bluuebunny/crossref_metadata_2025 huggingface dataset:
Inhibition of the enzyme purine nucleoside phosphorylase (PNP) reduces refractoriness to transfused platelets in a dog model
Authors: Slichter Sherrill J. , Deeg H. Joachim , Osborn William R. A.
Published: March 2008

Summary. To better define the role of T‐cells in alloimmunization, we induced purine nucleoside phosphorylase (PNP) deficiency in a dog platelet transfusion model. Short‐term administration of eight different drug schedules using several combinations of the PNP inhibitor 8‐aminoguanosine and/or deoxyguanosine did not induce significant toxicity in four treated animals as demonstrated by blood chemistries, cell counts, and autologous platelet recovery and survival measurements. However, continuous long‐term daily administration of these agents produced significant renal and/or hepatic toxicity leading to death in five of six animals. Modification of the drug schedule to early short‐term administration of both deoxyguanosine and 8‐aminoguanosine, followed by long‐term intermittent doses of only 8‐aminoguanosine, was not only well tolerated by all the animals but also resulted in significant immunosuppression. Overall, six of nine evaluable dogs (67%) treated with some combination of PNP inhibitors did not become refractory to eight weekly transfusions of platelets from a single random donor dog, P<0.005, compared to untreated controls, only 3/21 (14%) not immunized.

Furthermore, in four evaluable recipients, discontinuation of the 8‐aminoguanosine while continuing platelet transfusions from their original donors did not result in refractoriness. In addition, these four recipients were also unable to recognize platelets from two other random donors. This suggests that both specific and non‐specific tolerance to foreign platelet antigens had been induced by PNP‐inhibitor therapy. Other evidence for the efficacy of this immunosuppressive treatment was the almost normal post‐transfusion recovery and survival of donor platelets, both during and after treatment. This suggests failure to form even low levels of platelet alloantibodies in the immunosuppressed recipients. In contrast, B‐cell immunity to soluble antigens was intact as demonstrated by a normal antibody response to keyhole limpet haemocyanin (KLH) antigen.
Serum transferrin receptor as an index of iron absorption
Authors: Cook James D. , Dassenko Sandra , Skikne Barry S.
Published: March 2008

Summary. Recent studies indicate that serum transferrin receptor levels are a quantitative index of tissue receptor mass. To determine whether the latter plays a role in the regulation of iron absorption, we examined the relationship between serum receptor, serum ferritin and iron absorption in healthy subjects. Using radioisotopic techniques we measured absorption of inorganic iron in 174 subjects and dietary nonhaem iron in 60 subjects. With both forms of iron, the correlation with absorption was far lower for serum receptor than for serum ferritin and was no longer significant when subjects with depleted iron stores were excluded. These results indicate that in normal subjects the iron store is the main physiological determinant of iron absorption and that in the absence of iron deficiency, tissue receptor mass, reflected by serum transferrin receptor levels, has no discernible influence.
Different sensitivity of normal and leukaemic progenitor cells to Ara‐C and IL‐3 combined treatment
Authors: Lista P. , Brizzi M. F. , Rossi M. , Resegotti L. , Clark S. C. , Pegoraro L.
Published: March 2008

Summary. In the present study the effects of a combined treatment with cytosine‐arabinoside (Ara‐C) and inter‐leukin‐3 (IL‐3) on acute myeloblastic leukaemia clonogenic cells and on normal haemopoietic progenitors was investigated, with the aim of improving the tumoricidal effect of cycle specific drugs.

Blast cells from 24 acute myeloblastic leukaemia (AMT,) patients were screened with a short‐term proliferative assay based on 3H‐thymidine (3H‐TdR) uptake for their response to IL‐3. To evaluate the synergism between the growth factor and Ara‐C, the cells were pretreated for 3d in liquid culture in the presence or absence of IL‐3 (10 U/ml) and for the last 24 h with Ara‐C (3 μg/ml). The cells were then washed and seeded in semisolid media to assess their clonogenic ability. The results showed that, in those cases which were good responders to IL‐3 in the 3H‐TdR uptake assay (19 out of a), Ara‐C exposure eliminated a greater proportion of clonogenic cells if pretreated with IL‐3 than if untreated (P < 0.001), while in cases unresponsive to IL‐3 this effect was not significant. Moreover, when the same protocol was applied to bone marrow cells from normal donors, it was found that IL‐3 pretreatment did not significantly enhance the toxic effect of Ara‐C on day 14 granulocyte‐macrophage colony forming units (CFU‐GM) and erythroid burst forming units (BFU‐E). Finally IL‐3 pretreatment was also able to increase the cytotoxic effect of Ara‐C on leukaemic cells co‐cultured, to simulate clinical AML remission, with normal bone marrow cells. The results indicate that IL‐3 may improve the therapeutic index of cycle‐specific drugs in AML therapy.
The 14;18 translocation in European cases of follicular lymphoma: comparison of Southern blotting and the polymerase chain reaction
Authors: Pezzella Francesco , Ralfkiaer Elisabeth , Gatter Kevin C. , Mason David Y.
Published: March 2008

Summary. The 14;18 chromosomal translocation is widely recognized as a cytogenetic abnormality associated with follicular lymphomas, but estimates of its frequency in this type of lymphoma vary widely from less than 50% to almost 90%. Furthermore, no extensive data have been published on the frequency of t(14;18) in European cases of follicular lymphoma. Lymph nodes from 51 patients with follicular lymphomas obtained from two European centres (Oxford and Copenhagen) were examined for the presence of this translocation. Southern blotting and the polymerase chain reaction (PCR) were used in 26 cases and the PCR alone in 25 cases (from which only degraded DNA or formalin fixed samples were available). DNA probes capable of detecting rearrangement at both the major and the minor breakpoint regions were employed. We could detect t(14:18) in only 21 out of 51 cases (41%). However, a review of the literature showed that comparable results have been obtained previously using both cytogenetic and molecular biological techniques and our results support the view that the global incidence of t(14;18) in follicular lymphoma is no greater than 70%. Furthermore, this study has indicated that the PCR is a reliable method for identifying t(14:18) when only formalin‐fixed paraffin‐embedded tissue or degraded DNA is available.
Factor VIII heavy chain polypeptides in plasma and concentrates
Authors: Kemball‐Cook G. , Bevan S. A. , Barrowcliffe T. W.
Published: March 2008

Summary. Factor VIII polypeptides in plasma and FVIII concentrates have been analysed by an electrophoretic technique based on that of Weinstein et al (1981). Samples were complexed with radiolabelled anti‐FVIII Fab, and the immunocomplexes visualized by SDS‐polyacrylamide electrophoresis. The technique visualized FVIII heavy chain polypeptides in all types of samples, including plasma, without further purification.

Fresh or frozen normal plasma (collected into protease inhibitors) contained a range of polypeptides with the largest dominant band at an apparent Mr of 250–300 kDa, and the smallest at 80–90 kDa: no bands were produced from samples of severe haemophilic plasma.

Cryoprecipitate had a similar polypeptide distribution to normal plasma, but intermediate purity FVIII concentrates showed more degraded patterns which varied between products: the 250–300 kDa bands were reduced or absent, the 80–90 kDa bands were more pronounced than in plasma, and in one product a polypeptide was seen at approximately 40–50 kDa. In some products heat treatment for viral inactivation increased the proportion of smaller FVIII polypeptides.

Highly‐purified FVIII concentrate derived from plasma was also degraded relative to plasma FVIII, and two products obtained by recombinant DNA technology both showed degraded, though slightly different. profiles.

The native structure of FVIII in fresh plasma appears heterogeneous with a predominance of higher M, forms: these are degraded to a greater or lesser extent during concentrate production, dependent on the manufacturing processes used.
In vitro and in vivo inhibition of monocyte phagocytic function by factor VIII concentrates: correlation with concentrate purity
Authors: Pasi K. J. , Hill F. G. H.
Published: March 2008

Summary. Seven factor VIII concentrates of varying purity (specific activity 0.67–10 units factor VIII procoagulant activity (VIII: C)/mg protein) have been examined over a range of concentrations (0.5–0.005 u/ml factor VIII) to establish their effect on in vitro monocyte phagocytic function. All these products inhibited monocyte phagocytic function, but the monoclonally purified product tested was significantly less inhibitory than the others (P<0.05). The degree of inhibition observed was independent of the virus inactivation method used to increase product safety. However, comparison of three products of differing purity from the same manufacturer showed that the inhibitory effect decreased as product purity increased. In vivo studies of monocyte function following infusion of two of these factor VIII concentrates demonstrated inhibition of circulating monocytes. The possible role of product purity in modulating immune responses in haemophiliacs is discussed.
Blood findings in generalized mastocytosis: evidence of frequent simultaneous occurrence of myeloproliferative disorders
Authors: Horny H.‐P. , Ruck M. , And M. Wehrmann , Kaiserling E.
Published: March 2008

Summary Blood findings in 61 cases of generalized mastocytosis (GM) were evaluated. The cases were divided into two major variants: Systemic mastocytosis (SM; n= 34) with urticaria pigmentosa‐like skin lesions, and malignant mastocytosis (MM; n = 27), without skin involvement. The following results were obtained:

Significant differences between MM and SM were found in the main haematological parameters (erythrocyte, platelet and leucocyte counts and haemoglobin level): normal values were found in 16 of the SM cases, but never in MM.

The main pathological findings were: in SM, anaemia (9/34) and leucocytosis (5/34); and in MM, leucocytosis (19/ 27), monocytosis (14/271, eosinophilia (12/27), bicytopenia (12/27, mostly anaemia with thrombocytopenia), basophilia (10/27) and isolated anaemia (7/27).

The major finding was a significant difference between MM and SM in the incidence of myeloproliferative disorders (MPD), myelodysplasia and mast cell leukaemia (MCL): these disorders occurred in 23 (92%) MM patients, but only in two (6%) SM patients (P < 0.001). The four instances of MCI, and two of myelodysplasia all occurred with MM. Of the 19 cases of MPD, six (SM, 1; MM. 5) were acute variants (acute myeloid and myelomonocytic leukaemias) and 13 (SM. 1; MM, 12) were chronic variants. No case of malignant lymphoma was noted.

The blood picture in 10 of the 13 chronic MPD cases represented an atypical chronic myeloid leukaemia for which the preliminary descriptive term‘mastocytosis‐associated MPD’is proposed.

A survey of 103 published cases (SM, 77; MM. 26) yielded similar findings, including a high incidence of MPD and MCL in MM. These findings add further weight to the argument for recognizing SM and MM as two separate entities.
Surface expression of CD 11b/CD18 of pseudo‐Pelger granulocytes in chronic myeloid leukaemia
Authors: Mazzone Antonino , And Davide Pasotti , Ricevuti Giovanni
Published: March 2008

Summary The aim of the present study was to evaluate the surface membrane glycoproteins of pseudo‐Pelger granulocytes in six patients suffering from chronic myeloid leukaemia (CML). We studied the functional and immunochemical activities of five monoclonal antibodies (MoAb) minimally reactive with integrin familial antigens of pseudo‐Pelger granulocytes. The study conducted with cytofluorimetric and immunological alkaline phosphatase anti‐alkaline phosphatase (APAAP) analysis showed a decreased expression of CD11b/CD18 detected by antibodies OKM1, 60.1 and 60.3 (P<0.001). Lymphocyte function associated antigen (LFA‐1) was expressed in normal amounts in pseudo‐Pelger granulocytes. There was decreased expression of CD11b/ CD18 in pseudo‐Pelger granulocytes with respect to controls (P<0.001) after stimulation with formyl‐met‐leu‐phe (FMLP). We conclude that acquired pseudo‐Pelger granulocyte dysfunction may be correlated to decrease of surface glycoprotein expression of CD11b/CD18.
Cytogenetic analysis of 147 cases of non‐Hodgkin's lymphoma: non‐random chromosomal abnormalities and histological correlations
Authors: Juneja S. , Lukeis R. , Tan L. , Cooper I. , Szelag G. , Parkin J. D. , And P. Ironside , Garson O. M.
Published: March 2008

Summary A prospective cytogenetic study of patients with non‐Hodgkin's lymphoma (NHL) presenting to one institution was commenced in 1983 as part of a larger study including histology, immunophenotyping, cytokinetics and survival. 175 patients were studied over 5 years and G‐banded karyotypes were successfully obtained in 147. Chromosome abnormalities were detected in 135 cases (92%) with the commonest abnormality being t(14:18)(q32;q21) in 69 cases. Other non‐random translocations were much less frequent, i.e. t(11;14) in seven cases and t(8;14) in four cases. Other specific structural changes included partial deletions of 6q (breakpoints ranging within q13‐q23). 3q (breakpoints ranging within q21‐q27), lq and 10q22. Chromosome regions highlighted as being frequently involved in structural abnormalities were 11q13‐q25. 1p22‐p36, 3q21‐q27 and 6q13‐q23. Several specific recurring breakpoints were identified and these included 14q32, 18q21. 1p36 and 6q21. Frequently occurring numerical abnormalities were gains of chromosomes 3, 7, × and 12. Correlation with histological type showed, as expected, that t(14:18) was present in 89% of follicular lymphoma but also occurred in 30% of diffuse lymphoma. Abnormalities of 11q were correlated with the diffuse histologies as a group, whereas both numerical and structural abnormalities of chromosome 3 correlated with the diffuse large cell lymphoma (DLCL) subtype, and t(11;14) with diffuse small cleaved cell lymphoma (DSCCL). Although not statistically significant, abnormalities of 6q occurred twice as frequently in DLCL than in any other variety. However, several other commonly occurring abnormalities, such as extra copies of chromosomes 7, ×, 12 and most of the structural abnormalities of 1p, did not correlate with any histological type. Therefore this large cytogenetic study has confirmed some previously reported correlations between specific chromosome abnormalities and histological subtypes of non‐Hodgkin's lymphoma and has also identified some new correlations which may prove useful in the investigation of the biological basis of the disease.
Denaturing gradient gel electrophoresis and direct sequencing of PCR amplified genomic DNA: a rapid and reliable diagnostic approach to beta thalassaemia
Authors: Losekoot M. , Fodde R. , Harteveld C. L. , Heeren H. van , Giordano P. C. , Bernini L. F.
Published: March 2008

Summary The analysis of polymerase chain reaction (PCR)‐amplified β‐globin DNA with allele‐specific oligonucleotide (ASO) probes reveals a very heterogeneous spectrum of β‐thalassaemia in the Netherlands. However about 20% of the β‐thalassaemia mutations cannot be identified with this approach. The combination of specific amplification of certain regions of the β‐globin gene with denaturing gradient gel electrophoresis (DGGE) allowed us to rapidly localize several of these mutations to specific regions of the gene. which were again amplified and directly sequenced.

We believe that the combination of DGGE and the direct sequence determination of PCR amplified genomic DNA represents a valid alternative to the‘ASO probes’approach, especially in countries where a very heterogeneous spectrum of β‐thalassaemia mutations occurs.
Osteoblast stimulation in multiple myeloma lacking lytic bone lesions
Authors: Bataille Régis , Chappard Daniel , Marcelli Christian , Rossi Jean‐Franclois , Dessauw Philippe , Baldet Pierre , And Jacques Sany , Alexandre Christian
Published: March 2008

Summary The reasons why some patients with multiple myeloma (MM) do not develop severe bone loss, or even develop sclerotic bone lesions, remain unclear. In order to answer this question at the cellular and tissue level, we evaluated the histological bone condition of 10 patients with MM who never developed lytic bone lesions during the course of their disease (including two patients with sclerotic MM).

Myeloma‐induced bone changes in the close vicinity of myeloma cells were evaluated by quantitative histology (bone histomorphometry). All 10 patients presented a significantly increased osteoblastic activity. This was associated with an increased bone resorption in seven of the 10 cases. Three patients had a pure osteoblastic presentation. These features were the reverse of the pattern observed in seven patients with lytic bone lesions: increased bone resorption with decreased bone formation. Almost all of these 10 patients showing excessive osteoblastic activity had increased serum bone gla protein levels, a specific marker of bone formation. Finally, 90% of these patients were lambda MM (70% of them were IgG lambda MM), an immunoglobulin subtype previously associated with the sclerotic MM variants.

In conclusion, a subset of patients with MM never develop severe bone loss because of the stimulation of osteoblastic activity. These patients belong to the same family as osteosclerotic MM, presenting more frequently the IgG type and lambda subtype.
Purification and characterization of a pi class glutathione S‐transferase from human leukaemic cells
Authors: Hall Andrew , Foster Sally , Proctor Stephen J. , Cattan Alex R.
Published: March 2008

Summary The glutathione S‐transferases are a group of enzymes involved in the detoxification of a wide range of xenobiotics. Elevation of the level of activity of glutathione S‐transferases within the cytosol has been associated with the development of resistance to a number of cytotoxic drugs, including some commonly used in the treatment of leukaemia. In this paper we describe the purification and characterization of an anionic (p class) form of the enzyme from the peripheral blood of patients with acute myeloid leukaemia, chronic myeloid leukaemia, and acute lymphocytic leukaemia and the spleen of a patient with chronic lymphocytic leukaemia. We present evidence that the form of enzyme purified closely resembles pi class glutathione S‐transferase purified from human placenta. Immunoblotting performed on cytosol from the leukaemic cells from a range of cases of leukaemia at presentation, or on treatment, demonstrated that this form of glutathione S‐transferase was the predominant isoenzyme expressed in all cases studied. However, in the limited number of cases studied there was no correlation between the level of expression and response to chemotherapy, suggesting that increased expression of pi class GST is not the sole cause of resistance to bifunctional alkylating agent in human leukaemias.
Irradiation of platelets with UV‐B light exposes fibrinogen binding sites via an intracellular mechanism
Authors: Kooy M. van Marwijk , Borghuis L. , Prooijen H. C. van , Aarts‐Kiemens M. I. , Akkerman J. W. N.
Published: March 2008

Summary Previous studies have shown that ultraviolet irradiation (UVI) causes platelet aggregation. In the present study we exposed platelet suspensions to a relatively high dose of UV‐B (8 J/cm2) under conditions comparable to those of UVI of platelet concentrates in order to obtain more insight into the UV‐induced aggregation response and to evaluate the significance of this phenomenon for the clinical use of UV‐irradiated platelet concentrates.

This study provides evidence that UV‐B induced aggregation is mediated by a Ca2+‐dependent process of fibrinogen binding to an intact glycoprotein IIb—IIIa complex on platelet membranes. Although UV‐induced platelet aggregation is independent of thromboxane A2 formation and ADP secretion, it requires metabolic energy, cytosolic Ca2+ and a low cyclic‐AMP level. Thus, UV‐B irradiation causes platelet aggregation by exposing fibrinogen binding sites via an intracellular mechanism. Since the amount of bound fibrinogen following UVI is relatively low (about 2300 molecules/ platelet) and the binding remains reversible, its effect on platelet behaviour after transfusion may be minor.
Options and limitations of teicoplanin in febrile granulocytopenic patients
Authors: De Pauw Ben E. , Novakova Irena R. O. , Donnelly J. Peter
Published: March 2008

Summary During the years 1985–89 three studies on the efficacy and safety of teicoplanin in the treatment of febrile granulocytopenic patients suffering from haematological malignancies were assessed. In the first prospective study, teicoplanin at a dose of 400 mg/d was added to the initial empiric therapy of 65 febrile granulocytopenic episodes. When teicoplanin was given because of proven or presumed Gram‐positive infection, 67% of cases were treated successfully. Patients with skin and soft‐tissue infections achieved a 78% response rate. The second study on 120 patients was designed as a prospective randomized trial to compare the efficacy and toxicity of ceftazidime with and without teicoplanin. Response was achieved in 25/51 (49%) cases from the ceftazidime and in 33/52 cases (63%) from the combination groups. Seven of 18 (39%) cases with initial bacteraemia in the monotherapy group compared with 10/20 (50%) cases in the combination group responded to the empiric regimen. A retrospective analysis was carried out on the efficacy of teicoplanin in 125 cases of proven Gram‐positive Hickman line‐associated bacteraemia. Teicoplanin proved to be effective in eradicating 80% of the Staphylococcus epidermidis. 72% of the Staph. aureus, 90% of the ‘viridans’ streptococci and 67% of the enterococci. Removal of the catheter was required in 13% of cases in order to control the infection, and only one patient died of a catheter‐related septicaemia caused by Staph. aureus. The total success rate leaving the catheter in situ was 78%, and 92% if cases with catheter removal were included. Serum levels of teicoplanin were predictable giving a peak and trough concentration on the fourth day of 30.4 ± 5.0 mg/l and 9.8 ± 1.7 mg/l. Hearing loss of 20 dB at 8000 Hz was noted in one case and transient liver or kidney disturbances attributable to the drug were observed in 4% of cases.
A randomized study of teicoplanin plus ciprofloxacin versus gentamicin plus piperacillin for the empirical treatment of fever in neutropenic patients
Authors: Kelsey Stephen M. , Collins Peter W. , Delord Corrine , Weinhard Barbara , Newland Adrian C.
Published: March 2008

Summary We compared the combination of teicoplanin plus ciprofloxacin with gentamicin plus piperacillin for the empirical treatment of fever in 80 neutropenic patients. A favourable clinical response rate was achieved in 28/38 (74%) patients receiving teicoplanin plus ciprofloxacin and in 17/ 35 (49%) of those receiving gentamicin plus piperacillin (P = 0.05). Microbiologically documented infections accounted for 55% of febrile events. When these episodes were analysed separately, response to teicoplanin plus ciprofloxacin remained unchanged at 74% whereas only 35% patients responded to gentamicin and piperacilin (P=0.034). Gram‐positive organisms accounted for 78% bacterial isolates with Staphylococcus epidermidis the most common pathogen. Ten out of 12 (83%) Staph. Epidermidis infections resolved when treated with teicoplanin and ciprofloxacin as compared with a response rate of only two out of eight (75%) with gentamicin and piperacillin (P = 0.032). The combination of teicoplanin and ciprofloxacin was associated with no severe drug‐related adverse events: by contrast, two patients receiving gentamicin plus piperacilin were withdrawn owing to adverse drug reactions, one with acute renal failure and one following a severe allergic reaction to piperacillin. We conclude that teicoplanin with ciprofloxacin is more effective than gentamicin plus piperacillin for the empirical treatment of febrile neutropenic patients. The high incidence of Gram‐positive infection in our unit probably justifies the use of a specific anti‐Gram‐positive agent in the first‐line antibiotic regimen.
Prevention of Gram‐positive and Candida albicans infections using teicoplanin and fluconazole: a randomized study in neutropenic children
Authors: Schaison G. , And A. Baruchel , Arlet G.
Published: March 2008

Summary We have shown that the combination of teicoplanin (T) and ceftriaxone (C) given once daily as first‐line empirical antimicrobial therapy is very effective (85%) and safe in febrile neutropenic children (ICAAC. 1988). Like others (ICAAC. 1989) we have stressed the new life‐threatening problems with Streptococcus mitis and Strep. sanguis. Considering the regular sensitivity of Gram‐positive organisms to T. it is possible that the introduction of the glycopeptide at the onset of aplasia may be helpful. The high incidence of fungal infection or superinfection supports the routine use of an effective antifungal agent when fever is still present or relapse occurs.

Our study was designed to assess the prevention of (1) staphylococcal and streptococcal infections using T when the central catheter is being placed, and (2) candida infection by the simultaneous use of a new anti‐mycotic agent fluconazole (F) administered once‐daily. Patients were randomized into two arms of the study. Arm A received T 6 mg/kg/d and F 3 mg/kg/d starting when the catheter was put in place. If fever occurred. C 50 mg/kg/d and amikacin (A) 1 5 mg/kg/d were added and T increased to 10 mg/kd/d. If a febrile relapse occurred, amphotericin B was added. In arm B the combination of the three agents T 10 mg/kg/d, C 50 mg/kg/d and A 15 mg/kg/d was started only if there was fever. Amphotericin B was used in febrile relapses. Forty‐six patients were eligible (23 in each arm: no statistically significant differences between the two groups). The mean age of the patients was 8 years and mean duration of aplasia 23 and 24 d. Results were as follows: arm A the duration of T + F alone was 10 d, 22 patients had a febrile episode with 1 Gram‐positive, 11 Gram‐negative and one candida strain isolated in eight patients. C +A were effective in 72% of these patients. There were no deaths. In arm B, fever occurred in all patients during the first 6 d, with eight Gram‐positive and six Gram‐negative organisms isolated in nine patients.

C + T + A were successful in 83% of patients. A bacterial superinfection was documented in four patients, candida superinfection in seven patients and one patient died from Candida parakrusei septicaemia. When the total amount of C and A used in both arms was compared, there was a reduction in 20% of the two agents in arm A, but T was equivalent in both arms. Treatment was well tolerated in both arms.

Teicoplanin given very early is an effective agent in preventing Gram‐positive infection, fluconazole given simultaneously is a very promising agent in the prevention of C. albicans superinfection and the value of ceftriaxone in relapsing fevers is confirmed.
Priming of normal human neutrophils by recombinant human growth hormone
Authors: Wiedermann Christian J. , Niedermühlbichler Monika , Geissler Dietmar , Beimpold Hugo , Braunsteiner Herbert
Published: March 2008

Summary Growth hormone (GH) plays an important role in the development, maintenance and function of the immune system. Previous data has demonstrated that GH is also a newly defined macrophage‐activating factor. Activation of polymorphonuclear neutrophils (PMN) by GH has not yet been examined. This paper presents studies demonstrating the effects of GH on the migratory behaviour and respiratory burst of PMN. In a modified Boyden chamber chemotaxis assay, GH did not stimulate PMN locomotion when added directly to the cells but potently inhibited formylpeptide‐stimulated chemotaxis with effective concentrations in the picomolar range. The migration inhibition observed is known from studies on PMN priming‐factors to be due to enhanced adhesiveness of PMN to artificial surfaces such as nitrocellulose, suggesting that GH stimulates PMN adhesiveness. Priming of PMN by GH was confirmed by direct demonstration of a stimulatory effect on reduction of nitroblue tetrazolium. These findings suggest that GH may be involved in the regulation of PMN functions.
Characterization of the pseudogenic and genic homologous regions of von Willebrand factor
Authors: Marchetti G. , Patracchini P. , Volinia S. , Aiello V. , Schiavoni M. , Ciavarella N. , Calzolari E. , Schwienbacher C. , Bernardi F.
Published: March 2008

Summary The homologous pseudogenic and genic regions of von Willebrand factor (vWF) were studied in DNA from a patient with homozygous deletion of vWF genes and compared with a normal control. This analysis indicates informative restriction patterns for the investigation of restriction fragment length polymorphisms (RFLPs) and gene lesions, and for molecular cloning. A useful new genic Xbal RFLP was found and characterized.

A large BgIII fragment of the pseudogenic region was cloned and mapped, and single sequences (9 kb) were used as probes. Corresponding genic and pseudogenic fragments, which contain exons 23–28, and specific restriction patterns were identified, including a new polymorphic TaqI site that was mapped in the gene. A cloned fragment contains the 5’boundary of the pseudogene and recognizes an additional and unknown homologous sequence in the genome.

The chromosomal localization of the vWF pseudogene and of the breakpoint cluster region (BCR) gene were compared by ‘in situ’ hybridization: overlapping patterns were detected. The cloning, characterization and mapping of the pseudogenic region improves the analysis of this portion of chromosome 22 affected by several somatic and constitutional alterations, and also of the corresponding genic region on chromosome 12.
Synergistic action in platelet activation induced by an antiplatelet autoantibody in ITP
Authors: Yanabu Mutsumasa , Nomura Shosaku , Fukuroi Tsutomu , Soga Tetsuji , Kondo Koji , Sone Naoaki , Kitada Chikaho , Nagata Hirokazu , Kokawa Terutoshi , Yasunaga Kojiro
Published: March 2008

Summary. We detected an autoantibody which activated normal platelets in a patient with immune thrombocytopenic purpura and investigated the mechanism by which this autoantibody mediated platelet activation. The patient's IgG induced platelet aggregation and ATP secretion in normal platelet‐rich plasma (PRP). IgG‐induced aggregation was inhibited by aspirin (ASA), apyrase, a protein kinase C (PKC) inhibitor and two anti‐platelet glycoprotein (GP) IIb/IIIa monoclonal antibodies. The increase of aequorin‐detected intraplatelet Ca2+ induced by the patient's IgG was extremely slight. Phosphorylation of a 40 kDa protein was induced by the patient's IgG without any obvious phosphorylation of a 20 kDa protein, and was inhibited by a PKC inhibitor but not by ASA. With ASA‐treated normal PRP, the patient's IgG failed to induce aggregation itself, but enhanced ADP‐ or STA2‐induced aggregation. Western blotting and immuno‐precipitation experiments showed that the patient's IgG reacted to a protein of 36 kDa. These results suggest that the platelet activation induced by this autoantibody depended on both the selective activation of PKC and the slight Ca2+ mobilization induced by thromboxane A2 synthesis, while the aggregation depended on secretion induced by the synergistic action of the above two mechanisms and was mediated through GP IIb/IIIa.
Polymerase chain reaction for detection of human cytomegalovirus infection in a blood donor population
Authors: Bevan I. S. , Daw R. A. , Day P. J. R. , Ala F. A. , Walker M. R.
Published: March 2008

Summary. We have used the polymerase chain reaction (PCR) to analyse 420 normal donor blood samples taken at a city centre donation site. Three sets of human cytomegalovirus (HCMV) primers specific to the HXLF6, immediate early and late antigen gp64 genes, of the α, β and γ antigen coding regions respectively, were used to allow for the possibility of sequence variation. There was perfect correlation between the three sets of primers. Latex agglutination and enzyme‐linked immunosorbent assay (ELISA) were also employed to provide data for a comparative study. The complete data show that infection with human cytomegalovirus is not only age related but is also sex related. The female population examined using PCR reached a peak infection rate of 80% by the age of 40–50 years whereas the male population reached a 98% infection rate following an almost linear increase with age. Latex agglutination data shows a similar picture although the infection rate peaks around 20% lower than with PCR. The data shows an increase in sensitivity using the PCR rather than the serology although the clinical significance of this has yet to be determined. The work presented here also demonstrates the suitability of the polymerase chain reaction as a potential diagnostic and epidemiological tool.
Diminished AγT fetal globin levels in Sardinian haplotype II β°‐thalassaemia patients are associated with a four base pair deletion in the AγT promoter
Authors: Manca L. , Cocco E. , Gallisai D. , Masala B. , Gilman J. G.
Published: March 2008

Summary In Sardinia, the β‐39 nonsense mutation is the primary cause of β°‐thalassaemia. This mutation is found mainly on β‐globin gene cluster haplotypes I and II, which differ in their Aγ globin types (AγI and AγT, respectively). This report presents data on Gγ, AγI and AγT levels, and the presence or absence of a 4 base pair (bp) deletion at ‐225 to ‐222 of the Aγ globin promoter, in 55 poly‐transfused β°‐thalassaemia major patients. Six patients were homozygotes for the normal (N) Aγ promoter lacking the 4 bp deletion, had no AγT globin, and their mean Gγ:AγI:AγT ratio was 52·9:47·1:0. Twenty‐five patients were homozygotes for the mutant (M) Aγ promoter with the 4 bp deletion, had no AγI globin, and the mean Gγ:AγI:AγT ratio was 62·1:0:37·9. For M/M compared to N/N, the lower AγT than AγI was significant by the t‐test (P<0·001). Twenty‐four N/M cases had mean Gγ:AγI:AγT of 56:24·4:19·6, and the lower AγT than AγI was also significant (P<0·001). Partial haplotype analysis on these and 17 other β°‐thalassaemia patients suggested that the 4 bp deletion was strongly associated with haplotype II. Of 33 M/M, 32 were haplotype II/II and one was II/5a; of 31 N/M, 29 were I/II and two were II/IX; of eight N/N, seven were haplotype I/I and one was I/IX. These data show a strong association of the 4 bp promoter deletion with decreased expression of the AγT globin gene on haplotype II.
Splenic lymphoma with villous lymphocytes: natural history and response to therapy in 50 cases
Authors: Mulligan Stephen P. , Matutes Estella , Dearden Claire , Catovsky Daniel
Published: March 2008

Summary We studied the natural history and response to treatment in 50 patients with splenic lymphoma with villous lymphocytes followed for a minimum of 6 months and up to 15 years (median 3·7 years). The disease occurs in the elderly (median 68 years) and affects males more than females (M/F ratio 1·77). The median survival for the group was not reached but 82% were surviving at 3 years and 78% at 5 years. Twelve patients (24%) died, one‐third of deaths were disease related (four patients) and one‐half were due to cardiovascular disease or another malignancy (six patients). The outcome was worse for males (31% died) than females (11% died).

Fourteen patients were not treated and 10 remain alive between 1 and 6 years from diagnosis. The remaining patients were treated by chemotherapy, splenic irradiation or splenectomy. The response to chemotherapy was poor and only eight of 22 (36%) patients treated achieved a good response. Splenic irradiation was employed in seven patients and three benefited from it. Splenectomy seems to be the treatment of choice, with significant improvement seen in 19 of 20 patients with one post‐operative death. This response, lasting from 6 months to 7 years (median 4 years), was seen irrespective of whether splenectomy was the initial treatment or used later in management.
Establishment and characterization of a plasma cell leukaemia cell line dependent for growth on IL‐6 and a bi‐phenotypic subclone dependent upon both IL‐3 and IL‐6
Authors: Kobayashi M. , Miyagishima T. , Imamura M. , Maeda S. , Gotohda Y. , Iwasaki H. , Sakurada K. , Miyazaki T.
Published: March 2008

Summary A human plasma cell leukaemia cell line (HSM‐2) and a subclone (HSM‐2.3) have been established from the bone marrow of a patient with bi‐phenotypic leukaemia. Proliferation assays using a variety of cytokines demonstrated that HSM‐2 proliferated in response to recombinant interleukin‐6 (rIL‐6), but did not respond to rIL‐1, rIL‐2, rIL‐3, rIL‐4, rIL‐5, recombinant granulocyte‐colony stimulating factor (rG‐CSF), or recombinant granulocyte‐macrophagecolony stimulating factor (rGM‐CSF), and that HSM‐2.3 responded to rIL‐3 and rIL‐6. HSM‐2 expressed the CD38 (OKT10), PCA‐1, cytoplasmic‐IgM, and surface kappa light chain. HSM‐2.3 expressed the CD14 (My4), CD33 (My9). CD38 (OKT10), CD19 (B4), CD24 (OKB2), CD10 (J5), PCA‐1. HSM‐2 and HSM‐2.3 are useful tools for analysing the possible role of IL‐3 and IL‐6 in the oncogenesis of plasma cell leukaemia.
The spectrum of β‐thalassaemia mutations on the Indian subcontinent: the basis for prenatal diagnosis
Authors: Varawalla N. Y. , Old J. M. , Sarkar R. , Venkatesan R. , Weatherall D. J.
Published: March 2008

Summary The β‐thalassaemia mutations in 702 unrelated carriers originating from seven different regions of the Indian subcontinent have been characterized using allele specific priming of the polymerase chain reaction (PCR). It was possible to identify the mutations in 688 (98%) of the individuals studied. Eleven different mutations were identified, of which five common ones accounted for 93·6%; namely the ones at IVS‐1 position 5 (G‐C), codons 8/9 (+G), IVS‐1 position 1 (G‐T), codons 41/42 (‐CTTT) and the 619 bp deletion at the 3’end of the gene. The mutations at IVS‐2 position 1 (G‐A) and codon 30 (G‐C), previously undescribed in Asian Indians, were found in two and six individuals respectively. Some regional variation in the distribution of β‐thalassaemia alleles was noted. These findings should prove useful for the development of a first trimester prenatal diagnosis programme based on direct detection of mutations.
Congenital spherocytosis, B19 parvovirus infection and inherited interstitial deletion of the short arm of chromosome 8
Authors: Cohen H. , Walker H. , Delhanty J. D. A. , Lucas S. B. , Huehns E. R.
Published: March 2008

Summary We report two siblings with congenital spherocytosis, multiple phenotypic abnormalities and an inherited interstitial deletion of the short arm of chromosome 8 (8p). The propositus came to our attention with acute bone marrow hypoplasia secondary to B19 parvovirus infection. The bone marrow trephine biopsy appearances of intranuclear eosinophilic degeneration in the erythroblasts may be pathognomonic of B19 parvovirus induced acute bone marrow aplasia. The presence of B19 parvovirus DNA was demonstrated in erythroblasts by in situ hybridization. Chromosome analysis of peripheral blood lymphocytes from both siblings showed an interstitial deletion of the short arm of chromosome 8, del (8) (p11p21). This abnormal chromosome was inherited from their mother, who showed this deletion as well as a small fragment representing the deleted 8p chromosome portion, del (8) (p11p21), + f. Centromeric material from chromosome 8 was detected in this chromosome fragment by in situ hybridization using an alpha satellite probe (pJM 128), but not by C banding. Chromosome analysis of skin fibroblasts from the mother and a third sibling with a similar karyotype showed the deleted fragment in over 80% of cells. Cells in which the fragment was absent exhibited the deleted 8p, suggesting there was no mosaicism. The mother and the third sibling were phenotypically normal without spherocytosis. A fourth sibling and the father were normal. The chromosome abnormality was not observed in five of the mother's siblings, suggesting that it arose de novo in the mother. Our findings strongly support a locus for congenital spherocytosis on the short arm of chromosome 8. The frequency of defects at this locus is unknown.
Late onset immune pancytopenia following bone marrow transplantation
Authors: Bashey Asad , Owen Irene , Lucas Geoffrey F. , Amphlett Nigel W. , Jones Margaret M. , Lawal Adebayo , McMullin Mary‐Frances , Mahendra Prem , Tyfield Linda A. , Hows Jill M.
Published: March 2008

Summary A 17‐year‐old boy developed autoimmune pancytopenia in the absence of chronic graft‐versus‐host disease 170 d after allogeneic bone marrow transplantation (BMT) from his HLA identical brother. The anaemia and thrombocytopenia responded to conventional immunosuppressive treatment, but the neutropenia was refractory to this and to splenectomy and subsequent removal of splenic remnant.

Following total lymphoid irradiation the neutrophil count rose to low normal levels but thrombocytopenia and anaemia secondary to marrow hypoplasia required transfusion support. Bone marrow function was finally normalized by an additional transfusion of donor marrow without prior immunosuppressive therapy. We conclude that late onset immune pancytopenia post BMT caused by antibodies of probable donor origin may be life threatening in the absence of chronic graft‐versus‐host disease.
Patterns of CD16 and CD56 expression in persistent expansions of CD3+NKa+ lymphocytes are predictive for clonal T‐cell receptor gene rearrangements
Authors: Sivakumaran M. , Richards Stephen J. , Hunt Kaye M. , Steed Andrew J. , Bynoe A. Geraldine , Morgan Mary M. , Pyrah Roger , Roberts Bryon E. , Scott Colin Stephen
Published: March 2008

Summary. Phenotypic characteristics, and correlations between the expression of membrane NK‐associated (NKa) determinants (CD11b, CD16, CD56 and CD57) and T cell receptor (TCR) genotypic patterns, were examined in 25 patients with persistent (>6 months) expansions of CD3+WT31+NKa+ (CD8+ and CD8dim+) lymphocytes. These studies showed that distinct NKa phenotypic profiles were restricted to cases with rearranged TCR configurations and that clonal CD3+NKa+ components could be predicted in most cases by assessing relationships between membrane CD16 and CD56 expression. For all normal NKa subpopulations, there was a high correlation (P<0·0001: n= 31) between the expression of these two membrane determinants. Markedly increased CD16 expression by CD3+NKa+ cells, in relation to CD56 (i.e. a high CD16:CD56 ratio), was found exclusively in cases with rearranged TCR (13/16 cases); 2/3 of the remaining cases showing significantly reduced CD16:CD56 ratios and high (>2·0) CD3+CD56+ absolute numbers. In contrast, 7/9 of the germline TCR cases had a normal CD16:CD56 ratio and 2/9 a decreased ratio with low (<1·0) CD3+CD56+ absolute numbers. A high ratio of CD16:CD56 expression by CD3+NKa+ lymphocytes was therefore informative for 82% of TCR rearrangements in this series: and analysis of CD16 and CD56 expression was predictive for germline and rearranged TCR configurations in 24/25 persistent CD3+NKa+ expansions.
Hepatitis C antibody assay in a longitudinal study of haemophiliacs
Authors: Lim Seng G. , Lee Christine A. , Charman Heather , Tilsed Gillian , Griffiths Paul D. , Kernoff Peter B. A.
Published: March 2008

Summary. Stored sera from 28 patients with inherited coagulation disorders who had developed non‐A non‐B hepatitis (NANBH) following a first exposure to clotting factor concentrates and 15 similar, but unmatched, patients who had received blood products but had normal transaminases on sequential testing were tested using the Ortho enzyme‐linked immunosorbent assay (ELISA) anti‐HCV assay. Twenty‐seven of the 28 patients with NANBH were anti‐HCV positive after exposure. In 10 of those in whom dates of first exposure and seroconversion were well‐defined, the median time interval to NANBH was 4 weeks (range 1–7) and to anti‐HCV seroconversion was 11 weeks (range 7·5–14·5). None of the 15 patients without NANBH developed anti‐HCV. This first generation Ortho ELISA anti‐HCV assay showed 96% sensitivity and 100% specificity and has potential use as an adjunct in the surveillance of new clotting factor products.
Prothrombin fragment 1 + 2, thrombin–antithrombin III complexes and D‐dimers in acute deep vein thrombosis: effects of heparin treatment
Authors: Estivals M. , Pelzer H. , Sie P. , Pichon J. , Boccalon H. , Boneu B.
Published: March 2008

Summary. Plasma levels of prothrombin fragment 1 + 2 (F 1 + 2), of thrombin–antithrombin III complexes (TAT) and of D‐dimers were evaluated at several time intervals in 15 patients affected by acute proximal deep vein thrombosis, complicated or not by pulmonary embolism, and treated by conventional heparin therapy for 9 d. The mean levels of the three markers remained significantly increased throughout the period of observation, except for F 1 + 2 on day 9, when compared to normal values established in a population of normal healthy blood donors. However, whereas heparin significantly decreased the plasma levels of F 1 + 2 and of TAT complexes in less than 3 d, D‐dimer levels were not significantly altered. Significant correlations were observed between the plasma levels of the three markers but they were not correlated to the actual intensity of heparin treatment evaluated as the activated partial thromboplastin time prolongation. These results indicate that heparin improves the hypercoagulable state associated with a deep vein thrombosis within the first days of treatment as indicated by TAT and F 1 + 2. They also account for the performances of D‐dimer assay for the diagnosis of deep vein thrombosis in patients already receiving heparin, a common situation in routine hospital practice.
Production of soluble transferrin receptor by K562 erythroleukaemia cells
Authors: Baynes Roy D. , Shih Yuan J. , Cook James D.
Published: March 2008

Summary. The present study was undertaken to examine the production of soluble transferrin receptor by K562 erythroleukaemia cells under controlled experimental conditions. The concentrations of soluble and cellular transferrin receptor were measured by immunoassay employing monoclonal antibodies. Cellular ferritin was also measured as an index of iron supply. With incubation up to 48 h there was a progressive increase in the concentration of souble transferrin receptor. Manipulating iron supply by adding iron chelators or diferric transferrin to the incubation medium produced marked alterations in cellular receptor and ferritin content. Under all such conditions examined, the relationship between soluble and cellular receptor remained highly constant. These findings support clinical studies of serum receptor suggesting that over a broad spectrum of haematological disorders there is a fixed relationship between serum receptor and tissue receptor mass.