bluuebunny/crossref_metadata_2025 huggingface dataset:
Increased apoptosis in bone marrow B lymphocytes but not T lymphocytes in myelodysplastic syndrome
Authors: Amin Hesham M. , Jilani Iman , Estey Elihu H. , Keating Michael J. , Dey Amanda L. , Manshouri Taghi , Kantarjian Hagop M. , Estrov Zeev , Cortes Jorge E. , Thomas Deborah A. , Giles Francis J. , Albitar Maher
Published: May 2003

The hallmark of myelodysplastic syndrome (MDS) is enhanced apoptosis in myeloid, erythroid, and megakaryocytic cells in the bone marrow leading to ineffective hematopoiesis. Recent studies suggested that immunological and microenvironmental factors play a role in the pathophysiology of this disease. We report a significant increase in apoptosis in bone marrow B lymphocytes in MDS as compared to that found in acute myeloid leukemia and healthy controls. Furthermore, we demonstrate that patients with refractory anemia with excess blasts in transformation (RAEB-T) had apoptosis levels in lymphocytes similar to those seen in other subtypes of MDS. Our findings suggest that the alterations in B lymphocytes in the form of increased apoptosis can be seen in MDS and support the concept that immune modulation plays a role in the pathophysiology of MDS.
Classification of pediatric acute lymphoblastic leukemia by gene expression profiling
Authors: Ross Mary E. , Zhou Xiaodong , Song Guangchun , Shurtleff Sheila A. , Girtman Kevin , Williams W. Kent , Liu Hsi-Che , Mahfouz Rami , Raimondi Susana C. , Lenny Noel , Patel Anami , Downing James R.
Published: May 2003

Contemporary treatment of pediatric acute lymphoblastic leukemia (ALL) requires the assignment of patients to specific risk groups. We have recently demonstrated that expression profiling of leukemic blasts can accurately identify the known prognostic subtypes of ALL, including T-cell lineage ALL (T-ALL), E2A-PBX1, TEL-AML1, MLL rearrangements, BCR-ABL, and hyperdiploid karyotypes with more than 50 chromosomes. As the next step toward developing this methodology into a frontline diagnostic tool, we have now analyzed leukemic blasts from 132 diagnostic samples using higher density oligonucleotide arrays that allow the interrogation of most of the identified genes in the human genome. Nearly 60% of the newly identified subtype discriminating genes are novel markers not identified in our previous study, and thus should provide new insights into the altered biology underlying these leukemias. Moreover, a proportion of the newly selected genes are highly ranked as class discriminators, and when incorporated into class-predicting algorithms resulted in an overall diagnostic accuracy of 97%. The performance of an array containing the identified discriminating genes should now be assessed in frontline clinical trials in order to determine the accuracy, practicality, and cost effectiveness of this methodology in the clinical setting.
Systemic regulation of Hephaestin and Ireg1 revealed in studies of genetic and nutritional iron deficiency
Authors: Chen Huijun , Su Trent , Attieh Zouhair K. , Fox Tama C. , McKie Andrew T. , Anderson Gregory J. , Vulpe Chris D.
Published: May 2003

Hephaestin is a membrane-bound multicopper ferroxidase necessary for iron egress from intestinal enterocytes into the circulation. Mice with sex-linked anemia (sla) have a mutant form of Hephaestin and a defect in intestinal basolateral iron transport, which results in iron deficiency and anemia. Ireg1 (SLC11A3, also known as Ferroportin1 or Mtp1) is the putative intestinal basolateral iron transporter. We compared iron levels and expression of genes involved in iron uptake and storage in sla mice and C57BL/6J mice fed iron-deficient, iron-overload, or control diets. Both iron-deficient wild-type mice and sla mice showed increased expression of Heph and Ireg1 mRNA, compared to controls, whereas only iron-deficient wild-type mice had increased expression of the brush border transporter Dmt1. Unlike iron-deficient mice, sla mouse enterocytes accumulated nonheme iron and ferritin. These results indicate that Dmt1 can be modulated by the enterocyte iron level, whereas Hephaestin and Ireg1 expression respond to systemic rather than local signals of iron status. Thus, the basolateral transport step appears to be the primary site at which the small intestine responds to alterations in body iron requirements.
Molecular analyses of patients with hyperferritinemia and normal serum iron values reveal both L ferritin IRE and 3 new ferroportin (slc11A3) mutations
Authors: Hetet Gilles , Devaux Isabelle , Soufir Nadem , Grandchamp Bernard , Beaumont Carole
Published: May 2003

Unexplained hyperferritinemia is a common clinical finding, even in asymptomatic persons. When early onset bilateral cataracts are also present, the hereditary hyperferritinemia-cataract syndrome (HHCS), because of heterozygous point mutation in the L ferritin iron-responsive element (IRE) sequence, can be suspected. We sequenced the L ferritin exon 1 in 52 DNA samples from patients referred to us for molecular diagnosis of HHCS. We identified 24 samples with a point mutation/deletion in the IRE. For the 28 samples in which no IRE mutation was present, we also genotyped HFE mutations and sequenced both H ferritin and ferroportin genes. We found an increased frequency of His63Asp heterozygotes (12 of 28) but no H ferritin mutations. We identified 3 new ferroportin mutations, producing, respectively, Asp157Gly, Gln182His, and Gly323Val amino acid replacements, suggesting that these patients have dominant type 4 hemochromatosis. This study demonstrates that both L ferritin IRE and ferroportin mutations can account for isolated hyperferritinemia. The presence of cataract does not permit the unambiguous identification of patients with HHCS, although the existence of a family history of cataract was only encountered in these patients. This raises the intriguing possibility that lens ferritin accumulation might be a factor contributing to age-related cataract in the general population. Additional causes of isolated hyperferritinemia remain to be identified.
Sirolimus, tacrolimus, and low-dose methotrexate for graft-versus-host disease prophylaxis in mismatched related donor or unrelated donor transplantation
Authors: Antin Joseph H. , Kim Haesook T. , Cutler Corey , Ho Vincent T. , Lee Stephanie J. , Miklos David B. , Hochberg Ephraim P. , Wu Catherine J. , Alyea Edwin P. , Soiffer Robert J.
Published: May 2003

We studied the feasibility and activity of adding sirolimus to tacrolimus and low-dose methotrexate as graft-versus-host disease (GVHD) prophylaxis in recipients of alternative donor transplants. Forty-one patients with hematologic malignancies were conditioned with cyclophosphamide and total body irradiation. Marrow stem cells were from an HLA-A, -B, and -DR compatible, unrelated donor (n = 26, 68%), froma5of6 antigen-matched unrelated donor (n = 8, 20%), or from a 5 of 6 antigen-matched family member (n = 5, 12%). Therapeutic serum levels of sirolimus were attained in most patients. All evaluable patients engrafted. An absolute neutrophil count of 500/μL was achieved on day +18 (range, 11-32 days). Sustained platelet counts of more than 20 000/ μL were attained on day +29 (range, 14-98 days). Grades 0-I acute GVHD occurred in 75% of patients. Grades II, III, and IV acute GVHD occurred in 13%, 8%, and 5%, respectively (total grades II-IV GVHD, 26%). Median survival is 366 days (95% CI 185, not estimable) and actuarial survival at 1 year is 52%. Oral sirolimus is tolerable, adequate blood levels are achievable, and there is a low rate of acute GVHD compared with historical data in this high-risk population. This novel agent is worthy of further study in allogeneic transplantation.
CXCL12 expression by invasive trophoblasts induces the specific migration of CD16– human natural killer cells
Authors: Hanna Jacob , Wald Ori , Goldman-Wohl Debra , Prus Diana , Markel Gal , Gazit Roi , Katz Gil , Haimov-Kochman Ronit , Fujii Nobutaka , Yagel Simcha , Peled Amnon , Mandelboim Ofer
Published: May 2003

In the maternal decidua, natural killer (NK) cells, characterized by lack of CD16, are found in direct contact with the fetal extravillous trophoblasts (EVTs). It is yet unknown which factors contribute to the specific homing of this unique NK subset to the decidua. In this study we analyze the chemokine receptor repertoire on various NK populations derived from the peripheral blood and decidua. We show that CXCR4 and CXCR3 receptors are preferentially expressed on CD16– NK subsets derived either from the peripheral blood or the decidua and that these receptors are involved in migration of all NK subsets to their ligands. We further demonstrate in vivo that invading EVTs that eventually perform endovascular invasion express CXCL12, the ligand for CXCR4, but not ligands for CXCR3. Indeed, specific accumulation of the CD16– NK cells at the expense of CD16+ cells was observed only when in vitro migration was performed with ligands for CXCR4. Finally, incubation of the peripheral blood CD16– NK cells with cytokines present in the decidua, especially interleukin 15 (IL-15), resulted in the expression of chemokine receptor repertoire similar to that observed on decidual NK cells, suggesting an additional important regulatory effect of local decidual cytokines.
Syk activation is a leukotriene B4–regulated event involved in macrophage phagocytosis of IgG-coated targets but not apoptotic cells
Authors: Canetti Claudio , Hu Bin , Curtis Jeffrey L. , Peters-Golden Marc
Published: May 2003

Macrophages are called upon to ingest both IgG-coated targets and apoptotic cells. Important roles for tyrosine kinase Syk and leukotriene B4 (LTB4) are recognized in FcγR-mediated phagocytosis. Here we evaluated the roles of Syk and LTB4 in macrophage phagocytosis of apoptotic thymocytes versus IgG-coated erythrocytes. Macrophage ingestion of apoptotic thymocytes was not influenced by exogenous or endogenous LTB4 nor associated with Syk activation (phosphorylation). By contrast, LTB4 dose-dependently amplified FcγR-mediated phagocytosis as well as Syk activation. Furthermore, a role for endogenous LTB4 in Syk activation during FcγR-mediated phagocytosis was demonstrated using pharmacologic and genetic abrogation of 5-lipoxygenase. LTB4 was unique among 5-lipoxygenase products in this regard, since LTD4 and 5-hydroxyeicosatetraenoic acid (HETE) were unable to amplify Syk activation in response to FcγR engagement. Ca2+ chelation studies revealed that FcγR-mediated Syk activation as well as LTB4 amplification thereof was Ca2+ regulated. These 2 parallel phagocytic processes therefore exhibit initial divergence in signal transduction events, with Syk activation being an LTB4-regulated event in FcγR-mediated but not apoptotic cell ingestion. As LTB4 is an important proinflammatory product of macrophages, we speculate that this divergence evolved to permit FcγR-mediated phagocytosis to proceed in an inflammatory milieu, while apoptotic cell clearance is noninflammatory.
The APC-independent anticoagulant activity of protein S in plasma is decreased by elevated prothrombin levels due to the prothrombin G20210A mutation
Authors: Koenen Rory R. , Tans Guido , van Oerle René , Hamulyák Karly , Rosing Jan , Hackeng Tilman M.
Published: May 2003

Protein S exhibits anticoagulant activity independent of activated protein C (APC). An automated factor Xa–based one-stage clotting assay was developed that enables quantification of the APC-independent activity of protein S in plasma from the ratio of clotting times (protein S ratio [pSR]) determined in the absence and presence of neutralizing antibodies against protein S. The pSR was 1.62 ± 0.16 (mean ± SD) in a healthy population (n = 60), independent of plasma levels of factors V, VIII, IX, and X; protein C; and antithrombin, and not affected by the presence of factor V Leiden. The pSR strongly correlates with the plasma level of protein S and is modulated by the plasma prothrombin concentration. In a group of 16 heterozygous protein S–deficient patients, the observed mean pSR (1.31 ± 0.09) was significantly lower than the mean pSR of the healthy population, as was the pSR of plasma from carriers of the prothrombin G20210A mutation (1.47 ± 0.21; n = 46). We propose that the decreased APC-independent anticoagulant activity of protein S in plasma with elevated prothrombin levels may contribute to the thrombotic risk associated with the prothrombin G20210A mutation.
Long targeting arms do not increase the efficiency of homologous recombination in the β-globin locus of murine embryonic stem cells
Authors: Lu Zhi Hong , Books Jason T. , Kaufman Richard M. , Ley Timothy J.
Published: May 2003

The correction of mutant β-globin genes has long been a therapeutic goal for patients with β-thalassemia or hemoglobinopathies. The use of homologous recombination (HR) to achieve this goal is an attractive approach because it eliminates the need to include regulatory sequences in the therapeutic construct, and it eliminates mutagenesis induced by random integration. However, HR is a very inefficient process for gene correction, and its efficiency is probably locus dependent. The length of targeting arms is thought to be a determinant of targeting efficiency, so we compared the ability of standard (8-kb) versus very long (16-, 24-, and 110-kb) regions of homology to correct a mutant murine β-globin gene in embryonic stem cells. Increasing the length of the targeting sequences did not increase the efficiency of HR in this locus, suggesting that alternative approaches will be required to improve the efficiency of this approach for globin gene correction.
Comparison of molecular markers in a cohort of patients with chronic myeloproliferative disorders
Authors: Kralovics Robert , Buser Andreas S. , Teo Soon-Siong , Coers Jörn , Tichelli Andre , van der Maas Anthonie P. C. , Skoda Radek C.
Published: May 2003

Decreased expression of c-MPL protein in platelets, increased expression of polycythemia rubra vera 1 (PRV-1) and nuclear factor I-B (NFIB) mRNA in granulocytes, and loss of heterozygosity on chromosome 9p (9pLOH) were described as molecular markers for myeloproliferative disorders (MPDs). To assess whether these markers are clustered in subgroups of MPDs or represent independent phenotypic variations, we simultaneously determined their status in a cohort of MPD patients. Growth of erythropoietin-independent colonies (EECs) was measured for comparison. We observed concordance between EECs and PRV-1 in MPD patients across all diagnostic subclasses, but our results indicate that EECs remain the most reliable auxiliary test for polycythemia vera (PV). In contrast, c-MPL, NFIB, and 9pLOH constitute independent variations. Interestingly, decreased c-MPL and elevated PRV-1 also were observed in patients with hereditary thrombocythemia (HT) who carry a mutation in the thrombopoietin (TPO) gene. Thus, altered c-MPL and PRV-1 expression also can arise through a molecular mechanism different from sporadic MPD.
Decreased factor VIII levels during acetaminophen-induced murine fulminant hepatic failure
Authors: Doering Christopher B. , Parker Ernest T. , Nichols Christopher E. , Lollar Pete
Published: May 2003

During human fulminant hepatic failure (FHF) circulating levels of most hemostatic proteins fall dramatically. Concurrently, factor VIII (fVIII) procoagulant activity rises despite destruction of the hepatocytes considered responsible for fVIII synthesis. This observation suggests a role for cells other than hepatocytes in fVIII biosynthesis during FHF. We have attempted to identify nonhepatocytic sites of fVIII biosynthesis by inducing FHF in mice using acetaminophen overdose, a common cause of human FHF. Acetaminophen-treated mice consistently displayed signs characteristic of FHF, including elevated plasma aminotransferase activity. However, acetaminophen-treated mice demonstrated markedly reduced fVIII activity, contrary to the observation in human FHF. von Willebrand factor antigen levels were only mildly reduced, suggesting that the decrease in fVIII is not secondary to loss of von Willebrand factor. These results imply that there are fundamental differences in the regulation of plasma fVIII levels between humans and mice.
Unrelated donor stem cell transplantation compared with chemotherapy for children with acute lymphoblastic leukemia in a second remission: a matched-pair analysis
Authors: Borgmann Anja , von Stackelberg Arend , Hartmann Reinhard , Ebell Wolfram , Klingebiel Thomas , Peters Christina , Henze Günter
Published: May 2003

Allogeneic stem cell transplantation (SCT) is frequently considered as treatment for relapsed childhood acute lymphoblastic leukemia (ALL). For patients without a matched sibling donor, SCT from unrelated donors (UD-SCT) has been increasingly performed during the past years. However, UD-SCT–related mortality and morbidity is still considerable, and the question remains as to which patients are at such high risk of recurrence that UD-SCT is indicated and, conversely, which patients do not require transplantation for long-term disease control. A matched-pair analysis was performed among patients treated according to Acute Lymphoblastic Leukemia Relapse Berlin-Frankfurt-Münster (ALL-REZ BFM) Study Group protocols after first relapse with chemotherapy or UD-SCT. Altogether 81 pairs were identified that could be matched exactly for site of relapse and immunophenotype, and as closely as possible for duration of first remission, age, diagnosis date, and peripheral blast cell count at relapse. No significant difference in the probability of event-free survival (pEFS) between UD-SCT and chemotherapy existed regarding 28 pairs with an intermediate prognosis (0.39 ± 0.10 vs 0.49 ± 0.11,P = .105), whereas the pEFS was significantly different in the 53 pairs with a poor prognosis (0.44 ± 0.07 vs 0.00 ± 0.00, P < .001). The major reasons of treatment failure among patients who underwent UD-SCT were therapy-related death (TRD; 24/81) and relapses (20/81). In contrast, TRD rarely occurred in patients treated with chemotherapy alone (3/81), but relapse was much more common (62/81). In conclusion, UD-SCT provides better event-free survival for children with high-risk relapsed ALL. However, there is no clear advantage of UD-SCT in patients with intermediate prognosis.
Megakaryocytes require thrombospondin-2 for normal platelet formation and function
Authors: Kyriakides Themis R. , Rojnuckarin Ponlapat , Reidy Michael A. , Hankenson Kurt D. , Papayannopoulou Thalia , Kaushansky Kenneth , Bornstein Paul
Published: May 2003

Mice that lack the matricellular angiogenesis inhibitor, thrombospondin-2 (TSP2), display a bleeding diathesis, despite normal blood coagulation and the lack of thrombocytopenia. Although platelets do not contain detectable levels of TSP2, TSP2-null platelets are compromised in their ability to aggregate in vivo in response to denudation of the carotid artery endothelium, and in vitro following exposure to adenosine diphosphate (ADP). Megakaryocytes (MKs) show high levels of TSP2 by immunohistochemical analysis of bone marrow. However, when cultured in vitro, MKs contain little TSP2 protein or mRNA. These findings suggest that most TSP2 is acquired from the bone marrow microenvironment. Consistent with this hypothesis, MKs take up recombinant TSP2 in an integrin-dependent manner when it is supplied in the culture medium. Furthermore, uptake of TSP2 in vitro affects MK differentiation and proplatelet formation. The functional significance of this process is supported by the presence of ultrastructural abnormalities in TSP2-null bone marrow, including extensive fragmentation of the peripheral zone in MKs and failure of this zone to form close associations with vascular sinuses. We conclude that the uptake of TSP2 by MKs from the marrow milieu is required for proper MK function and the release of functionally competent platelets.
Diagnosing occupational asthma: how, how much, how far?: Table 1
Authors: Moscato G. , Malo J-L. , Bernstein D.
Published: May 2003

Diagnosing occupational asthma is still a challenge because it is based on a stepwise approach in which the depth of investigative means may vary depending on resources. The authors herewith review the existing investigative means from the approach of outlining controversies and queries.

There is no validated clinical questionnaire for diagnosing occupational asthma. Immunological investigation is limited by the lack of standardised extracts for skin-prick testing and specific immunoglobulin E assessments. Criteria for interpretation of changes in peak expiratory flow rates and bronchial responsiveness to pharmacological agents are still open to discussion.

It is worth improving the methodology of specific inhalation challenges, either in the laboratory or in the workplace, to facilitate more extensive use of these tests. Validation of new means that assess airway inflammation, such as exhaled nitric oxide and induced sputum, needs to be performed. There is a need to increase the use of these diagnostic tests because the diagnosis is still too often based on “clinical impression”.
Monoclonal anti-interleukin‐5 treatment suppresses eosinophil but not T‐cell functions
Authors: Büttner C. , Lun A. , Splettstoesser T. , Kunkel G. , Renz H.
Published: May 2003

Influx of eosinophils in airway mucosa and airway lumen is a hallmark of bronchial asthma.In-vitrodata and animal studies indicate that the T‐helper (Th) type‐2 cell cytokine, interleukin (IL)‐5, plays an important role in eosinophil maturation, differentiation, recruitment, and survival. The objective of this study was to determine whether intravenous treatment with monoclonal anti-IL‐5 would affect the number of peripheral blood eosinophils, their activation status, T‐cell activation or the pattern of Th1 and Th2 cytokine production.

Over a period of 6 months, 19 asthmatics were investigated in a double-blind, placebo-controlled, multicentre study with mepolizumab (SB 240563) anti-IL‐5 antibody administered three times. Before each infusion and 12 weeks after the last infusion, peripheral blood leukocytes were examined, qualitative and quantitative distribution of eosinophils and lymphocyte subpopulations, frequencies of IL‐2, ‐3, ‐4, ‐5, -10, -13, interferon‐γ‐producing CD4 T‐cells and serum eosinophil cationic protein (ECP) levels were determined.

Treatment with mepolizumab resulted in a marked, rapid and sustained decrease of eosinophil numbers (median values from 300 to 45 per µL) paralleled by decreased levels of serum ECP (median values from 15 to 5 µg·L−1). Distribution of T‐cell subsets and T‐cell cytokine production were not altered during antibody treatment.

In conclusion, administration of mepolizumab to asthmatic patients markedly reduces peripheral blood eosinophils without altering the distribution and activation status of lymphocytes.
Effects of tobacco smoking on findings in chest computed tomography among asbestos-exposed workers
Authors: Vehmas T. , Kivisaari L. , Huuskonen M.S. , Jaakkola M.S.
Published: May 2003

This study examined the effects of smoking on the findings in chest computed tomography (CT) in 587 asbestos-exposed construction workers (11 females, 576 males, mean age 62 yrs; 18 never-smokers, 406 exsmokers and 163 current smokers). The workers were imaged with spiral CT and high-resolution CT. A total of 13 radiological signs were scored by three radiologists independently. The workers' medical data, smoking habits and occupational exposures were collected at an interview. The effects of smoking status and smoked pack-yrs (0–87.5) on the CT signs were studied using multivariate analysis adjusted for confounding factors.

Smoking increased all emphysema signs and contributed to bronchial wall thickening. Smoking was negatively associated with curvilinear and septal lines as well as with parenchymal bands. In persons who had smoked <10 pack-yrs, smoking was positively related to paraseptal emphysema and to bronchial wall thickening and negatively related to septal lines, subpleural nodules and honeycombing.

Smoking was related to several abnormal computed tomography signs even among those with relatively small exposure. Computed tomography can detect changes due to smoking at an early stage.
Transient early wheeze is not associated with impaired lung function in 7‐yr-old children
Authors: Lau S. , Illi S. , Sommerfeld C. , Niggemann B. , Völkel K. , Madloch C. , Grüber C. , Nickel R. , Forster J. , Wahn U. ,   the Multicentre Allergy Study Group
Published: May 2003

The aim of the present study was to analyse determinants of lung function in 7‐yr-old children with different wheezing patterns (early, persistent and late onset) in a prospective cohort study.

The German Multicentre Allergy Study (MAS) followed 1,314 children from birth onwards. Annual assessments included clinical check-ups, a structured interview and repeated measurement of specific immunoglobulins Ig(E) directed against food and inhalant allergens. At the age of 7 yrs, lung function was measured by body plethysmography in 800 children.

Episodes of wheezing in the past 12 months (“current wheeze”) were strongly associated with reduced lung function at age 7 yrs. Children with wheezing episodes only during the first 3 yrs of life showed a slight impairment in maximal expiratory flow when 50% of the forced vital capacity remains to be exhaled (98.9±24.2versus103.2±22.8% of the predicted value in children who never wheeze). Separate analysis of determinants of pulmonary function within these subgroups resulted in distinctly different patterns. Determinants of impaired lung function in the group of current wheezers were: time in years since first wheeze, a parental history of atopy, current sensitisation to indoor allergens, elevated cord blood IgE levels and a low ponderal index at birth. In the group of transient early wheezers, frequent lower respiratory tract infections early in life and maternal smoking during pregnancy were significant but weak determinants of impaired lung function.

The present results indicate that determinants of pulmonary function in 7‐yr-old children differ with respect to different wheezing phenotypes, demanding different therapeutic strategies. Although transient early wheezers were found to have normal-to-subnormal lung function, children with asthmatic symptoms (persistent and late-onset disease) at age 7 yrs already show significant impairment of expiratory flow volumes.
Increased tumour necrosis factor‐α plasma levels during moderate-intensity exercise in COPD patients
Authors: Rabinovich R.A. , Figueras M. , Ardite E. , Carbó N. , Troosters T. , Filella X. , Barberà J.A. , Fernandez-Checa J.C. , Argilés J.M. , Roca J.
Published: May 2003

Post-training downregulation of muscle tumour necrosis factor (TNF)‐α messenger ribonucleic acid (mRNA) expression and decrease in cellular TNF‐α levels have been reported in the elderly. It is hypothesised that chronic obstructive pulmonary disease (COPD) patients may not show these adaptations due to their reduced ability to increase muscle antioxidant capacity with training.

Eleven COPD patients (forced expiratory volume in one second 40±4.4% of the predicted value) and six age-matched controls were studied. Pre- and post-training levels of TNF‐α, soluble TNF receptors (sTNFRs: sTNFR55 and sTNFR75) and interleukin (IL)‐6 in plasma at rest and during exercise and vastus lateralis TNF‐α mRNA were examined.

Moderate-intensity constant-work-rate exercise (11 min at 40% of pretraining peak work-rate) increased pretraining plasma TNF‐α levels in COPD patients (from 17±3.2 to 23±2.7 pg·mL−1; p<0.005) but not in controls (from 19±4.6 to 19±3.2 pg·mL−1). No changes were observed in sTNFRs or IL‐6 levels. After 8 weeks' endurance training, moderate-intensity exercise increased plasma TNF‐α levels similarly to pretraining (from 16±3 to 21±4 pg·mL−1; p<0.01). Pretraining muscle TNF‐α mRNA expression was significantly higher in COPD patients than in controls (29.3±13.9versus5.0±1.5 TNF‐α/18S ribonucleic acid, respectively), but no changes were observed after exercise or training.

It is concluded that moderate-intensity exercise abnormally increases plasma tumour necrosis factor‐α levels in chronic obstructive pulmonary disease patients without exercise-induced upregulation of the tumour necrosis factor‐α gene in skeletal muscle.
To clip or not to clip? Noseclips for spirometry
Authors: Chavasse R. , Johnson P. , Francis J. , Balfour-Lynn I. , Rosenthal M. , Bush A.
Published: May 2003

The use of noseclips for open-circuit spirometry is sporadic, despite guidelines encouraging their use. The authors aimed to evaluate whether noseclips significantly affected measurements of forced vital capacity (FVC) and forced expiratory volume in one second (FEV1) in children attending a tertiary, paediatric respiratory centre.

Children attending the asthma and cystic fibrosis (CF) clinics were asked to perform two sets of spirometry, one with and one without noseclips in random order, 20-min apart.

Paired data was obtained on 62 patients (32 asthma, 30 CF) with a median age of 11.4 yrs (range 7.2–17.2 yrs). There were no systematic differences in FEV1or FVC measured with and without noseclips, although seven children (11%) had clinically significant differences in FEV1of >190 mL.

There is no clear advantage to wearing noseclips when performing open-circuit spirometry. Individuals should be assessed to ascertain their optimal technique, which should then be used consistently in clinical practice. Noseclips should probably be retained for research protocols.
Spirometry in young children: should computer-animation programs be used during testing?
Authors: Gracchi V. , Boel M. , van der Laag J. , van der Ent C.K.
Published: May 2003

Currently, computer-animation programs are frequently used to instruct and stimulate young children in performing maximal expiratory flow/volume (MEFV) curves. The reproducibility and maximal performance of MEFV manoeuvres with and without the use of two computer-animation programs (the “candles” and the “balloon” programs) were evaluated.

Eighty-eight children, aged 4–8 yrs, were randomly assigned to one of the two animation programs. All children performed two series of at least three technically acceptable curves, one series with the incentive and one without, in random order.

With the use of computer-animation programs, a lower proportion of children were able to fulfil international criteria for forced vital capacity (FVC) and forced expiratory volume in one second (FEV1) reproducibility. The use of incentives improved reproducibility and performance of peak expiratory flow (PEF). Performance of FVC decreased significantly in 6–8‐yr-old children using the animation programs.

Training with a program for a short period of time before the formal lung-function test may be valuable. According to the results, however, the use of these programs during tests under the guidance of an experienced lung-function technician cannot be routinely recommended because of possible deteriorating effects on reproducibility and performance of forced expiratory manoeuvres.
N-acetylcysteine reduces the risk of re-hospitalisation among patients with chronic obstructive pulmonary disease
Authors: Gerrits C.M.J.M. , Herings R.M.C. , Leufkens H.G.M. , Lammers J-W.J.
Published: May 2003

The aim of this study was to evaluate the effect of oralN‐acetylcysteine in the prevention of re-hospitalisation for chronic obstructive pulmonary disease (COPD) exacerbations.

Using the PHARmacoMOrbidity linkage (PHARMO) system the authors included all patients aged ≥55 yrs who had been dispensed medication, labelled for respiratory indications (anatomical therapeutic chemical (ATC) classification system: R03), between 1986–1998 and who had also been hospitalised for COPD (International Classification of Diseases (ICD)‐9: 491, 492, 496) in this time frame. These subjects were subsequently divided into two groups, those who had receivedN‐acetylcysteine following discharge from their first admission between 1986–1998 and those who had not. All the patients were studied starting from their initial discharge, until their first readmission, death or end of data collection period. The maximum follow-up period was 1 yr.

A total of 1,219 patients, who were hospitalised for COPD between 1986–1998, were included in this study. After adjustment for disease severity, it was observed that the use ofN‐acetylcysteine was significantly associated with a reduced risk of readmission. The readmission risk was significantly lower in patients with high average daily doses ofN‐acetylcysteine.

In conclusion it was observed thatN‐acetylcysteine reduces the risk of re-hospitalisation for chronic obstructive pulmonary disease by ∼30% and that this risk reduction is dose-dependent.
Uptake of18fluorodeoxyglucose in the cystic fibrosis lung: a measure of lung inflammation?
Authors: Labiris N.R. , Nahmias C. , Freitag A.P. , Thompson M.L. , Dolovich M.B.
Published: May 2003

Positron emission tomography is a three-dimensional imaging technique that measures physiological effects, including metabolism.18Fluorodeoxyglucose has been extensively used as a tracer of cellular energy metabolism in the brain and in tumour detection. As neutrophils utilise glucose as an energy source during their respiratory burst, it was hypothesised that18fluorodeoxyglucose uptake, by these cells, could be interpreted as a measure of neutrophil activation in cystic fibrosis (CF).

Ten adult CF patients were given a bolus intravenous injection of18fluorodeoxyglucose, followed by a 90-min dynamic mid-lung acquisition scan. Right-lung18fluorodeoxyglucose uptake was assessed using a Patlak plot and values were converted to glucose utilisation. Three clinically inactive pulmonary sarcoidosis patients served as controls.

From the 10 CF patients with baseline sputum neutrophils of 14×106cells·mL−1who were investigated, seven were found to have sputum at a normal or slightly depressed glucose utilisation rate (mean 1.33 µmol·g−1·h−1) compared with a mean of 2.82 µmol·g−1·h−1for the sarcoidosis patients. In eight patients, receiving inhaled tobramycin therapy, no change in lung glucose utilisation or sputum neutrophil counts were found.

Despite high-sputum neutrophil levels, lung glucose utilisation was not elevated in patients with cystic fibrosis.
Bronchodilator tolerance: the impact of increasing bronchoconstriction
Authors: Wraight J.M. , Hancox R.J. , Herbison G.P. , Cowan J.O. , Flannery E.M. , Taylor D.R.
Published: May 2003

Chronic exposure to β‐agonists causes tolerance to their bronchodilator effects, which is best demonstrated during acute bronchoconstriction. The aim of the present study was to assess whether tolerance becomes more evident with increasing bronchoconstriction, as might occur in acute asthma.

In a randomised, double-blind, placebo-controlled, crossover study comprising 15 patients, the treatments were salbutamol 400 µgq.i.d. or placebo givenviaDiskhaler® for 28 days with a 2‐week washout between treatments. Patients attended on days 14, 21 and 28. Bronchoconstriction was induced on two of these three occasions to achieve a reduction in the forced expiratory volume in one second (FEV1) of 0 (no methacholine), 15 and 30% (using methacholine) in a randomised order. Immediately after this, salbutamol 100 µg, 100 µg and 200 µg was inhaled at 0, 5, and 10 min. FEV1was measured over 40 min. Dose/response curves were plotted and values for the area under the curve (AUC)0–40FEV1were compared between treatments and by degree of bronchoconstriction.

Regular salbutamol resulted in attenuation of the acute response to β‐agonist, which was increasingly evident with greater bronchoconstriction. With a reduction in FEV1of 0, 15 and 30%, the AUC0–40FEV1with salbutamol were 11.2, −14.6 and −35.7% respectively, compared to placebo. There was a linear relationship between the magnitude of bronchoconstriction and the between-treatment differences in AUC0–40FEV1.

Increasing bronchoconstriction conferred greater susceptibility to the effects of bronchodilator tolerance.
Rigid bronchoscopy induces bacterial translocation: an experimental study in rats
Authors: Nayci A. , Atis S. , Talas D.U. , Ersoz G.
Published: May 2003

Bronchoscopy has the potential to propagate infections. Bacterial translocation was hypothesised to be the cause of infections observed following bronchoscopy and this study was designed to assess the risk of bacterial translocation following rigid bronchoscopy in rats.

A total of 30 rats were evaluated. The study group (n=15) underwent rigid bronchoscopy. Arterial blood gas analysis was performed in all rats. Blood and tissue cultures from the ileum, caecum, mesenteric lymph nodes, liver, spleen, mediastinal lymph nodes and lung were obtained 24 h following bronchoscopy.

Bacterial translocation to the mesenteric lymph nodes was found in seven of 15 rats (46.7%) that underwent bronchoscopy, compared with none of the controls. Of the seven positives, three rats (42.8%) also demonstrated other organ involvement, such as the liver and spleen.Escherichia coli,Salmonella typhymirium,S. enteritidisandPseudomonasspp. were found as translocating bacteria. In the study group, pH and arterial oxygen tension were significantly lower and arterial carbon dioxide tension was higher, compared with controls.

This study shows that rigid bronchoscopy may induce bacterial translocation in rats. Further investigations aimed at understanding the clinical consequences of this phenomenon are warranted.
Dexamethasone treatment does not inhibit fibroproliferation in chronic lung disease of prematurity
Authors: Dik W.A. , Versnel M.A. , Naber B.A. , Janssen D.J. , van Kaam A.H. , Zimmermann L.J.I.
Published: May 2003

Pulmonary fibrosis results from excessive fibroblast proliferation and increased collagen deposition and occurs in chronic lung disease of prematurity (CLD). Platelet-derived growth factor (PDGF)-BB is mitogenic for fibroblasts and levels are increased in fibrotic lung disorders. Systemic dexamethasone (DEX) treatment improves pulmonary function and reduces inflammation in infants with or at risk of CLD. However, the effect of DEX treatment on fibroblast activity, PDGF-BB and collagen synthesis in the lungs of CLD patients is uncertain.

Bronchoalveolar lavage (BAL) fluids, obtained from 15 infants at risk of CLD before and after DEX treatment, were analysed for fibroblast mitogenicity, PDGF-BB, N‐terminal propeptide of collagen type III (PIIINP) and interleukin (IL)-1β levels and inflammatory cell numbers.

After DEX treatment, the mitogenic activity of BAL fluid for fibroblasts was not reduced but increased. The change in mitogenicity correlated with a change in BAL fluid PDGF-BB levels. Furthermore, BAL fluid-induced fibroblast proliferation was blocked using an inhibitor of the PDGF receptor. DEX treatment did not influence PIIINP levels, but reduced IL-1β levels and inflammatory cell numbers in BAL fluid.

This study suggests that dexamethasone treatment does not reduce fibroblast proliferation despite apparent downregulation of inflammation. The present findings do not support the use of dexamethasone for prevention of the fibrotic response in infants at risk of chronic lung disease of prematurity.
Transcription factor activation and mitogenic synergism in airway smooth muscle cells
Authors: Ediger T.L. , Schulte N.A. , Murphy T.J. , Toews M.L.
Published: May 2003

Simultaneous treatment of human airway smooth muscle (HASM) cells with lysophosphatidic acid (LPA) and epidermal growth factor (EGF) leads to strikingly synergistic stimulation of mitogenesis. The purpose of this study was to explore potential sites for signal integration mediating synergism, focusing on extracellular signal-regulated kinase (ERK) and transcription factors involved in proliferation and inflammation as likely candidates.

Activation of ERK was analysed by immunoblotting. Transcription factor activation was assessed using HASM cells transduced with luciferase reporter gene constructs.

LPA and EGF both activated ERK but had no synergistic effect when combined. LPA and EGF both activated activator protein (AP)‐1, cyclic adenosine monophosphate response element-binding protein, nuclear factor of activated T‐cells and the serum response element; however, only AP‐1 activation exhibited synergism. Activation of the inhibitory guanine nucleotide-binding protein and of ERK signalling pathways were required for most transcription factor responses to LPA. In contrast, nuclear factor (NF)‐κB was activated by LPA but not EGF and NF‐κB activation was completely blocked only when Rho was inhibited. Rapid activation of Rho was observed in response to LPA but not to EGF. Importantly, inhibition of Rho selectively blocked synergism in both AP‐1 activation and mitogenesis.

In summary, extracellular signal-regulated kinase activation is required for many transcription factor responses to lysophosphatidic acid and epidermal growth factor, however it is not synergistic. Activation of activator protein‐1 is synergistic, and Rho activation by lysophosphatidic acid is required for synergism in both activator protein‐1 activation and mitogenesis.
The role of adherence in tuberculosis HIV‐positive patients treated in ambulatory regimen
Authors: Rocha M. , Pereira S. , Ferreira L. , Barros H.
Published: May 2003

Adherence to antituberculosis treatment, a major determinant of outcome, is of special concern in human immunodeficiency virus (HIV)‐positive patients. However, tuberculosis death in HIV‐positive patients remains higher than in HIV‐negative patients, regardless of adherence.

To assess determinants for an unfavourable tuberculosis outcome (defined as no cure or death), and determinants for nonadherence to antituberculosis treatment, 70 HIV‐positive patients with tuberculosis referred to an outpatient centre were studied. Patient outcome was compared according to HIV risk factors, other opportunistic diseases, antiretroviral drugs use, currenti.v.drugs use, a methadone programme participation, tuberculosis features, treatment characteristics and adherence. Adherent and nonadherent patient characteristics were also compared.

An unfavourable outcome occurred in 22.9% of patients and 32.9% were nonadherent with therapy. Nonadherence was the only independent determinant for an unfavourable outcome. Adherence was independently associated with currenti.v.drug use, treatment complications and use of methadone.

This study confirms that human immunodeficiency virus-infected patients, treated as outpatients, have high rates of nonadherence and that adherence is the strongest determinant for tuberculosis outcome. Independently of the current use of drugs, a methadone programme improves adherence to treatment.
Increased immunoreactivity of stromal cell-derived factor‐1 and angiogenesis in asthma
Authors: Hoshino M. , Aoike N. , Takahashi M. , Nakamura Y. , Nakagawa T.
Published: May 2003

Stromal cell-derived factor‐1 (SDF‐1) acts as a chemoattractant for leukocytes and can induce neovascularisation. To examine the role of SDF‐1 in the development of angiogenesis, immunohistochemical studies were performed on bronchial biopsy specimens from asthmatic and control subjects.

Bronchial biopsy specimens were obtained from 13 asthmatic and eight control subjects. The number of vessels and the percentage area they occupied were estimated after staining for type‐IV collagen. In addition the number of SDF‐1‐positive cells was determined.

There was a significant increase in the number of vessels and the percentage vascularity in the submucosa of asthmatic subjects compared with control subjects. Asthmatic subjects exhibited a greater number of SDF‐1‐positive cells in the airway mucosa than control subjects. The degree of vascularity was associated with the number of SDF‐1‐positive cells. Furthermore, the number of SDF‐1‐positive cells was inversely correlated with airway calibre and airway hyperresponsiveness. Colocalisation studies revealed that endothelial cells, macrophages and T‐lymphocytes were the major sources of SDF‐1.

These findings suggest that increased vascularity of bronchial mucosa in asthmatic subjects is closely related to the expression of stromal cell-derived factor‐1 positive cells, which may play a role in remodelling of airwaysviaangiogenesis.
Dehydroepiandrosterone sulphate reduces chronic hypoxic pulmonary hypertension in rats
Authors: Hampl V. , Bíbová J. , Povýšilová V. , Herget J.
Published: May 2003

Pathogenesis of pulmonary hypertension includes vascular smooth muscle cell membrane depolarisation and consequent calcium influx. Usually, calcium-gated potassium channels are activated under such conditions and repolarise the membrane. However, in pulmonary hypertension they are downregulated. The authors hypothesised that pharmacological augmentation of these channels would reduce pulmonary hypertension.

Dehydroepiandrosterone sulphate (DHEA‐S, 0.1 mg·mL−1), a recently characterised activator of calcium-gated potassium channels, was given to rats in drinking water.

Pulmonary arterial blood pressure, increased by 4 weeks of hypoxia (from 15±0.2 to 29.4±2.5 mmHg), was selectively attenuated in rats treated with DHEA‐S for the whole duration of the hypoxic exposure (23.9±0.9 mmHg) and in rats given DHEA‐S only after pulmonary hypertension had fully developed (last 2 weeks of hypoxia; 24.4±1.4 mmHg). Pulmonary vascular remodelling and right ventricular hypertrophy associated with pulmonary hypertension were also reduced by DHEA‐S. Cardiac index and systemic arterial blood pressure did not differ among the groups.

The authors conclude that treatment with an activator of calcium-gated potassium channels, dehydroepiandrosterone sulphate, known to be well tolerated by humans, reduces hypoxic pulmonary hypertension in rats.
Increased vascular endothelial growth factor in acute eosinophilic pneumonia
Authors: Nishigaki Y. , Fujiuchi S. , Yamazaki Y. , Matsumoto H. , Takeda A. , Fujita Y. , Okamoto K. , Fujikane T. , Shimizu T. , Kikuchi K.
Published: May 2003

Acute eosinophilic pneumonia (AEP) is associated with the presence of diffuse pulmonary infiltrates on the chest radiograph and an increased number of eosinophils and an elevation of interleukin (IL)‐5 levels in bronchoalveolar lavage (BAL) fluid. Vascular endothelial growth factor (VEGF) is a constitutively expressed protein encoded by messenger ribonucleic acid in human eosinophils and is released following stimulation with IL‐5. However, the roles of IL‐5 and VEGF in the pathogenesis or activity of this disease have not been clarified.

The authors investigated the cells and the levels of these two factors in BAL fluid in five AEP patients and five normal controls before and after corticosteroid treatment.

The absolute number of eosinophils·mL−1, IL‐5 and VEGF levels in patients before treatment were higher than in controls (53.8versus0.3×104·mL−1, 490.1versus5.2 pg·mL−1and 643.0versus133.9 pg·mL−1, respectively). IL‐5 and VEGF rapidly decreased to the control level in parallel with clinical improvement. The relationship between eosinophilia and IL‐5 and VEGF levels was strongly significant.

Elevated interleukin‐5 in the lung may initiate the recruitment of eosinophils and enhance the release of mediators, such as vascular endothelial growth factor from eosinophils, which, in turn, increases the permeability of blood vessels.