bluuebunny/crossref_metadata_2025 huggingface dataset:
Molecular basis of haemophilia A
Authors: Oldenburg J. , Ananyeva N. M. , Saenko E. L.
Published: October 2004

Summary.  Technologies in molecular biology have greatly advanced the knowledge regarding the origin of haemophilia A and the physiology of the factor VIII (FVIII) protein. A variety of different mutations in the FVIII gene have been identified and their effects on the FVIII protein described. It has been shown that the frequency of haemophilia A is due to a high mutation rate predominantly in male germ cells. A significant proportion is originating de novo in early embryogenesis from somatic mutations, a finding that has implications for genetic counselling. The life‐cycle of the FVIII protein and its structure‐function relationships are continuously clarified. Most recently it has been shown that FVIII clearance from the circulation is mediated by the low‐density lipoprotein receptor‐related protein (LRP) and cell‐surface heparan sulphate proteoglycans (HSPGs). These findings raise hope for novel recombinant FVIII molecules with prolonged half‐life that may improve therapies for haemophlia A.
Emerging and receding risks of therapeutic regimens for haemophilia
Authors: Farrugia A. , Manno C. S. , Evatt B. L.
Published: October 2004

Summary.  During the past two decades, the improvement of therapeutic agents for the management of haemophilia has created the opportunity for individuals with haemophilia to live normal lives. However, in some instances, the progress made has been accompanied by emergence of unexpected risks and other new complications. A number of viruses have either emerged, or become greater risks to people with haemophilia. In addition, the drive of many countries towards self‐suffiency in blood products may in fact be endangering people with haemophilia by restricting blood donation to a pool of donors with high infection risk, discouraging commercial interests from developing safer products, and discouraging use of ‘foreign’ products even where that may be the safer option. Gene therapy for haemophilia, although an encouraging new treatment, has brought with it a number of adverse events, including risk of virus infection and development of carcinomas. The risk of inhibitors is still the most important problem for people with haemophilia, and a recent report showed that the type of factor concentrate does not impact significantly on this risk. Despite the advent of new and promising treatments for haemophilia, heathcare providers must be alert to new risks posed by them.
Product delivery in the developing world: options, opportunities and threats
Authors: Farrugia A.
Published: October 2004

Summary.  While many developed countries are moving to recombinant coagulation factors as their preferred modality for delivering haemophilia care, the cost of these products currently impedes their access by developing countries. A number of options are available to these countries for the provision of plasma‐derived therapeutic products. The decreasing market for plasma‐derived coagulation factors in the developed world is leading to the generation of a surplus of these products and an ability to offer them outside their traditional markets if prices are affordable. Current indications are that the commercial fractionation industry of the developed world has an excess capacity in both available plasma and fractionation plants. It would seem that accessing this capacity might have attractions for the developing world. Countries wedded to achieving self‐sufficiency in haemophilia products may elect to develop a strategy for fractionating domestically sourced plasma. This may be achieved by the generation of a capacity to fractionate within the country or by contracting the fractionation to an external agency overseas. However, reliance on domestic plasma should not be allowed to impede access to sufficient and safe coagulation products. Irrespective of the route chosen, products need to attain satisfactory compliance to standards for safety, quality and efficacy. This is best done through alignment of the products with the requirements of credible regulatory agencies. While the approval of the mainstream regulators of the developed world affords considerable assurance regarding product quality, the increasing efforts made by fractionation agencies in the developing world to attain best practice is commendable and augurs well for the enhancement of haemophilia care in these countries.
Acquired bleeding disorders: the impact of health problems in the developing world
Authors: Isarangkura P. , Mahasandana C. , Chuansumrit A. , Angchaisuksiri P.
Published: October 2004

Summary.  Several acquired bleeding disorders in the developing world have impacts on health, including late vitamin K deficiency bleeding (VKDB) in infants, dengue haemorrhagic fever (DHF), and malaria. This paper describes their clinical manifestations, mechanisms involved, and treatment.
Comprehensive care for haemophilia around the world
Authors: Evatt B. L. , Black C. , Batorova A. , Street A. , Srivastava A.
Published: October 2004

Summary.  Comprehensive haemophilia care has been defined as the continuing supervision of all medical and psychosocial factors affecting the person with haemophilia family. Services offered by haemophilia treatment centres (HTCs) adopting the comprehensive care model include establishing prophylaxis and other treatment protocols, development of psychosocial, education and research programme, maintenance of a patient registry, genetic and reference diagnostic services and orchestration and management of a wide variety of multidisciplinary interventions. Most centres practising this model of care are based in developed countries and can meet costs for plentiful treatment products through government or insurance‐company funding. Not all the programmes are dependent on the level of product supply, however, and many have been supported in countries with emerging economies as part of national healthcare systems, particularly in relation to blood management. In this paper we present perspectives from different areas of the world on how to adopt, adapt and achieve economically appropriate models of comprehensive care.
Mutation Analysis in F9 Gene of 17 Families with Haemophilia B from Iran
Authors: Enayat M. S. , Karimi M. , Chana G. , Farjadian S. , Theophilus B. D. M. , Hill F. G. H.
Published: October 2004

Summary.  Seventeen haemophilia B families from Iran were investigated to determine the causative mutation. All the essential regions of the F9 gene were initially screened by conformational sensitive gel electrophoresis and exons with band shift were sequenced. Seven of the 15 mutations identified in these families were novel mutations. The mutations were authenticated in nine families as other affected members or heterozygous female carriers were available for verification.
Preimplantation genetic diagnosis: new reproductive options for carriers of haemophilia
Authors: Lavery S.
Published: October 2004

Summary.  Preimplantation genetic diagnosis for haemophilia offers couples at risk for transmitting the condition the opportunity to embark on a pregnancy knowing that the embryo is unaffected by the disease. The technique aims to increase the range of reproductive options available to these couples and remove the need for invasive prenatal diagnosis and the difficult decision on whether to terminate an affected pregnancy. This aims to reduce the anxiety associated with reproduction often seen in these couples. Patients undergo a cycle of in vitro fertilization followed by embryo biopsy. The single blastomeres are then analysed using fluorescent in situ hybridization to detect the sex of the embryo, and only female embryos are transferred to the uterus. Recently a PCR based approach has allowed specific mutation detection, and therefore the transfer of unaffected male and female embryos.
Dose and outcome of care in haemophilia – how do we define cost‐effectiveness?
Authors: Fischer K. , Van Den Berg H. M. , Thomas R. , Kumar S. , Poonnoose P. , Viswabandya A. , Mathews V. , Kavitha M. L. , Bhattacharji S. , Srivastava A.
Published: October 2004

Summary.  Severe haemophilia (factor [F]VIII/FIX activity < 0.01 IU mL−1) is characterized by repeated haemarthroses resulting in severe arthropathy in adulthood. In 1958, Professor Nilsson in Sweden introduced prophylactic infusions with clotting factor concentrates at regular intervals in order to maintain clotting factor levels above 0.01 IU mL−1 and to prevent bleeding. Since then, evidence of the long‐term beneficial effects of prophylactic treatment for severe haemophilia has been increasing and it has become the recommended treatment strategy for children with severe haemophilia by both the World Health Organization and the US National Hemophilia Foundation Medical and Scientific Advisory Committee. However, the implementation of this recommendation has been hampered by issues of cost and venous access. The high costs of prophylaxis have largely prevented its use in major parts of the world. The question therefore is whether the current models of replacement of clotting factor concentrates, while certainly being effective, are also optimal. Can the data on outcome at different levels of factor replacement be used to assess their cost‐effectiveness?
Membrane isoforms of human immunoglobulins of the A1 and A2 isotypes: structural and functional study
Authors: LEDUC I. , DROUET M. , BODINIER M.‐C. , HELAL A. , COGNE M.
Published: October 2004

As for IgM, human IgA occurs either as soluble molecules in plasma and various other body fluids, or as membrane‐bound molecules on differentiated B cells, where they are part of the B‐cell receptor for antigen (BCR). We studied the structure of transcripts encoding the membrane‐anchored α‐chain of the human BCRα, which may be present in two different forms resulting from alternate splicing of the α‐chain mRNA (type I or type II). The ratio of type I versus type II did not vary upon stimulation of a B‐cell line with various cytokines. Rather, it differed strikingly in cells expressing either the IgA1 or IgA2 isotype of the BCRα, with virtually no type II α‐chain in the latter. Co‐modulation experiments also yielded different results for both isotypes, since they demonstrated a physical association of both membrane (m)IgA1 and mIgA2 with CD79b, the β component of the BCR Igα/Igβ heterodimer, but only of mIgA1 with CD19. Whatever the isotype, the BCR of the IgA class was able to carry out signal transduction upon cross‐linking by specific monoclonal antibodies but, in contrast to mIgM, it relied mainly on the entry of extracellular Ca2+ rather than on the release of intracellular stocks.
Modulation of immune dysfunction during murine leukaemia retrovirus infection of old mice by dehyroepiandrosterone sulphate (DHEAS)
Authors: ARAGHI‐NIKNAM M. , LIANG B. , ZHANG Z. , ARDESTANI S. K. , WATSON R. R.
Published: October 2004

Ageing, leukaemia and acquired immune deficiency syndrome (AIDS) are conditions with dysregulated cytokine production. As dehydroepiandrosterone sulphate (DHEAS) restored normal cytokine production in old mice its effects on retrovirally infected old mice were investigated. Retrovirus infection and ageing‐induced immune dysfunction. Murine retrovirus‐infected old C57BL/6 female mice consumed 0·22 or 0·44 μg of DHEAS/mouse/day beginning 2 weeks postinfection for 10 weeks. DHEAS largely prevented the retrovirus‐induced reduction in T‐cell and B‐cell mitogenesis. DHEAS supplement prevented loss of cytokines [interleukin‐2 (IL‐2) and interferon‐γ] secretion by mitogen‐stimulated splenocytes representing T helper 1 (Th1) cell phenotypes. It also suppressed the retrovirus‐induced, excessive production of cytokines (IL‐6 and IL‐10) by Th2 cells. The highest dose of DHEAS reduced IL‐6 production by splenocytes from uninfected old mice by 75% while increasing their IL‐2 secretion by nearly 50%. Thus immune dysfunction induced by ageing, even when exacerbated by murine retrovirus infection, was largely prevented by DHEAS.
Involvement of Stat3 in interleukin‐6‐induced IgM production in a human B‐cell line
Authors: FARIS M. , KOKOT N. , STAHL N. , NEL A. E.
Published: October 2004

Interleukin‐6 (IL‐6) is an important B‐cell growth and differentiation factor. IL‐6 treatment of the human lymphoblastoid cell line, SKW6.4, leads to increased IgM production. We have previously shown that IL‐6 induces activation of JAK1 and JAK2 in human B cell lines. A chimeric IL‐6 receptor, comprised of the intracellular tail of the IL‐6 receptor subunit gp130 fused to the extracellular domain of the epidermal growth factor (EGF) receptor, was stably transfected into SKW6.4 cells. EGF treatment induced IgM production in cells transfected with an intact gp130 cytoplasmic tail, but not in untransfected cells or cells transfected with a cytoplasmic tail lacking all four signal transducers and activators of transcription (Stat) binding sites. Moreover, EGF treatment induced Stat3 phosphorylation in cells transfected with the intact chimeric EGF–gp130 receptor along with induction of DNA‐mobility shift of a classical interferon‐γ‐activated site. To define further the relation between Stat3 activation and enhanced IgM production, we determined the effect of chimeric gp130 on the transcriptional activation of a genetic element linked to immunoglobulin production, namely the immunoglobulin heavy chain enhancer (IgH‐enhancer). Parental as well as transfected SKW6.4 cells were transiently transfected with an IgH‐enhancer–luciferase construct. The transcriptional activity of the IgH–luciferase construct was induced upon ligation of the full‐length chimeric receptor but not by truncated gp130 receptors. Moreover, the gp130‐induced activity of this reporter gene was abrogated by Stat3EE, a mutant Stat3 incapable of binding DNA. These results indicate that IL‐6‐induced B‐cell differentiation, as measured by IgM production, may be controlled by Stat3 proteins.
Tumour necrosis factor‐α up‐regulates decay‐accelerating factor gene expression in human intestinal epithelial cells
Authors: ANDOH A. , FUJIYAMA Y. , SUMIYOSHI K. , SAKUMOTO H. , OKABE H. , BAMBA T.
Published: October 2004

The increased expression of decay‐accelerating factor (DAF) has been detected in intestinal epithelial cells at the inflamed mucosa. In this study, we examined the effects of tumour necrosis factor (TNF)‐α on DAF expression in three intestinal epithelial cell lines. DAF mRNA expression was evaluated by Northern blot analysis, and DAF protein expression was analysed by biotin labelling and immunoprecipitation. TNF‐α induced a marked increase in DAF mRNA and protein expression in HT‐29, T84 and Caco‐2 cells. In HT‐29 cells, the effects of TNF‐a on DAF mRNA accumulation were observed in a dose‐dependent manner; DAF mRNA accumulation reached a maximum at 3–6 hr, and then gradually decreased. These effects of TNF‐α required de novo protein synthesis. Messenger RNA stability studies suggested that TNF‐α partially regulated DAF gene expression by a posttranscriptional mechanism. Moreover, the combination of TNF‐α and interleukin (IL)‐4 induced an additive increase in DAF mRNA accumulation in HT‐29 and T84 cells. In human intestinal epithelial cells, TNF‐α acts as a potent inducer of DAF mRNA expression, indicating an important role for TNF‐α in the regulation of DAF expression at the inflamed mucosa.
Flexibility in T‐cell receptor ligand repertoires depends on MHC and T‐cell receptor clonotype
Authors: GELUK A. , VAN MEIJGAARDEN K. E. , OTTENHOFF T. H. M.
Published: October 2004

T‐cell receptors (TCR) recognize peptides complexed to self‐major histocompatibility complex (MHC) molecules. Recognition of peptide/MHC ligands by the TCR is highly peptide specific. However, certain TCRs can also recognize sequence‐related and ‐unrelated (‘mimicry’) epitopes presented by homologous MHC molecules. Using two human, human leucocyte antigen‐DR1 (HLA‐DR1)‐restricted T‐cell clones specific for HA p307–319, we identified several diverse combinations of peptide‐MHC complexes that are functionally equivalent in their ability to trigger T‐cell stimulation. These findings demonstrate that a single TCR can productively interact with different peptides complexed to self‐ as well as non‐self‐MHC molecules. This extended reactivity is human leucocyte antigen (HLA) allele and TCR clonotype dependent, as the peptide repertoire recognized depends on the presenting HLA‐DR molecule and varies among different TCRs that both recognize the HA p307–319/DR1 complex. Importantly, certain peptide analogues can completely change the HLA‐restriction pattern of the TCR: T‐cell recognition of the wild‐type peptide that was absent in the context of a non‐self HLA‐DR molecule, was restored by complementing substitutions in altered peptide ligands, that could not be presented by the original restriction element. This mechanism may play an important role in allorecognition.
The antigen receptor complex on cord B lymphocytes
Authors: MACARDLE P. J. , WEEDON H. , FUSCO M. , NOBBS S. , RIDINGS J. , FLEGO L. , ROBERTON D. M. , ZOLA H.
Published: October 2004

The neonatal immune system responds to a restricted range of antigens, producing largely IgM antibody of low affinity. Comparison of the components of the B‐cell antigen receptor complex shows significantly elevated membrane levels of IgM in neonatal B cells, compared with adult cells. CD79, which acts as the signal transducer for membrane immunoglobulin, is elevated in parallel with IgM, while IgD is elevated to a lesser degree. CD19, CD21, CD22 and CD81, which are all involved in transmitting activation signals when immunoglobulin is engaged, are not elevated. CD32, which is involved in negative regulation of activation, is present at reduced levels on cord B cells. The elevation of B‐cell membrane IgM persists during infancy. Neonatal B cells respond in vitro to interleukin‐4 (IL‐4) by further elevation of membrane IgM levels. The elevated level of membrane IgM may make neonatal B cells easier to trigger by low concentrations of antigen, but in vitro activation and immunoglobulin modulation experiments did not show significant differences between cord and adult B‐cell responses to anti‐IgM.
Apoptosis in a Fas‐resistant, T‐cell receptor‐sensitive human leukaemic T‐cell clone
Authors: DELEHANTY L. L. , PAYNE J. A. , FARROW S. N. , BROWN R. , CHAMPION B. R.
Published: October 2004

The Fas (CD95) antigen plays a key role in regulating T‐cell activation and survival. We have generated a Fas‐resistant subclone of the human T‐cell leukaemia line, H9, which is still able to undergo apoptosis in response to T‐cell receptor ligation. Molecular analyses revealed that resistance to Fas‐mediated apoptosis was due to a heterozygous mutation in the death domain of the Fas gene which generates a stop codon, and thus encodes a truncated Fas molecule. Fas ligation was able to induce apoptosis in the presence of cycloheximide, indicating that the mutant Fas molecule retained some signalling capability, which is death‐domain independent. These cells will provide a useful tool for dissecting the complexities of Fas signalling pathways.
Mouse hepatitis viral infection induces an extrathymic differentiation of the specific intrahepatic αβ‐TCRintermediate LFA‐1high T‐cell population
Authors: LAMONTAGNE L. , MASSICOTTE E. , PAGE C.
Published: October 2004

Mouse hepatitis virus type 3 (MHV3), a coronavirus, is an excellent model for the study of thymic and extrathymic T‐cell subpopulation disorders induced during viral hepatitis. It was recently reported that, in addition to the intrathymic T‐cell diVerentiation pathway, an extrathymic differentiation pathway of αβ‐T‐cell receptor (TCR) T lymphocytes exists in the liver, and becomes important under pathological situations such as autoimmune diseases, malignancies or hepatic bacterial infections. In the present study, we compared the phenotypes of resident hepatic, splenic or thymic T‐cell subpopulations during the acute viral hepatitis induced by MHV3 in susceptible C57BL/6 mice. The number of liver‐resident mononuclear cells (MNC) increased during the viral infection, while cellularity decreased. Single positive (SP) CD4+ cells strongly increased in both the liver and thymus, while double positive (DP) (CD4+CD8+) cells, present in the liver and thymus of mock‐infected mice, decreased in C57BL/6 mice during the viral infection. A shift of αβ‐TCRintermediate T cells toward αβ‐TCRhigh was evidenced in the liver and thymus of infected mice, but not in the spleen. The few αβ‐TCRint double negative (DN) (CD4‐CD8‐) cells also decreased following viral infection. αβ‐TCRint or high lymphocytes expressing high levels of leucocyte function antigen‐1 (LFA‐1) increased in the liver of MHV3‐infected mice. In addition, liver‐resident T cells expressed strongly the CD44 (Pgp‐1) activation marker, suggesting that they were either activated or antigen experienced during the viral infection. No significant change in T‐cell subpopulations was detected in the spleen, suggesting that MHV3 infection could induce an early in situ differentiation of resident hepatic T cells rather than a recruitment of lymphocytes from peripheral lymphoid organs.
Suppression of murine thyroiditis via blockade of the CD40–CD40L interaction
Authors: CARAYANNIOTIS G. , MASTERS S. R. , NOELLE R. J.
Published: October 2004

The CD40 ligand (gp39) is transiently expressed on activated CD4+ T cells and mediates cognate helper function by interacting with CD40 on B cells. Increasing evidence suggests, however, critical involvement of gp39 not only in antibody‐mediated responses but also in the development of effector T cells. Here, we have investigated the effect of in vivo gp39 blockade on the induction of murine experimental autoimmune thyroiditis (EAT), a T‐cell‐mediated disease. Over a 5‐week period, EAT was induced in SJL mice with thyroglobulin (Tg) and adjuvant. Concomitantly, mice received intraperitoneal (i.p.) injections of MR1, a gp39‐specific hamster monoclonal antibody (mAb), at 4‐day intervals. Control mice were challenged with Tg but received equivalent doses of hamster immunoglobulin (HIg). It was observed that the control mice developed severe thyroiditis whereas the MR1‐treated mice exhibited very low levels of infiltration that were mostly focal in nature. Blockade of gp39 was effective since the Tg‐specific IgG titres were low or undetectable in all MR1‐treated animals compared with the controls. In addition, upon restimulation with Tg in vitro, lymph node cells (LNC) from Tg‐primed, MR1‐treated mice proliferated less strongly and secreted significantly lower amounts of interleukin‐2 (IL‐2) and interferon‐γ (IFN‐γ) than LNC from untreated or HIg‐treated controls. These results strongly suggest that in vivo blockade of gp39 suppresses EAT by inhibiting the priming of inflammatory Tg‐specific T‐helper type 1 cells.
Demonstration of cytoplasmic CD32 (FcγRII) within human lymphocytes following microwave treatment
Authors: SANDILANDS G. P. , BURNETT E. R. , MACPHERSON S. A. , DOWNIE I. , MORE I. A. R. , MACSWEEN R. N. M.
Published: October 2004

We have recently described a cytoplasmic form of CD32 (FcγRII) within the vast majority of normal human peripheral blood lymphocytes (PBL) including T cells. The function of cytoplasmic CD32 is not known. These flow cytometric studies were conducted using single cell suspensions of PBL that had been prefixed and permeabilized using methanol/triton‐X‐100. In this study we have attempted to visualize cytoplasmic CD32 by immunocytochemistry using normal PBL processed in various ways and have also looked for CD32 within tissue lymphocytes. Weak cytoplasmic CD32 staining was observed in paraffin sections of normal lymphocytes but only when sections were microwave treated. The intensity of staining for CD32 did however, appear to be much stronger within infiltrating lymphocytes found in autoimmune diseases or in rejecting allografts: an observation that suggests that up‐regulation of cytoplasmic CD32 may occur when T cells become activated in vivo. Microwave treatment of PBL suspensions was shown to disrupt the outer cell membrane, thus effectively permeabilizing the cell and allowing for the detection of cytoplasmic components, like CD32, by flow cytometry. Microwave treatment may, therefore, afford an alternative method for cell permeabilization and may prove to be a useful method for the study of cytoplasmic molecules in cell suspensions and in paraffin‐embedded tissues.
Pentoxifylline in vivo and in vitro down‐regulates the expression of the intercellular adhesion molecule‐1 in monocytes
Authors: NEUNER P. , KLOSNER G. , POURMOJIB M. , KNOBLER R. , SCHWARZ T.
Published: October 2004

Since pentoxifylline (PTX) was recently recognized as a substance with antiinflammatory capacities, we studied the in vivo and in vitro effect of PTX on the expression of the intercellular adhesion molecule‐1 (ICAM‐1) on human monocytes. For this purpose four healthy volunteers were treated with PTX (5×400 mg/day) for 2 days. Monocytes were isolated before and after PTX treatment and ICAM‐1 expression was investigated. As shown by fluorescence‐activated cell sorter (FACS) analysis, cultured monocytes isolated after oral application of PTX expressed significantly decreased amounts of ICAM‐1 when compared with monocytes collected prior to oral PTX application. Northern blot analysis revealed reduced amounts of ICAM‐1 mRNA in monocytes derived from volunteers after oral PTX treatment in comparison with monocytes isolated before oral PTX administration. Similarly, in monocytes treated with PTX (200 μg/ml) in vitro ICAM‐1 was found decreased both at the protein and mRNA level in comparison with untreated cells. The inhibitory effect of PTX on ICAM‐1 expression in monocytes could be reversed by the addition of exogenous tumour necrosis factor‐α (TNF‐α; 200 U/ml) suggesting that ICAM‐1 down‐regulation is mediated secondary to TNF‐α suppression by PTX. The specific role of TNF‐α in mediating ICAM‐1 expression in cultured monocytes could be confirmed by the finding that a neutralizing anti‐TNF‐α antibody partially down‐regulated ICAM‐1 expression. The observed suppressive in vivo and in vitro effects of PTX on ICAM‐1 expression in monocytes may contribute to the recently described antiinflammatory effects of PTX, e.g. in sepsis or allergic contact dermatitis.
Modulatory effect of interleukin‐10 on the production of platelet‐activating factor and superoxide anions by human leucocytes
Authors: BUSSOLATI B. , MARIANO F. , MONTRUCCHIO G. , PICCOLI G. , CAMUSSI G.
Published: October 2004

We observed that human monocytes (MO) and polymorphonuclear neutrophils (PMN) stimulated by lipopolysaccharide (LPS) produce platelet‐activating factor (PAF) in a pattern characterized by an early and a delayed peak of synthesis. The early peak of PAF synthesis was due to a direct stimulation of these cells through mCD14 receptor as it was inhibited by anti‐CD14 monoclonal antibody. The delayed and sustained peak of PAF synthesis was dependent on protein synthesis and cytokine production as shown by the inhibitory effect of cycloheximide on both MO and PMN, and of anti‐tumour necrosis factor‐α (anti‐TNF‐α) and of anti‐interleukin‐8 (anti‐IL‐8) neutralizing antibodies on MO and PMN respectively. IL‐10 completely prevented this second, cytokine‐dependent peak of PAF synthesis. In contrast, IL‐10 markedly enhanced the first peak of PAF synthesis both in MO and PMN. Moreover, IL‐10 was shown to modulate the production of superoxide anions (O‐2) on both MO and PMN. As suggested by previous studies, IL‐10 inhibited the delayed production of O‐2. In the present study, we observed that IL‐10 directly stimulated an early production of O‐2. In addition, IL‐10 enhanced the synthesis of O‐2 by MO and PMN challenged with LPS. The IL‐10‐induced O‐2 production was dependent, at least in part, from its effect on PAF synthesis, as it was inhibited by the PAF receptor antagonist WEB 2170. These results suggest that IL‐10 may upregulate the early synthesis of PAF and O‐2 triggered by direct LPS stimulation, whereas it may downregulate the delayed production of these mediators.
The β1 integrin, very late activation antigen‐4 on human neutrophils can contribute to neutrophil migration through connective tissue fibroblast barriers
Authors: GAO J.‐X. , ISSEKUTZ A. C.
Published: October 2004

Polymorphonuclear leucocyte (PMNL) accumulation in extravascular tissues and inflammatory exudates is dependent on their migration through blood vessel endothelium and then through connective tissue. Previously we utilized a barrier of human synovial and dermal fibroblasts (HSF or HDF) grown on microporous filters, as a model of PMNL migration through connective tissue. Those studies showed that β2 (CD18) and the β1 integrins, very late activation antigen‐5 (VLA‐5) and VLA‐6, in part mediate this PMNL migration. Here we report that VLA‐4, which can also be expressed at low levels on activated PMNL, is also involved in PMNL migration induced by C5a through fibroblast (HSF and HDF) barriers, because monoclonal antibody (mAb) to VLA‐4 significantly inhibited (by 20–30%) PMNL migration. Blocking the function of CD18, VLA‐5 or VLA‐6 was not required for detection of the VLA‐4‐mediated migration. Combination treatment with mAb to VLA‐4 and with mAb to VLA‐5 or to VLA‐6 further inhibited PMNL migration, irrespective of whether CD11/CD18 mechanisms were blocked with anti‐CD18 mAb or not. Treatment of PMNL with a peptide based on the VLA‐4‐binding domain in the CS‐1 fragment of fibronectin, but not a control peptide, inhibited PMNL migration to a comparable extent to treatment with mAb to VLA‐4. A low level of VLA‐4 was expressed on C5a‐activated PMNL, detected by immunofluorescence flow cytometry. These results suggest that VLA‐4 can be mobilized by human peripheral blood PMNL and can, in addition to VLA‐5, VLA‐6 and CD11/CD18 integrins, mediate PMNL migration through connective tissue. This is in marked contrast to PMNL transendothelial migration, where β1 integrins appear to play no significant role.
Cellular redox status influences both cytotoxic and NF‐κB activation in natural killer cells
Authors: VALLE BLAZQUEZ M. , LUQUE I. , COLLANTES E. , ARANDA E. , SOLANA R. , PEÑA J. , MUÑOZ E.
Published: October 2004

The role of cellular redox status in both cytotoxic activity and NF‐κ B activation in natural killer (NK) cells was investigated. The results indicate that stimulation of NK cells, either freshly isolated from peripheral blood lymphocytes (PBL) or long‐term cultured NK clones, with specific cell targets results in an increased binding activity of NF‐κB and AP‐1 transcription factors measured by gel retardation. Pretreatment of NK cells with the antioxidant pyrrolidine dithiocarbarmate (PDTC) leads to the inhibition of NF‐κB activation but the AP‐1 binding to DNA was superinduced. The inhibition of NF‐κB by PDTC paralleled with an inhibition of spontaneous cytotoxicity mediated by NK cells. Moreover, the inhibitors of serine proteases, N‐α‐tosyl‐l‐lysine chloromethyl ketone and N‐α‐tosyl‐l‐phenylalanine chloromethyl ketone, also blocked the cytolytic activity of NK cells against the sensitive target K562. In contrast, NK activity was not affected by pretreatment of the effector cells with the proteasome inhibitor N‐acetyl‐leu‐leu‐norleucinal which selectively inhibits NF‐κB activation. Altogether, these results support the hypothesis that the activation of NK cells involved transcriptional and post‐transcriptional events, and that reactive intermediates may play an important role in the molecular processes related with the generation of a cytotoxic response by NK cells.
Effectiveness of interferon beta treatment in relapsing‐remitting multiple sclerosis: an Italian cohort study
Authors: Russo Pierluigi , Paolillo Andrea , Caprino Luciano , Bastianello Stefano , Bramanti Placido
Published: October 2004

Rationale, aims and objectives  Randomized clinical trials (RCTs) have provided evidence for the efficacy of interferon beta (IFNβ) in the treatment of relapsing‐remitting multiple sclerosis (RRMS). The aim of this study was to evaluate the effectiveness of IFNβ treatment in clinical practice.

Methods  This was a national, multicentre, observational study of patients with confirmed RRMS. Demographic, clinical and therapeutic data were retrospectively collected for each patient enrolled in the study.

Results  The study cohort consisted of 427 patients exposed to and 245 never exposed to IFNβ treatment during the study period (for a total 2297 patient‐years of follow‐up). Among the exposed patients, 215 were initially untreated and then began IFNβ later in the follow‐up period; 137 of these patients were exposed to IFNβ for more than 2 years. In these patients, IFNβ treatment reduced the mean relapse rate by 24.2%[95% confidence interval (CI): 5.8–42.5%]. For 640 of the 672 patients enrolled in the study, it was possible to calculate the area under the disability/time curve compared to that present at baseline. A total of 117 (18.3%) patients displayed disability progression. Adjustment of the disability progression rates for potential confounders and/or for propensity scores by Poisson regression model resulted in relative risks for patients exposed to IFNβ treatment compared to those never exposed to IFNβ of 0.87 (95% CI: 0.56–1.34) after an exposure of ≤ 2 years, and of 0.35 (95% CI: 0.21–0.60) after an exposure of > 2 years.

Conclusions  These findings suggest that the evidence from RCTs on the treatment of RRMS with IFNβ has been effectively translated into routine clinical practice.
Application of hepatitis serology testing algorithms to assess inappropriate laboratory utilization
Authors: Ozbek Ozgen A. , Oktem Mehmet A. , Dogan Guliz , Abacioglu Yusuf H.
Published: October 2004

Rationale, aims and objectives  Many studies pointed out inappropriate utilization of laboratory caused by excessive amounts of tests ordered by doctors. To prevent and to eliminate the ordering of unhelpful tests, introducing diagnostic algorithms, which are also a suitable practice for application to hepatitis serology, have been suggested. This study aimed to determine inappropriate test ordering rates with respect to the commonly approved  algorithms  for  serological  diagnosis  of  viral  hepatitis.

Methods  To assess the number of inappropriate test orders, laboratory records of samples sent for hepatitis A, B, and D serology were reviewed and evaluated retrospectively with respect to algorithms for serological diagnosis of viral hepatitis. Orders including serological marker groups with inadequate clinical information to determine whether or not the order was inappropriate were excluded from the analysis.

Results  Application of diagnostic algorithms showed that 50% of anti‐HAV IgM and anti‐HAV total; 12.7% of anti‐HBs, 12.7% of anti‐HBc total, 78.5% of anti‐HBc IgM, 87.3% of HBe Ag, 78.8% of anti‐HBe, 58.7% of anti‐HD total orders were made inappropriately.

Conclusions  Our study provides information for inappropriate laboratory utilization for hepatitis serology testing and we suggest to use diagnostic algorithms applied by the serology laboratory to decrease the rate of unhelpful test orders.
An evaluation of the adequacy of outpatient monitoring of thyroid replacement therapy
Authors: Stelfox Henry Thomas , Ahmed Sofia B. , Fiskio Julie , Bates David W.
Published: October 2004

Objectives  Hypothyroid patients managed with excessive or insufficient thyroid replacement therapy are often difficult to clinically recognize. Monitoring may prevent or minimize the consequences of adverse drug events (ADEs). We sought to develop an explicit model of medication monitoring and to evaluate monitoring processes and ADEs in patients taking levothyroxine.

Methods  A retrospective chart review of 400 outpatients receiving levothyroxine therapy between 1 January 2000 and 1 January 2001 at a large North American tertiary care hospital. We measured the proportion of patients satisfying minimum monitoring criteria, experiencing specific monitoring errors and having levothyroxine‐related ADEs. Explicit monitoring criteria were derived from the literature and through expert opinion. Adverse drug events were identified using structured implicit reviews.

Results  Overall, only 56% (95% confidence interval [95% CI] 51–62%) of the patients prescribed levothyroxine received the minimal recommended monitoring. Errors were identified at all stages of the monitoring model. Patients who received the recommended monitoring had fewer levothyroxine‐related ADEs (1% vs. 6%, P = 0.013) than those who did not. Minority status (white people 2% vs. black people 4% vs. Hispanics 14%, P = 0.023) and primary language (English 3% vs. Non‐English 20%, P = 0.002) were the patient characteristics associated with levothyroxine‐related ADEs.

Conclusion  Only half of outpatients taking levothyroxine at one tertiary care hospital received the recommended monitoring during one year of follow‐up. Levothyroxine‐related ADEs were more frequent in patients with lower‐quality monitoring and in minorities and non‐English speakers.
First episode psychosis: a novel methodology reveals higher than expected incidence; a reality‐based population profile in Northumberland, UK
Authors: Proctor S. E. , Mitford E. , Paxton R.
Published: October 2004

Rationale, Aims and Objectives   To document the natural history/clinical course of an unselected population presenting with first episode psychosis across a Mental Health Trust.

Method  An observational database was set up covering all patients over 15 years of age. Data were collected at pre‐sentation  and annual follow‐up  intervals.

Results   A  total of 227 patients presented during the 3 years initial study period with a first episode psychosis. The commonest diagnoses were psychotic depression 19%, paranoid schizophrenia 11%, persistent delusional disorder 7% and bipolar affective disorder 7.5% giving an annual incidence of 30.36 per 100 000 population. At presentation, half had been admitted to hospital, nearly half of whom were detained under the Mental Health Act, and only a quarter were currently employed. Twenty‐six per cent had an episode of deliberate self‐harm and 14% had harmed others. A recurring pattern emerged of over half the patients being no  longer  in  contact  with  mental  health  services  1  year after presentation.  Data c ollection  is  ongoing.

Conclusions    Clinical and managerial cooperation achieved a practical framework for data collection. This approach in an unselected population of patients yielded new insights into the course of first episode psychosis. The higher incidence than expected from the literature has implications for local strategic planning and provides a framework for detailed evaluation of a complex patient group.
An audit of clinical outcomes and client and referrer satisfaction with a Mood and Anxiety Disorders Unit
Authors: Lewis Raylene , Musella Emma , Berk Michael , Dodd Seetal , McKenzie Helen , Hyland Mary
Published: October 2004

Objective  To evaluate outcome and client and referrer satisfaction with the service provided by a Mood and Anxiety Disorders Unit (MADU).

Method  MADU was a specialized clinical service for the assessment and management of individuals suffering with affective and anxiety disorders. Clients were referred to MADU from a variety of health service providers. A telephone survey of 30 clients and 20 referrers who have used the services of MADU was conducted, investigating outcome satisfaction with the service provided by MADU.

Results  Clients and referrers reported a high level of satisfaction with the service provided by MADU. There was a high degree of adherence to treatment recommendations. The mean Patient Global Impression of Improvement (PGI) rating by the clients before the MADU assessment was 2.74 (SD = 1.27). In comparison the mean PGI rat‐ing at the  time  of  follow‐up  was  6.64  (SD = 1.91).

Conclusions  Specialist mood disorders units are a useful and potentially cost‐effective additional service included as a part of a mental health service.
Patient satisfaction profiling of individual physicians: impact of panel status
Authors: Murff Harvey J. , Orav E. John , Lee Thomas H. , Bates David W. , Fairchild David G.
Published: October 2004

Rationale, aims and objectives  Evaluation of physician performance is increasingly based on patient satisfaction. However, few data are available regarding the extent to which individual physician profiles might be influenced by factors such as whether a physician's practice is open or closed. We evaluated whether panel status (whether or not a physician is accepting new patients) is associated with patient satisfaction with their primary care physician (PCP).

Methods  Cross‐sectional analysis of patient satisfaction surveys. Surveys were available for 1750 patients cared for by 69 PCPs. Patient satisfaction with their PCP was determined based on a composite of six questions derived from the Medical Outcomes Study. We used Generalized Estimating  Equations  to  adjust for physician level variation.

Results  Patients of closed‐panel physicians were more likely to rate their satisfaction with the provider as ‘Excellent’ or ‘Very Good’ compared to patients of open‐panel physicians (78% vs. 69%, P < 0.0001). After adjusting for satisfaction with the practice site, provider years in practice, managed care coverage, provider productivity, and patient race, the association between a closed panel and satisfaction remained significant (odds ratio 1.60, 95% confidence  interval  1.10–2.31).

 Conclusions  Individual physicians’ patient satisfaction data are confounded by factors not likely to be adjusted for in available profiles. After adjusting for other variables, physicians with closed panels still had better patient satisfaction compared to physicians with open panels. Further research is necessary to determine if panel status might also confound patient satisfaction.
Gender bias in cardiovascular advertisements
Authors: Ahmed Sofia B. , Grace Sherry L. , Stelfox Henry Thomas , Tomlinson George , Cheung Angela M.
Published: October 2004

Rationale Women with cardiovascular disease are treated less aggressively than men. The reasons for this disparity are unclear. Pharmaceutical advertisements  may  influence  physician  practices  and  patient  care. 

Aims and objective To determine if female and male patients are equally likely to be featured  in  cardiovascular  advertisements. 

Methods We examined all cardiovascular advertisements from US editions of general medical and cardiovascular journals published between 1 January 1996 and 30 June 1998. For each unique advertisement, we recorded the total number of journal appearances and the number of appearances in journals’ premium positions. We noted the gender, age, race and role of both the primary figure and the  majority  of  people  featured  in  the  advertisement. 

Results Nine  hundred and nineteen unique cardiovascular advertisements were identified of which 254 depicted a patient as the primary figure. A total of 20%[95% confidence interval (CI) 15.3–25.5%] of these advertisements portrayed a female patient, while 80% (95% CI 74.5–84.7%) depicted a male patient,P < 0.0001. Female patient advertisements appeared 249 times (13.3%; 95% CI 8.6–18.9%) while male patient advertisements appeared 1618 times (86.7%; 95% CI 81.1–91.4%),P < 0.0001. Female patient advertisements also had significantly fewer mean appearances than male patient advertisements in journals’ premium positions (0.82 vs. 1.99,P = 0.02). Similar results were seen when the advertisements were analysed according to predominant gender.

Conclusions Despite increasing emphasis on cardiovascular disease in women, significant under‐representation of female patients exists in cardiovascular advertisements. Physicians should be cognizant of this gender bias.
An evaluation of risk factors for adverse drug events associated with angiotensin‐converting enzyme inhibitors
Authors: Morimoto Takeshi , Gandhi Tejal K. , Fiskio Julie M. , Seger Andrew C. , So Joseph W. , Cook E. Francis , Fukui Tsuguya , Bates David W.
Published: October 2004

Rationale, aims and objectives  To identify potential factors leading to discontinuation of angiotensin‐converting enzyme (ACE) inhibitors because of adverse drug events.

Methods  Retrospective cohort study was conducted at outpatient clinics affiliated with an urban tertiary care hospital. ACE inhibitors were administered to 2225 consecutive outpatients.

Results  In 19% of the total cohort, ACE inhibitors were discontinued because of adverse drug events. Cox proportional hazard model identified the following independent risk factors for discontinuation because of adverse drug events: age, female gender, ethnicity other than African American or Latino, no history of previous ACE inhibitor use, history of cough caused by another ACE inhibitor, hypertension, anxiety or depression, no hemodialysis, and elevated creatinine. History of smoking was shown to be a risk factor for cough [hazard ratio (HR): 2.5; 95% confidence interval (CI): 1.1–5.7], angioedema (HR: 2.7; 95% CI: 1.1–7.0), and hyperkalaemia (HR: 5.4; 95% CI: 1.3–23.2). History of ACE inhibitor‐induced cough was not only a risk factor for cough (HR: 12.9; 95% CI: 7.5–22.3) but also for angioedema (HR: 9.1; 95% CI: 2.1–39.9). Patients with creatinine ≥ 1.6 mg dL−1 were likely to discontinue ACE inhibitors because of renal dysfunction (HR: 4.7; 95% CI: 1.5–12.7) and hyperkalaemia (HR: 10.9; 95% CI: 3.1–39.0). East Asians were more likely to develop cough (HR: 2.5; 95% CI: 1.1–5.7) and hyperkalaemia (HR: 80.3; 95% CI: 5.4–1190) and African Americans to develop angioedema (HR: 3.5;  95%  CI:  1.3–8.9). 

Conclusions  Although further validation is necessary, these risk factors should help doctors identify patients with elevated risk for adverse drug events because of ACE inhibitors.
Use of porcine tumor necrosis factor receptor 1‐Ig fusion protein to prolong xenograft survival
Authors: Costa C. , Bell N. K. , Stabel T. J. , Fodor W. L.
Published: October 2004

Abstract:  Background:  Delayed rejection of xenografts is a major hurdle that needs to be addressed to achieve long‐term engraftment in the pig‐to‐primate transplant setting. Both vascular and avascular xenografts are susceptible to a delayed rejection process that comprises humoral and cellular responses. Tumor necrosis factor (TNF) is believed to play a role in this process by promoting cell activation, apoptosis and the recruitment of inflammatory cells. To address this problem, we engineered the donor cell in such a way that it could block both human and porcine TNF.

Methods:  We produced a recombinant fusion protein containing the extracellular domain of the porcine TNF‐Receptor 1 and an IgG Fc moiety (pTNFR1Ig). We first evaluated by flow cytometry the pTNFR1Ig capacity to prevent TNFα‐induced expression of SLAI, SLAII, VCAM‐1, ICAM‐1 and E‐selectin on the cell surface of porcine aortic endothelial cells (PAEC). The effect on TNFα‐mediated cell death was also assessed by propidium iodide staining after incubating PAEC with TNFα plus cycloheximide for 24 h. PAEC and porcine fibroblasts were subsequently engineered by retroviral infection to express and secrete pTNFR1Ig and their resistance to the TNFα effects was tested in vitro. Finally, we transplanted mock‐control and pTNFR1Ig‐expressing PAEC under the kidney capsule of BALB/c mice in the absence of immunosuppression and examined the degree of rejection at 2 and 3 weeks post‐transplantation.

Results:  Treatment with pTNFR1Ig resulted in a very potent blockade of human, porcine and murine TNFα activity on porcine cells. It inhibited the upregulation of all cell surface markers of activation tested as well as the TNFα‐mediated cell death. Moreover, pTNFR1Ig‐expressing PAEC showed prolonged engraftment in a pig‐to‐mouse xenotransplant model.

Conclusions:  Incorporation of strategies that block TNF may prove useful in the development of xenografts resistant to delayed rejection.