From October 1987 to June 1992, 359 renal transplantations were performed, while, over the same period, 25 patients with a follow-up of more than six months underwent repeat renal transplantation: 23 for a second transplant and 2 for a third transplant. The initial disease was glomerular for 17 patients and interstitial for 6 patients. The mean age of the patients at the time of the repeat transplantation was 36.9 years (range: 20 to 53 years). The mean survival after the first transplantation was 3 years and 10 months (range: 1 week to 10 years). The reasons for loss of the first transplant can be classified as follows: acute rejection (n = 5), chronic rejection (n = 14), surgical failure (n = 5) or sepsis (n = 1). 9 patients received conventional immunosuppressant therapy, while 16 patients (64%) received four-drug therapy including cyclosporin. The actuarial one-year survival of the patients and the transplants was 100% and 92%, respectively. The mean serum creatinine was 136.4 +/- 65 mumol/l (range: 59 to 298 mumol/l). Ten patients developed rejection after a mean of 18.6 days (range: 6 to 30 days) and 2 patients suffered from 2 episodes of rejection within 4 months. These results illustrate the low postoperative surgical and immunological complication rate in this group of patients whose long-term results are at least comparable to those of first transplantations in our group.

Actuarial Analysis↗

PubMedDate not supplied

High-dose cytotoxic therapy with autologous bone marrow or peripheral blood progenitor cell transplantation in malignant lymphomas.

The present paper is an attempt to assess the efficiency of high-dose cytotoxic therapy followed by autologous bone marrow or peripheral progenitor cell rescue with hematopoietic growth factor support given in a group of 27 patients (16 men, 11 women) at the Department of Hematology of the Mont Godinne University Clinics, mainly in the same interval 1990-1994. The reasons for introducing such a therapy in these patients (6 with Hodgkin's disease, 14 with intermediate or high grade, aggressive non Hodgkin lymphomas and 7 with low grade follicular non Hodgkin lymphomas) were relapse of disease after conventional therapy (11 cases), resistance to initial therapy (5 patients) or because of histologically proven transformation to a more aggressive form (one case); in 10 patients with extended, poor prognosis forms, the procedure was used as part of the first line therapy. The conditioning high dose chemotherapy was given according to various regimens, most of them containing Cyclophosphamide, BCNU and Etoposide, with or without total body irradiation. In 14 patients, bone marrow (BM) graft was used, while peripheral blood progenitor cells (PBPC) were infused in the remaining 13 patients. The number of infused granulocyte-macrophage colony forming units (CFU-GM) ranged between 7,650 and 3,900,000/kg, with a mean value of 461,000/kg. The median time intervals required to reach an absolute neutrophil count > 500/microliter, a platelet count > 50,000/microliter and a hematocrit > 30% were 13 days, 20 days and 23 days respectively. Growth factors (GM-CSF and G-CSF) and PBPC use shortened the time for neutrophil recovery as well as neutropenia-related complications. No procedure-related death was observed and complete remission was achieved in 22 cases (81.4%); after a mean follow-up of 32.6 months, 14 patients (55.5%) are alive and free of disease, while in 7 patients (31% of the complete responders) relapse occurred at an average time interval of 8.2 months since the procedure.

Adult↗

PubMedDate not supplied

[Kidney transplantation from living related donors: experiences from one center].

Living related donor (LRD) renal transplantation has remained underdeveloped in France up until now. The reduction of the number of available grafts and especially the superiority of the results of LRD transplants recently led us to develop this type of transplantation. We present the retrospective analysis of our experience of 63 cases from March 1973 to June 1995. The actuarial graft survival rate was 91% at 1 year and 87% at 3, 5 and 10 years. The 5- and 10-year survival rates of HLS-identical transplants (n = 17) was 100%. The donor morbidity was minimal (2 cases of parietal suppuration, 1 pulmonary atelectasis). These results emphasize the superiority of LRD transplantation, which also has the advantage of allowing transplantation of hyperimmunized patients in whom the waiting time for a brain-dead donor kidney is long and unpredictable.

Actuarial Analysis↗

PubMedDate not supplied

[Monseur's urethroplasty in extensive sclero-inflammatory stenoses of the urethra: an operation of the future or already outdated? Apropos of 21 cases].

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Evaluation Studies as Topic↗

PubMedDate not supplied

Inhibition of actin polymerization by cytochalasin B induces polyploidization and increases the number of nucleolar organizer regions in human megakaryocyte cell lines.

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.

Actins↗

PubMedDate not supplied

Inhibition of tubulin polymerization in megakaryocyte cell lines leads to polyploidization which affects the metabolism of actin.

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.

Actins↗

PubMedDate not supplied

[Evaluation of a dot-blot technique for the detection of Epstein-Barr virus antibodies (IgG anti-viral capsid antigen)].

Infectious mononucleosis syndrome is caused by several infectious agents, including Epstein-Barr virus (EBV), cytomegalovirus (CMV), and Toxoplasma gondii. The ImmunoDot EBV VCA-IgG test (Biomedical Diagnostics) is a dot-blot enzyme-immunoassay, which determines the presence of heterophile antibodies and IgG antibodies directed against Epstein-Barr viral capsid antigen (anti-VCA), CMV and T. gondii. This test is simple, unitary, ready-to-use, and rapid (40 minutes), requiring approximately 10 microliters of serum or plasma or 20 microliters of whole blood. The aim of this study was to compare the results of this technique for the determination of anti-VCA IgG antibodies with those obtained by the reference technique using indirect immunofluorescence on 185 serum specimens. A sensitivity of 99.2%, a specificity of 92.4%, a correlation with indirect immunofluorescence of 97.3%, a the absence of cross-reactions with CMV, T. gondii, and herpes simplex viruses and the absence of interference with antinuclear antibodies were the main results of this comparative study. In association with another test (Monolert 2TM) detecting IgM and IgG antibodies against Epstein-Barr nuclear antigen, the dot-blot method represents a useful screening strategy for EBV response.

Antibodies, Viral↗

PubMedDate not supplied

Endomitotic index of megakaryocytes measured by flow cytometry helps to diagnose hematological disorders with abnormal platelet counts.

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.

Adolescent↗