bluuebunny/crossref_metadata_2025 huggingface dataset:
Corticosteroid Therapy for Acquired F VIII:C Inhibitors
Authors: Spero J. A. , Lewis J. H. , Hasiba U.
Published: July 2010

Summary.

The appearance of an acquired inhibitor to factor VIII clotting activity (F VIII:C) in a non‐haemophiliac is an uncommon phenomenon; but, when it occurs, represents a difficult therapeutic problem. Cyclophosphamide has been the most frequently used form of therapy while corticosteroids, when used alone, have been felt to be ineffective. During the past 25 years, 18 of these patients have been evaluated in Pittsburgh. Their mean age was 63·7 years. Nine were female and nine male. Sixteen of the 18 received corticosteroids as their primary form of therapy. Eleven of the 16 (69%) showed clinical and laboratory improvement. Seven of the 11 had a complete response and the remaining four a partial response. Five patients demonstrated no improvement with corticoid therapy. The time to response varied from 4 to 41 d (x̄ 15·8). It was concluded that corticosteroids when used alone provide therapeutic results comparable to immunosuppressive therapy in the treatment of acquired inhibitors to F VIII:C and therefore represent a therapeutic alternative to immunosuppressive agents.
Assessment of bone marrow histology in Hodgkin's disease: correlation with clinical factors
Authors: Bartl R. , Frisch B. , Burkhardt R. , Huhn D. , Pappenberger R.
Published: July 2010

Summary.

Bone marrow biopsies of 491 untreated and 170 treated patients with Hodgkin's disease (HD) were investigated. Marrow involvement was found in 10% and 25% respectively. Positive biopsies were rare in clinical stages I and II (1% and 2%), but the incidence rose to 25% and 45% in stages III and IV. HD patients with nodular sclerosis in lymph node histology had a low incidence of bone marrow involvement (4%), while those with lymphocytic depletion had a high incidence (22%). Of nine clinical and six histological parameters tested, bone marrow involvement proved to be the most significant predictive factor indicating an unfavourable course. Moreover, classification of the bone marrow manifestations according to the degree of lymphocytic infiltration, proved to be simple, reproducible and prognostically significant. Normal haematopoietic tissue was found in only 20% of the negative biopsies of untreated patients. The remaining 80% were characterized by a variety of non‐specific reactions. These included marrow hypoplasia and leukaemoid and exudative reactions each of which indicated a poor prognosis; and epithelioid‐cell granulomas and lymphoid nodules which predicted favourable survival curves. Adequate bone marrow biopsy is a valuable part of the investigation of patients with HD, as both positive and negative biopsies provide information of prognostic significance.
In vitro suppression of erythropoiesis by bone marrow adherent cells from some patients with fungal infection
Authors: Zanjani Esmail D. , McGlave Philip B. , Davies Scott F. , Banisadre Mussa , Kaplan Manuel E. , Sarosi George A.
Published: July 2010

We present evidence that alterations in marrow adherent cell (Mø) function may play a role in the suppression of erythropoiesis in some patients with fungal infection. Bone marrow (BM) cells from 12 normals and 10 patients with histoplasmosis were cultured in plasma clots before and after removal of Mø. BM from five patients (Group A) produced normal numbers of erythroid colonies (EC). In the remaining patients (Group B), smaller numbers of EC were detected (P<0.01). Removal of Mø from BM of normals and Group A patients resulted in decreased growth of EC. In contrast, Mø depletion of BM from patients in Group B resulted in greater EC formation (P< 0.01). When normal Mø were admixed with normal or patients’BM‐Mø, enhanced EC formation resulted. Whereas, at similar concentrations, Mø from group B patients caused inhibition of EC formation (P< 0.005). The erythro‐regulatory function of Mø, including the inhibitory action of patients’Mø, was mediated via a soluble agent(s) since media conditioned by Mø mimicked the action of these cells. Three patients in Group B were restudied 14 months after treatment with amphotericin B, when blood parameters had returned to normal. At this time, normal patterns of EC formation and Mø activity were observed.
Measurement of the density of human platelets and its relationship to volume
Authors: Martin J. F. , Shaw T. , Heggie J. , Penington D. G.
Published: July 2010

Summary. New methods are described for platelet isolation and buoyant density determination using low‐speed centrifugation in continuous density gradients of Percoll. The conditions used do not induce loss of granule or cytoplasmic markers and enable reproducible platelet frequency distributions to be obtained in linear density gradients. Such frequency distributions are normal with a mode of 1·0645 ± 0·0015 g cm−3 (mean ± SD, n= 20). Platelets fixed in 0·1% glutaraldehyde show a modal density of 1·0712 ± 0·0005 g cm−3. Content of protein, lactate dehydrogenase, beta‐thromboglobulin and 3H‐serotonin correlate closely with platelet numbers throughout the density distribution.

The frequency distribution of platelet volume between 2·2 and 21 fl fits a log normal model and cell volume in density subfractions from the most dense to the least dense also approximate log normality. There is a positive correlation between mean platelet volume and buoyant density with a small increment between the least and the most dense extremes. Platelet subfractions separated by volume using a FACS II cell sorter differ substantially from each other in cell volume but the difference in mean density of four different volume fractions is negligible. In discontinuous density gradients of Stractan factors other than platelet density must influence the separation of platelets, as rebanding of platelets from interfaces shows a wide variation in buoyant density when analysed in continuous gradients.

It is concluded that analysis of platelet buoyant density in continuous Percoll gradients supports the view that platelet density, like platelet volume, is determined primarily during thrombocytopoiesis and that volume and density are largely independent elements of platelet heterogeneity.
Improved survival following bone marrow transplantation for aplastic anaemia
Authors: Feig Stephen A. , Champlin Richard , Arenson Edward , Yale Coralee , Ho Winston , Tesler Alan , Gale Robert Peter ,   for the UCLA Bone Marrow Transplant Team
Published: July 2010

Summary. We transplanted 46 patients with severe aplastic anaemia with a new pretransplant immunosuppressive regimen consisting of cyclophosphamide (200 mg/kg) and low‐dose total body irradiation (3 Gy). This regimen (CY‐TBI‐2) was designed to decrease the high risk of graft rejection associated with the use of cyclophosphamide alone, without increasing the incidence of graft‐versus‐host disease (GHVD) or interstitial pneumonia (IPn). Two‐year actuarial disease‐free survival of patients conditioned with CY‐TBI‐2 was 62% (95% CI: 47–77%). Only one patient rejected her graft and the incidence and severity of GVHD and IPn were not increased compared to previous studies. Patients < 25 years of age had excellent 2‐year survival of 82% (95% CI: 69–95%). These data indicate that CY‐TBI‐2 is an effective means of preventing graft‐rejection and achieving long‐term disease‐free survival in multiply transfused patients with severe aplastic anaemia.
Paraproteinaemia plus osteolytic lesions in typical hairy‐cell leukaemia
Authors: Jansen J. , Bolhuis R. L. H. , Nieuwkoop J. A. van , Schuit H. R. E. , Kroese W. F. Stenfert
Published: July 2010

Summary. Most cases of hairy‐cell leukaemia (HCL) involve proliferations of neoplastic B lymphocytes. In rare cases, M‐proteins or osteolytic lesions have been documented in patients with HCL.

In this study two patients with typical HCL are reported in whom both paraproteinaemia and osteolytic lesions of the femoral neck developed. In one of the patients the production of the M‐protein by hairy cells could be established. In the other patient, at autopsy no signs of myeloma were found. The hairy cells from inside the osteolytic lesion had the same immunological phenotype as hairy cells from the peripheral blood, the spleen, and other parts of the bone marrow.

These cases once more confirm the B‐cell nature of many cases of HCL, and show that hairy cells can have functional capacities usually attributed to much more mature B lymphocytes, i.e. plasma cells.
Iron metabolism in murine erythroleukaemic cells
Authors: Peto T. E. A. , Rutherford T. R. , Thompson J. L. , Weatherall D. J.
Published: July 2010

Summary. In an attempt to develop a model system for analysing iron metabolism in a relatively homogeneous population of early red cell precursors, the intracellular distribution of 59Fe was examined in Friend murine erythroleukaemic cells after induction of haemoglobin synthesis with dimethylsulphoxide. After incubation of the cells with 59Fe‐labelled transferrin, 59Fe was incorporated into haemoglobin, various ferritin fractions, and into the pellet obtained by centrifugation. No intracellular transferrin or low molecular weight compounds were found. In a series of ‘chase’ experiments 59Fe accumulated in haem, and some of this radioactivity appeared to be derived from the ferritin fraction. Extra iron could be mobilized from ferritin during chase experiments using iron deficient incubation medium. These studies indicated that, at least under these experimental conditions, ferritin iron in early red cell precursors can be utilized for haemoglobin synthesis.
Female marrow donors increase the risk of acute graft‐versus‐host disease: effect of donor age and parity and analysis of cell subpopulations in the donor marrow inoculum
Authors: Atkinson Kerry , Farrell Caroline , Chapman Graeme , Downs Kathryn , Penny Ronald , Biggs James
Published: July 2010

Summary. We evaluated 27 factors for their influence on acute graft‐versus‐host disease (GVHD) in 40 recipients of HLA‐identical sibling marrow transplants. These factors included the doses of mononuclear cell subpopulations present in the donor marrow inoculum quantitated using a panel of monoclonal antibodies. Female donors were associated with increased severity of acute GVHD, and the older the female donor the greater this effect. Increasing donor parity was also associated with an increased risk of acute GVHD. The number of T cells, T cells subsets, natural killer cells and monocytes infused did not influence the incidence or severity of acute GVHD in this study, and we could not explain the influence of female donors and of female donor age on acute GVHD by the cellular content of their marrow inocula. We postulate that non‐HLA histocompatibility antigen disparity is a more important determinant for acute GVHD than the number of infused donor T cells, especially when female donors are used. The association between acute GVHD and increasing parity suggests that some female marrow donors have been pre‐sensitized to their respective recipients by preceding pregnancies.
Antigenic determinants in idiopathic thrombocytopenic purpura
Authors: Lynch Dona M. , Howe Stephen E.
Published: July 2010

Summary. Serum platelet bindable immunoglobulin (SPBIg) was determined in a group of 23 idiopathic thrombocytopenic purpura (ITP) patients and compared to 20 normal, healthy controls. The mean SPBIg of the ITP group was 16.1 (±17.9 SD) fg/platelet, while the normals were substantially lower, 40 (± 1.2) fg/platelet. Sera from patients of both groups were then incubated with platelet fractions immobilized on nitrocellulose membrane strips (Western Blotting) to detect platelet antigen specificity using a peroxidase labelled indicator antibody. The normal patient sera did not react with platelet fractions on the nitrocellulose strips. However, 21 of 2 3 ITP sera bound to one or more platelet fractions with large variations in the number and molecular weights of the platelet fractions identified by ITP antibody. These observations suggest the presence of multiple antigenic binding sites for platelet specific immunoglobulin in ITP sera. This variation may reflect heterogeneous antibodies binding to diverse antigens or homogeneous antibodies to a limited number of antigenic determinants shared by several discrete platelet molecules.
Cytogenetic follow‐up studies of recipients of T‐cell depleted allogeneic bone marrow
Authors: Lawler Sylvia D. , Harris Helen , Millar J. , Barrett Ann , Powles R. L.
Published: July 2010

Summary. Serial cytogenetic studies of bone marrow and blood cells were made in leukaemic patients who had received an allogeneic bone marrow graft from a donor of unlike sex. The donor marrow was treated with the monoclonal antibody Campath‐1 before infusion. The persistence of a significant proportion of dividing recipient cells in marrow and blood was observed after grafting. These recipient cells showed evidence of radiation damage of both the stable and unstable types. More than one transient clone of chromosomally abnormal recipient cells was observed in three cases. The differences between the cytogenetic findings in patients receiving donor marrow from which T cells have been removed and those cases previously studied in our Leukaemia Unit who had received untreated donor marrow are discussed.
The use of monoclonal antibodies UCH/β and UCHγ for the antenatal diagnosis of /β‐thalassaemia
Authors: Allen C. , Weereratne H. , Gale R. , Beverley P. C. L. , Huehns E. R.
Published: July 2010

Summary. Monoclonal antibodies for the β‐globin chain of human HbA (UCHβ) and the γ‐globin chain of human HbF (UCHβ) have been made. UCHβ indirectly labelled with rhodamine‐labelled goat anti‐mouse lg and directly fluores‐ceinated UCHy have been used, via double labelling immuno‐fluorescence microscopy, to assay for the presence ofβ‐chains in fetal erythrocytes obtained at fetoscopy from fetuses at risk for β‐thalassaemia major. The results from 111 cases demonstrate that β‐chain synthesis of as low as 1.8% can be detected in fetal erythrocytes. The immunochemical labelling can be rendered more sensitive by the use of biotinylated UCHβ and avidin‐FITC conjugates. This method is rapid and can be used for a sample that is highly contaminated with maternal erythrocytes.
Polycythaemia vera in young people: an analysis of 58 cases diagnosed before 40 years
Authors: Najean Yves , Mugnier Pascale , Dresch Catherine , Rain Jean‐Didier
Published: July 2010

Over 20 years, 58 cases of PV in young people (46 meeting the full PVSG criteria, 12 with elevated red cell volume and leucocytosis or thrombocytosis, without splenomegaly) were studied and have been followed for periods of 3–24 years. These cases represent approximately 5% of the cases of PV referred to the Department of Nuclear Medicine of St Louis Hospital during this period. They differ from older patients in the initial clinical severity, the short interval between the first symptoms and the diagnosis, frequent presentation with a life‐threatening complication (two cases of hepatic vein thrombosis, six thrombotic or haemorrhagic events, six splenectomies, two abortions) and a very enlarged spleen in half the cases. However, after the initial complications, the overall survival is very long (exceeding 70%, even when including the initial complications, at 15 years).

The vascular accidents occur exclusively in the phlebotomized patients, the main risk factor being the poor stability of the haematocrit. Only one acute leukaemia was observed among the 14 cases treated by radioactive phosphorus and/ or alkylating chemotherapy. The most frequent late complication was evolution towards myelofibrosis. This spent phase seemed to occur earlier in patients treated by phlebotomy.

On the basis of this data, we would advise the following therapeutic strategy: phlebotomies, as soon as the diagnosis is established, and a systematic long‐term treatment by hydroxyurea with the hope of reducing the number of vascular complications and of delaying the evolution towards the spent phase and the myelofibrosis.
Aberrant expression of immunoglobulin light chain isotype in B lymphocytes from patients with monoclonal gammopathies: isotypic discordance and clonal B‐cell excess
Authors: Oritani Kenji , Katagiri Shuichi , Tominaga Nobuhiko , Iida Masato , Amano Toshio , Karasuno Takahiro , Matsumura Itaru , Mitsui Hideki , Kanayama Yoshio , Yonezawa Takeshi , Tarui Seiichiro
Published: July 2010

A study was made of 29 patients with monoclonal gammopathies to detect aberrations in immunoglobulin (Ig) light chain isotype expression in lymphocytes at various levels of B‐cell differentiation, namely, circulating surface Ig positive (SIg +) cells, Ig‐secreting cells (plaque forming cells, PFC) and mitogen‐induced PFC. By using kappa‐lambda analysis, two major phenotypes of aberrant Ig light chain isotype expression were found in circulating B cells at these three levels of differentiation: an absolute increase in B cells bearing the same Ig light chain isotype as that of monoclonal protein (clonal B‐cell excess), and a relative decrease in those B cells (isotypic discordance). Isotypic discordance (ID) was found to be essentially negative in patients with monoclonal gammopathy of undetermined significance (MGUS) provided that they were in a stable condition. In myeloma patients, ID was found only in stage I, except for a remission case of stage III (4/7 in stage I, 0/8 in stage II, and 1/6 in stage III). ID was not restricted to a circulating SIg+ cell level but was also demonstrable at a spontaneous or pokeweed mitogen‐induced PFC level. However, ID was negative at a PFC level induced by Staphylococcus aureus Cowan I. Clonal B‐cell excess (CE) was frequently found in patients with active myeloma but not in stable patients (0/8 in MGUS, 1/7 in stage I, 8/8 in stage II, and 4/6 in stage III). CE was positive not only at a circulating SIg+ cell level but also at a circulating PFC level. Furthermore, patients with CE at a PFC level were found to have a higher proliferating capacity, defined as a percentage labelling index of marrow myeloma cells, than those without CE at a PFC level (P<0.02). ID and CE can therefore be considered as useful markers for discriminating between MGUS and myeloma, evaluating the clinical stability and predicting the clinical course.
PML/RARα fusion gene is expressed in both granuloid/macrophage and erythroid colonies in acute promyelocytic leukaemia
Authors: Takatsuki Hiroshi , Sadamura Shingo , Umemura Tsukuru , Abe Yasunobu , Goto Tatsuro , Yufu Yuji , Katsuno Makoto , Hirata Jouji , Nishimura Junji , Nawata Hajime
Published: July 2010

Summary. Acute promyelocytic leukaemia (APL) associated with a t(15:17) translocation generates a PML/KARa chimaeric gene which is transcribed as a fusion PML/RARα mRNA. To clarify the pathophysiologic role of PML/RARα in APL patients, we examined the expression of PML/RAKα in haemopoietic colonies in five patients with APIA by reverse transcriptase polymerase chain reaction (RT‐PCR) analysis. By the two‐step RT‐PCR method, we demonstrated that PML/ RARα positive clones were present in progenitor cells including both CFU‐GM'and BFU‐E in two cases. This result suggests that the translocation of PML/RARα occurred in a pluripotent stem cell in some APL patients. In four patients we detected two amplified cDNA fragments of 780 and 640 bp which presumably arose by alternative splicing of the PML gene. Interestingly of CFU‐GM and BFU‐E colonies examined in four patients, there were three different types of colonies: those expressing only the 780 bp fragment, those expressing only the 640 bp fragment, and those expressing both fragments. This suggests that alternative splicing was clonally determined in each colony. We describe a useful RT‐PCR technique for the study of gene expression in a limited number of haemopoietic precursor cells.
Pure red cell aplasia of pregnancy: a distinct clinical entity
Authors: Baker R. I. , Manoharan A. , De E. , Luca  , Begley C. G.
Published: July 2010

Summary. We describe a 31‐year‐old patient with pure red cell aplasia of pregnancy, successfully managed with regular blood transfusions. In vitro studies showed specific inhibition of day 14 erythroid colonies (BFU‐E) using serum and purified immunoglobulin G (IgG) obtained from the patient at diagnosis (before blood transfusion). The inhibition of BFU‐E disappeared when haematological remission occurred 3 weeks after delivery and she remains clinically well with a normal haemoglobin 4 years later.
Soluble tumour necrosis factor receptors, tumour necrosis factor and interleukin‐6 in serum in granulocytopenic patients with fever
Authors: Steinshamn Sigurd , Brekke Ole‐Lars , Waage Anders
Published: July 2010

Serum levels of TNF, IL‐6 and soluble TNF receptors p55 and p75 (sTNFR‐p55 and sTNFR‐p75) were examined in 14 patients with acute myeloid leukaemia during 43 courses of chemotherapy. The patients experienced 30 episodes of fever which occurred during granulocytopenia (defined as granulocyte counts <0.2×109/1) and six fever episodes when granulocyte counts were >1.0×109/1. Febrile episodes were classified as microbiologically defined infection, clinically defined infection, and unexplained fever. Levels of bioactive IL‐6 and immunoreactive TNF increased in response to fever during granulocytopenia, whereas bioactive TNF was not detected in any sample in this study. During granulocytopenia, both sTNFR rose significantly in microbiologically defined infection (P<0.01 for sTNFR‐p55 and P<0.05 for sTNFR‐p75), but not in the other two categories. The ratio of sTNFR‐p55 to sTNFR‐p75 was higher during febrile periods in granulocytopenia than in a non‐granulocytopenic situation with granulocyte counts >1.0×109/1 (P<0.01). We conclude that granulocytopenia affects release of the two sTNFR differently during febrile periods, and that release of sTNFR‐p75 in response to fever is reduced during granulocytopenia, suggesting a role for the granulocytes in systemic release of sTNFR‐p75.
Low expression of p27 and low proliferation index do not correlate in hairy cell leukaemia
Authors: Chilosi Marco , Chiarle Roberto , Lestani Maurizio , Menestrina Fabio , Montagna Licia , Ambrosetti Achille , Prolla Gabriel , Pizzolo Giovanni , Doglioni Claudio , Piva Roberto , Pagano Michele , Inghirami Giorgio
Published: July 2010

The molecular basis accounting for the peculiar clinical and biological features of hairy cell leukaemia (HCL) is currently unknown. Deregulation of cell cycle genes plays a significant role in oncogenesis and there is considerable evidence suggesting that Cdk inhibitors (Ckis) function as tumour suppressors. We and others have recently demonstrated low expression of Cki p27 in very aggressive neoplasms and high‐grade lymphomas. To investigate whether HCL cases express normal p27 protein, as in other low‐grade lymphomas with a low proliferation index, 58 cases of HCL were characterized using a sensitive biotin‐streptavidin‐immunoperoxidase technique and specific antibodies against p27. All HCL cases showed either no or very weak reactivity, in contrast to other types of low‐grade B‐cell lymphoma [22 cases of chronic lymphocytic leukaemia (CLL), 12 cases of gastric marginal B‐cell lymphoma (MALT), 16 cases of follicular lymphomas and two cases of splenic marginal zone lymphomas]. To investigate the possible mechanism(s) accounting for the low p27 expression observed in hairy cells, multiple approaches were used. According to these molecular studies, low levels of p27 are not as a result of (1) increased ubiquitin‐mediated degradation, (2) decreased levels of p27 transcription or (3) p27 somatic mutations and/or allelic loss. These findings suggest that low p27 protein expression in HCL may be achieved through post‐transcriptional regulation. Finally, our data demonstrate that p27 expression in HCL does not correlate with either cell cycle progression or proliferation index, suggesting that low levels of p27 in hairy cells may be associated with their unique stage of B‐cell differentiation and/or the activation of as yet unknown pathways.
Identification of mutations in 15 Hungarian families with hereditary protein C deficiency
Authors: Dávid M. , Losonczy H. , Sas G. , Nagy Á. , Kutscher G. , Meyer M.
Published: July 2010

Hereditary protein C deficiency represents about 2–9% of inherited thrombophilias. Our aim was to elucidate the molecular basis of protein C deficiency in 25 members of 15 Hungarian families with venous thromboembolic diseases and to identify hitherto undescribed genetical defects in the protein C (PROC) gene. The exons of the PROC gene were screened for mutations with the combination of polymerase chain reaction (PCR) and denaturing gradient gel electrophoresis (DGGE), and/or PCR and single‐strand conformation polymorphism (SSCP) analysis. The amplified DNA fragments with aberrant migration during DGGE and SSCP were sequenced. Mutations were determined in the PROC gene in all of the examined families: one nonsense mutation, one rare frameshift deletion and nine different missense mutations, of which three were novel (1493 A→G, 35Asp→Gly; 6231 G→A, 173Gly→Glu; 8476 C→T, 254Thr→Ile). The combination of hereditary protein C deficiency with other hereditary thrombophilias was rather common. DGGE and SSCP were proved to be efficient and cost‐effective screening methods in the genetic analysis of hereditary protein C deficiency.
Graft‐versus‐tumour effect in non‐small‐cell lung cancer after allogeneic peripheral blood stem cell transplantation
Authors: Moscardó Federico , Martínez Jesús A. , Sanz Guillermo F. , Jiménez Carmen , Cervera José , Sanchís José , Vera Francisco J. , Sanz Miguel A.
Published: July 2010

Clinical evidence of a graft‐vs.‐tumour effect in solid tumours after haematopoietic stem cell transplantation is lacking. We report for the first time a complete and durable regression of a stage IB non‐small‐cell lung carcinoma in a patient who had received an allogeneic peripheral blood haematopoietic stem cell transplant for acute myeloblastic leukaemia in first complete remission. Disappearance of the tumour coincided with development of graft‐vs.‐host disease. This suggests that simultaneous generation of cytotoxic T lymphocytes against lung carcinoma cells could have been responsible for the regression. This unique clinical observation broadens the possibility of using allogeneic haematopoietic stem cell transplantation in treating neoplasias lacking significant sensitivity to chemotherapy.
Improving the diagnosis of coeliac disease in anaemic women
Authors: Unsworth D. J. , Lock R. J. , Harvey R. F.
Published: July 2010

The majority of patients with newly diagnosed coeliac disease have iron deficiency anaemia. Occult gluten enteropathy is common. We looked at blood donor volunteers, unable to donate because they were unexpectedly found to be anaemic, to determine the incidence of coeliac disease and whether this diagnosis is routinely considered. In a 4 month period, 110 973 blood donor volunteers were seen and 1% (1197 women and 183 men) were found to be anaemic. Of 483 anaemic samples selected for testing, 32 out of 483 were positive for IgA anti‐endomysial antibodies. Microcytic anaemia was found in all but three of the 32 seropositive cases. Twenty‐five out of 32 volunteers agreed to have endoscopic small bowel biopsies and 22 out of 25 (88%) had the typical histological appearances of coeliac disease. Twenty‐one out of 22 cases were women. In no case prior to our intervention had these women been investigated for the possibility of coeliac disease. By selecting anaemic subjects for screening, there was an improved detection rate (over 6%) compared with non‐anaemic volunteers (0%). There were far more anaemic women in the study population (ratio of anaemic women to anaemic men 5·5:1). We show that, especially in anaemic menstruating women, coeliac disease is unrecognized and under‐investigated.
Megakaryocyte maturation is associated with expression of the CXC chemokine connective tissue‐activating peptide CTAP III
Authors: Deutsch Varda , Bitan Menachem , Friedmann Yael , Eldor Amiram , Vlodavsky Israel
Published: July 2010

Connective tissue‐activating peptide III (CTAP III), a CXC chemokine derived from the chemokine precursor platelet basic protein (PBP) by proteolytic cleavage, has been identified in platelets, activated macrophages, neutrophils and T lymphocytes. CTAP III can support stem cell‐derived haematopoiesis yet inhibits the proliferation of committed megakaryocyte (MK) progenitors. This investigation was aimed at characterizing CTAP III expression in human MKs and determining it's role in MK differentiation. We report high expression of CTAP III in mature human bone marrow (BM) MKs and megakaryoblast cell lines following differentiation induction with phorbol ester 12‐myristate 13‐acetate (PMA). Immunostaining with anti‐CTAP III antibodies demonstrated its prominent presence in platelet producing zones in the cytoplasm and intense staining around the periphery of the large BM MKs. In cultures of logarithmically growing megakaryoblast cell lines DAMI, CHRF‐288 or MEG01, which contain primarily 2N cells, only 15% of the cells expressed CTAP III. The addition of PMA stimulated high levels of CTAP III after 24 h in more than 75% of the cells, being expressed in both the 2N and large polyploid MKs. Reverse transcription polymerase chain reaction (RT‐PCR) revealed upregulation of CTAP III mRNA after only 1 h of exposure to PMA that was sustained for 24 h. In the bone marrow of idiopathic thrombocytopenic pupura (ITP) patients undergoing accelerated MK maturation and thrombopoiesis, 99% of large MKs and 95% of small MKs expressed high levels of CTAP III. While the biological function of this chemokine in MKs is not known, these studies demonstrate that molecular upregulation of CTAP III in MKs is associated with maturation and, as with other chemokines, may be involved in proliferation arrest and cellular interactions with extracellular matrix and platelet production.
Thalidomide maintenance following high‐dose therapy in multiple myeloma: a UK myeloma forum phase 2 study
Authors: Feyler Sylvia , Rawstron Andy , Jackson Graham , Snowden John A. , Cocks Kim , Johnson Roderick J.
Published: July 2010

Thalidomide maintenance has unresolved issues regarding dosage and toxicity. We evaluated this in five dose cohorts in 100 patients. At a median follow‐up of 32·3 months, 23 patients had stopped thalidomide for disease progression, 54 for side effects. 3‐year overall and progression‐free survival was 76% and 41% respectively. Dosage did not influence disease outcome but greatly affected toxicity. Fifteen patients converted from partial remission to complete remission on thalidomide at a median of 13·5 months. Maintenance doses >200 mg were largely unachievable and peripheral neuropathy was the main toxicity. Lower doses enabled more patients to stay on the drug for a useful period of time.
Efficacy and tolerability of high‐dose methotrexate in central nervous system positive or relapsed lymphoproliferative disease following liver transplant in children
Authors: Taj Mary M. , Messahel Boo , Mycroft Julie , Pritchard‐Jones Kathy , Baker Alistair , Height Susan , Hadzic Nedim , Pinkerton C. Ross
Published: July 2010

Childhood post‐transplant lymphoproliferative disease (PTLD) is a heterogeneous condition in which treatment varies, from the reduction of immunosuppression to moderately intensive chemotherapy. While low‐dose chemotherapy/rituximab has been found to be effective, moderately intensive chemotherapy is required for patients who relapse, have classic non‐Hodgkin lymphoma or have fulminant PTLD. Methotrexate (Mtx) is highly effective in lymphomas and crosses the blood‐brain barrier. However, there are no data in the literature regarding its safety in post‐liver transplant patients. We describe four cases of high‐grade lymphomas (three diffuse large B cell and one T‐cell lymphoblastic), post‐liver transplant, for which chemotherapy including high‐dose Mtx (HDMTX) was the treatment of choice. In total, 20 doses of HDMTX (1–5 g/m2) were given. The treatment was well tolerated and all four patients had a good response. One case of central nervous system (CNS) diffuse large B‐cell lymphoma was treated with HDMTX alone. We conclude that, in the absence of significant organ damage, HDMTX can safely be given to liver transplant patients, but should only be administered in specialist oncology units. Proof of effectiveness as a single agent in CNS lymphoma needs further studies.
The human side of hypoxia‐inducible factor
Authors: Smith Thomas G. , Robbins Peter A. , Ratcliffe Peter J.
Published: July 2010

When humans are exposed to hypoxia, systemic and intracellular changes operate together to minimise hypoxic injury and restore adequate oxygenation. Emerging evidence indicates that the hypoxia‐inducible factor (HIF) family of transcription factors plays a central regulatory role in these homeostatic changes at both the systemic and cellular levels. HIF was discovered through its action as the transcriptional activator of erythropoietin, and has subsequently been found to control intracellular hypoxic responses throughout the body. HIF is primarily regulated by specific prolyl hydroxylase‐domain enzymes (PHDs) that initiate its degradation via the von Hippel‐Lindau tumour suppressor protein (VHL). The oxygen and iron dependency of PHD activity accounts for regulation of the pathway by both cellular oxygen and iron status. Recent studies conducted in patients with rare genetic diseases have begun to uncover the wider importance of the PHD‐VHL‐HIF axis in systems‐level human biology. These studies indicate that, in addition to regulating erythropoiesis, the system plays an important role in cardiopulmonary regulation. This article reviews our current understanding of the importance of HIF in human systems‐level physiology, and is modelled around the classic physiological response to high‐altitude hypoxia.
Detection of tumour necrosis factor/cachectin in pleural effusion of patients with lung cancer
Authors: ISHII Y , UCHIYAMA Y , HASEGAWA S , KINOSHITA T , MITSUI K , KOJIMA H , FUJITA T
Published: July 2010

We have found that the pleural effusion obtained from a patient with lung cancer (adenocarcinoma) has cytotoxic activity against the patient's lung cancer cells. This finding occurred in the course of establishing a lung cancer cell line from the patient's pleural effusion. The cytotoxic factor was partially purified from the pleural effusion and characterized. It had cytotoxicity against L-929 mouse fibroblasts in the standard 18-h killing assay of tumour necrosis factor (TNF). By molecular sieving chromatography, the activity appeared at molecular weight of 50 000. This activity was completely blocked by a monoclonal antibody to TNF. From these results, we conclude that the cytotoxic factor in the pleural effusion is TNF. The concentration of TNF in the pleural effusion was 34.5 pg/ml by radioimmunoassay. In addition, we detected TNF activity and protein in two other cases of carcinomatous pleural effusion. Therefore, it would appear that in vivo TNF displays cytotoxic activity against cancer cells.
Neonatal myasthenia gravis: antigenic specificities of antibodies in sera from mothers and their infants
Authors: TZARTOS S J , EFTHIMIADIS A , MOREL E , EYMARD B , BACH J-F
Published: July 2010

Transient neonatal myasthenia gravis (MG) is a human model or passively transferred MG, In an effort to understand the characteristics of the most pathogenic antibodies in MG, we studied the fine antigenic specificities of anti-AChR antibodies in sera from 21 MG mothers (nine of which had transiently transferred the disease) and 17 of their infants. Although in a few cases significant differences in antibody specificities were observed between mothers and infants, whether myasthenic or not, generally the antigenic specificities of the antibodies in sera from infants were very similar to those of their mothers. Furthermore, no characteristic differences were detected between the antibody repertoires of mothers who transferred the disease and those who did not.
Distribution of the MHC antigens after liver transplantation: relationship with biochemical and histological parameters
Authors: ROUGER P , GUGENHEIM J , GANE P , CAPRON-LANDEREAU M , MICHEL F , REYNES M , BISMUTH J
Published: July 2010

The aim of this study was to analyse the change in tissue distribution of HLA class I and class II molecules in patients during the course of liver transplantation. Sixty-one liver biopsies were analysed in 24 patients with or without rejection (acute or chronic) episode. In 93% of cases with chronic rejection and in 48% of cases with acute rejection episodes, HLA class I antigens are expressed in the membrane and/or the cytoplasm of hepatocytes. The expression of these molecules was significantly correlated with the ASP (aspartate amino transferase), AP (alkaline phosphatase), the bile duct damage and the centrolobular lesions. In contrast, the expression of HLA class II molecules is not correlated with either the type of rejection, the biological or histological findings. A strong expression of HLA class I antigen on hepatocytes after day 60 may be one of the best signs of chronic rejection.
Jacalin: a lectin mitogenic for human CD4 T lymphoctyes
Authors: PINEAU N , AUCOUTURIER P , BRUGIER J C , PREUD'HOMME J L
Published: July 2010

The major protein component of seeds from jackfruit is the lectin jacalin. Jackfruit crude extracts are known to stimulate human lymphocytes, but the mitogenic properties of purified jacalin have not been studied in detail so far. Study of the proliferative response of cell populations from normal human peripheral blood to purified jacalin showed it to be mitogenic through an interaction with lymphocytes by its lectin-binding site, as shown by inhibition by IgA. Jacalin failed to stimulate B cells to proliferate and to undergo plasma cell maturation. It induced a proliferation of CD4 (and not CD8) lymphocytes, as shown by phenotypic analysis of cells recovered after culture and by studies of the response of isolated T cell subpopulations. The proliferative response to jacalin was autologous monocyte-dependent. The kinetics of jacalin-induced DNA synthesis, expression of CD25 and interleukin-2 secretion was shifted by comparison with that induced by phytohaemagglutinin. The reason for the restricted responsiveness of CD4T cells is presently unclear; jacalin bound to all blood cells and did not significantly co-cap with CD1, CD2, CD3, CD4, CD8 and CD38, and jacalin response was neither enhanced nor inhibited by antibodies to these surface antigens.
Cytokine regulation of human monocyte interleukin-1 (IL-1) productionin vitro. Enhancement of IL-1 production by interferon (IFN) gamma, tumour necrosis factor-alpha, IL-2 and IL-1, and inhibition by IFN-alpha
Authors: DANIS V A , KULESZ A J , NELSON D S , BROOKS P M
Published: July 2010

IL-1 production (secreted and cell-associated) was measured in monocyte cultures stimulated by a variety of agents in vitro. Monocytes either adherent to conventional plastic culture plates in serum-free conditions, or in suspension in culture medium containing serum were stimulated to produce IL-1 during culture. In non-adherent, serum-free conditions, monocytes produced very low or undetectable amounts of IL-1 during 20 h of culture. Lipopolysaccharide (LPS) induced equivalent amounts of secreted and cell-associated IL-1, although at very low concentrations more cell-associated IL-1 was produced. IL-1 production in response to LPS could be augmented by crude lymphokine, IFN-γ, or tumour necrosis factor (TNF) alpha. TNF-α preferentially augmented the production of cell-associated IL-1 in LPS-stimulated cultures. TNF-α induced a significant amount of IL-1 (mainly cell-associated) directly but could also induce IL-1 secretion when combined with IL-2 or IFN-γ, or when in the presence of serum. IL-2 acted synergislically with low concentrations of IFN-γ or IL-1 to induce significant levels of IL-1 production. IFN-α did not induce any IL-1 production, but was a potent inhibitor of IL-1 production induced by a variety of stimuli. These results suggest that IL-1 production may be enhanced or reduced by different cytokines at concentrations likely to be found in chronic inflammatory lesions.
Prevention of adjuvant arthritis in rats by a nonapeptide from the 65-kD mycobacterial heat-shock protein
Authors: YANG X-D , GASSER J , FEIGE U
Published: July 2010

Adjuvant arthritis in Lewis rats is a model of T cell-mediated autoimmune arthritis resembling human rheumatoid arthritis. A nonapeptide from the 65-kD heat-shock protein of Mycobacterium bovis BCG, amino acid sequence 180–188, has been described to carry the dominant immunogenic epitope(s) for both arthritis-protective and arthritogenic T cell clones. Here we demonstrate that immunizations with the synthetic nonapeptide completely protected rats against adjuvant arthritis induced by M. tuberculosis. Interestingly, deletion of the N-teminal threonine of the nonapeptide resulted in loss of the protective activity. Pretreatments with the nonapeptide resulted in an immune response to the nonapeptide and to M. tuberculosis. After immunizations with the synthetic nonapeptide, only low titres of nonapcptidc-spccific antibodies were produced, whereas a significant cellular immune response to the nonapeptide was observed. In addition, the protection was transferable to naive rats by spleen T cells. These findings document the requirement of a T cell-specific immune response to the dominant epitope of the 65-kD mycobacterial heat-shock protein for the protection against adjuvant arthritis and suggest the feasibility of immune intervention in autoimmune arthritis through the use of synthetic peptides.
Effect of phorbol myristate acetate on T cell colony formation, interleukin-2 (IL-2) receptor expression and IL-2 production by cells from patients at all stages of HIV infection
Authors: ALLOUCHE M , LUNARDI-ISKANDAR Y , VARELA-MILLOT C , ITZHAKI M , CORNUET P , MEYER P , JASMIN C , GEORGOULIAS V
Published: July 2010

We and others have shown that several T cell responses induced by the mitogen phytohaemagglutinin (PHA), including T cell colony formation, IL-2 receptor (IL-2R) expression, and IL-2 production are impaired in patients with AIDS and lymphadenopathy syndrome (LAS). We investigated whether phorbol myristate acetate (PMA) could act in synergy with PHA (as it does in healthy subjects) to enhance in vitro T cell responses of patients at all stages of infection by HIV. In AIDS patients with opportunistic infections (AIDS/OI), PHA + IL-2 + PMA led to a total disappearance of T cell colonies in 10/11 patients, among whom six already displayed very low numbers of colonies induced by PHA + IL-2 (less than 50 colonies/5 × 104 cells). In contrast, T cell colony formation induced by PHA + IL-2 + PMA was maintained or increased, compared with that induced by PHA + IL-2, in five out of six AIDS patients with Kaposi's sarcoma (AIDS/KS), 10/14 LAS and six out of seven HIV-seropositive asymptomatic (HIV+/AS) homosexuals. In these three groups of patients, a low percentage of colony cells induced by PHA + IL-2 + PMA expressed CD3 and CD4 molecules, but 50-89% of cells were IL-2R (Tac) positive, as in healthy controls. Studies on T cell activation and IL-2 production were performed on a selected group of 12 HIV-infected patients for whom sufficient numbers of lymphocytes could be obtained. PMA induced CD4 down-modulation in controls and in HIV-infccted patients. However. CD3 down-modulation and induction of the Tac chain of IL-2R by PMA were significantly impaired in patients, compared with controls, and these two parameters were correlated. Although PHA alone induced virtually normal levels of Tac antigen on patients’ cells, Tac induction by PHA + PMA was significantly decreased in patients versus controls. Cells from five out of 10 patients tested failed to produce detectable amounts of IL-2 after PHA stimulation, whereas IL-2 production increased significantly in all patients tested (n = 9) after PH A + PMA, with a level of IL-2 activity significantly higher than in controls. No correlation was found in this group of patients between the effects of PMA + PHA on T cell colony formation, Tac expression, or IL-2 production, as compared with PHA alone. Taken together, our results indicate that in vitro T cell functional studies with PMA may be useful to evaluate better the defects of T cell activation in HIV-infected patients.
Natural killer cell activity during measles
Authors: GRIFFIN D E , WARD B J , JAUREGUI E , JOHNSON R T , VAISBERG A
Published: July 2010

Natural killer cells are postulated to play an important role in host anti-viral defences. We measured natural killer cell activity in 30 individuals with acute measles (73 ± 21 lytic units (LU)/107 cells) and 16 individuals with other infectious diseases (149 ± 95 LU) and found it reduced compared with values for adults (375 ± 70 LU; P<0.001) or children (300 ± 73 LU, P<0.01) without infection. Reduced natural killer cell activity was found in measles patients with (84 ± 30 LU) and without (55 ± 18 LU) complications and was present for at least 3 weeks after the onset of the rash. Activity was increased by in vitro exposure of cells to interleukin-2. Depressed natural killer cell activity parallels in time the suppression of other parameters of cell-mediated immunity that occurs during measles.
Evidence that the Fc region of autologous rabbit IgG isolated before and after hyperimmunization is structurally different: recognition by rheumatoid factor and monoclonal antibodies
Authors: MURRAY J S , BROWN J C
Published: July 2010

Autologous hyperimmune (HI) and pre-inoculation (PI) rabbit Fcγ populations were found to be conformationally different by spectroscopic measurements, and antigenically different by measurements which examined rheumatoid factor (RF) and monoclonal antibody (MoAb) binding specificity for both populations in ELISA. Circular dichroism spectra of HI rabbit Fcγ (prepared from animals after hyperimmunization with streptococcal vaccine) were both qualitatively and quantitatively different, particularly in the 225–228 nm range, in comparison to both homologous normal and autologous PI Fcγ. Binding studies in ELISA showed that affinity constants obtained for reactions of both rabbit RF and various murine MoAb with HI IgG and Fc were approximately 10-fold higher relative to those observed for PI IgG and Fc. Enzymatic deglycosylation of HI and PI Fcγ led to elimination of CD spectral differences. Further, association constants obtained for RF and MoAb reactions with deglycosylated (sialic acid and galactose removed) PI Fcγ were equivalent to those obtained in the presence of untreated HI Fcγ. Together, these results suggest the complex oligosaceharide structure of rabbit IgG may play a significant role in the expression of Fcγ determinants, and alteration of this structure under hyperimmune or other conditions may be related to induction of an RF response.
Immune studies in infants with congenital syphilis
Authors: SAMSON G R , BEATTY D W , MALAN A F
Published: July 2010

Seventeen neonates with congenital syphilis were studied to determine the immune response of the fetus following intra-uterine infection with Treponema pallidum. The results were compared with those from healthy controls matched for gestational age, birth weight and sex. B cells, IgM, and circulating immune complexes were significantly elevated in the infected newborns. There were no differences in lymphocyte transformation to phytohaemagglutinin (PHA) and in the CD3, CD4, and CD8 lymphocytes between infants with congenital syphilis and controls. Newborns with congenital syphilis have a heightened humoral response but no quantitative abnormality in cell-mediated immunity. Speculation on the role of the circulating immune complexes is presented.