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Biomedical subjects

D Hollard

Publications and source records attributed to D Hollard.

At least 37 records · Page 2Linked to original sources

Splenectomy during chronic lymphocytic leukemia.

The combination of discontinuous high-dose chlorambucil therapy with splenectomy greatly increased the prognosis of aggressive forms of chronic lymphocytic leukemia (CLL). The median survival for 43 patients was 84 months from diagnosis and 48 months from splenectomy. For 15 stages O, according to Rai classification, obtained after splenectomy, duration ranged from 3 to 105 months. The median survival of a group of patients showing "nodular splenic infiltration" was 104 months and superior to that of a group of patients showing "diffuse splenic infiltration" (72 months). In four of 15 cases studied, the peripheral blood lymphocytic clone disappeared after splenectomy.

Adult↗

Megakaryocytic development in liquid cultures of cryopreserved leukocyte stem cell concentrates from chronic myelogenous leukemia patients.

The proliferation and differentiation of human megakaryocytes in liquid culture has been obtained using cryopreserved light-density blood cell concentrates from chronic myelogenous leukemia (CML) patients. A large number of megakaryocytes, representing 20%-60% of total cells cultured, developed after 12-14 days in liquid cultures supplemented with human plasma, while fetal calf serum supported the development of cells of the megakaryocytic lineage poorly. Ploidy studies showed the presence of 8N and 16N cells in human plasma-supplemented cultures while very few cells with DNA content greater than 4N were found in those supplemented with fetal calf serum. Using the FACS IV cytofluorometer, 1-2 X 10(6) megakaryocytes/h were sorted after immunolabeling of the human plasma-cultured cells with a monoclonal antibody reacting against the platelet glycoprotein complex IIb-IIIa. Thus, cryopreserved CML blood stem cell concentrates seem to offer a reproducible source of human megakaryocytes that retain their capacity to proliferate and differentiate in liquid cultures. These megakaryocytes can be used for the study of platelet glycoprotein biosynthesis as well as the regulation of megakaryocytopoiesis.

Blood Specimen Collection↗

[Myelofibrosis. Pathology of the microenvironment].

Myelofibrosis is the most representative model of the association between the hemopoiesis failure end the abnormalities of the myeloid microenvironment. Long term culture in Dexter's liquid system is a good model for the in vitro study of myeloid differentiation. An experimental in vitro system using an irradiated under layer and aplastic patients plasma has yielded some preliminary results allowing the production of megakaryocytic colonies. Close contact between these colonies and the fibroblasts or stromal cells in the adherent layer could probably be used for the study of the interrelation between the stromal abnormalities and myeloid pathology.

Hematopoietic Stem Cells↗

[Malignant non-Hodgkin's lymphoma associated with a serum monoclonal immunoglobulin].

Among the 394 non-Hodgkin's lymphomas (NHL) observed at our hospital over a thirteen year period, we have found 17 associated monoclonal gammopathies (4.3%). Fourteen gammopathies could be correlated to the lymphoid tumor due to a study of immunological membrane markers and to the comparative evolution of the disease and the monoclonal gammopathy. Two different groups could be distinguished: Six patients out of the fourteen studied (43%) had a gammopathy similar to the surface immunoglobulins of the tumour lymphoid cells. Seven of the seventeen showed, after treatment, a parallel evolution between the tumor and the gammopathy. This originates directly from the tumoral secretion and deserves to be classified among the B lymphoid excretory tumors. Six were IgM and one was IgG. Eight of the fourteen patients studied (57%) did not show any immunological relation between the lymphoma and the monoclonal gammopathy. The evolutions (10 of the 17 patients) of the tumor and of the gammopathy were independent. Among the eleven gammopathies (1 double gammopathy), 5 were IgG, 4 were IgA and 2 were IgM. The incidence of these gammopathies related to the total group tf lymphomas is 2.5%. Given the age of these patients, the association seems fortuitous.

Adult↗

A prospective randomized study comparing the efficacy of Timentin alone or in combination with amikacin in the treatment of febrile neutropenic patients.

One hundred febrile episodes in neutropenic (PMN less than 500/mm3) patients were treated with Timentin alone or in combination with amikacin. The overall response rate in 87 episodes was 82.9% with Timentin alone and 84.5% with the combination. Eleven out of 15 patients with septicaemia were cured by Timentin alone and 12 out of 13 by the combination, a response rate of 73.3% and 92.4% respectively (not-statistically significant P = 1.307). The rates of superinfection were low. Few side effects occurred. Timentin is a useful antibiotic in the treatment of febrile neutropenic patients and was, in this study, as effective alone as in combination with an aminoglycoside in the initial therapy.

Adolescent↗

Monoclonal antibodies against human hemopoietic cells and the separation of progenitor cells from bone marrow.

Monoclonal antibodies against myeloid cell surface antigens were selected according to their reactivity against normal human bone marrow cells. Each of the antibodies detected a different population of maturing bone marrow cells, but almost none of the progenitor cells assayed (seven-day GM-CFC, 14-day GM-CFC, CFU-E, BFU-E). Myeloid cells, erythroid cells, and the majority of lymphocytes and monocytes were simultaneously depleted with the mixture of these three antibodies using fluorescence-activated cell sorting (FACS) or panning for cell separation; 14-day GM-CFC and BFU-E were enriched six- to 16-fold when negative cells were sorted. After a negative selection step using panning, BFU-E and 14-day GM-CFC were enriched four- to seven-fold and five- to 13-fold, respectively. Negative cells obtained after panning or cell sorting were also enriched in blast cells (24.4% and 27.2%, respectively) and depleted in maturing bone marrow cells, with the exception of variable numbers of lymphocytes, monocytes, and plasma cells. FACS of negative cells using both forward light-scatter and perpendicular light-scatter parameters resulted in a cell population that contained a majority of undifferentiated blasts and 5.5%-9.6% BFU-E and 14-day GM-CFC.

Antibodies, Monoclonal↗

Enrichment of high proliferation potential colony forming cells from mouse marrow by selecting low-density cells expressing receptors for wheat germ agglutinin.

Suspensions enriched for high proliferation potential colony forming cells (HPP-CFC's) were prepared from mouse marrow by selecting low-density cells stained with fluoresceinated wheat germ agglutinin on a fluorescence activated cell sorter. HPP-CFC's were 12-fold more concentrated in the final suspensions and co-enriched with a population of primitive CFUs detected in the spleens of irradiated mice reconstituted 13 days earlier with marrow. This is consistent with previous observations suggesting that these populations are closely related. The degree of enrichment for other haemopoietic progenitors was in the order HPP-CFC greater than day 8 CFUs greater than BFUe greater than GM-CFC greater than CFUe. "Extra" colonies developed when human placental conditioned medium (HPCM) was added to cultures of enriched suspensions already containing pregnant mouse uterus extract (PMUE). HPP-CFC's probably formed most of these and we discuss why counting these extra colonies may be more reliable than the conventional "size" assay for HPP-CFC's.

Animals↗

[Refractory anemia with an excess of blasts: evidence for 2 evolving groups in a group of 56 patients].

Cytological and kinetic findings in 56 patients with refractory anemia with excess blasts (RAEB) were studied. The different statistical methods used produce similar results: our group of patients can be separated by cytological parameters in two populations (overt leukemic or hematologically stable evolution). Different functional lesions may be responsible for these two types; pluripotential stem cell dysfunction in the first case and abnormality of the granulocytic committed stem cell in the second. A score, based on a small number of simple and reliable parameters, is able to distinguish these two populations.

Adolescent↗

Cryopreservation of human bone marrow cells by a modification of the two-step cooling technique.

The first attempt to freeze human bone marrow cells with a two-step cooling method is reported. A simple and reliable way of obtaining stable first-step subzero freezing baths is described. One-milliliter samples each containing 20 X 10(6) bone marrow cells and 10% Me2SO were frozen in polypropylene cryotubes. Using these experimental conditions, the optimal freezing temperature was found to be in the range of -36 to 37.5 degrees C for BM progenitor cell (GM-CFC, CFUE, and BFUE) survival. Such temperatures were easily obtained in stable sludges of anisole or K2CO3 eutectic solution in water. The optimal holding time was 20 min before plunging tubes into liquid nitrogen. Similar or improved progenitor cell recoveries were observed compared with the conventional cooling technique. Adaptation of this two-step technique for the freezing of large volumes of BM cells for autografting is under investigation.

Anisoles↗

[Non-gonadic complications of chemoradiotherapy in Hodgkin's disease. A study of eighty-three patients (author's transl)].

Eighty-three patients with Hodgkin's disease were treated with a combination of chemotherapy and radiotherapy. 43 were included in protocol 1 (from january 1970 to january 1974) and 40 in protocol 2 (february 1974 to december 1977). In protocol 1, staging laparotomy was not systematically performed (20 cases). Treatment consisted of 2 intravenous injections in Vinblastine and total nodal irradiation. In protocol 2, laparotomy was systematic in patients over 50 (35 cases). Patients with stages 1 and II treated as mentioned above. Patients with stage III received two Mopp courses followed by total nodal irradiation. Patients older than 50 with stages I and II and poor prognosis factors received chemotherapy only. Laparotomy was associated with a 0% mortality rate and a 3,6% morbidity rate. No myelitis or pericarditis were observed. Herpes zoster occurred in 24% of the patients, pulmonary apex fibrosis in 6%, hypothyroiditis in 2,4%, and leucopenia in 3,6%. Two late infectious complications were fatal. No solid tumor was apparent. Acute leukemia and non-Hodgkin malignant lymphoma developed in two patients. Good tolerance, shortness of treatment, and remission rate, warrant the pursuit of protocol 2 in which systematic laparotomy for patients under 50 allows total nodal irradiation and therefore reduction of chemotherapy.

Adolescent↗

Non-Hodgkin's malignant lymphomas: a cytologic classification based on statistical analysis and correlated with prognosis.

On the basis of cytologic studies of 144 patients with non-Hodgkin's malignant lymphomas (NHML), a cytologic classification was established that was composed of three groups (1, 2 and 3) containing nine classes (1A, 1B, 1C; 2A, 2B, 2C; 3A, 3B, 3C). Statistical analyses were carried out using 52 well-defined elementary cytologic characteristics. All the results given by the data-gathering procedures (hierarchical and dynamic clustering methods), by the descriptive analyses (multifactorial and canonical analysis) and by the stepwise discriminant analysis were in agreement with the proposed cytologic classification system. The most discriminating cytologic characteristics of the classes were the cell size, the nuclear-cytoplasmic ratio, the degree of cytoplasmic basophilia, the homogeneity of the cytoplasm and the number of mitoses. The use of these properties renders this classification system reproducible and applicable to clinical practice. A comparison was made between the cytologic classes and the NHML patients as grouped according to their clinical courses. Of the patients in classes 2A, 2B and and 2C, 94% showed acute leukemic characteristics. Of those contained in classes 3A, 3B and 3C, 78% showed poor prognosis or metastatic patterns. Of those contained in classes 1A, 1B and 1C, 76% showed a good prognosis pattern.

Adolescent↗

Purification and cryopreservation of granulomonocytic colony forming cells (GM-CFC) from the blood of patients with chronic granulocytic leukemia (CGL) for autologous transplantation.

A simple density-cut separation technique is described for obtaining GM-CFC concentrates from the peripheral blood of patients with chronic granulocytic leukemia (CGL) for autografting at the acute blastic phase of the disease. Large numbers of granulomonocytic colony forming cells (GM-CFC) were recovered from a single procedure (68.4 X 10(6) GM-CFC). The cryopreservation of these concentrates in liquid nitrogen allowed the recovery of 46.6 +/- 33.8% viable GM-CFC. An improved yield of GM-CFC was obtained by avoiding washing the cells (65.2 +/- 41.1%). The separation technique resulted in a concentrated suspension of progenitors in a small volume of medium (mean = 15 ml) allowing a great reduction in the amount of the cryoprotective agent injected into the patient at autografting. Preliminary data obtained in 4 patients transfused in blastic crisis after chemotherapy, with or without TBI, indicated the capacity of stored concentrates to repopulate these patients.

Adult↗

Similarity of CSF of media conditioned by different human tissues.

Conditioned media prepared using human placenta, spleen, bone marrow and peripheral blood leucocytes revealed a common pattern of two distinct species of colony-stimulating factors (CSF) separable by gel filtration. The peak of greatest activity, active against both human and mouse marrow, had an apparent molecular weight (MW) of 24,000-28,000 Daltons. A peak of low activity detected only against mouse marrow had an apparent MW of approximately 150,000 Daltons. The type of progenitor cells stimulated by the crude medium, by the low MW CSF species, and by the high MW species were similar for the four conditioned media despite their different origins. No difference was found in either the MW of the human active CSF present or the type of progenitor cells stimulated by media conditioned with cells of leukemic origin.

Animals↗

Use of the tritiated thymidine-labelling index of the myeloblast-promyelocyte pool for the identification of the leukemic population in oligoblastic leukemia.

The bone marrow myeloblast-promyelocyte-labelling index of 32 patients with oligoblastic leukemia was studied at diagnosis and during the course of the disease. A low value (less than 0.17) or a decrease in the labelling index is suggestive of a poor prognosis with development into acute myeloblastic leukemia. Leukemic myeloblasts are characterized by their inability to differentiate and their low labelling index. The low labelling index or the decrease in this labelling index are, thus, an indication of an increase in the leukemic population.

Acute Disease↗

High rate of long-term survivals in AML treated by chemotherapy and androgenotherapy: a pilot study.

An unexpectedly high rate of long-term survivors in acute myeloid leukemia (AML) has been observed in a group of patients who achieved a first complete remission (CR) after induction of a therapeutic regimen including daunorubidomycine, vincristine, prednisone and stanozolol, and anabolizing steroid. The rate of CR was 52%. Maintenance therapy was very simple and the only association with stanozolol during this period suggested to us that an androgen, at low dosages, might be responsible for the unusual long-term survival time (45 months with a 95% confidence limit from 30 to 86 months). On the basis of our first observation, high dosages of the androgen were used during the induction phase of treatment but failed to demonstrate any advantage when associated with a drug regimen, including cytosine arabinoside (ARA-C). The reevaluation of each parameter for our group of patients did not allow selection of patients in terms of their age or the hematologic data obtained at presentation. Effects of androgen on the socalled normal hemopoietic cells and on the leukemic cells are discussed, particularly the possible antagonistic effect with ARA-C. A prospective statement is made concerning the possible condition of the prolongation of the complete remission in AML according to some experimental data which enforce the stimulative activity of androgen on the myeloid proliferation.

Adolescent↗

Normal and leukaemic human haemopoietic cells in diffusion chamber. A morphological and functional CFU-C study.

Leukaemic human bone marrow and peripheral blood cells were cultured for 25 d in diffusion chambers implanted into cyclophosphamide treated mice. Normal bone marrow cells were cultured simultaneously. These cells were studied both morphologically and functionally (CFU-C). The leukaemic cells behaved heterogeneously, 2 groups being distinguishable in accordance with their initial in vitro growth pattern (1: no growth or microcluster growth. 2: macrocluster growth). Group I showed progressive cellular death with a diminution of granulocytic progenitors and the appearance of a predominantly macrophagic population. This behaviour resembled that of the control group. The initial microcluster growth pattern remained identical throughout the entire culture period. Group 2, after considerable cellular death up to d 5, showed an explosive proliferation of the granulocytic progenitors and incomplete differentiation (up to myelocyte). The initial macrocluster growth pattern remained identical.

Adolescent↗