bluuebunny/crossref_metadata_2025 huggingface dataset:
The role of interferons in the management of carcinoid tumours
Authors: ÖBerg K. , Eriksson B.
Published: March 2008

Malignant carcinoid tumours with the carcinoid syndrome has over the years presented a therapeutic challenge. The patients might not only die from tumour progression but also from symptoms relating to hormone overproduction and the specific cardiac disease, e. g. right heart fibrosis and failure. Surgery has been the treatment of choice in local disease, but when liver metastases have developed other treatment procedures must be considered. Conventional chemotherapy has been of little beneficial value, with response rates of only 10–30%, whereas a new somatostatin analogue, octreotid, is effective in controlling clinical symptoms but not tumour progression. Interferon treatment was introduced in 1982 by our group, and we are now presenting treatment results of 130 patients with histologically verified malignant carcinoid tumours and liver metastases. One hundred and eleven patients were treated with a median dose of 6 mega units (MU) of interferon alpha, five times weekly (dose range 3–9 MU), whereas 29 patients received conventional chemotherapy. Forty‐seven out of 111 patients (42%) treated with interferon alpha demonstrated a significant biochemical response and 15% demonstrated more than 50% reduction in tumour size. In another 43 (39%) patients stabilization of the carcinoid disease have been noted, whereas 21(19%) showed progressive disease. The median duration of response was 34 months. Subjective responses with improvement of diarrhoea, flush and/or bronchocon‐striction were noticed in 76 patients (68%). The 19 patients treated with chemotherapy demonstrated only 10% biochemical response, lasting for only 3–5 months. The survival analysis demonstrates a median survival of only 8 months in the group of patients treated with chemotherapy, compared to 80+ months (P<0·001) in the groups treated with interferon alpha. Interferon adverse reactions of fatigue, weight loss and anaemia were manageable. Neutralizing interferon antibodies were documented in 5–15% of the patients.

Interferon alphas are active in patients with malignant carcinoid tumours. Clinical symptoms are significantly reduced following reduction of circulating hormones. Interferon might also have an impact on survival in this group of patients. The side‐effects are moderate and managed by dose adjustments.
Interferon in combination with antitumourigenic phenyl derivatives: potentiation of IFNα activity in‐vitro
Authors: Samid Dvorit , Yeh Tze‐Jou , Shack Sonsoles
Published: March 2008

Any attempt to eradicate the heterogeneous cell population of a tumour mass would require the use of appropriate combination treatment protocols. The antitumour effects of interferon alpha (IFNα) in combination with AS2‐1, the hydrolysis product of 3‐phenylacetyl‐amino‐2,6‐piperidinedione, were examined using several human tumour cell lines as a model. These included the malignant melanoma A375, adenocarcinoma of the prostate PC3 (hormone‐insensitive bone metastasis), and the erythroleuk‐aemia line K562. AS2‐1 suppressed tumour growth through non‐toxic mechanisms, with 1 mg/ml causing approximately 50% inhibition of the melanoma and prostate tumour cell proliferation. By contrast, primary normal human skin fibroblasts were significantly less sensitive to the antiprolifer‐ative effect of AS2‐1. Suppression of tumour growth was seen also with AS2‐1 treatment of the erythroleukaemia K562; in these cultures the drug also induced dose‐dependent differentiation, as indicated by the increased haemoglobin production. Interestingly, addition of low doses of IFNα markedly enhanced the antitumour and differentiating effects observed with AS2‐1. Treatment with 200–300 IU/ml of IFN (which caused about 20% inhibition of growth) together with 1 mg/ml of AS2‐1 resulted in over 80% inhibition of the melanoma and prostate cancer cell proliferation, suggesting a synergistic activity of the two agents. This was substantiated by quantitative analysis of the differentiation induced in K562 erythroleukaemia. It appears, therefore, that IFNα and AS2‐1 may act through synergistic mechanisms to effectively inhibit tumour growth and promote differentiation in a variety of human malignant cell lines.
Retinoids and interferon: a new promising combination?
Authors: Bollag W.
Published: March 2008

The retinoids: all‐transretinoic acid (tretinoin), 13‐cis retinoic acid (isotretinoin) and the aromatic retinoids etretinate and acitretin have a preventive and therapeutic effect on chemically‐induced tumours. Clinically, retinoids have shown variable effectiveness in therapy and/or prevention of oncological diseases of skin, head and neck, lung, bladder, vulva and bone marrow, With a few exceptions, monotherapy with retinoids has not been satisfactory. Similarly, monotherapy with interferon alpha has been used successfully only for some specific indications. Retinoids have a marked differentiation‐inducing effect which may contribute to their therapeutic effect. Experiments were carried out in transformed cell lines to test the combination of retinoids with interferon alpha and other cytokines on differentiation. In HL‐60 cells, an acute promyelocytic leukaemia cell line, induction of differentiation was determined by induction of an oxidative burst potential. Retinoids showed the following order of activity: tretinoin > isotretinoin > acitretin. Cytokines had no differentiation‐inducing effect by themselves. However, the addition of the following cytokines to retinoids potentiated the retinoid‐induced differentiation: IFNα, IFNβ, IFNγ, TNFα, G‐CSF, IL‐1α and IL‐4. In experiments with HL‐60 or other cell lines, the pattern of differentiation‐induction was always dependent on the particular retinoid/cytokine combination. IFNα provoked a marked potentiation of retinoid‐induced differentiation. The combination of the antipro‐liferative and differentiation‐inducing effect of the retinoids together with the antiproliferative, immunostimulatory and differentiation‐potentiating effects of IFNα suggests that this combination might be a particularly promising treatment for neoplastic diseases.
Lactoferrin inducible monocyte cytotoxicity defective in esterase deficient monocytes
Authors: McCormick Julie A. , Markey Geraldine M. , Morris T. C. M. , Auld P. W. , Alexander H. D.
Published: March 2008

We recently demonstrated a higher incidence of monocyte esterase deficiency in patients with lymphoproliferative neoplasia and gastrointestinal carcinoma than in the normal population. Using lactoferrin to stimulate monocyte cytotoxicity we have now compared the abilities of esterase positive and esterase deficient monocytes to lyse K562 cells. The esterase deficient monocytes were from normal subjects whose monocytes have consistently failed to show cytoche‐mical esterase staining over 30–72 months and which lack specific monocyte isoenzymes. The esterase positive monocytes were from age and sex matched control subjects. Esterase positive monocytes responded to lactoferrin stimulation by a five‐fold increase in cytotoxicity whereas deficient monocytes failed to produce any response.

These results indicate a possible defect in cytotoxicity of esterase deficient monocytes and together with the epidemiological findings suggest a link between monocyte esterase deficiency and neoplasia.
Autologous blood stem cell transplantation for chronic granulocytic leukaemia in transformation: a report of 47 cases
Authors: Reiffers J. , Trouette R. , Marit G. , Montastruc M. , Fabères C. , Cony‐Makhoul P. , David B. , Bourdeau M. J. , Bilhou‐Nabera C. , Lacombe F. , Feuillatre‐Fabre F. , Vezon G. , Bernard Ph. , Broustet A.
Published: March 2008

Forty‐seven patients with chromosome Philadelphia‐positive (Ph1) chronic granulocytic leukaemia (CGL) in transformation underwent autologous transplantation of peripheral blood stem cells (ABSCT) collected at the original diagnosis before any treatment. They were treated with three consecutive strategies: single transplant (group I=17 patients), double transplant (group II = 13 patients), double transplant followed by recombinant alpha interferon (group III=17 patients). Forty‐three patients were restored to a second chronic phase with a cytogenetic conversion (more than 10%, Ph1‐negative marrow metaphases) occurring in 14 of the 29 evaluable patients. Most patients had a recurrent transformation occurring 2‐43 months after ABSCT and finally eight patients are still alive in second chronic phase 4‐49 months after ABSCT (median = 24 months). The actuarial median duration of second chronic phase was 3 months, 10 months and 18 months for group I. group II and group III patients (P < 0·0001). The encouraging results observed for group III patients prompt us to propose ABSCT for patients in chronic phase with initial prognostic factors, suggesting that recombinant alpha interferon will not be effective if administered as front‐line therapy.
Cell‐mediated cyclic thrombocytopenia treated with azathioprine
Authors: Dan Kazuo , Inokuchi Kohichi , An Emi , Nomura Takeo
Published: March 2008

We studied a male patient with cyclic thrombocytopenia whose bone marrow megakaryocyte count showed cyclic fluctuations in synchrony with cyclic changes of platelet count. He failed to respond to either prednisolone or bolus methylprednisolone therapy, but subsequently he was successfully treated with azathioprine. To investigate the underlying pathogenesis of the cyclic fluctuations in the platelet count. we studied the kinetics of megakaryocyte progenitor cells (CFU‐Meg) and the effects of the patient's peripheral blood mononuclear cells on CFU‐Meg. In one cycle of the platelet fluctuation, the increase in the CFU‐Meg number preceded an increase in the bone marrow megakaryocyte count and then the platelet count. In the latter half of the cycle, CFU‐Meg, bone marrow megakaryocytes and platelets began to decrease in that order. Peripheral blood mononuclear cells obtained from the patient in the thrombo‐cytopenic phase suppressed megakaryocyte colony formation from normal bone marrow cells in a dose‐dependent manner. In contrast, these cells obtained in the phase of a normal platelet count did not suppress megakaryocyte colony formation at all. These findings indicate that the cause of platelet fluctuation is periodic failure of megakaryocytopoiesis at the stage of CFU‐Meg and that the patient's peripheral mononuclear cells are responsible for periodically suppressing the CFU‐Meg.
Reduction of platelet glycoprotein Ib in uraemia
Authors: Sloand E. M. , Sloand J. A. , Prodouz K. , Klein H. G. , Yu M. W. , Arvath L. H. , Ricke W. F.
Published: March 2008

Patients with uraemia have abnormal platelet function that may be partially corrected by haemodialysis, cryoprecipitate or 1‐desamino‐8‐d‐arginine vasopressin (DDAVP). We studied the platelet von Willebrand factor receptor. glycoprotein Ib (GPIb). and plasma von Willebrand factor (vWF) in uraemic patients undergoing chronic haemodialysis. Using the slope of agglutination of formalin‐fixed platelets as an index of response to ristocetin (with a constant amount of normal plasma as a source of vWF), we found the response of platelets from uraemic patients, both before (2·7± 1·5. n= 40) and after dialysis (1·2 ± 1·2, n= 40) to be significantly less than that for normal controls (14·1 ± 10·2. n= 20: P < 0·001). In addition, the agglutination response of platelets obtained after dialysis was less than that of platelets obtained before dialysis (P < 0·001). Immunoblotting demonstrated decreased or absent staining of glycocalicin, a subunit of GPIb, in platelet lysates from 25 patients. All platelet samples with reduced glycocalicin also had decreased responses to ristocetin. Tritium‐labelled platelets from seven patients showed decreased labelling of a protein with an electrophoretic mobility equivalent to that of GPIb (140000 daltons). In addition, platelets with the lowest levels of surface GPIb, as demonstrated by flow cytometry. also had decreased ristocetin agglutination and decreased staining on immunoblot. Levels of von Willebrand factor antigen and ristocetin cofactor in plasma from 10 patients were generally within the normal range, although post‐dialysis levels tended to be higher than pre‐dialysis levels. The pre‐ and post‐dialysis plasma vWF multimeric patterns were normal.
The haematology of homozygous sickle cell disease after the age of 40 years
Authors: Morris Joanne , Dunn David , Beckford Marjorie , Grandison Yvonne , Mason Karlene , Higgs Douglas , Ceulaer Karel De , Serjeant Beryl , Serjeant Graham
Published: March 2008

Haematological indices have been studied in 181 patients with homozygous sickle cell (SS) disease aged 40–73 years. Cross‐sectional analyses in 5‐year age bands indicated age‐related decreases in HbF (males only), total haemoglobin and platelet counts. Longitudinal studies within individuals confirmed the downward age‐related trend in haemoglobin and platelets and also revealed a falling reticulocyte count, most significant when expressed as absolute values. Total nucleated cells also fell although the decline was significant only in females. These observations are consistent with a progressive bone marrow failure which is not explained by the commonly occurring renal impairment in older SS patients since the changes persisted in analyses confined to patients with normal creatinine levels. The mechanism of this bone marrow failure is currently unknown.
Fanconi's anaemia and pregnancy
Authors: Alter Blanche P. , Frissora Christine L. , Halpéirin Daniel S. , Freedman Melvin H. , Chitkara Usha , Alvarez Emanuel , Lynch Lauren , Adler‐Brecher Barbara , Auerbach Arleen D.
Published: March 2008

We have identified six new cases of Fanconi's anaemia (FA) who had pregnancies, and reviewed 11 others from the literature. At least 110 FA females have reached 16 years of age or more, of whom 15% became pregnant. There were a total of 26 pregnancies, resulting in 19 births and 18 surviving children. Anaemia and/or thrombocytopenia worsened during pregnancy in 10 patients, but five subsequently improved: seven had no haematological problems. Seven of the FA patients who had pregnancies died subsequently from cancer, and two from thrombocytopenic bleeding 3 and 20 years later. There were no peripartum deaths. Pregnancy in FA is clearly possible, with increased risks that are manageable from both the haematological and the obstetric aspects.
P53 gene mutations in acute myelogenous leukaemia
Authors: Hu Guiying , Zhang Wei , Deisseroth Albert B.
Published: March 2008

A polymerase chain reaction–single strand conformation polymorphism (PCR‐SSCP) assay was used to identify the exons which contained point mutations in the conserved regions (exons 4–8) of the p53 gene in 49 acute myelogenous leukaemia (AML) patients. SSCP analysis in our study was consistent with the results of subsequent direct DNA sequencing in detecting point mutational change in exons 5 and 8 of one AML patient and in exons 7 and 8 of two additional AML patients. The mutations were located at codons 245 and 273, which have been found in many other tumours, and codons 178 and 290, which have not been reported previously. All of the p53 proteins in which we detected point mutations were immunoprecipitated by the p53 monoclonal antibody PAb 240, which has been shown to recognize a mutant conformation of p53 protein. Thus, our results indicate that functional inactivation of the p53 gene by point mutational change might be one of the mechanisms underlying disease progression of AML.
Two myelodysplastic syndrome cases with the inv(ll)(pl5q23) as a sole chromosomal abnormality
Authors: Mitani Kinuko , Sato Yuko , Hayashi Yasuhide , Miura Yasusada , Miyagawa Kiyoshi , Yazaki Yoshio , Hirai Hisamaru
Published: March 2008

Two myelodysplastic syndrome cases (one with acute nonlymphocytic leukaemia (M2) transformed from myelodysplastic syndrome (MDS), and the other with chronic myelomonocytic leukaemia following refractory anaemia with excess of blasts in transformation) showed the inv(11)(p15q23) as a sole chromosomal abnormality. Gene probes for c‐Ha‐ras‐1 and c‐ets‐1 were hybridized to meta‐phase cells from bone marrow of these patients, c‐ets‐1 gene, which is mapped to 11q23, was demonstrated to have translocated to the short arm in the rearranged chromosome 11 in both cases. On the other hand, c‐Ha‐ras‐1 gene, which is located at 11p15, was translocated to the long arm in the rearranged chromosome 11 in patient 1, and deleted in patient 2. Our findings suggest that there may be heterogeneity in molecular events involved in the chromosomal rearrangement among the inv(11)‐carrying MDS.
A monoclonal antibody (NMC‐VIII/10) to factor VIII light chain recognizing Glu1675–Glu1684 inhibits factor VIII binding to endogenous von Willebrand factor in human umbilical vein endothelial cells
Authors: Shima Midori , Yoshioka Akira , Nakajima Mitsuru , Nakai Hiroaki , Fukui Hiromu
Published: March 2008

The monoclonal antibody NMC‐VIII/10 is a neutralizing antibody which recognizes the Glu1675–Glu1684 sequence of the factor VIII light chain and inhibits factor VIII (FVIII) binding to immobilized von Willebrand factor (vWf). In this study we immunohistochemically determined, using human umbilical cord tissue, whether or not NMC‐VIII/10 has an inhibitory effect on FVIII binding to endogenous vWF in endothelial cells. Tissue sections were reacted with purified FVIII followed by peroxidase‐conjugated monoclonal antibody (C5) recognizing the 54 kD fragment of the FVIII heavy chain. The labelling pattern of bound FVIII was similar to that of endogenous vWF and appeared as a fine granular deposit in the endothelial cells. Addition of purified vWF completely inhibited the binding of FVIII to endothelial cells. Furthermore, FVIII did not bind to endothelium in the presence of 0·25 M CaCl2, and similarly, thrombin‐treated FVIII did not bind to the vascular site. These findings suggested that FVIII was bound to endogenous vWF in the endothelial cells. The binding reaction was completely inhibited by NMC‐VIII/10, confirming that the monoclonal antibody recognizes the specific epitope responsible for FVIII binding to endogenous vWF.
Amino acid metabolism during platelet storage for transfusion
Authors: Murphy Scott , Munoz Santiago , Parry‐Billings Mark , Newsholme Eric
Published: March 2008

Previous studies indicated that the concentration of ammonia rises during storage of platelet concentrates (PC) at 22°C for transfusion and that fuels other than glucose are important for metabolism. Therefore, in the current study, we measured the concentrations of 17 plasma amino acids during PC storage; 16 of these either rose or were unchanged while the concentration of glutamine fell to zero by day 4. As the concentration of glutamine fell, the concentration of glutamate rose with a relationship suggesting that 65–75% of the glutamine was metabolized no further than glutamate. Phosphate‐dependent glutaminase activity was present in platelets at 22·3 ± 6·3 nmol/min/mg protein, a level similar to that seen in lymphocytes and macrophages. Leucodepletion studies excluded a significant contribution of contaminating leucocytes to these measurements. Thrombin stimulation did not increase the rate of glutamine metabolism. Analysis of the rates of glutamine metabolism suggests that it accounts for most of the ammonia produced during PC storage. However, it appears to be relatively insignificant as a metabolic fuel. The role of glutamine metabolism for platelets is uncertain. It may be a vestige of a pathway in the megakaryocyte. The ammonia which it produces may be deleterious for platelets and for patients with liver disease who receive PC infusions.
Childhood polyclonal T cell lymphocytosis with neutropenia: effects of antilymphocyte globulin and granulocyte colony stimulating factor in vitro and in vivo
Authors: Laurie A. S. , Gibson F. M. , Josten K. M. , Talbot P. J. , Rutherford T. R. , Lilleyman J. S. , Gordon‐Smith E. C.
Published: March 2008

Summary The pathogenesis of the neutropenia that occurs in some patients with chronic T cell lymphocytosis is not well understood. We have investigated a 15‐year‐old girl with this syndrome. Initial committed bone marrow progenitor numbers (CFUgm) were low but markedly increased in vitro following T cell depletion. Similarly a transient correction of neutropenia was observed following in vivo lymphocyte depletion with antilymphocyte globulin. A sustained neutrophil recovery was achieved with daily therapy using recombinant human granulocyte colony stimulating factor (rhG‐CSF) despite persistence of the lymphocytosis; during successful therapy CFUgm numbers remained low, and were not increased by the in vitro addition of rhG‐CSF. These observations suggest the possibility of an inhibitory regulatory mechanism specifically acting on neutrophil granulopoiesis.
B‐cell chronic lymphocytic leukaemia cells express and release transforming growth factor‐β
Authors: Kremer Jean‐Pierre , Reisbach Gilbert , Nerl Christoph , Döurmer Peter
Published: March 2008

Summary Transforming growth factor‐β (TGF‐β) has been described as a potent inhibitor of various cell types, among others of primitive haematopoietic progenitors in vitro, and as a negative autocrine regulator of normal B lymphocyte growth and differentiation. In the present study we investigated TGF‐β gene expression in peripheral blood mononuclear cells (PBMC) and in B cells from patients with B‐cell chronic lymphocytic leukaemia (B‐CLL) and from normal controls. Monocyte depleted B‐CLL cells expressed constitutively mRNA for TGF‐β1 and secreted low amounts of TGF‐β activity into the culture medium. Stimulation of cells by phorbol ester noticeably enhanced mRNA levels as well as protein secretion in most cases. TGF‐β activity was of the same magnitude as in normal controls. We next analysed TGF‐β in highly enriched B lymphocytes from B‐CLL (95100% CD19+), and found that TGF‐β secretion was up to 3 times higher than in the original PBMC population. It is discussed that B‐CLL cells might escape from negative regulation by TGF‐β and, on the other hand, inhibit normal haematopoietic cell proliferation and thereby achieve a growth advantage in the haematopoietic tissues.
Insights into heparin‐induced thrombocytopenia
Authors: Anderson George P.
Published: March 2008

Summary Heparin‐induced thrombocytopenia (HIT) is characterized by the presence of an antibody which can activate platelets in the presence of heparin. The in vitro Ca2+ mobilization induced by purified IgG from patients with HIT was evaluated to elucidate the platelet activation mechanism in this syndrome. HIT‐IgG induced platelet activation in a heparin‐dependent manner. This activation was inhibited by high viscosity, consistent with a previously documented cell‐cell‐mediated mechanism. We found that F(ab')2 fragments from an anti‐GPIIb/IIIa monoclonal antibody in some cases, and dextran sulphate or salmon sperm DNA in all cases, could substitute for heparin, suggesting that heparin exposes a neo‐antigen on the platelet surface rather than serving as the primary antigen in the process. Furthermore, heparin, dextran sulphate and salmon sperm DNA all augmented the platelet activation induced by aggregated IgG, suggesting an additional charge shielding effect. These data suggest the following mechanism for platelet activation in HIT: heparin binds to the platelet surface, exposing a neoantigen which elicits an antibody response in some patients. Antibody binding in the presence of heparin leads to platelet activation through Fc receptor mediated platelet‐platelet interaction, a process augmented by the charge shielding effect of heparin.
Hb H disease in a Turkish family resulting from the interaction of a deletional α‐thalassaemia‐1 and a newly discovered poly A mutation
Authors: Yuuregir G. T. , Aksoy K. , ÇUuruuk M. A. , Dikmen N. , Fei Y.‐J. , Baysal E. , Huisman T. H. J.
Published: March 2008

Summary We have analysed the α‐globin gene defects present in several members of a large family from Southern Turkey. One deletional α‐thalassaemia‐1 (type MED‐II) was found in 10 subjects; this deletion is in excess of 26.5 kb and includes all and α‐globin genes. Besides the common types of deletional α‐thalassaemia‐2 (—3.7 kb and —4.2 kb) we observed a nondeletional α‐thalassaemia‐2 that results from an AG mutation (AATAAAÁTGAA) in the polyadenylation signal of the α2‐globin gene; the same AG replacement is present in the Ψαl gene. The mutation must cause a considerable α‐chain deficiency as is evidenced by the haematological data for five members with Hb H disease due to a compound heterozygosity for α‐thalassaemia‐1 (MED‐II) and the newly discovered poly A mutation. Several members had additional β‐chain abnormalities (Hb S, Hb D‐Los Angeles, β‐thalassaemia); the 11 persons with a Hb S heterozygosity and various α‐globin gene defects (—α/αα; αTα/αα, ——/αα, —α/—α and ——/αTα) showed a decrease in the level of Hb S that was directly related to the severity of the α‐chain deficiency.
Type IIB von Willebrand's disease: gene mutations and clinical presentation in nine families from Denmark, Germany and Sweden
Authors: Donnér Mikael , Kristoffersson Ann Charlotte , Lenk Harald , Scheibel Elma , Dahlbäck Bjöurn , Nilsson Inga Marie , Holmberg Lars
Published: March 2008

Summary Type IIB of von Willebrand's disease (vWD) is a variant in which the structurally abnormal von Willebrand factor (vWF) shows an increased affinity for the platelet vWF receptor, glycoprotein Ib (GPIb). This may sometimes give rise to platelet aggregation and thrombocytopenia in vivo. In 20 patients from nine unrelated families with type IIB vWD from Denmark, Germany and Sweden we studied the molecular defect by amplification and direct sequencing of parts of exon 28 which encode for the vWF domain that interacts with platelet GPIb. Three different point mutations were identified one of which has not previously been reported. Fifteen patients from five families were heterozygous for the Arg543Trp substitution. The mutation had occurred independently in all five families and in two of them represented a de novo mutation. In one of these families the father, though asymptomatic and with normal laboratory test results, carried the mutation in heterozygous form. In three families, four affected members were found to be heterozygous for the Arg543°Cys substitution. The mutations were of different origin at least in two of the families. The third substitution, Val554Leu, which has not previously been described, was found in one patient and was due to a de novo mutation. In most of the patients spontaneous thrombocytopenia had been recorded on at least one occasion. Five of the patients with the Arg543Trp substitution and the one with the Val555Leu substitution had all had bleeding associated with thrombocytopenia in the neonatal period of early infancy.
Improved post‐transfusion quality of density separated AS‐3 red cells after extended storage
Authors: Keegan T. E. , Holme A. Heaton S. , And M. Owens , Nelson E. J.
Published: March 2008

Summary A centrifugal method of red cell density separation was utilized for unit processing in these studies to determine the quality of the lighter fraction (neocytes) after storage for up to 42 d and to evaluate whether the heavier fraction (gerocytes) deteriorated more rapidly than neocytes during storage. Each unit was passed through a Leukotrap filter to remove white cells prior to density separation. Red cell recovery and survival were evaluated using double label technetium‐99m with either chromium‐51 or nonradio‐isotopic chromium which permitted concurrent paired analysis. In vivo neocyte red cell recovery, as tested on the same 11 donors on days 1. 7 and 42 of storage, was effectively unchanged. Recovery and survival half‐life (that is, the number of days after transfusion at which half of the cells infused remain in the circulation) of 42 d stored gerocytes were significantly lower than similarly stored neocytes (75.5 ± 7.2% and 20.1 ± 6 5 d for gerocytes versus 84.4±4.9% and 39.0±9.0 d for neocytes). One‐day stored neocytes showed a 16.5% increase in red cell availability over the combined average for 42 d stored neocytes and gerocytes. Statistically, while there were significantly higher ATP, 2,3‐DPG, and lactate levels pre‐storage by paired t‐test for neocytes compared to gerocytes, by day 42 there were no significant differences detected between the two red cell fractions by any of the in vitro variables measured. These studies suggest that this simple separation technique for leucocyte‐poor red cell units provides a neocyte population with improved viability and the potential for increased transfusion intervals in chronically transfused patients.
Band 3‐Memphis is associated with a lower transport rate of phosphoenolpyruvate
Authors: Ideguchi Hiroshi , Okubo Kenshi , Ishikawa Atsuko , And Yuko Futata , Hamasaki Naotaka
Published: March 2008

Summary Band 3‐Memphis is the most common variant of erythrocyte band 3 protein, in which a single amino acid substitution (Lys56→Glu) in a cytoplasmic domain of band 3 has been found. We showed here that the prevalence of the variant was particularly higher in the Japanese population than that in other populations. The calculated gene frequency of the variant in Japanese was 0.156, which was about 4 times higher than that in Caucasian. Although it has been generally accepted that the cytoplasmic domain of band 3 bears no relation to anion transport activity, we found that the transport rate of phosphoenolpyruvate in erythrocytes of homozygotes was decreased to about 80% of that in control cells, and that of heterozygotes was at an intermediate level. The evidence indicates that some structural changes in the cytoplasmic domain of band 3 may have influence on the conformation of anion transport system.
Proliferation of non‐Hodgkin‐lymphoma lymphocytesin vitrois dependent upon follicular dendritic cell interactions
Authors: Petrasch Stephan , Kosco Marie , Perez‐Alvarez Carmelo , Schmitz Jörn , Brittinger Günter
Published: March 2008

Summary.Follicular dendritic cells (FDC) are located within the B‐cell follicles of non‐malignant lymphatic tissues and within non‐Hodgkin‐lymphomas (NHL) derived from the germinal centre or the mantlezone. The interactions between FDC and non‐neoplastic B‐cells have been extensively investigated but so far no data on functional studies with FDC isolated from lymphoma tissue are available. Using an enzyme cocktail to digest lymph nodes from patients with NHL followed by density centrifugation, single cell suspensions enriched in FDC and B‐lymphocytes were obtained. In these preparations FDC formed small cellular clusters with an average of five neoplastic lymphocytes for every FDC. Immunocytochemistry with Ki67 revealed that after 24 h of culture 23.7% of the cells within the cluster were in late G1to M phase. In contrast, only 10.2% of the lymphoma cells scattered outside the clusters were in these activated stages. As visualized by autoradiography, after 72 h of incubation the rate of proliferation was 16.8 times higher for the lymphoma cells involved in cluster formation as compared to those lymphocytes not associated with FDC. The data indicate thatin vitroFDC from NHL lymph nodes form a microenvironment favourable for the activation and proliferation of lymphoma cells. The search for cytokines secreted by FDC in lymphoma tissue is under way.
Quinidine‐induced thrombocytopenic purpura: clinical presentation in relation to drug‐dependent and drug‐independent platelet antibodies
Authors: Nieminen Urpo , Kekomäki Riitta
Published: March 2008

Summary. We have studied the clinical course of quinidineinduced thrombocytopenia in relation to the presence of drug‐dependent (dd‐ab:s) and drug‐independent antibodies in 14 patients. Thrombocytopenia was reversible in 9 d after discontinuation of quinidine treatment in 10 patients. In four it lasted more than 1 month. Drug‐dependent antibodies of IgG class were detectable in seven patients: in six by an immunofluorescence test applying flow cytometry and in one patient by a monoclonal antibody‐immobilized platelet protein assay (MAIPA) only. The dd‐ab:s of this patient had glycoprotein (GP) IIIb/IIIa specificity. Five of the six patients with dd‐ab:s by immunofluorescence test had GPIb/IX‐specific dd‐ab:s by MAIPA. They recovered within 5 d after discontinuation of the drug. All four patients with prolonged thrombocytopenia had elevated platelet‐associated IgG (PAIgG) in the acute phase as studied by a direct platelet immunofluorescence test. The remaining five patients displayed a relatively rapid clinical recovery but less uniform pattern of immunological findings. The results suggest that patients with GPIb/IX‐specific dd‐ab:s recover promptly despite an acute and profound thrombocytopenia. Another sub‐group with prolonged thrombocytopenia had persistently elevated PAIgG during the convalescent phase.
Is there a mechanical factor of haemolysis in patients with positive IgG‐type direct antiglobulin test?
Authors: Delamaire M. , Durand F. , Grosbois B. , Le Dantec G. , Dauriac C. , Le Goff M.‐C. , Le Blay R. , Genetet B.
Published: March 2008

Summary. In autoimmune haemolytic anaemia, the presence of antibodies on the erythrocyte membrane results in haemolysis through an immune process, but does it not alter the rheological properties of red blood cells (RBC), thus adding a mechanical factor to haemolysis? This study was designed to examine the rheological properties of erythrocytes sensitized with IgG‐type antibodies. The study involved 20 patients with anaemia and positive direct antiglobulin test, including 12 with straightforward haemolysis, 10 samples sensitized in vitro, and 20 controls.

The following haemorheological parameters were studied: erythrocyte filtration, blood and plasma viscosities, titration of adenosine triphosphate (ATP) and 2–3‐DPG, erythrocyte morphology under scanning electron microscopy.

The results showed increased erythrocyte rigidity (P<0.025) as well as higher blood viscosity compared to controls with similar haematocrit values, and unaltered ATP and 2–3‐DPG (consistently with scanning electron microscope observations). These haemorheological disorders were more noticeable in patients with clear‐cut haemolysis, and there was a correlation between the increase in erythrocyte rigidity indices and the haemolytic parameters, especially haptoglobin (P<0.001). The in vitro study confirmed the results obtained ex vivo.

To conclude, the mechanical properties of antibody‐coated erythrocytes are impaired, which may promote the immunological mechanism favouring haemolysis in the spleen.
Lymphoid lineage‐associated features in acute myeloid leukaemia: phenotypic and genotypic correlations
Authors: Paietta Elisabeth , Van Ness Brian , Bennett John , Racevskis Janis , Gucalp Rasim , Cassileth Peter , Wiernik Peter H.
Published: March 2008

Summary. This study is intended to establish biological correlation between the expression of lymphoid associated features in acute myeloid leukaemia (AML). In 62 AML patients, predominantly enrolled on Eastern Cooperative Oncology Group (ECOG) treatment protocols, in whom immunoglobulin (Ig) as well as T‐cell receptor beta chain (TCR‐beta) gene rearrangement analyses had been performed, morphology, cytochemistry, antigen profile and karyotype were reviewed retrospectively. Nuclear reactivity with anti‐TdT antibody was demonstrated in 34 patients (55%) and confirmed by ribonuclease protection assay in all patients tested. Five TdT‐protein negative patients were TdT‐transcript positive. Lymphoid antigens (lyA) were detected in 24 of 51 cases tested (47%) with B‐cell antigens (CD19. CD10) being restricted to TdT+ AML (P= 0.03). Only two patients had Ig heavy, none had Ig light chain or TCR‐beta gene rearrangements. Although both patients with rearranged Ig loci were TdT+, either by protein or RNA analysis, the low incidence of such rearrangement within the TdT* AML group (6%) argues against a significant association between the presence of TdT and crosslineage Ig gene rearrangements in AML. While FAB‐diagnoses did not differ between TdT+ and TdT or lyA+ and lyA− AML, particular immunophenotypic features correlated with TdT positively, e.g. the presence of early antigens, CD34 and HLA‐DR, and the absence of the more mature myelo‐monocytic antigens, CDw65 and CD14. Certain cytogenetic abnormalities were associated with TdT+ AML such as inv(16) (p13q22) or t(16:16) (p12;q22) (five patients; P= 0.03) and t(8;21) (q22;q22) (three patients). A greater number of TdT− than TdT* AML patients had only normal karyotypes (P= 0.06). Neither immunophenotypic nor karyotypic correlations could be established for lyA4 AML.
Low‐dose etoposide: a potential therapy for myelodysplastic syndromes
Authors: Ogata K. , Yamada T. , Ito T. , Gomi S. , Tanabe Y. , Ohki I. , Dan K. , Nomura T.
Published: March 2008

Summary. Four patients with refractory anaemia with excess blasts in transformation (RAEB‐t) and seven patients with acute leukaemia (ALL transformed from myelodysplastic syndromes (MDS) were treated with etoposide (50 mg, 2 h infusion, two to seven times per week) for at least 4 weeks. Of 10 assessable patients, three RAEB‐t patients achieved partial response and one AL patient achieved complete remission. Three of the four responders were resistant to prior repeated low‐dose cytarabine therapy. The responders did not require transfusions for 2–9 months while continuing on etoposide therapy. The side‐effects were mild and well tolerated. Three possible mechanisms, i.e. a cytotoxic effect, differentiation‐induction of malignant cells, and prolongation of blood cell survival by destroying the reticuloendotheliai system, may explain the effects of etoposide. We conclude that low‐dose etoposide is a potential therapy for MDS and atypical leukaemia.
Bone marrow cells from A/J mice do not proliferate in interleukin‐3 but express normal numbers of interleukin‐3 receptors
Authors: Hapel Andrew J. , Fung Ming‐Chiu , Mak Nai‐Ki , Morris Carolyn , Metcalf Donald , Nicola Nicos
Published: March 2008

Summary. Haemopoietic cells from A/J mice do not form colonies (proliferate) in response to interleukin‐3 (multi‐CSF. IL‐3). We have examined different populations of cells from A/J mice and shown that, despite their failure to proliferate in response to IL‐3, cells from bone marrow, spleen and the peritoneum all bound 125I‐labelled IL‐3. A wide variety of cell types bound IL‐3 as determined by autoradiography, including promyelocytes, myelocytes, metamyelocytes, polymorphs, promonocytes, monocytes, eosinophils and lymphocytes, but not nucleated erythroid cells, and the proportion of each cell type binding label was similar when cells from A/J mice were compared with those of C57B1/6 and Balb/c mice. Bone marrow cells from A/J mice internalized interleukin‐3 with normal kinetics and mRNA extracted from these cells contains the same species of IL‐3 receptor and IL‐3 receptor‐like mRNAs as are found in the other strains. Collectively the data suggest that the failure of haemopoietic cells from A/J mice to proliferate in response to IL‐3 is related to a selective defect in signalling to proliferation specific genes. This defect is apparently not related to internalization or processing of the IL‐3/IL‐3‐receptor complex, but may be due to failure to activate appropriate accessory molecules in the cell.
Inactivation of the retinoblastoma gene in acute myelogenous leukaemia
Authors: Tang Jih‐Luh , Yeh Shiou‐Hwei , Chen Pei‐Jer , Lin Ming‐Tseh , Tien Hwei‐Fang , Chen Yao‐Chang
Published: March 2008

Summary. To investigate the role of retinoblastoma susceptibility (RB) gene inactivation in leukaemogenesis, we evaluated 36 bone marrow specimens of acute leukaemia for RB protein expression by immunoprecipitation and Western blot analysis. 15 patients had no detectable RB protein at initial screening. However, nine of them were subsequently excluded due to evidence of protein degradation. Of 27 valid cases, six (22%) were repeatedly shown to lack expression of the RB protein with three different anti‐RB antibodies. Five were patients with acute myelogenous leukaemia (AML) and one, mixed‐lineage acute leukaemia. The RB inactivation was noted more frequently in AML (5/18, 28%) than in acute lymphoid leukaemia (0/7, 0%). By karyotyping, none of these six patients exhibited cytogenetic changes involving chromosome 13q14, the RB locus. There is no correlation between inactivation of the RB gene and FAB subtypes or cytogenetic changes. Four patients achieved complete remission with standard chemotherapy for 6, 12, 20 and 26+ months, respectively. Southern and Northern blot analyses further indicated that the RB genes were grossly intact and the level of RB transcripts did not decrease in the majority of these six patients. These results suggest that the absence of RB products in some of acute leukaemia might be regulated at the post‐transcriptional level, and it imposes no significant effect on treatment response and prognosis.
Some ‘antiphospholipid antibodies’ bind to β2‐glycoprotein I in the absence of phospholipid
Authors: Keeling D. M. , Wilson A. J. G. , Mackie I. J. , Machin S. J. , Isenberg D. A.
Published: March 2008

Summary. Some antiphospholipid antibodies (aPL) only bind to anionic phospholipids in the presence of a serum cofactor, β2‐glycoprotein I (β2GPI). Whether these aPL can bind to β2GPI in the absence of phospholipid is controversial. We have purified anticardiolipin antibodies (aCL) from the plasma of four patients and β2GPI from normal plasma by solid phase affinity methods. All four aCL bound to cardiolipin and phosphatidylserine in the presence of β2GPI but not in its absence. The binding of two of the antibodies to cardiolipin and phosphatidylserine at various concentrations of human β2GPI was compared with that obtained using 10% bovine serum. The two antibodies responded differently to increasing β2GPI concentrations, and binding to phosphatidylserine was relatively greater than to cardiolipin using human β2GPI. All four aCL bound to plastic plates coated with β2GPI in the absence of phospholipid, and β2GPI in the fluid phase had no effect on binding. Binding to β2GPI coated plates was increased equally when bovine serum or bovine albumin were used as the sample diluent in place of gelatine. These findings and those of others have important implications for the design of assays for antiphospholipid antibodies.
Serum immunoreactive erythropoietin in patients with idiopathic aplastic and Fanconi's anaemias
Authors: Das R. E. Gaines , Milne Alison , Rowley Megan , Smith E. C. Gordon , Cotes P. Mary
Published: March 2008

Summary. In patients with idiopathic aplastic anaemia (n = 34) and Fanconi's anaemia (n = 8), sampled once or on several occasions, serum erythropoietin (Epo) increased with increasing severity of anaemia with apparently similar rates of increase in each group. However, after adjustment for Hb, log Epo values for the Fanconi's anaemics tended to be greater than those for the idiopathic aplastic anaemics (P < 0·01).

Erythropoietin concentrations in serum samples from patients with Fanconi's and idiopathic aplastic anaemias tended to be greater than in samples from patients with anaemias from protein energy malnutrition, myelodysplasia and iron deficiency.

The results suggest that there is no deficiency of erythropoietin in Fanconi's and idiopathic aplastic anaemias and that if exogenous erythropoietin is of any benefit it would need to be administered in doses large enough to induce a significant increase in log Epo.

Results of the study illustrate the need to take account of the assumptions which underlie interpretation of the statistical analysis. Use of erythropoietin values in place of log Epo gives misleading conclusions demonstrable as invalid as the conditions for normality of distribution of the data and homogeneity of variances were not satisfied.
Serum erythropoietin levels in anaemic patients with advanced human immunodeficiency virus infection
Authors: Camacho J. , Poveda F. , Zamorano A. F. , Valencia M. E. , Vázquez J. J. , Arnalich F.
Published: March 2008

Summary. This study attempts to evaluate the adequacy of the erythropoietin (EPO) response in 42 anaemic patients with advanced human immunodeficiency virus (HIV) infection [30 with acquired immunodeficiency syndrome (AIDS) and 12 with AIDS‐related conditions] by comparing their serum EPO levels with those found in a non‐HIV reference population consisting of 3 6 patients with anaemia of chronic disorders (ACD) and 57 with iron deficiency anaemia (IDA). Although the average Hb concentration was similar in the three groups, the EPO level for HIV patients (mean ± SEM, 64·3 ± 7·7 mU/ml) did not differ significantly from that in ACD patients (45·3 ± 8·3 mU/ml, P > 0·1), and both groups had a lower mean EPO level (P < 0·05 and P < 0·01 respectively) than IDA subjects (133·5 ± 18·7 mU/ml). Thirteen HIV patients on zidovudine therapy showed similar mean Hb and EPO levels to those in the untreated patients. A significant inverse correlation between the log of serum EPO and the Hb values was observed in the three groups. However, this relationship was found to be stronger in IDA patients than in either HIV or ACD subjects (P < 0·001), with no difference between the two latter groups (P > 0·2). These data suggest that the EPO response is blunted in the anaemia associated with advanced HIV infection.
Multiple independent immunoglobulin class‐switch recombinations occurring within the same clone in myeloma
Authors: Palumbo Antonio , Battaglio Silvano , Astolfi Monica , Frieri Roberto , Boccadoro Mario , Pileri Alessandro
Published: March 2008

Multiple myeloma (MM) is characterized by the expansion of terminally differentiated plasma cells. It is still uncertain whether the clonogenic fraction is confined to the plasma cell or pre‐plasma cell compartment. We examined the immunoglobulin (Ig) rearrangement of myeloma heavily infiltrated bone marrow cells with a probe from the heavy chain J region (JH) and the BamHI, EcoRI and HindIII restriction enzymes which are appropriate for the detection of clonal VDJ recombination. In 23/39 MMs. clonal Ig gene rearrangement was detected with BarnHI, EcoRI and HindIII enzymes. Unexpectedly, in 14/39 patients both BamHI and EcoRI failed to detect Ig rearrangement, whereas HindIII consistently demonstrated VDJ recombination. The 5’sites of BamHI, EcoRI and HindIII restriction fragments are precisely defined by the VDJ rearrangement. Since the 3’ends of BamHI and EcoRI restriction fragments are downstream from the switch μ region and change in size during switch recombination, the absence of rearranged bands is determined by several autonomous recombinations affecting the switch region. By contrast, the 3’ends of HindIII restriction fragments are upstream, their size does not vary during isotype switch allowing the constant detection of clonality. Accordingly, in 35% of patients the clonogenic fraction seems to originate from a pre‐switch B cell. This B cell will differentiate to a mature plasma cell developing multiple independent switch recombinations, as the variable mechanism of switch recombination suggests.