bluuebunny/crossref_metadata_2025 huggingface dataset:
Osmotic properties of human erythrocytes treated by phospholipase A2 from bee venom
Authors: Vaysse Jenny , Pilardeau Paul , Gattegno Liliane , Garnier Michel
Published: March 2008

During treatment of human red cells with phospholipase A) from bee venom, a linear increase of the MCV and of the osmotic fragility occurs in parallel with the cleavage of the accessible phospholipids. However, even after maximal hydrolysis, i.e. degradation of up to 65% of the phosphatidylcholines and up to 6% of the phosphatidylethanolamines, almost no haemolysis is observed and the median corpuscular fragility is only 7% higher than that of control cells incubated without enzyme. Addition of albumin to the medium results in an important rise of the susceptibility to hypotonic saline solutions. Osmotic fragility curves obtained with red cells submitted to mild phospholipase action show evidence of subpopulations of cells with various sensitivities to osmotic lysis. This phenomenon can be partly explained by the heterogeneity of the cleavage intensity among the cell population. This hypothesis is supported by the studies of the lipid composition of phospholipase treated red cells fractionated according to their sensitivity to hypotonic lysis or to their size.
Peripheral blood stem cell autografts in the treatment of lymphoid malignancies: initial experience in three patients
Authors: Bell A. J. , Figes Ann , Oscier D. G. , Hamblin T. J.
Published: March 2008

Large numbers of circulating haemopoietic progenitor cells were collected from three patients with lymphoid neoplasms for the purpose of haemopoietic reconstitution following intensive chemotherapy or chemoradiotherapy. Each patient underwent four to six leukaphereses at a time when the circulating stem cell pool was expanded, 2‐3 weeks after the end of myelosuppressive chemotherapy. The peripheral blood mononuclear cells (PBMNC) thus obtained were assayed for granulocyte‐macrophage colony‐forming cells (CFU‐GM) and were cryopreserved in liquid nitrogen. Two patients were autografted with PBMNC containing 60 x 104 and 76 x 104 CFU‐GM/kg body weight respectively. Both these patients showed prompt engraftment which is stable at + 10 and + 6 months. A third patient who was autografted with PBMNC containing 33 x 104 CFU‐GM showed only temporary and incomplete engraftment. These observations confirm that the peripheral blood may be used as a source of haemopoietic stem cells for autografting. but criteria for predicting engraftment are at present uncertain.
Post‐transfusion purpura secondary to passive transfer of anti‐P1A1 by blood transfusion
Authors: Ballem Penny J. , Buskard Noel A. , Decary Francine , Doubroff Phillip
Published: March 2008

A patient developed severe post‐transfusion purpura (PTP) following transfusion of two units of packed red blood cells. The timing of the patient's thombocytopenia suggested passive immunization rather than the typical anamnestic response associated with classical PTP. Investigation of the blood donors revealed one with evidence of a platelet specific antibody of high titre. This donor was typed as P1A1 negative and the antibody was shown to have anti‐P1A1 specificity.
A microplate ELISA for the detection of platelet alloantibodies: comparison with the platelet immunofluorescence test
Authors: Sintnicolaas Kees , Van Der Steuijt Koos J. B. , Van Putten Wim L. J. , Bolhuis Reinder L. H.
Published: March 2008

An enzyme‐linked immunoassay (ELISA) for the detection of platelet alloantibodies has been compared in detail with the platelet immunofluorescence test (PIFT). The ELISA appeared much simpler to perform than the PIFT. Both tests were comparable with regard to reproducibility and sensitivity. Alloantibodies were detected with ELISA and PIFT in 13 out of 14 patients who were refractory to random donor platelets. The value of these tests as a platelet crossmatch assay was determined in a retrospective comparison of the test results and the clinical transfusion responses expressed as the 1 h post‐transfusion platelet recovery. 39/41 (95%) negative ELISA crossmatches and 30/33 (91%) negative PIFT crossmatches appeared to be associated with a successful platelet transfusion, whereas 7/10 positive ELISA cross‐matches and 2/4 positive PIFT crossmatches appeared to be associated with transfusion failures. The high frequency of‘correct’negative tests indicates the importance of both assays in the prospective selection of compatible platelet donors for alloimmunized patients. However, because of its simplicity, the ELISA appears the method of choice for this purpose.
Increased sensitivity to complement of megakaryocyte progenitors in paroxysmal nocturnal haemoglobinuria
Authors: Dessypris Emmanuel N. , Gleaton John H. , Clark Douglas A.
Published: March 2008

Paroxysmal nocturnal haemoglobinuria (PNH) is a haemolytic disorder characterized by an increased sensitivity of peripheral blood cells including platelets to the lytic action of complement (C). Previous studies have demonstrated that in PNH bone marrow erythroid colony and burst forming units, as well as granulocytic‐monocytic colony forming units, have an increased sensitivity to complement‐induced injury as compared with normal erythroid and myeloid progenitors. The purpose of this study was to investigate the effect of complement on PNH and normal marrow megakaryocytic progenitors (CFU‐M). Bone marrow non‐adherent and T‐cell depleted light density mononuclear cells from three patients with PNH and six normal volunteers were exposed to fresh or heat‐inactivated AB human serum in the presence of medium or isotonic aqueous sucrose solution for 30 min at 37°C. After being washed the cells were assayed for CFU‐M by the plasma clot method in a complement free medium containing 6% medium conditioned by T‐lymphocytes stimulated by phytohaemagglutinin. The number of megakaryocytic colonies grown from PNH marrow cells exposed to isotonic sucrose and C’was reduced to one third of those grown from PNH cells exposed to isotonic sucrose and heat‐inactivated C, or to medium with and without C. In contrast, the number of megakaryocytic colonies grown from normal marrow cells exposed to isotonic sucrose and C’was unchanged. These findings indicate that PNH marrow CFU‐M express an increased sensitivity to C‐mediated injury similar to that detected on PNH‐erythrocytes by the sucrose haemolysis test, and support the hypothesis that the PNH defect is expressed at the level of pluripotent haematopoietic stem cell.
Late haematological complications in severe aplastic anaemia
Authors: Tichelli A. , Gratwohl A. , Würsch A. , Nissen C. , Speck B.
Published: March 2008

137 patients with severe aplastic anaemia (SAA) were treated in Basel from 1976 to 1986. 34 underwent bone marrow transplantation (BMT) and 103 received antilymphocyte globulin (ALG) therapy. We have analysed the incidence of late haematological complications in both groups of patients.

20 patients treated with ALG developed a late haematological complication. A myelodysplastic syndrome or frank leukaemia occurred in eight and paroxysmal nocturnal haemoglobinuria (PNH) in 13 patients. Nine of the 13 patients with PNH had clinical signs of haemolysis, four only had positive laboratory tests. One patient had PNH and acute leukaemia. The risk of developing a haematological complication increased continuously and reached 57% at 8 years. Neither PNH nor leukaemia occurred in patients treated with BMT. The increased survival rate and the long observation time after ALG therapy have revealed a new perspective of the prognosis of aplastic anaemia. Patients treated with BMT appear to be cured whereas those treated with ALG remain at risk for late complications.
Natural killer cell‐mediated inhibition of bone marrow colony formation (CFU‐GM) in refractory anaemia (preleukaemia): evidence for patient‐specific cell populations
Authors: Kerndrup Gitte , Hokland Peter
Published: March 2008

The role of peripheral blood mononuclear cells (PB‐MNC) on the growth of bone marrow (BM) CFU‐GM was investigated in refractory anaemia (RA) patients. Whereas normal donor PB‐MNC were found to inhibit autologous day 7 CFU‐GM, PB‐MNC from RA patients exhibited little modulatory effect on autologous or allogeneic day 7 CFU‐GM. In contrast, patient PB‐MNC inhibited autologous CFU‐GM at day 10 at a time where no significant inhibition was seen in the PB‐MNC/RA CFU‐GM combination. The identity of the inhibitory cells was investigated using anti‐T8 + and anti‐N901 + subsets purified by immune‐rosette depletion with a panel of monoclonal antibodies. The activity of these subsets was tested on immature myeloid cells enriched for MY7 + cells, and it was found that cells highly enriched for NK cells were responsible for the inhibition. Further support for NK cells as the inhibitory cells was obtained in experiments where a positive correlation between the level of PB NK cytotoxicity against K562 cells and the degree of CFU‐GM inhibition was demonstrated. Thus, these data suggest the presence of a specialized subset of NK cells with a capacity to inhibit autologous CFU‐GM. Since RA is a potentially premalignant disease, in which a significant number of cases transform into AML, these findings also suggest a physiological role for NK cells in suppression of newly arisen clonogenic cells at least in early stages of the disease.
Acute leukaemia with t(4;11) in patients previously exposed to carcinogens
Authors: Archimbaud Eric , Charrin Christiane , Guyotat Denis , Magaud Jean‐Pierre , Gentilhomme Odile , Fiere Denis
Published: March 2008

We report three cases of acute leukaemia with t(4;11) (q21;q23), one of them of undifferentiated and the other two of lymphoid phenotype, occurring after adjuvant radiochemotherapy for breast cancer (two cases) or occupational exposure to radiation (one case). Although the myeloid phenotype and characteristic chromosomal anomalies usually observed in secondary leukaemia were lacking, our observations raise the possibility of causal relationship between exposure to carcinogens and the occurrence of a leukaemia with t(4;11).
Histology of neuroblastoma involving bone marrow: the problem of detecting residual tumour after initiation of chemotherapy
Authors: Reid M. M. , Hamilton P. J.
Published: March 2008

One hundred and eighty bone marrow trephine biopsies obtained during 68 staging procedures before and during treatment of 20 children with Stage IV neuroblastoma were reviewed. A primitive cell infiltrate with prominent fibrous stromal reaction and increased reticulin characterized the marrow at presentation. Once treatment had begun, primitive cells became difficult to find but fibrosis and a marked increase in reticulin persisted in most cases. Distortion in marrow architecture and the continuation of abnormal stromal reactions may reflect failure to eradicate tumour despite absence of detectable primitive cells. If current therapeutic options are to be assessed properly, uniformly accepted criteria to define continuance of marrow infiltration in children with neuroblastoma are needed. A scheme for classifying marrow histological appearances is presented which may prove to be of more value than a simple distinction between ‘involved’and ‘uninvolved’.
In vitro interferon‐gamma production by cultured T‐cells in severe aplastic anaemia: correlation with granulomonopoietic inhibition in patients who respond to anti‐thymocyte globulin
Authors: Laver Joseph , Castro‐malaspina Hugo , Kernan Nancy A. , Levick Janet , Evans Robert L. , O'Reilly Richard J. , S. Malcolm A.
Published: March 2008

T‐cell‐mediated inhibition of granulomonopoietic progenitors (CFU‐GM) was studied in vitro in 27 patients with severe aplastic anaemia (AA). In nine out of 13 responders to anti‐thymocyte globulin (ATG), cultured T‐cells obtained prior to therapy, as well as their conditioned medium strongly suppressed both normal allogeneic and autologous CFU‐GM (the latter obtained after marrow recovery). Addition of anti‐interferon‐gamma to the cultures abolished the suppressive effect on CFU‐GM. After ATG therapy, no similar inhibitory effect was detected. Employing the panning method with monoclonal antibodies (CD4+ for inducer/helper and CD8+ for cytotoxic/suppressor T‐cells) we were able to show that the cells responsible for in vitro CFU‐GM inhibition were included in the cytotoxic/suppressor T‐cell subpopulation. Cultured T‐cells and their conditioned medium obtained from 14 non‐responders to ATG did not show CFU‐GM suppression. The mean interferon (IFN) levels in the T‐cell conditioned media of ATG‐responders was 625 ± 125 u/ml while in non‐responders the level was 45 ± 15 u/ml (normal control levels 43 ± 24 u/ml). Freshly isolated peripheral blood lymphocytes from either group did not show any in vitro inhibitory effect. The response rate to ATG was statistically significant when the generation in culture of high versus low IFN production was compared (P=0.0001). Experiments with T‐cells obtained from heavily transfused thalassaemia major, and myelodysplastic syndrome patients, as well as normal volunteers, also did not demonstrate any suppression of CFU‐GM. Our results indicate that the response rate to ATG is significantly higher in patients with AA who have an abnormal regulation of interferon‐gamma (g‐IFN) production.
The syndrome of abnormal chromatin clumping in leucocytes: a myelodysplastic disorder with proliferative features?
Authors: Felman Pascale , Bryon Paul‐Andre , Gentilhomme Odile , Ffrench Martine , Charrin Christiane , Espinouse Daniel , Viala Jean‐Jacques
Published: March 2008

Five patients presenting extremely exaggerated chromatin clumping in leucocytes, associated with a loss of segmentation, are described. Peripheral cytopenia, a high percentage of circulating immature granulocytes (chiefly myelocytes) with variable leucocytosis, marrow hypercellularity with granulocytic hyperplasia and moderate dysplastic changes in erythroblastic and megakaryocytic lines, constitute, together with the granulocytic nuclear anomaly, the characteristic features of the studied cases. Cytogenetic analysis showed a clonal 12p anomaly in one of the three cases available for study. Survival was poor with a median of 15 months, bleeding and infections being responsible for the majority of deaths. All these findings point to abnormal chromatin clumping in leucocytes as the marker of a true entity with both myelodysplastic and myeloproliferative features. We propose that it take its place among myelodysplastic syndromes (MDS) beside chronic myelomonocytic leukaemia (CMML), with which it shares many similarities in clinical and biological behaviour.
Human parvovirus B19‐induced transient pancytopenia in a child with hereditary spherocytosis
Authors: Hanada Takashi , Koike Kazutoshi , Takeya Tosiki , Nagasawa Toshiro , Matsunaga Yasuko , Takita Hitoshi
Published: March 2008

We report a child with hereditary spherocytosis who developed human parvovirus B19 (HPV B19)‐induced transient pancytopenia. Symptoms of acute‐phase illness subsided promptly after infusion of gammaglobulin containing HPV B19 antibody. The HPV B19‐containing serum on admission inhibited not only erythroid colony growth but also myeloid and megakaryocyte colony growths. The inhibitory effect was abrogated by convalescent‐phase serum or gammaglobulin solution containing HPV B19 antibody. These findings suggest that HPV B19‐induced inhibition of haematopoiesis may be pathogenetic for transient bone marrow failure of the patient. The passive immunization may be effective in shortening the duration of acute‐phase illness of HPV B19 infection.
Inhibition of bone marrow myelopoiesis and erythropoiesis in vitro by anti‐retroviral nucleoside derivatives
Authors: Johnson Margaret , Caiazzo Teresa , Molina Jean‐Michel , Donahue Robert , Groopman Jerome
Published: March 2008

We have studied the in vitro effects of nucleoside inhibitors of reverse transcriptase on bone marrow myeloid and erythroid progenitor development. Both 3′‐azido‐3′‐deoxythymidine (AZT) and 2′,3′‐dideoxycytidine (DDC) potently inhibited in vitro haematopoiesis, while 2′,3′‐di‐deoxyadenosine (DDA) was less inhibitory in this culture system. Studies of candidate antiretroviral agents in bone marrow culture may predict clinical toxicity as well as lead to strategies for combination therapy which minimize impairment of haematopoiesis.
Effects of type β transforming growth factors on haematopoietic progenitor cells
Authors: Hino Masayuki , Tojo Arinobu , Miyazono Kohei , Urabe Akio , Takaku Fumimaro
Published: March 2008

The effects of type β transforming growth factors (TGF‐βs) on normal human and murine haematopoietic progenitor cells were examined using bone marrow colony assays. In erythroid colony assays, TGF‐β1 inhibited human CFU‐E derived colony formation, BFU‐E derived burst formation, and murine BFU‐E derived burst formation in a dose dependent manner between 0.1 and 5.0 ng/ml. However, murine CFU‐E derived colony formation was unaffected even at a concentration of 5.0 ng/ml TGF‐β1. In myeloid colony assays, different sensitivity of progenitor cells to the inhibitory effects of TGF‐βs was observed between both species. TGF‐β1 inhibited murine granulocyte‐macrophage colony (GM‐colony) formation and granulocyte colony (G‐colony) formation in a dose dependent manner between 0.1 and 5.0 ng/ml, but had no remarkable effects on human GM‐colony and G‐colony formation. TGF‐β2 also had similar inhibitory effects on haematopoietic progenitor cells, while its inhibitory effect was less potent than that of TGF‐β1. Thus our data suggest that TGF‐β may be involved in negative regulation of haematopoiesis and that its inhibitory action may be restricted in lineage and/or species specific manner.
Effects of preparative procedures and of cell activation on flow of white cells through micropore filters
Authors: Nash G. B. , Jones J. G. , Mikita J. , Christopher B. , Dormandy J. A.
Published: March 2008

Using newly developed filtration techniques to analyse the flow resistance of white blood cells (WBC), the effects of preparative procedures and of various treatments (including activation) have been tested. Flow rates of WBC were measured using both 5 and 8 μm pore filters. It was found that EDTA was the anticoagulant of choice over heparin and sodium citrate, and that calcium‐containing media should be avoided because they promoted cell‐cell aggregation. Exposure to density separation media did not significantly alter filtration rates, but storage of prepared WBC for even 1 h caused deterioration in flow. Storage of whole blood before preparation of the WBC had a much lesser effect. Storage appeared to be linked to cell activation, and use of the activating agents phorbol ester (PE, 4.5 × 106 M/l) and f‐methyl‐leucyl‐phenylalanine (FMPL, 10‐7 and 10‐9 M/l) also increased WBC resistance to pore transit. The effect of PE was greater, causing rapid pore blockage, but was nonspecific, while the lesser effect of FMLP was restricted to granulocytes and was concentration dependent. Cell volume was increased by these agents, but measurement of the filterability of hypotonically swollen granulocytes showed that volume changes alone only partly explained the activation effects. These results suggest that activation in vivo would have a significant rheological effect, detectable by filtration methods.
Bone marrow transplantation (BMT) versus immunosuppression for the treatment of severe aplastic anaemia (SAA): a report of the EBMT* SAA Working Party
Authors: Bacigalupo A. , Hows J. , Gluckman E. , Nissen C. , Marsh J. , Van Lint M. T. , Congiu M. , De Planque M. M. , Ernst P. , McCann S. , Ragavashar A. , Frickhofen N. , Wursch A. , Marmont A. M. , Gordon‐Smith E. C.
Published: March 2008

This is an analysis of 509 patients with severe aplastic anaemia (SAA) treated in Europe between 1981 and 1986; 218 patients were treated by allogeneic bone marrow transplantation (BMT) from HLA identical sibling donors and 291 with immunosuppressive therapy (IS) with antilymphocyte globulin (ALG). The overall actuarial survival was 63% after BMT and 61% after IS therapy at 6 years. All patients fulfilled the criteria of SAA; however, most patients with a neutrophil count of <0.2 × 109/l also had infections and haemorrhages. Therefore a further subclassification was defined by pretreatment peripheral blood neutrophil count: very severe aplastic anaemia (vSAA) (<0.2 × 109/l neutrophils) and moderately severe aplastic anaemia (mSAA) (0.2‐0.5 × 109/l neutrophils).

A Cox regression analysis showed that the only significant pre‐treatment variables were a low neutrophil count (P= 0.001) and increasing age (P= 0.05). Thus it seemed reasonabie to analyse survival data after combined stratification for neutrophils (vSAA versus mSAA) and age (cut off at 20 years). BMT was superior to IS in patients with vSAA under 20 years of age (64%v.38%;P= 0.01). IS was superior to BMT in patients with mSAA aged 20 or more (82%v.62%;P= 0.002). The two treatments gave comparable results in young patients with mSAA (BMT = 58%, IS = 62%;P= 0.1), and in older patients with vSAA (BMT = 44%. IS = 43%;P= 0.06).

Overall 75/218 and 87/291 patients, given BMT or IS respectively, died. The major cause of failure in BMT patients was graft rejection (n= 22) or problems associated with graft‐versus‐host disease. For ALG patients the major problem was persistence of the aplasia with haemorrhage (n= 32) or infections (n= 46).

This study indicates that over 60% of patients with SAA can be successfully treated with either BMT or IS. Overall survival does not differ in the two groups, though significant differences emerge after stratification for severity of the aplasia and age.
Blast cell vacuoles in childhood lymphoblastic leukaemia
Authors: Lilleyman J. S. , Hann I. M. , Stevens R. F. , Richards S. M. , Eden O. B.
Published: March 2008

As part of a central review of cell morphology in childhood lymphoblastic leukaemia (ALL), marrow smears from entrants to the Medical Research Council trial UKALL VIII, other than those from children with B‐ALL, were studied prospectively for the presence or absence of blast cell vacuoles and for any clinical or biological relevance this feature might have.

Adequate slides were available from 733 patients (88% of the trial entrants) after five with B ALL were excluded. Vacuolated blast cells (> 10%) were present in 204 (28%). The presence of vacuoles was associated with PAS positivity (%2= 27.8; P<0.0001), a diagnostic white cell count (WBC) <50 × 109/l (%2= 13.1; P<0.0001), and the immunophenotype of ‘common’ALL (CD10 positive) (%2= 9.1; P<0.01). There was no clear association with French‐American‐British (FAB) type L1 or L2. The 204 patients with vacuoles had a significantly superior disease free survival compared to the remainder (2P= 0.01), a difference which remained significant when the analysis was stratified by FAB type (2P= 0.01), age (2P= 0.02) or sex (2P= 0.02), but which was lost when stratified by WBC (2P= 0.06).

These findings provide further evidence that, outside the context of B‐ALL, vacuoles are indicative of a relatively benign disease which responds well to therapy. The French‐American‐British (FAB) classification should be modified to take this into account.
Protein S and C4b‐binding protein in fetal and neonatal blood
Authors: Melissari E. , Nicolaides K. H. , Scully M. F. , Kakkar V. V.
Published: March 2008

The levels of protein C, free protein S, C4b‐binding protein and the distribution of total protein S between a free form and a conjugated form with C4b‐binding protein, were measured in fetuses between 15 and 35 weeks of gestation, in newborns and in maternal blood throughout pregnancy. Fetal and newborn umbilical blood samples were obtained by cordocentesis; in the case of newborns, immediately after delivery. C4b‐binding protein was detected in five out of seven newborns and in only three out of 20 fetuses. The fetuses (21, 24, 29 weeks, respectively) whose blood contained C4b‐binding protein (10%, 29%, 12% of adult level, respectively), did not survive: the possible cause of death for one was given as viral infection; all three were severely deformed. Only free protein S was detected in fetal blood and the mean value was 40%. By contrast, the mean levels of protein C in the fetuses increased from 11% to 15.5% of adult levels with advancing gestation. We conclude that the relatively high levels of free protein S in fetal blood may contribute to its non‐coagulability and compensate for the low protein C levels.
Activity of ribonuclease H in cells of chronic B‐lymphocytic leukaemia: correlation with clinical stage
Authors: Papaphilis A. D. , Kamper E. F. , Kattamis C. , Pangalis G. A.
Published: March 2008

Peripheral blood mononuclear cells (PBMNC) from 23 healthy subjects and 39 patients with B‐cell chronic leukaemia (B‐CLL) were assayed for ribonuclease H activity using as substrate the filter‐immobilized synthetic homopolymer hybrid 3H‐poly(rA):poly(dT). In 69% of the leukaemia patients examined enzyme activities were above those estimated in cells from the healthy controls. The mean enzyme levels for the normal and the leukaemic samples group were (per cent substrate hydrolysis): 8.1 ± 8.9 and 58.7 ± 40.8, respectively, their difference being statistically highly significant (P < 0.0001). This result does not represent homogeneity within the cells but is due to a subclass of cells within the leukaemic clone containing the enzyme, thus contributing through pool size fluctuation to the wide variations of enzyme activity observed among the patients. These cells containing high activity could not be identified with either the prolymphocytes or the large lymphocytes. The activity of ribonuclease H in the examined CLL patients correlated with disease stage (Binet) (P= 0.011) and appeared to serve as a sensitive indicator of disease progression when compared with a number of other known prognostic parameters.
The value of immunocytochemical staining of lymph node aspirates in diagnostic cytology
Authors: Oertel J. , Oertel B. , Kastner M. , Lobeck H. , Huhn D.
Published: March 2008

This study investigates the applicability of immunocytochemical techniques towards improving the cytological diagnosis of lymph node disorders. Cytocentrifuge preparations of fine needle aspirates were examined using an indirect immunoperoxidase method and the alkaline phosphatase–antialkaline phosphatase method.

36 reactive lymph nodes were evaluated. 23 patients showed T cell predominance as assessed by the presence of CD3+ cells. 13 patients showed an excess of CD24+ cells over CD3+ cells. In all patients the B cell population showed no light chain class restriction.

48 aspirates from patients with B‐non‐Hodgkin's lymphoma of low malignancy were investigated. The majority of the cells in each of these cases expressed CD24 and 47 cases were monoclonal with respect to their light chain determinants. In 37/48 cases the antibody OKT9 reacted with <15% of cells.

18 aspirates were obtained from patients with B‐non‐Hodgkin's lymphoma of high malignancy. 17 patients showed light chain class restriction and a high percentage of CD24+ (>80%) and OKT9+ (>40%) cells.

8 malignancies were considered to be of T‐cell origin. A low percentage (<15%) of Ig+CD24+ cells with a high number of CD3+ and/or CD4+ cells suggests a T‐cell lymphoma. The majority of neoplastic cells in peripheral T‐cell lymphomas (excluding mycosis fungoides) expressed OKT9 and CD3 expression was found to be variable.

In 15 cases of Hodgkin's disease, there were 11 correct cytologic diagnoses. A significant number of BerH2 (CD30)+ cells having the morphology of Sternberg‐Reed cells supports this diagnosis. 25 aspirates were obtained from patients with metastatic malignant tumours. Marker studies in eight cases helped in distinguishing metastatic malignant tumours from malignant lymphomas.

Our findings indicate that the immunocytochemical technique is applicable to cytological material and that the simplicity of the procedure merits application to routine diagnostic cytology.
Erythroid progenitor cell kinetics in chronic haemodialysis patients responding to treatment with recombinant human erythropoietin
Authors: Reid C. D. L. , Fidler J. , Oliver D. O. , Cotes P. Mary , Pippard M. J. , Winearls C. G.
Published: March 2008

The response of bone marrow and peripheral blood erythroid progenitors to human recombinant erythropoietin (rHuEPO) was studied in nine haemodialysed renal failure patients receiving this hormone for the correction of their anaemia. The haematocrit rose in all patients in response to thrice weekly injections of escalating rHuEPO doses (12–192IU/kg). Both the numbers of CUF‐e and BFU‐e and their proliferative state in the bone marrow as well as BFU‐e numbers in the peripheral blood were estimated before treatment and again after correction of the anaemia, at 16 h following an intravenous dose of rHuEPO.

Following treatment bone marrow BFU‐e numbers fell to a mean of 24.5% (P <0.01) of the pre‐treatment values although there was no significant change in CFU‐e or circulating BFU‐e numbers. The mitotic rate (percentage S‐phase cells) estimated by tritiated thymidine suicide rose from 45.2% to 68.4% (P <0.05) in the case of CFU‐e and from 16.4% to 45.1% (P <0.05) for BFU‐e following treatment with rHuEPO thus indicating in‐vivo sensitivity of both the primitive as well as the mature erythroid progenitors to the hormone. The fall in BFU‐e numbers in the bone marrow after several months of treatment may be due to a loss of cells from this progenitor pool by maturation that is uncompensated by replacement from the pluripotential stem cell compartment.
Identification of a CpG mutation in the coagulation factor‐IX gene by analysis of amplified DNA sequences
Authors: Siguret V. , Amselem S. , Vidaud M. , Assouline Z. , Kerbiriou‐Nabias D. , Piétu G. , Goossens M. , Larrieu M. J. , Bahnak B. , Meyer D. , Lavergne J. M.
Published: March 2008

In a family with no previous bleeding history, the sister of a single, severely affected haemophilia B patient requested carrier detection and prenatal diagnosis. In Southern blots, using Taq I digested DNA and a factor‐IX cDNA probe, a normal invariant band at 1.6 kb was missing in the haemophiliac suggesting the loss of the Taq I site at the 5’end of exon h. A 162 bp sequence which includes the suspected mutant region was amplified by the polymerase chain reaction in each DNA. Two oligonucleotide probes were synthesized and differed by only one base pair which substituted a T for C in the normal Taq I recognition sequence. The amplified DNA was dot‐blotted and hybridized with the labelled probes. The altered sequence hybridized to DNA from the affected individual, his sister and her fetus and not to DNA from the normals. The mutation, involving the haemophiliac, his mother, his sister and and her fetus, transforms a CGA codon that encodes for arginine in the catalytic domain of the protein into a UGA stop codon.
Effects of cis‐diamminedichloroplatinum (II) upon haemopoietic progenitors and the haemopoietic microenvironment in mice
Authors: Nikkels Peter G. J. , De Jong Johannes P. , Ploemacher Rob E.
Published: March 2008

Summary. We studied the short‐ and long‐term effects of a fractionated injection of cis‐diamminedichloroplatinum (II) (CDDP) upon the haemopoietic stroma and the haemopoietic precursor cell compartment of young and adult mice. The integrity of the stromal microenvironment was investigated using three different assays including quantification of (a) the fibroblastoid progenitor cell compartment, (b) the regenerative capacity after subcutaneous implantation of spleen and femur, and (c) the growth of normal bone marrow progenitors in lethally irradiated CDDP‐treated mice.

CDDP treatment induced a slight anaemia which lasted for the observation period of 1 year, and could not be restored by infusion of normal bone marrow cells. The population size of haemopoietic progenitors was severely decreased immediately after CDDP treatment and the CFU‐S recovery in the bone marrow was slow and temporary. Stromal function was significantly decreased and normalization occurred within approximately 40 d, depending on the stromal parameter measured. Subsequently, the regenerative capacity of the stroma showed a second decrease which was still detected at 1 year. This pattern of stromal damage has not been reported for any other cystostatic agent. Since the other two assays did not detect a second decrement in stromal integrity it is implied that the three stromal assays used detect different stromal functions. We conclude that CDDP treatment of both young and adult mice results in severe short‐term damage and a late occurring secondary regenerative defect of the haemopoietic organ stroma.
Monocyte‐derived macrophages as helper cells in monocyte‐mediated cytolysis
Authors: Dallegri Franco , Ballestrero Alberto , Frumento Guido , Goretti Riccardo , Patrone Franco
Published: March 2008

Summary. Human peripheral blood monocytes (M), incubated with opsonized zymosan particles (OPZ), lysed human erythrocyte (RBC) targets, as detected by a 51Cr release method. Conversely, cells derived in vitro from M (monocyte‐derived macrophages, MDM) were ineffective. When added to the M‐RBC system, MDM enhanced the lysis. The lysis by M and M plus MDM was prevented by catalase, azide and amino acids (alanine, taurine), consistent with the requirement for hypochlorous acid (HOCl). Moreover, MDM per se incapable of generating HOCl augmented the HOCl recovery from the M‐RBC system. The results provide evidence for a previously unrecognized form of interaction between two distinct populations of mononuclear phagocytes.
Inability to detect transferrin receptors on P. falciparum parasitized red cells
Authors: Pollack Simeon , Schnelle Vera
Published: March 2008

Summary The mechanism by which P. falciparum takes up iron from transferrin has been explored. Binding of 125I labelled transferrin to parasitized red cells at 37°C is two‐fold greater than to control cells; at 0°C there is no significant difference. The binding is non‐specific as judged from the following: it is not saturable: it is not limited to transferrin as lactoferrin (which has iron binding domains) and bovine serum albumin (which does not) also bind in excess to parasitized red cells. A transferrin receptor complex could not be demonstrated when parasitized red cells, to which 125I transferrin was bound, were solubilized in Triton X100. Previous observation showed that uptake of transferrin iron by parasitized red cells is not accompanied by equimolar uptake of transferrin protein. We therefore suggest that non‐specifically bound transferrin is endocytosed, that the protein is degraded and the iron selectively retained.
Effect of red cell age on vesiculation in vitro
Authors: Greenwalt T. J. , Dumaswala U. J.
Published: March 2008

Summary Microvesiculation of RBC occurs in vivo and during in vitro storage. Vesiculation of young and old populations of RBC was measured under blood bank conditions. The RBC in whole units of blood were divided by density into two approximately equal fractions in a closed system. After storage for 35 d the haemoglobin‐containing vesicles were quantitatively recovered. Quantitation of protein, cholesterol and phospholipids showed significantly larger amounts in the vesicles shed by the younger RBC. Otherwise vesicles shed by the two populations of RBC did not differ.
Von Willebrand disease investigated by two novel RFLPs
Authors: Bernardi F. , Guerra S. , Patracchini P. , Volinia S. , Buzzoni D. , Ballerini G. , Casonato A. , Marchetti G.
Published: March 2008

Two partial cDNAs for von Willebrand factor (vWF) were used to investigate gene lesions and restriction fragment length polymorphisms (RFLPs) in vW disease (vWd) and normal controls. No gene alteration was detected but two TaqI RFLPs, likely to be intronic and originating from point mutations, were found in the 3’ part of vWF gene. The two TaqI RFLPs, identified by the same probe, are informative in approximately 50% of the subjects. Used in combination with two other known RFLPs, they define several haplotypes similarly distributed in vWd and normals. Linkage disequilibrium between loci identified by the RFLPs is present.

In a family study the RFLP patterns demonstrate homozygosity for the affected vWF gene in a severe (type III) patient and identify several heterozygous subjects. The RFLPs analysis has been related to the haemostatic values and multimer distribution. In two of the four unrelated patients with severe vWd examined the RFLPs study indicates double heterozygosity for the affected vWF genes.
Two different theta (θ) globin gene deletions observed among black newborn babies
Authors: Fei Y. J. , Fujita S. , Huisman T. H. J.
Published: March 2008

Two different θ1 globin gene deletions have been detected in the Black population of the southeastern U.S.A. Restriction mapping of the α globin gene cluster indicated that one deletion removes ∼ 1.8 kb of DNA including most of the θ1 globin gene, while the second removes ∼ 6 kb of DNA including the entire θ1 globin gene. These defects do not cause haematological alterations in the newborn or in the adult (established for the smaller deletion only), nor were changes observed in polypeptide chain patterns at birth.
A comparative study of combination chemotherapy versus marrow transplant in first remission in adult acute lymphoblastic leukaemia
Authors: Proctor S. J. , Hamilton P. J. , Taylor Penny , Carey P. , Hargrave S. , Evans R. G. B. , Summerfield G. , Finney R. , Saunders P. , Goff D. , Reid M. M.
Published: March 2008

Summary. The results of conventional chemotherapy in adult acute lymphoblastic leukaemia (ALL) have not improved substantially in recent years. The present study is based on a flexible policy of marrow transplantation (allograft and autograft without marrow purging) in first remission compared with a group treated with standard maintenance therapy after a common induction sequence. The actuarial disease free survival (DFS) and actuarial overall survival (OS) at 3 years for autologous marrow grafted patients was 30% and 65% respectively. The allogeneic transplant group had DFS of 30% and OS at 3 years of 38% compared with DFS (12%) and OS (12%) for patients on 6‐mercaptopurine and methotrexate maintenance. The actuarial disease free survival calculations include patients on protocol not entering remission, therefore, giving the worst possible result.

We conclude that high dose chemo/radiotherapy with autologous marrow rescue in first remission followed by no maintenance provides better results in terms of overall survival and quality of life than standard ALL maintenance in adult patients. Results for allogeneic transplant in ALL are less good in terms of duration and quality of survival and the majority of deaths are related to causes other than leukaemic relapse.
Decay‐accelerating factor (DAF) on the blood cell membranes in patients with paroxysmal nocturnal haemoglobinuria (PNH): measurement by enzyme‐linked immunosorbent assay (ELISA)
Authors: Ninomiya Haruhiko , Abe Tsukasa , Shichishima Tsutomu , Terasawa Takashi , Fujita Teizo
Published: March 2008

Summary. We developed a quantitative enzyme‐linked immunosorbent assay (ELISA) for decay‐accelerating factor (DAF) on blood cell membranes using monoclonal anti‐DAF antibodies. DAF is an integral membrane protein of several blood cells. It regulates the C3 and C5 convertase of the classical and alternative pathways of complement activation. It is partially or completely deficient in the membranes of blood cells of patients with paroxysmal nocturnal haemoglobinuria (PNH).

The ELISA we developed for DAF measurement indicated a reliable range of measurement from 2.25 to 11.25 ng of DAF. In particular, ELISA proved to be a technically simple method and its sensitivity was enhanced by using avidin‐biotin complex. In this study, DAF levels in extracts of erythrocytes from 30 healthy donors and in extracts of PMN and platelets from 15 healthy donors were measured by ELISA. The DAF content of blood cells from eight patients with PNH and erythrocytes from 13 patients with anaemia were also measured. The DAF levels of normal erythrocytes, PMN and platelets were 3110.960, 28000.13900 and 3100.1370 (mean±SD) molecules/cell, respectively. In general, the DAF content of PNH cells was below the normal range, although it was within the normal range in some cases of PNH. The DAF levels of PNH‐I and ‐II erythrocytes were estimated from the ratio of PNH‐I, ‐II and ‐III erythrocytes. And, in four cases of PNH, the DAF levels of PNH‐I erythrocytes separated by acidified serum lysis were measured by ELISA. In most cases of PNH, DAF was found to be partially deficient in PNH‐I and PNH‐II erythrocytes.
Further characterization of human myeloid antigens (gp160,95; gp150; gp67): investigation of epitopic heterogeneity and non‐haemopoietic distribution using panels of monoclonal antibodies belonging to CD‐11b, CD‐13 and CD‐33
Authors: Favaloro E. J. , Bradstock K. F. , Kabral A. , Grimsley P. , Zowtyj H. , Zola H.
Published: March 2008

Summary. We have investigated the binding of over 30 different monoclonal antibodies (MAB) belonging to three distinct clusters of differentiation (CD‐11b; CD‐13; CD‐33; as defined by the Third International Workshop on Leucocyte Differentiation Antigens (ILWS), 1986), and which are reactive with three distinct myeloid restricted surface antigens (‘gp160,95′; ‘gp150′; ‘gp67′). By investigating reactivity with non‐haemopoietic cells, we have confirmed that CD‐11b and CD‐33 MAB reactivity is largely restricted to haemopoietic cells, whilst CD‐13 MAB showed additional binding to a wide range of non‐haemopoietic cells. Epitopic heterogeneity was also investigated within each cluster of differentiation. Tested anti‐CR3 (CD‐11b) MAB varied in their ability to block the binding of complement coated sheep red blood cells and zymosan particles. A more detailed analysis of MAB binding heterogeneity was performed by competitive inhibition assays. It was demonstrated that MAB from both CD‐11b and CD‐13 bind to several distinct epitopes (at least six and five respectively) on their respective antigen molecules. In contrast, CD‐33 MAB appear to bind to only a single site on ‘gp67′. These data may allow for a clearer appreciation of the disparate functional effects obtained using different MAB reagents to individual myeloid antigens, as reported by a number of workers.
5‐Aminolaevulinic acid synthase activity in developing human erythroblasts
Authors: Fitzsimons E. J. , May A. , Elder G. H. , Jacobs A.
Published: March 2008

Summary. 5‐Aminolaevulinic acid (ALA) synthase activity was measured in highly purified preparations of age‐matched human erythroblasts. Enzyme activity in immature normoblasts was four‐fold higher than that found in late orthochromatic normoblasts. ALA synthase activity in the immature erythroblasts in primary acquired sideroblastic anaemia (PASA) was reduced and remained unchanged during further erythroid differentiation. The pattern of erythroblast ALA synthase activity in two patients with congenital dyserythropoietic anaemia (CDA) and in one patient with β‐thalassaemia intermedia was similar to that found in PASA.

This study has clearly demonstrated reduced erythroblast ALA synthase activity in PASA but has also found reduced enzyme activity in conditions in which ring sideroblasts are not prominent. This would suggest that haem synthesis is abnormal in PASA but that reduced erythroblast ALA synthase activity does not inevitably lead to ring sideroblast formation.
Abnormal megakaryopoiesis in patients with myelodysplastic syndromes: analysis of cellular and humoral defects
Authors: Geissler D. , Zwierzina H. , Pechlaner C. , Gaggl S. , Schmalzl F. , Konwalinka G. , Braunsteiner H.
Published: March 2008

Summary. In 13 patients with myelodysplastic syndrome (MDS) mature and immature erythropoietic (CFU‐E, BFU‐E), granulopoietic (CFU‐GM) and megakaryopoietic (CFU‐Meg) colony formation from human bone marrow mononuclear cells was evaluated in a microagar culture system. All but three patients exhibited abnormal CFU‐Meg. The defect of CFU‐Meg paralleled the reduction of BFU‐E, whereas CFU‐GM number declined to a lesser extent. Not only the CFU‐Meg number, but also the number of megakaryocytes (Mk) per colony was reduced suggesting an additional functional CFU‐Meg defect.

Megakaryocytic growth factor (Meg‐CSF) abnormalities in MDS patients were detected using normal nonadherent T‐lymphocyte depleted bone marrow cells as target cells for serum testing. Even for sera from patients with a reduction of platelets and bone marrow megakaryocytes Meg‐CSF levels were not increased.

No cellular or humoral inhibition could be detected in an MDS patient with a 5q – karyotype, who had an isolated defect of the megakaryocytic cell lineage at presentation.

Some patients revealed a spontaneous formation of mixed erythrocytic, granulocytic and megakaryocytic clusters in the presence of fetal calf serum or autologous patient serum, probably representing autonomous proliferation of the malignant clone. In conclusion, both cellular and humoral factors can cause abnormalities of the megakaryocytic cell lineage in MDS patients.
Bone marrow transplantation for adult poor prognosis lymphoblastic lymphoma in first complete remission
Authors: Milpied N. , Ifrah N. , Kuentz M. , Maraninchi D. , Colombat P. , Blaise D. , Harousseau J. L.
Published: March 2008

Summary. Twenty‐five adult patients, 19 males, six females, age 16–43 years (median 23), with lymphoblastic lymphoma received allogeneic or autologous bone marrow transplantation in first complete remission. Twelve patients were Murphy stage IV with bone marrow and/or CNS involvement and 13 were stage III of whom nine had thoracic involvement. Complete remission was achieved with an intensive anthracycline containing multiagent chemotherapy protocol. Twelve patients with an HLA identical sibling received an allogeneic marrow and 13 without a donor received their own marrow harvested a median of 2 months (0–4 months) after complete remission and purged in vitro with either mafosfamide (eight patients) or anti T‐cell monoclonal antibodies and complement (three patients). Bone marrow transplantation was performed 1–7 months (median 3 months) after achieving first complete remission. The conditioning regimen consisted of cyclophosphamide or high dose melphalan and total body irradiation. The actuarial 4‐year disease‐free survival is 68% (±9% SE). The actuarial probability of relapse was 26% (±3% SE) with a median follow up to 22 months. There was no difference between allogeneic and autologous transplantation with eight out of 12 allo patients in first continuous complete remission 26–45 months after transplant and nine out of 13 auto in continuous complete remission 15–75 months after transplant. These results compare favourably with those achieved with the best chemotherapeutic regimen used for such patients.