bluuebunny/crossref_metadata_2025 huggingface dataset:
Expression of ribosomal and translation-associated genes is correlated with a favorable clinical course in chronic lymphocytic leukemia
Authors: Dürig Jan , Nückel Holger , Hüttmann Andreas , Kruse Elisabeth , Hölter Tanja , Halfmeyer Katja , Führer Anja , Rudolph Roland , Kalhori Naser , Nusch Arnd , Deaglio Silvia , Malavasi Fabio , Möröy Tarik , Klein-Hitpass Ludger , Dührsen Ulrich
Published: March 2003

B-cell chronic lymphocytic leukemia (B-CLL) is a heterogenous disease with a highly variable clinical course. Recent studies have shown that CD38 surface expression on the malignant cell clone may serve as a prognostic marker in that CD38+ patients with B-CLL are characterized by advanced disease stage, lesser responsiveness to chemotherapy, and shorter survival than CD38− patients. To further investigate the molecular phenotype of these 2 clinical subgroups, we compared the gene expression profiles of CD38+ (n = 25) with CD38− (n = 45) B-CLL patients using oligonucleotide-based DNA chip microarrays representative of approximately 5600 genes. The results showed that B-CLLs display a common gene expression profile that is largely independent of CD38 expression. Nonetheless, the expression of 14 genes differed significantly between the 2 groups, including genes that are involved in the regulation of cell survival. Furthermore, unsupervised hierarchical cluster analysis of 76 B-CLL samples led to the separation of 2 major subgroups, comprising 20 and 56 patients. Clustering to the smaller group was due in part to the coordinate high expression of a large number of ribosomal and other translation-associated genes, including elongation factors. Importantly, we found that patients with high expression of translation factors were characterized by a more favorable clinical course with significantly longer progression-free survival and reduced chemotherapy requirements than the remaining patients (P < .05). Our data show that gene expression profiling can help identify B-CLL subtypes with different clinical characteristics. Furthermore, our results suggest a role of translation-associated genes in the pathogenesis of B-CLL.
Biochemical and functional characterization of Rab27a mutations occurring in Griscelli syndrome patients
Authors: Ménasché Gaël , Feldmann Jérôme , Houdusse Anne , Desaymard Catherine , Fischer Alain , Goud Bruno , Basile Geneviève de Saint
Published: March 2003

Rab27a is a member of the Rab family of small GTPase proteins, and thus far is the first member to be associated with a human disease (ie, the Griscelli syndrome type 2). Mutations in the Rab27a gene cause pigment as well as cytotoxic granule transport defects, accounting for the partial albinism and severe immune disorder characteristics of this syndrome. So far, 3 Rab27a missense mutations have been identified. They open a unique opportunity to designate critical structural and functional residues of Rab proteins. We show here that the introduction of a proline residue in the α4 (Ala152Pro) or β5 (Leu130Pro) loop, observed in 2 of these spontaneous mutants, dramatically affects both guanosine triphosphate (GTP) and guanosine diphosphate (GDP) nucleotide-binding activity of Rab27a, probably by disrupting protein folding. The third mutant, Trp73Gly, is located within an invariant hydrophobic triad at the switch interface, and was previously shown in active Rab3A to mediate rabphilin3A effector interaction. Trp73Gly is shown to display the same nucleotide-binding and GTPase characteristics as the constitutively active mutant Gln78Leu. However, in contrast to Gln78Leu, Trp73Gly mutant construct neither interacts with the Rab27a effector melanophilin nor modifies melanosome distribution and cytotoxic granule exocytosis. Substitutions introduced at the 73 position, including the leucine residue present in Ras, did not restore Rab27a protein functions. Taken together, our results characterize new critical residues of Rab proteins, and identify the Trp73 residue of Rab27a as a key position for interaction with the specific effectors of Rab27a, both in melanocytes and cytotoxic cells.
Distinct HLA associations by stroke subtype in children with sickle cell anemia
Authors: Hoppe Carolyn , Klitz William , Noble Janelle , Vigil Lara , Vichinsky Elliott , Styles Lori
Published: March 2003

Children with sickle cell anemia (SCA) carry a 200-fold increased risk for cerebral infarction. Stroke can be the result of small-vessel (SV) or large-vessel (LV) disease. However, it is unknown whether these subtypes result from the same pathophysiologic processes. Complete HLA genotyping was performed on 231 eligible children previously enrolled in the Cooperative Study of Sickle Cell Disease (CSSCD). Cerebral infarction on magnetic resonance imaging (MRI) was documented in 71 patients, and 160 patients had negative findings on MRI. Based on MRI/magnetic resonance angiography (MRA) findings, infarct size, and location, 36 patients were classified as having LV stroke and 35 as having SV stroke. When comparing the total MRI+ group with the MRI− group, HLA DPB1*0401 was associated with increased stroke risk (P = .01), whereas DPB1*1701 (P = .02) conferred protection from stroke. These DPB1 associations with stroke were attributed to the SV stroke group, in whom DPB1*0401 was associated with susceptibility (P = .003) and DPB1*1701 with protection from stroke (P = .06). In the LV stroke subgroup, HLA-A*0102 (P = .02) and -A*2612 (P = .007) conferred susceptibility, whereas -A*3301(P = .04) protected from stroke. These results suggest that specific HLA alleles influence stroke risk and appear to contribute differently to SV and LV stroke subtypes. The distinct HLA associations with SV and LV stroke suggest that different pathologic processes may be involved in the development of stroke in children with SCA. If these results are confirmed in a larger study, HLA type may serve as a useful marker for the early identification of SCA patients at high risk for stroke.
CD40 induces human multiple myeloma cell migration via phosphatidylinositol 3–kinase/AKT/NF-κB signaling
Authors: Tai Yu-Tzu , Podar Klaus , Mitsiades Nicholas , Lin Boris , Mitsiades Constantine , Gupta Deepak , Akiyama Masaharu , Catley Laurence , Hideshima Teru , Munshi Nikhil C. , Treon Steven P. , Anderson Kenneth C.
Published: March 2003

Multiple myeloma (MM) is characterized by clonal expansion of malignant plasma cells in the bone marrow and their egress into peripheral blood with progression to plasma cell leukemia. Our previous study defined a functional role of CD40 activation in MM cell homing and migration. In this study, we examine signaling events mediating CD40-induced MM cell migration. We show that cross-linking CD40, using either soluble CD40L (sCD40L) or anti-CD40 monoclonal antibody (mAb), induces phosphatidylinositol 3–kinase (PI3K) activity and activates its downstream effector AKT in MM.1S cells. CD40 activation also activates the MAP kinase (MEK) pathway, evidenced by phosphorylation of extracellular signal-regulated mitogen-activated protein kinase (ERK), but not c-jun amino-terminal kinase (JNK) or p38, in a dose- and time-dependent manner. Using pharmacologic inhibitors of PI3K and MEK, as well as adenoviruses expressing dominant-negative and constitutively expressed AKT, we demonstrate that PI3K and AKT activities are required for CD40-induced MM cell migration. In contrast, inhibition of ERK/MEK phosphorylation only partially (10%-15%) prevents migration, suggesting only a minor role in regulation of CD40-mediated MM migration. We further demonstrate that CD40 induces nuclear factor (NF)–κB activation as a downstream target of PI3K/AKT signaling, and that inhibition of NF-κB signaling using specific inhibitors PS1145 and SN50 completely abrogates CD40-induced MM migration. Finally, we demonstrate that urokinase plasminogen activator (uPA), an NF-κB target gene, is induced by CD40; and conversely, that uPA induction via CD40 is blocked by PI3K and NF-κB inhibitors. Our data therefore indicate that CD40-induced MM cell migration is primarily mediated via activation of PI3K/AKT/NF-κB signaling, and further suggest that novel therapies targeting this pathway may inhibit MM cell migration associated with progressive MM.
A novel I-branching β-1,6-N-acetylglucosaminyltransferase involved in human blood group I antigen expression
Authors: Inaba Niro , Hiruma Toru , Togayachi Akira , Iwasaki Hiroko , Wang Xiao-Hui , Furukawa Yusuke , Sumi Ryoichi , Kudo Takashi , Fujimura Katsuya , Iwai Toshie , Gotoh Masanori , Nakamura Mitsuru , Narimatsu Hisashi
Published: March 2003

The human blood group i and I antigens are determined by linear and branched poly-N-acetyllactosamine structures, respectively. In erythrocytes, the fetal i antigen is converted to the adult I antigen by I-branching β-1,6-N-acetylglucosaminyltransferase (IGnT) during development. Dysfunction of the I-branching enzyme may result in the adult i phenotype in erythrocytes. However, the I gene responsible for blood group I antigen has not been fully confirmed. We report here a novel human I-branching enzyme, designatedIGnT3. The genes for IGnT1 (reported in 1993),IGnT2 (also presented in this study), and IGnT3consist of 3 exons and share the second and third exons. Bone marrow cells preferentially expressed IGnT3 transcript. During erythroid differentiation using CD34+ cells,IGnT3 was markedly up-regulated with concomitant decrease in IGnT1/2. Moreover, reticulocytes expressed theIGnT3 transcript, but IGnT1/2 was below detectable levels. By molecular genetic analyses of an adult i pedigree, individuals with the adult i phenotype were revealed to have heterozygous alleles with mutations in exon 2 (1006G>A; Gly336Arg) and exon 3 (1049G>A; Gly350Glu), respectively, of the IGnT3gene. Chinese hamster ovary (CHO) cells transfected with each mutatedIGnT3 cDNA failed to express I antigen. These findings indicate that the expression of the blood group I antigen in erythrocytes is determined by a novel IGnT3, not byIGnT1 or IGnT2.
Src-family kinase signaling modulates the adhesion ofPlasmodium falciparum on human microvascular endothelium under flow
Authors: Yipp Bryan G. , Robbins Stephen M. , Resek Mary E. , Baruch Dror I. , Looareesuwan Sornchai , Ho May
Published: March 2003

The pathogenicity of Plasmodium falciparum is due to the unique ability of infected erythrocytes (IRBCs) to adhere to vascular endothelium. We investigated whether adhesion of IRBCs to CD36, the major cytoadherence receptor on human dermal microvascular endothelial cells (HDMECs), induces intracellular signaling and regulates adhesion. A recombinant peptide corresponding to the minimal CD36-binding domain from P falciparum erythrocyte membrane protein 1 (PfEMP1), as well as an anti-CD36 monoclonal antibody (mAb) that inhibits IRBC binding, activated the mitogen-activated protein (MAP) kinase pathway that was dependent on Src-family kinase activity. Treatment of HDMECs with a Src-family kinase–selective inhibitor (PP1) inhibited adhesion of IRBCs in a flow-chamber assay by 72% (P < .001). More importantly, Src-family kinase activity was also required for cytoadherence to intact human microvessels in a human/severe combined immunodeficient (SCID) mouse model in vivo. The effect of PP1 could be mimicked by levamisole, a specific alkaline-phosphatase inhibitor. Firm adhesion to PP1-treated endothelium was restored by exogenous alkaline phosphatase. In contrast, inhibition of the extracellular signal–regulated kinase 1/2 (ERK 1/2) and p38 MAP kinase pathways had no immediate effect on IRBC adhesion. These results suggest a novel mechanism for the modulation of cytoadherence under flow conditions through a signaling pathway involving CD36, Src-family kinases, and an ectoalkaline phosphatase. Targeting endothelial ectoalkaline phosphatases and/or signaling molecules may constitute a novel therapeutic strategy against severe falciparum malaria.
Functional and comparative analysis of globin loci in pufferfish and humans
Authors: Gillemans Nynke , McMorrow Tara , Tewari Rita , Wai Albert W. K. , Burgtorf Carola , Drabek Dubravka , Ventress Nicki , Langeveld An , Higgs Douglas , Tan-Un Kian , Grosveld Frank , Philipsen Sjaak
Published: March 2003

To further our understanding of the regulation of vertebrate globin loci, we have isolated cosmids containing α- and β-globin genes from the pufferfish Fugu rubripes. By DNA fluorescence in situ hybridization (FISH) analysis, we show thatFugu contains 2 distinct hemoglobin loci situated on separate chromosomes. One locus contains only α-globin genes (α-locus), whereas the other also contains a β-globin gene (αβ-locus). This is the first poikilothermic species analyzed in which the physical linkage of the α- and β-globin genes has been uncoupled, supporting a model in which the separation of the α- and β-globin loci has occurred through duplication of a locus containing both types of genes. Surveys for transcription factor binding sites and DNaseI hypersensitive site mapping of the Fugu αβ-locus suggest that a strong distal locus control region regulating the activity of the globin genes, as found in mammalian β-globin clusters, may not be present in the Fugu αβ-locus. Searching the human and mouse genome databases with the genes surrounding the pufferfish hemoglobin loci reveals that homologues of some of these genes are proximal to cytoglobin, a recently described novel member of the globin family. This provides evidence that duplication of the globin loci has occurred several times during evolution, resulting in the 5 human globin loci known to date, each encoding proteins with specific functions in specific cell types.
Impaired granulocytopoiesis in patients with chronic idiopathic neutropenia is associated with increased apoptosis of bone marrow myeloid progenitor cells
Authors: Papadaki Helen A. , Eliopoulos Aristides G. , Kosteas Theodoros , Gemetzi Claudia , Damianaki Athina , Koutala Helen , Bux Juergen , Eliopoulos George D.
Published: March 2003

To probe the pathophysiologic mechanisms underlying neutropenia in patients with chronic idiopathic neutropenia (CIN) with hypoplastic and left-shifted granulocytic series in the bone marrow (BM), we have studied granulocytopoiesis in 32 adults with CIN by evaluating the number and survival characteristics of cells in several stages of granulocyte differentiation using flow cytometry and BM culture assays. We found that patients with CIN displayed a low percentage of CD34+/CD33+ cells, defective granulocyte colony-forming unit (CFU-G) growth potential of BM mononuclear or purified CD34+ cells, and low CFU-G recovery in long-term BM cultures (LTBMCs), compared with controls (n = 46). A low percentage of CD34+/CD33+ cells in patients was associated with accelerated apoptosis and Fas overexpression within this cell compartment compared with controls. No significant difference was documented in the percentage of apoptotic cells or the Fas+ cells within the fractionated CD34+/CD33−, CD34−/CD33+, and CD34−/CD33−/CD15+ BM subpopulations or the peripheral blood neutrophils, suggesting that the underlying cellular defect in CIN probably concerns the committed granulocyte progenitors. LTBMC stromal layers from patients produced abnormally high amounts of tumor necrosis factor α and cytokine levels in culture supernatants inversely correlated with the number of myeloid progenitor cells and positively with the proportion of apoptotic CD34+ cells. Patient LTBMC stromal layers displayed pathologic interferon γ and Fas-ligand mRNA expression and failed to support normal myelopoiesis. These data suggest that impaired granulocytopoiesis in CIN is probably due to overproduction of inflammatory cytokines by immune cells within the BM microenvironment that may exert an inhibitory effect on myelopoiesis by inducing Fas-mediated apoptosis in the granulocyte progenitors.
Paracrine interactions of basic fibroblast growth factor and interleukin-6 in multiple myeloma
Authors: Bisping Guido , Leo Regine , Wenning Doris , Dankbar Berno , Padró Teresa , Kropff Martin , Scheffold Christian , Kröger Matthias , Mesters Rolf M. , Berdel Wolfgang E. , Kienast Joachim
Published: March 2003

Myeloma cells express basic fibroblast growth factor (bFGF), an angiogenic cytokine triggering marrow neovascularization in multiple myeloma (MM). In solid tumors and some lymphohematopoietic malignancies, angiogenic cytokines have also been shown to stimulate tumor growth via paracrine pathways. Since interleukin-6 (IL-6) is a potent growth and survival factor for myeloma cells, we have studied the effects of bFGF on IL-6 secretion by bone marrow stromal cells (BMSCs) and its potential reverse regulation in myeloma cells. Both myeloma-derived cell lines and myeloma cells isolated from the marrow of MM patients were shown to express and secrete bFGF. Cell-sorting studies identified myeloma cells as the predominant source of bFGF in MM marrow. BMSCs from MM patients and control subjects expressed high-affinity FGF receptors R1 through R4. Stimulation of BMSCs with bFGF induced a time- and dose-dependent increase in IL-6 secretion (median, 2-fold; P < .001), which was completely abrogated by anti-bFGF antibodies. Conversely, stimulation with IL-6 enhanced bFGF expression and secretion by myeloma cell lines (2-fold;P = .02) as well as MM patient cells (up to 3.6-fold; median, 1.5-fold; P = .002). This effect was inhibited by anti–IL-6 antibody. When myeloma cells were cocultured with BMSCs in a noncontact transwell system, both IL-6 and bFGF concentrations in coculture supernatants increased 2- to 3-fold over the sum of basal concentrations in the monoculture controls. The IL-6 increase was again partially, but significantly, inhibited by anti-bFGF. The data demonstrate a paracrine interaction between myeloma and marrow stromal cells triggered by mutual stimulation of bFGF and IL-6.
A phase 2 trial of combination low-dose thalidomide and prednisone for the treatment of myelofibrosis with myeloid metaplasia
Authors: Mesa Ruben A. , Steensma David P. , Pardanani Animesh , Li Chin-Yang , Elliott Michelle , Kaufmann Scott H. , Wiseman Gregory , Gray Leigh A. , Schroeder Georgene , Reeder Terra , Zeldis Jerome B. , Tefferi Ayalew
Published: March 2003

Single-agent thalidomide (THAL) at “conventional” doses (> 100 mg/d) has been evaluated in myelofibrosis with myeloid metaplasia (MMM) based on its antiangiogenic properties and the prominent neoangiogenesis that occurs in MMM. THAL monotherapy at such doses produces approximately a 20% response rate in anemia but is poorly tolerated (an adverse dropout rate of > 50% in 3 months). To improve efficacy and tolerability, we prospectively treated 21 symptomatic patients (hemoglobin level < 10 g/dL or symptomatic splenomegaly) with MMM with low-dose THAL (50 mg/d) along with a 3-month oral prednisone (PRED) taper (beginning at 0.5 mg/kg/d). THAL-PRED was well tolerated in all enrolled patients, with 20 patients (95%) able to complete 3 months of treatment. An objective clinical response was demonstrated in 13 (62%) patients, all improvements in anemia. Among 10 patients who were dependent on erythrocyte transfusions, 7 (70%) improved and 4 (40%) became transfusion independent. Among 8 patients with thrombocytopenia (platelet count < 100 × 109/L), 6 (75%) experienced a 50% or higher increase in their platelet count. In 4 of 21 patients (19%), spleen size decreased by more than 50%. Responses observed were mostly durable after discontinuation of the PRED. The dose of THAL in this study (50 mg/d) was better tolerated than the higher doses used in previous studies. Adverse events associated with corticosteroid therapy were mild and transient. Clinical responses did not correlate with improvements in either intramedullary fibrosis or angiogenesis. THAL-PRED is well tolerated and preliminarily appears to be a promising drug regimen for treating cytopenias in patients with MMM.
The influence of immunosuppressive drugs on tolerance induction through bone marrow transplantation with costimulation blockade
Authors: Blaha Peter , Bigenzahn Sinda , Koporc Zvonimir , Schmid Maximilian , Langer Felix , Selzer Edgar , Bergmeister Helga , Wrba Friedrich , Kurtz Josef , Kiss Christopher , Roth Erich , Muehlbacher Ferdinand , Sykes Megan , Wekerle Thomas
Published: March 2003

We recently developed a murine protocol for the induction of allogeneic mixed chimerism and tolerance employing nonmyeloablative total body irradiation (TBI), standard-dose bone marrow transplantation (BMT), and costimulation blockade (cobl) with an anti-CD154 monoclonal antibody (mAb) plus CTLA4Ig. We now evaluated whether a short course (1 month) of immunosuppressive drugs, which would be ethically required in the clinical setting of organ transplantation to prevent graft loss in case tolerance is not achieved, interferes with tolerance induced with this regimen. Our results show that calcineurin inhibitors (cyclosporin A [CyA] or tacrolimus [FK]) inhibit development of long-term chimerism and abrogate tolerance induction in this model. Rapamycin (rapa), methylprednisolone (MP), FTY720, and mycophenolate mofetil (MMF), in contrast, have no negative effect on chimerism or tolerance development. Peripheral deletion of donor-reactive T cells, which usually occurs in the weeks following BMT in this model, is blocked by CyA and FK, but not by the other drugs tested. Furthermore, we found that the additional use of compatible immunosuppressive drugs (rapa plus MMF plus MP) allows the dose of TBI to be reduced, so that mixed chimerism and donor skin-graft acceptance can be achieved with 1 Gy using clinically feasible cell numbers. Thus, this protocol of BMT with costimulation blockade can be safely combined with a clinically tested immunosuppressive regimen to permit success with a lower dose of irradiation. These results should facilitate clinical application of this tolerance strategy.
Genomically complex lymphomas undergo sustained tumor regression upon MYC inactivation unless they acquire novel chromosomal translocations
Authors: Karlsson Åsa , Giuriato Sylvie , Tang Flora , Fung-Weier Jingly , Levan Göran , Felsher Dean W.
Published: March 2003

The targeted inactivation of oncogenes may be a specific and effective treatment for cancer. However, because human cancers are the consequence of multiple genetic changes, the inactivation of one oncogene may not be sufficient to cause sustained tumor regression. Moreover, cancers are genomically unstable and may readily compensate for the inactivation of a single oncogene. Here we confirm by spectral karyotypic analysis that MYC-induced hematopoietic tumors are highly genetically complex and genomically unstable. Nevertheless, the inactivation of MYC alone was found to be sufficient to induce sustained tumor regression. After prolongedMYC inactivation, some tumors exhibited a distinct propensity to relapse. When tumors relapsed, they no longer required the overexpression of MYC but instead acquired novel chromosomal translocations. We conclude that even highly genetically complex cancers are reversible on the inactivation of MYC, unless they acquire novel genetic alterations that can sustain a neoplastic phenotype.
Hepcidin, a putative mediator of anemia of inflammation, is a type II acute-phase protein
Authors: Nemeth Elizabeta , Valore Erika V. , Territo Mary , Schiller Gary , Lichtenstein Alan , Ganz Tomas
Published: March 2003

Hepcidin is a liver-made peptide proposed to be a central regulator of intestinal iron absorption and iron recycling by macrophages. In animal models, hepcidin is induced by inflammation and iron loading, but its regulation in humans has not been studied. We report that urinary excretion of hepcidin was greatly increased in patients with iron overload, infections, or inflammatory diseases. Hepcidin excretion correlated well with serum ferritin levels, which are regulated by similar pathologic stimuli. In vitro iron loading of primary human hepatocytes, however, unexpectedly down-regulated hepcidin mRNA, suggesting that in vivo regulation of hepcidin expression by iron stores involves complex indirect effects. Hepcidin mRNA was dramatically induced by interleukin-6 (IL-6) in vitro, but not by IL-1 or tumor necrosis factor α (TNF-α), demonstrating that human hepcidin is a type II acute-phase reactant. The linkage of hepcidin induction to inflammation in humans supports its proposed role as a key mediator of anemia of inflammation.
Genetic and nutritional factors contributing to hyperhomocysteinemia in young adults
Authors: Kluijtmans Leo A. J. , Young Ian S. , Boreham Colin A. , Murray Liam , McMaster Dorothy , McNulty Helene , Strain J. J. , McPartlin Joseph , Scott John M. , Whitehead Alexander S.
Published: March 2003

A modestly elevated total plasma homocysteine concentration (tHcy) is generally accepted as an independent and graded risk factor for various pathologies, including vascular diseases, neural tube defects, Alzheimer disease, and pregnancy complications. We analyzed 5 common functional polymorphisms in enzymes involved in homocysteine metabolism (ie, methylenetetrahydrofolate reductase [MTHFR] 677C>T and 1298A>C, methionine synthase [MTR] 2756A>G, cystathionine β-synthase [CBS] 844ins68, and methionine synthase reductase [MTRR] 66A>G) in 452 young adults, and quantified their independent and interactive effects on tHcy concentrations. Serum folate, red cell folate, vitamin B12, and tHcy concentrations were significantly influenced by MTHFR 677C>T genotypes. A particularly strong interaction was observed between theMTHFR 677TT genotype and serum folate, which led to a high tHcy phenotype that was more pronounced in males. The genetic contribution to the variance in tHcy was estimated to be approximately 9%, compared with approximately 35% that could be attributed to low folate and vitamin B12. Our study indicates that dietary factors are centrally important in the control of tHcy levels in young adults with additional, but somewhat weaker, genetic effects. These data underscore the potential benefits that may be gained by improving the dietary status of young adults, and provide support for the implementation of folate/B-vitamin food fortification programs.
Muscle weakness is related to utilization of health care resources in COPD patients
Authors: Decramer M , Gosselink R , Troosters T , Verschueren M , Evers G
Published: March 2003

The factors determining utilization of health care resources in patients with chronic obstructive pulmonary disease (COPD) are poorly understood. In order to obtain insight into these factors, we studied the utilization of health care resources in 57 stable COPD patients with a forced expiratory volume in one second (FEV1) of 36 +/- 9% predicted. Patients were divided into two groups: admitted at least twice in the last year (high medical consumption; n = 23) or not admitted in the last year (low medical consumption; n = 34). Other variables related to utilization of health care resources studied were; the number of hospital days; the number of out-patient visits to a pulmonary department in the last year; and the average daily dose (ADD) of corticosteroids taken in the last 6 months. The actual cost of utilization of health care resources, however, was not studied. In addition, pulmonary function, serum electrolytes, blood gas values, 6 min walking distance, respiratory and peripheral muscle force, and appraisal of self-care agency (ASA score) were studied. Pulmonary function, serum electrolytes, blood gas values, ASA score and walking distance were not different between the two groups (e.g. FEV1 36 +/- 8 vs 36 +/- 10% pred). Respiratory muscle forces tended to be lower in the high medical consumption group, this tendency almost reaching statistical significance for maximal expiratory pressure (PE,max) (p = 0.08). Peripheral muscle force, however, was clearly reduced in the high medical consumption group (quadriceps force 63 +/- 20 vs 82 +/- 26% pred; p < 0.05). The number of admissions, the number of hospital days, the number of out-patient visits, and ADD were interrelated and also related to ventilatory and peripheral muscle force (r -0.18 to -0.38). This relationship was statistically significant for PE,max, whilst a similar tendency was present for maximal inspiratory pressure (PI,max). In stepwise multiple regression analysis, only quadriceps force was a significant determinant of utilization of health care services. We conclude that utilization of health care services in patients with chronic obstructive pulmonary disease is related to ventilatory and peripheral muscle force. Whether or not reduced muscle force is simply an expression of disease severity remains to be determined.
Patterns of inspiratory muscle shortening during hypoxia and hypercapnia in dogs
Authors: Suzuki M , Suzuki S , Akahori T , Miyashita A , Yoshioka T , Sato M , Okubo T
Published: March 2003

The shortening of parasternal intercostal muscles (Para) and crural (Cru) and costal diaphragms (Cos) are not precisely understood. We therefore examined shortening patterns of these inspiratory muscles by using chronically implanted sonomicrometers in dogs. To avoid acute effects of surgery, measurements were performed 3 weeks after implanting the sonomicrometers. Patterns of length changes of Para, Cru, and Cos were measured during hypoxia and hypercapnia under two levels of anaesthesia. Respiratory length change (delta L) was assessed as a percentage change relative to the resting length at functional residual capacity (LFRC). Peak tidal shortening was defined as the maximal change from LFRC (delta L/LFRC). Under light anesthesia, the delta L/LFRC was the same among the three muscle groups at all tidal volumes (VT). Under deep anaesthesia, the delta L/LFRC both of Cru and Cos exceeded that of Para. Under light anaesthesia, the maximal shortening velocity ((delta L/LFRC)/delta t) of Cru was greater than that of Para. Under deep anaesthesia, the (delta L/LFRC)/delta t of Para was exceeded by that both of Cru and Cos. Furthermore, the (delta L/LFRC)/delta t of each inspiratory muscle was greater during hypoxia than during hypercapnia at equal volume. We conclude that: 1) the contribution of the diaphragm to ventilation increases during deep anaesthesia; 2) the muscle shortening velocity during hypoxia or hypercapnia is lower in parasternal intercostal muscles than in the diaphragm; and 3) there is no difference in the shortening pattern between crural and costal diaphragms.
Induction of nitric oxide synthase activity in pulmonary arteries from normoxic and chronically hypoxic rats
Authors: Carville C , Adnot S , Eddahibi S , Teiger E , Rideau D , Raffestin B
Published: March 2003

Chronic hypoxia has recently been shown to upregulate inducible nitric oxide synthase (iNOS) gene expression in rat lung. In the present study, we questioned whether induction of NO synthesis could alter the reactivity of pulmonary arteries (PA) from chronically hypoxic (CH) rats. Dose-response curves to phenylephrine (PE) 10(-9) to 5 x 10(-6) M) were examined in PA rings as well as response to L-arginine analogues in isolated lungs from CH or normoxic (N) rats after various incubation times. Although maximal contraction to PE did not differ in PA from CH rats compared to N rats at time 0 (361 +/- 53 vs 506 +/- 52 mg, respectively), it was markedly decreased after prolonged incubation (149 +/- 28 vs 386 +/- 47 mg, respectively, at 4 h; p < 0.001). This phenomenon persisted after endothelial-denudation, but was reversed by NG-monomethyl-L-arginine (L-NMMA) (5 x 10(-4) M) and prevented by actinomycin D (2 x 10(-6) M). In contrast, maximal contraction to PE in aorta from CH rats was similar at time 0 and 4 h. After a short incubation, PA contraction to L-NMMA was greater in CH than in N rats (96 +/- 17 vs 33 +/- 9 mg at 90 min; p < 0.05), was abolished after endothelial denudation, but persisted in CH rats in the presence of calmidazolium (5 x 10(-4) M). At 4 h, contraction to L-NMMA was abolished in endothelium-denuded PA from N rats but only attenuated in those from CH rats. In salt solution perfused lungs, L-NMMA added 30 or 90 min after isolation did not alter baseline pressure in N rats but caused its increase in CH rats. Whereas iNOS messenger ribonucleic acid (mRNA) was detectable by reverse-transcriptase polymerase chain reaction in the PA wall of N or CH rats after 4 h of incubation, it was absent in both at the time of isolation. In contrast, there was evidence of iNOS mRNA in lungs from CH rats at the time of isolation but no signal in those from N rats. In conclusion, there is induction of nitric oxide synthase activity in pulmonary arteries from normoxic and chronically hypoxic rats after prolonged incubation, but this effect is more pronounced in pulmonary arteries from chronically hypoxic rats.
Severe endobronchial obstruction in a girl with relapsing polychondritis: treatment with Nd YAG laser and endobronchial silicon stent
Authors: Sacco O , Fregonese B , Oddone M , Verna A , Tassara E , Mereu C , Rossi GA
Published: March 2003

Relapsing polychondritis (RP) is an uncommon disorder of unknown aetiology characterized by inflammation and destruction of the cartilaginous structures of many organs, including the tracheobronchial tree. When untreated, there is a high mortality rate, usually from respiratory obstructive complications. An 8 year old white girl, with a previous diagnosis of RP, was referred to our department for evaluation of worsening dyspnoea. Bronchoscopy showed localized inflammatory and fibrotic alterations of the mucosa, leading to severe obstruction of the left mainstem bronchus at its origin. The condition was successfully treated by endoscopic neodymium yttrium aluminium garnet (Nd YAG) laser. Re-evaluation of the patient, 7 months later, demonstrated bronchial stenosis and malacia requiring mechanical dilatation and positioning of an endobronchial silicon stent, which was well-tolerated by the patient.
Methimazole protects from experimental autoimmune uveitis (EAU) by inhibiting antigen presenting cell function and reducing antigen priming
Authors: Wang Peng , Sun Shu-Hui , Silver Phyllis B , Chan Chi-Chao , Agarwal Rajeev K , Wiggert Barbara , Kohn Leonard D , Jamieson Gordon A , Caspi Rachel R
Published: March 2003

Methimazole (methyl-mercapto-imidazole, MMI), a compound used clinically in therapy of Graves’ thyroiditis, was found to inhibit development of several autoimmune diseases in animal models. It was suggested on the basis of in vitro data that inhibition is through down-regulation of interferon-γ (IFN-γ)-induced expression of major histocompatibility complex class I and class II molecules. Here, we investigate the effect of MMI on experimental autoimmune uveoretinitis (EAU) and study its mechanism(s). Treatment of EAU with MMI administered in drinking water inhibited induction of the disease and associated antigen (Ag)-specific proliferation and cytokine production by draining lymph node cells (LNCs). The treatment was protective only if administered during the first but not during the second week after immunization, suggesting an effect on the induction phase of EAU. It is interesting that MMI inhibited disease in IFN-γ knockout mice, indicating that the in vivo protective effect is IFN-γ-independent. Flow cytometric analysis of draining LNCs extracted 5 days after immunization showed that MMI partly to completely reversed the increase in Mac-1+/class I+/class II+ cells induced by immunization and reduced the proportion of B7-1 and CD40-positive cells, suggesting a deficit in the Ag-presenting cell (APC) population. APC from untreated mice largely restored antigen-specific proliferation of MMI-treated LNCs. We suggest that MMI inhibits EAU at least in part by preventing the recruitment and/or maturation of APC, resulting in reduced generation of Ag-specific T cells.
Cerivastatin and atorvastatin inhibit IL-3-dependent differentiation and IgE-mediated histamine release in human basophils and downmodulate expression of the basophil-activation antigen CD203c/E-NPP3
Authors: Majlesi Yasamin , Samorapoompichit Puchit , Hauswirth Alexander W , Schernthaner Gerit-Holger , Ghannadan Minoo , Baghestanian Mehrdad , Rezaie-Majd Abdolreza , Valenta Rudolf , Sperr Wolfgang R , Bühring Hans-Jörg , Valent Peter
Published: March 2003

Recent data suggest that the statins, apart from their lipid-lowering activity, exhibit profound anti-inflammatory effects. Basophils are major proinflammatory effector cells in diverse pathologic reactions. We have examined the in vitro effects of five different statins on primary human basophils, their progenitors, and the basophil cell line KU-812. Preincubation of blood basophils with cerivastatin or atorvastatin (0.1–100 μM) for 24 h reduced their capacity to release histamine on immunoglobulin E (IgE)-dependent stimulation in a dose-dependent manner. These statins also inhibited IgE-dependent up-regulation of the basophil-activation antigen CD203c. Moreover, both statins suppressed interleukin-3-induced differentiation of basophils from their progenitors as well as 3H-thymidine uptake in KU-812 cells. All inhibitory effects of cerivastatin and atorvastatin were reversed by mevalonic acid (200 μM). The other statins tested (lovastatin, simvastatin, pravastatin) did not show significant inhibitory effects on basophils. Together, these data identify cerivastatin and atorvastatin as novel inhibitors of growth and activation of human basophils.
Role of MCP-1 and MIP-1α in retinal neovascularization during postischemic inflammation in a mouse model of retinal neovascularization
Authors: Yoshida Shigeo , Yoshida Ayako , Ishibashi Tatsuro , Elner Susan G , Elner Victor M
Published: March 2003

Macrophages are important participants in neovascularization. This study was designed to examine the role of the monocyte/macrophage chemotactic proteins, monocyte chemotactic protein-1 (MCP-1), and macrophage inflammatory protein-1α (MIP-1α) in a mouse model of oxygen-induced ischemic retinopathy and to determine whether the morphology and distribution of macrophages/microglia are concomitantly altered. The MCP-1, MIP-1α mRNA levels increased at 3 h after ischemia. MCP-1, MIP-1α, and vascular endothelial growth factor protein levels were also increased markedly and were maximal on days 1,0.5, and 1, respectively, after ischemia. In situ hybridization showed that MCP-1 and MIP-1α were localized in the hypoxic inner retina. Immunostaining demonstrated that the macrophages/microglia in the retina had morphological changes with enlarged processes, and some were closely associated with neovascular tufts at postnatal day 17. Coadministration of the neutralizing antibodies against MCP-1 and MIP-1α inhibited retinal neovascularization by 30%. Our data suggest that MCP-1 and MIP-1α are involved in the induction of retinal neovascularization and play a role in the inflammation induced by the ischemic retinopathy, possibly by modulating or attracting macrophages/microglia.
Tryptophan availability selectively limits NO-synthase induction in macrophages
Authors: Chiarugi Alberto , Rovida Elisabetta , Dello Sbarba Persio , Moroni Flavio
Published: March 2003

We studied the effects of tryptophan (TRP) availability on the synthesis and release of nitric oxide (NO) and tumor necrosis factor α (TNF-α) in interferon-γ (IFN-γ)-activated murine macrophages of the BAC1.2F5 cell line. IFN-γ (100 U/ml) not only increased the synthesis and release of NO and TNF-α from these cells but also induced indoleamine-2,3-dioxygenase, the rate-limiting enzyme of TRP catabolism. This led to an increased metabolic flow through the kynurenine pathway and significantly decreased TRP levels in macrophage incubation media. Low TRP concentrations in the media, however, modified IFN-γ effects. In TRP-“starved” cultures, in fact, the IFN-γ-mediated NO synthase induction was significantly reduced, and the increased TNF-α synthesis and release were not affected. Our results suggest hat a reduced local TRP availability may modify macrophage function and possibly the outcome of immune responses.
A unique role for IL-23 in promoting cellular immunity
Authors: Lankford Carla S R , Frucht David M
Published: March 2003

Recent discoveries of interleukin (IL)-23, its receptor, and its signal-transduction pathway add to our understanding of cellular immunity. IL-23 is a heterodimer, comprising IL-12 p40 and the recently cloned IL-23-specific p19 subunit. IL-23 uses many of the same signal-transduction components as IL-12, including IL-12Rβ1, Janus kinase 2, Tyk2, signal transducer and activator of transcription (Stat)1, Stat3, Stat4, and Stat5. This may explain the similar actions of IL-12 and IL-23 in promoting cellular immunity by inducing interferon-γ production and proliferative responses in target cells. Additionally, both cytokines promote the T helper cell type 1 costimulatory function of antigen-presenting cells. IL-23 does differ from IL-12 in the T cell subsets that it targets. Whereas IL-12 acts on naïve CD4+ T cells, IL-23 preferentially acts on memory CD4+ T cells. This review summarizes recent advances regarding IL-23, providing a functional and mechanistic basis for the unique niche that IL-23 occupies in cellular immunity.
Basophils express a type 2 cytokine profile on exposure to proteases from helminths and house dust mites
Authors: Phillips Clair , Coward William R , Pritchard David I , Hewitt Colin R A
Published: March 2003

The proteolytic activities frequently associated with sources of allergens and parasite secretions have been suggested as important immunomodulators. We have investigated whether the protease activity of the house dust mite allergen Der p1 and the secreted proteases of the hookworm Necator americanus are able to directly induce type 2 cytokine production by basophils. Der p1 and the secretions of N. americanus induced interleukin (IL)-4, IL-5, and IL-13 but not interferon-γ mRNA in KU812 basophils. Enzyme-linked immunosorbent assay confirmed that IL-4 and IL-13 were secreted. A nonproteolytic antigen failed to induce cytokine expression, and preincubation of Der p1 or N. americanus secretions with protease inhibitors inhibited cytokine expression. Data were confirmed using basophils purified from human peripheral blood. We speculate that tis innate mechanism may contribute to the development of a cytokine milieu that could promote immunoglobulin E synthesis, eosinophil recruitment, and the development of type 2 T cells.
Cross-talk between tumor cells and neutrophils through the Fas (APO-1, CD95)/FasL system: human glioma cells enhance cell viability and stimulate cytokine production in neutrophils
Authors: Hor Wei-Shio , Huang Wei-Lune , Lin Yee-Shin , Yang Bei-Chang
Published: March 2003

Many tumor cells are resistant to Fas-mediated killing, which has been primarily used as a mechanism to evade immune attack. In this study, we found a new action of Fas on tumors where activation of the Fas signal may force tumor cells to produce survival factors for neutrophils. Human peripheral circulating neutrophils in coculture with glioma cells showed significant delays in spontaneous apoptosis. Interleukin (IL)-6 and IL-8 partially mediated the glioma cell-associated, protective effect on neutrophils. The Fas agonistic antibody CH-11 dose-dependently stimulated the expression of IL-6 and IL-8 in glioma cells. Accordingly, blocking the Fas/FasL interaction reduced IL-6 and IL-8 production in glioma cells and impaired their protective effect on neutrophils. Coculture with glioma cells also affected the expression of cytokines in neutrophils, including IL-8, interferon-γ, and tumor necrosis factor α to various extents. Collectively, our results demonstrate bi-directional cross-talk between tumor and immune cells. Although Fas activation alone cannot induce apoptosis in tumor cells, it may potentially initiate an effective anti-tumor response through a circumvented mechanism.
Activity and subcellular distribution of cathepsins in primary human monocytes
Authors: Greiner Andrea , Lautwein Alfred , Overkleeft Herman S , Weber Ekkehard , Driessen Christoph
Published: March 2003

Cathepsins (Cat) in antigen presenting cells (APC) control antigen processing as well as major histocompatibility complex class II transport and function. The set of active Cat and the subcellular architecture of the class II antigen presentation compartment are largely unknown in primary human APC, including peripheral blood monocytes. We used novel chemical tools to visualize Cat in an activity-dependent manner. Primary human monocytes cntained active CatS, -B, and -H, while CatL was absent. Expression and activity patterns of Cat in human myelo-monocytoid cell lines were distinct from those found in primary cells. On a subcellular scale, the bulk of active Cat was concentrated in lysosomes in primary monocytes. In late endosomes, only active CatS was found in sizable amounts, colocalizing with C-terminal processing of the class II invariant chain and with cystatin C, the major endogenous Cat inhibitor. Late endosomes of human peripheral blood monocytes contain a well-controlled proteolytic machinery distinct from lysosomes, which is likely to play a key role in class II function.
Differential mRNA expression in circulating γδ T lymphocyte subsets defines unique tissue-specific functions
Authors: Hedges Jodi F , Cockrell Diane , Jackiw Larissa , Meissner Nicole , Jutila Mark A
Published: March 2003

To elucidate the functions of circulating γδ T cells, in the absence of antigen stimulation, the differential gene expression of two circulating γδ T cell subsets was analyzed. The two subsets, with distinct trafficking phenotypes in young calves, were GD3.5+, CD8−, WC1+ or GD3.5−, CD2+, WC1−, and 90–100% CD8+ and were sorted based on GD3.5 and γδ T cell receptor expression. Results from two different human arrays probed with cDNA from these γδ T cell subsets indicated that they have markedly different tissue-specific functions. The genes preferentially expressed by GD3.5+ (CD8−) γδ T cells demonstrated that they were highly activated, proliferative, and inflammatory, whereas those expressed by GD3.5− (primarily CD8+) γδ cells were involved in promoting quiescence, consistent with a role for γδ T cells as sentinel mucosal cells, and several were interferon-regulated genes. Gene expression and phenotypic assays indicated that CD8+ γδ T cells were apoptotic, whereas CD8− γδ T cells were apoptosis-resistant. Differential expression of multiple genes was confirmed in both arrays: That of 14 genes was confirmed by quantitative reverse transcriptase-polymerase chain reaction and that of seven proteins was confirmed by flow cytometry. This novel, genomic analysis of circulating γδ T cell subsets, without confounding effects of the tissue microenvironment, offers new insight into the biology and development of neonatal γδ T cells.