/patologia , Policitemia Vera/metabolismo , Policitemia Vera/patologia , Púrpura Trombocitopênica/metabolismo , Púrpura Trombocitopênica/patologia
9.
Anticancer Res ; 19(5B): 4193-8, 1999.
Artigo em Inglês | MEDLINE | ID: mdl-10628374

RESUMO

Megakaryocytes undergo a peculiar and irreversible program by which they become polyploid through repeated cycles of DNA synthesis without concomitant cell division. In order to study the possible concomitant role of protein kinase C and actin in megakaryocyte polyploidization, three cell lines (DAMI, HEL and K562), expressing some properties of the megakaryocytic lineage and known to differentiate into the megakaryocytic pathway in the presence of phorbol esters, were cultivated in the presence of phorbol myristate acetate alone (PMA, 5 x 10(-9) M, activator of protein kinase C, PKC) or concomitantly with cytochalasin B (2 micrograms/ml, inhibitor of actin polymerization). We have previously shown that DAMI, HEL and K562 cells in which actin polymerization was inhibited by cytochalasin B, acquired megakaryocytic properties in the way that they became polyploid, acquired a megakaryocytic phenotype and arrested proliferation (4). After four days of culture in the presence of PMA and cytochalasin B, the number of polyploid cells (estimated by flow cytometry) increased in comparison with control or PMA-treated cells. However, it was lower than in cytochalasin B-treated cells. Indeed, control cells predominantly diploid (2N) became polyploid with the appearance of 8N, 16N and 32N cells after addition of PMA, cytochalasin B or PMA + cytochalasin B. The endomitotic index (EI, as described in 5) which corresponds to the mean of (¿log2 DNA content expressed in N¿-1) was 0.5 +/- 0.1, 0.7 +/- 0.1 and 0.3 +/- 0.1 in control DAMI, HEL and K562 cells, respectively. The EI increased to 0.9 +/- 0.2; 1.0 +/- 0.2 and 0.4 + 0.1 in cells treated with PMA and to 1.6 +/- 0.3; 1.4 +/- 0, and 0.9 +/- 0.2 when PMA was added concomitantly to cytochalasin B. Total DNA estimated from the cell content and the percentage of cells present at each ploidy stage did not change in cytochalasin B-treated cells in comparison to control conditions. However, treatment of DAMI, HEL and K562 cells with PMA alone or concomitantly with cytochalasin B revealed that the total DNA content significantly decreased in these conditions. At last, treatment of the three cell lines with PMA alone or concomitantly with cytochalasin B for 4 days caused a complete inhibition of proliferation. In conclusion, the concomitant addition of PMA and cytochalasin B to the three cell lines lead to an augmentation of cell ploidy and to a cessation of proliferation. However, we did not observe any synergistic effect of the two compounds. The possible interaction between actin and protein kinase C is discussed in the paper.


Assuntos
Actinas/metabolismo , Megacariócitos/patologia , Proteína Quinase C/metabolismo , Carcinógenos/farmacologia , Divisão Celular/efeitos dos fármacos , Citocalasina B/farmacologia , Citometria de Fluxo , Humanos , Leucemia Megacarioblástica Aguda/enzimologia , Leucemia Megacarioblástica Aguda/genética , Leucemia Megacarioblástica Aguda/patologia , Megacariócitos/enzimologia , Microscopia de Contraste de Fase , Poliploidia , Acetato de Tetradecanoilforbol/farmacologia , Células Tumorais Cultivadas
10.
Anticancer Res ; 19(4B): 3229-35, 1999.
Artigo em Inglês | MEDLINE | ID: mdl-10652616

RESUMO

In order to investigate the protein synthesis in megakaryocyte polyploidization, phorbol myristate acetate (PMA, 5 x 10(-9) M), a differentiation marker known to induce megakaryocyte polyploidization, was added to human megakaryocytic cell lines (DAMI, HEL and K562) and the expression of platelet/megakaryocytic integrins, the numbers of nucleolar organizer regions (AgNORs) and the total protein content were estimated. Following exposure of PMA, the expression of the platelet membrane glycoprotein GPIIIa and thrombospondin and transferrin receptors was augmented in the three cell lines. The number of AgNORs shifted from 16.4 +/- 4.3, 24.4 +/- 2.5 and 13.6 +/- 3.1 for unstimulated cells to 20.0 +/- 5.3, 38.7 +/- 7.9 and 16.8 +/- 2.3 for PMA-treated DAMI, HEL and K562 cells, respectively. Furthermore, after treatment with PMA, the numbers of AgNORs clusters or nucleoles increased significantly to 179%, 238% and 154% of controls in DAMI, HEL and K562 cell lines, respectively. Finally, addition of PMA culture for four days, significantly increased the protein contents to 153%, 171% and 254% of controls for DAMI, HEL and K562 cell lines, respectively (p < 0.05 by t-test). In conclusion, the increase in the total protein content and in the number of AgNORs by PMA, suggests that PMA-induced-megakaryocyte polyploidization occurs by enhanced protein production.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Megacariócitos/efeitos dos fármacos , Região Organizadora do Nucléolo , Glicoproteínas da Membrana de Plaquetas/metabolismo , Acetato de Tetradecanoilforbol/farmacologia , Células Cultivadas , Humanos , Megacariócitos/citologia
11.
Anticancer Res ; 19(4B): 3259-64, 1999.
Artigo em Inglês | MEDLINE | ID: mdl-10652621

RESUMO

Megakaryocyte polyploidization responds to platelet demand and results from the lack of cytoplasmic separation while the nucleus keeps dividing. In order to investigate the role of actin in the megakaryocyte polyploidization, phorbol myristate acetate (PMA, 5 x 10(-9) M), a differentiation marker known to induce megakaryocyte polyploidization, was added to human megakaryocytic cell lines (DAMI and HEL) and G, F and total actins were estimated by DNase I inhibition. After four days of culture in the presence of PMA, G actin contents in pg per 10(6) cells were 13.0 pg +/- 2.8 and 1.0 pg +/- 0.1 for unstimulated DAMI and HEL cells. F actin contents per 10(6) cells were 5.8 pg +/- 1.5 and 0.1 pg +/- 0.0 for DAMI and HEL cells. Addition of PMA for four days to culture significantly increased G actin contents (235% and 268% of controls) and F actin contents (234% and 394%), for DAMI and HEL cell lines, respectively (p < 0.05 by t-test). In contrast, G/F actin ratio was not affected (p < 0.05 by t-test) by PMA. DAMI cells from each ploidy classes were then sorted on an ELITE Coulter and assayed for actin content. While total actin, G actin and F actin per cell increased in polyploid cells cultured with PMA, there was a reduction in G, F and total actin contents per diploid equivalent when cells became polyploid. In conclusion, megakaryocyte polyploidization of these cell lines is not related to an unbalance between G and F actins but would be rather due at least partly to a defect in total actin production that could lead to a prevention of the formation of the constriction ring in telophase.


Assuntos
Actinas/metabolismo , Megacariócitos/efeitos dos fármacos , Acetato de Tetradecanoilforbol/farmacologia , Células Cultivadas , Citometria de Fluxo , Humanos , Megacariócitos/metabolismo
12.
Arch Pediatr ; 17(10): 1451-4, 2010 Oct.
Artigo em Francês | MEDLINE | ID: mdl-20851581

RESUMO

Subgaleal hematoma in the newborn infant is rare, occurs early, and often bears serious consequences. We report on 2 subacute cases of bruising of the scalp that occurred following the use of a suction cup. Emergency treatment consisted of a transfusion of packed red blood cells and fresh frozen plasma. Children born by use of vacuum extractor or forceps require careful monitoring by the nursing staff throughout their stay in the maternity unit.


Assuntos
Hematoma/etiologia , Couro Cabeludo/irrigação sanguínea , Encéfalo/diagnóstico por imagem , Humanos , Recém-Nascido , Masculino , Forceps Obstétrico/efeitos adversos , Couro Cabeludo/diagnóstico por imagem , Tomografia Computadorizada por Raios X , Vácuo-Extração/efeitos adversos
15.
Haematologia (Budap) ; 29(3): 213-28, 1998.
Artigo em Inglês | MEDLINE | ID: mdl-10069447

RESUMO

A flow cytofluorometric measurement of megakaryocyte ploidy has been adapted from Tomer's method. Briefly, bone marrow is aspirated through a medium containing theophyllin, prostaglandin-E1 and other antiaggregant agents. Megakaryocytes-enriched buffy coats are recovered. Megakaryocytes are then stained for GP IIIa coupled with FITC and for DNA (propidium iodide). A Becton Dickinson FACScan flow cytometer is used for measuring the ploidy distribution of GP IIIa positive cells. The method we developed presents several advantages. Firstly, the time required is greatly decreased in comparison with other studies. Secondly, the washing steps are limited in number allowing a diminution of the cell loss. Thirdly, the method ensures a better megakaryocyte preservation. Finally, selection of megakaryocytes by ploidy and expression of GP IIIa can be made easily because of the simultaneity of the two stainings as well as by the use of a precise gating on the flow cytometer. Based on these results, we conclude that the present method provides a better means for the isolation and analysis of human normal megakaryocytes. This technique has been applied to the analysis of megakaryocyte populations from patients with abnormal platelet counts. In chronic myeloid leukemia patients, analysis of ploidy distribution shows a shift toward the low ploidy while in patients with immune thrombocytopenic purpura, polycythemia vera and essential thrombocythemia the ploidy distributions are shifted toward the high ploidy.


Assuntos
Antígenos CD/análise , Citometria de Fluxo/métodos , Megacariócitos , Glicoproteínas da Membrana de Plaquetas/análise , Ploidias , Adolescente , Adulto , Idoso , Feminino , Humanos , Integrina beta3 , Masculino , Pessoa de Meia-Idade
16.
Eur J Clin Invest ; 28(2): 138-44, 1998 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-9541128

RESUMO

BACKGROUND: Megakaryocyte polyploidization is an advantageous and regulated mechanism that leads to an increase in platelet production. In megakaryocytic cell lines, polyploidization can be obtained by using colchicine, an inhibitor of the tubulin spindle. The nucleolar organizer regions (AgNORs) are parts of nucleolar DNA transcribed into ribosomal RNA and are detected by the silver-staining technique. Their number is proportional to protein synthesis. RESULTS: To estimate protein synthesis in polyploid megakaryocytes, AgNORs are measured in three cell lines with megakaryocyte properties (DAMI, HEL and K562) after a 4-day culture in the presence or absence of colchicine. The mean number of AgNORs per cell was 16+/-4 (mean+/-SEM), 24+/-3 and 14+/-3 for DAMI, HEL and K-562 cell lines respectively. The addition of colchicine (10 ng mL[-1]) significantly increased the number of AgNORs per cell (DAMI 556%, HEL 338% and K-562 300% of controls, P < 0.05 using the t-test). Moreover, the number of nucleoles per cell after the addition of colchicine was augmented significantly (DAMI 246%; HEL 237% and K-562 148% of controls, P < 0.05 using the t-test). The total protein content estimated by Bradford's method increased significantly to 226%, 215% and 304% of controls in DAMI, HEL and K562 respectively (P < 005 using the t-test). After treatment with colchicine, the endomitotic index (EI) [mean of (log2 DNA content expressed in N)-1] measured by flow cytometry (and reflecting ploidy) increased to 234%, 255% and 301%, respectively, in DAMI, HEL and K-562 cell lines (P < 0.05 using the t-test). Concomitantly, the number of AgNORs per unit of DNA increased in the DAMI and HEL cell lines (P < 0.05 using the t-test) from 48+/-8 and 39+/-5, respectively, to 79+/-11 and 61+/-10. In contrast, the number of nucleoles and the total protein content per endomitotic index were not affected by colchicine (P > 0.05 using the t-test), but the number of nucleoles per endomitotic index of DAMI cells was affected. CONCLUSION: The increase in the number of the NORs induced by an agent known to stimulate polyploidization of megakaryocytic cell lines suggests that polyploidization occurs by a proportional increase in protein synthesis per DNA unit.


Assuntos
Colchicina/farmacologia , Megacariócitos/efeitos dos fármacos , Microtúbulos/efeitos dos fármacos , Região Organizadora do Nucléolo/efeitos dos fármacos , Poliploidia , Contagem de Células/efeitos dos fármacos , Nucléolo Celular/efeitos dos fármacos , Humanos , Leucemia Megacarioblástica Aguda , Leucemia Mielogênica Crônica BCR-ABL Positiva , Megacariócitos/metabolismo , Megacariócitos/patologia , Microtúbulos/genética , Região Organizadora do Nucléolo/genética , Proteínas/efeitos dos fármacos , Coloração pela Prata , Células Tumorais Cultivadas
17.
Eur Respir J ; 1(6): 498-504, 1988 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-2458961

RESUMO

The present study evaluates different cellular and soluble components in the bronchoalveolar lavage (BAL) from patients with interstitial lung disease. We observed an increased T4/T8 lymphocyte ratio in BAL but not in blood from 24 patients with active pulmonary sarcoidosis compared to sixteen normal individuals and to eleven patients with inactive pulmonary sarcoidosis. Seven patients with hypersensitivity pneumonitis had a normal T4/T8 ratio. In the active sarcoidosis and hypersensitivity pneumonitis groups, alpha 1-Protease Inhibitor (alpha 1 PI) in BAL is significantly higher than in the normal group and a significant correlation between the two antiproteases (alpha 2-macroglobulin and alpha 1 PI) is observed. These data demonstrate that antiprotease levels (alpha 1 PI and alpha 2 M) are increased in the lower respiratory tract of patients with interstitial lung disease and that among cellular and soluble components of BAL, alpha 2 M represents a sensitive marker of the alveolitis.


Assuntos
Alveolite Alérgica Extrínseca/diagnóstico , Proteínas Sanguíneas/análise , Líquido da Lavagem Broncoalveolar/análise , Pneumopatias/diagnóstico , Inibidores de Proteases/análise , Sarcoidose/diagnóstico , alfa-Macroglobulinas/análise , Líquido da Lavagem Broncoalveolar/citologia , Humanos , Linfócitos T Auxiliares-Indutores/classificação , Linfócitos T Reguladores/classificação , alfa 1-Antitripsina
18.
Eur J Clin Invest ; 28(10): 845-55, 1998 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9792999

RESUMO

BACKGROUND: Megakaryocyte polyploidization results from the lack of cytoplasmic separation while the nucleus keeps dividing. METHODS: To investigate the role of actin in the megakaryocyte polyploidization, three human cell lines with megakaryocytic properties (DAMI, HEL and K562) were incubated in the presence of cytochalasin B, an inhibitor of actin polymerization. These data were then compared with normal megakaryocytes. RESULTS: Compared with control conditions, cells cultured in the presence of cytochalasin B revealed an augmentation of cell size and ploidy and an arrest of cell proliferation. The expression of platelet membrane glycoproteins Ib, IIb/IIIa, IIIa and thrombospondin and transferrin receptors was augmented after treatment with cytochalasin B. Physiologically, the role of actin in inducing polyploidization could be related to an imbalance between G- and F-actins. To test this hypothesis, we measured G-, F- and total actin in cytochalasin B-treated cells. Actin was found to be increased significantly in cytochalasin B-treated DAMI and HEL cell lines. In contrast, the G/F-actin ratio was not affected by cytochalasin B. To confirm these actin changes in physiological megakaryocytopoiesis, G- and F-actin contents were then estimated in normal megakaryocytes. The G- and F-actin contents of megakaryocytes from eight normal patients exponentially decreased from 2 to 128n, whereas the total actin content per cell kept increasing. The G/F ratio was unaffected. CONCLUSION: Polyploidization of human megakaryocytes results from either a diminution of actin synthesis or an increased actin turnover, which in turn possibly abrogates the formation of the actin cleavage furrow in telophasis.


Assuntos
Actinas/metabolismo , Megacariócitos/ultraestrutura , Poliploidia , Actinas/análise , Actinas/genética , Células da Medula Óssea/química , Divisão Celular , Linhagem Celular , Tamanho Celular , Humanos , Megacariócitos/química , Megacariócitos/fisiologia , RNA Mensageiro/análise
19.
Acta Psychiatr Scand ; 92(4): 294-300, 1995 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-8848955

RESUMO

Decreased immunity in depressive as compared with control subjects has been well documented, although some depressed patients have severe alterations whereas others have milder ones or not at all. Since for equal severities of depression, there may be individual differences in the degree of perceived control over one's condition, we investigated the interaction of perceived control with immunological variations. Immune function (T and B lymphocytes, lymphocyte proliferation and natural killer cell activity (NKCA)) were evaluated in 34 adult major depressives and in 18 healthy controls. Lymphocyte proliferation did not differ between the two groups, but NKCA was significantly lower in the depressed patient group. Among the depressed subjects, those who experienced less subjective control also showed significantly lower NKCA. An internal locus of control appears to act as a buffer against the decrease in cellular immunity observed in major depression. Further studies should focus on methods of coping and on degree of perceived control rather than on diagnostic and nosographic variables alone.


Assuntos
Transtorno Depressivo/imunologia , Controle Interno-Externo , Células Matadoras Naturais/imunologia , Adulto , Linfócitos B/imunologia , Movimento Celular , Feminino , Humanos , Células Matadoras Naturais/fisiologia , Masculino , Pessoa de Meia-Idade , Fenótipo , Linfócitos T/imunologia
20.
Eur Respir J ; 7(6): 1111-9, 1994 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-7925881

RESUMO

Immunoglobulin A (IgA) and alveolar macrophages are two important components of the immune system in the respiratory tract. Fc alpha-receptors (Fc alpha R) are present on neutrophils, eosinophils and a series of human mononuclear phagocytes, including monocytes, alveolar macrophages and leukaemia cell lines (U-937). In the present study, using idiotypes and anti-idiotypic antibodies, we report that THP-1 cells, a myelomonocytic cell line, constitutively express Fc alpha R and that all IgA preparations used bind the receptor. Of the stimuli used (phorbol myristate acetate, retinoic acid, calcitriol), only calcitriol can induce differentiation of THP-1 cells, as assessed by CD14 expression. The expression of Fc alpha R appears to be independent of cell differentiation, since calcitriol pretreatment has no effect on IgA-binding. Finally, My43, a monoclonal antibody recognizing the Fc alpha R on U-937 cells, does not bind to THP-1 cells or to human alveolar macrophages. In addition, preincubation of THP-1 cells or human alveolar macrophages with My43 does not diminish IgA-binding to these cells. Ribonucleic acid (RNA) encoding the Fc alpha R isolated from U-937 is expressed, although possibly at a lower level, in alveolar macrophages and THP-1 cells. In conclusion, Fc alpha R are constitutively expressed on THP-1 cells and share some characteristics with the Fc alpha R described in human alveolar macrophages. THP-1 cells, therefore, may represent a reasonable model for further investigation of the interaction of immunoglobulin A and tissue macrophages.


Assuntos
Imunoglobulina A/imunologia , Macrófagos Alveolares/imunologia , Monócitos/imunologia , Receptores Fc/biossíntese , Sequência de Bases , Ligação Competitiva , Diferenciação Celular/imunologia , Linhagem Celular , Citometria de Fluxo , Imunofluorescência , Humanos , Imunoglobulina G/imunologia , Dados de Sequência Molecular , Reação em Cadeia da Polimerase
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