bluuebunny/crossref_metadata_2025 huggingface dataset:
Two novel mutations in the αIIb calcium-binding domains identify hydrophobic regions essential for αIIbβ3 biogenesis
Authors: Mitchell W. Beau , Li Ji Hong , Singh Fiza , Michelson Alan D. , Bussel James , Coller Barry S. , French Deborah L.
Published: February 2003

The recently published crystal structure of the external domains of αVβ3 confirms the prediction that the aminoterminal portion of αV, which shares 40% homology with αIIb, folds into a β-propeller structure and that the 4 calcium-binding domains are positioned on the bottom of the propeller. To gain insight into the role of the calcium-binding domains in αIIb biogenesis, we characterized mutations in the second and third calcium-binding domains of αIIb in 2 patients with Glanzmann thrombasthenia. One patient inherited a Val298Phe mutation in the second domain, and the other patient inherited an Ile374Thr mutation in the third domain. Mammalian cell expression studies were performed with normal and mutant αIIb and β3 cDNA constructs. By flow cytometry, expression of αIIb Val298Phe/β3 in transfected cells was 28% of control, and expression of αIIbIle374Thr/β3 was 11% of control. Pulse-chase analyses showed that both mutant pro-αIIb subunits are retained in the endoplasmic reticulum and degraded. Mutagenesis studies of the Val298 and Ile374 residues showed that these highly conserved, branch-chained hydrophobic residues are essential at these positions and that biogenesis and expression of αIIbβ3 is dramatically affected by structural variations in these regions of the calcium-binding domains. Energy calculations derived from a new model of the αIIb β-propeller indicate that these mutations interfere with calcium binding. These data suggest that the αIIb calcium-binding domains play a key structural role in the β-propeller, and that the structural integrity of the calcium-binding domains is critical for integrin biogenesis.
A significant diffuse component predicts for inferior survival in grade 3 follicular lymphoma, but cytologic subtypes do not predict survival
Authors: Hans Christine P. , Weisenburger Dennis D. , Vose Julie M. , Hock Lynette M. , Lynch James C. , Aoun Patricia , Greiner Timothy C. , Chan Wing C. , Bociek Robert G. , Bierman Philip J. , Armitage James O.
Published: February 2003

Grade 3 follicular lymphoma (FL3) is thought to have an aggressive clinical course. On the basis of possible biologic differences, the new World Health Organization (WHO) classification of lymphoma suggests further subdivision of FL3 into grades 3a and 3b and states that the percentage of involvement by diffuse large B-cell lymphoma (DLBCL) should also be reported. However, the clinical implications of these features are unclear. Therefore, we studied 190 newly diagnosed patients with lymph node–based FL3 who received anthracycline-containing combination chemotherapy. The follicular component was subclassified as grade 3a (FL3a) or grade 3b (FL3b) according to the WHO criteria, or as follicular large cleaved cell type (FLC). The percentage of a diffuse component, if present, was also recorded. Of the 190 cases, there were 107 FL3a (56%), 53 FL3b (28%), and 30 FLC (16%) cases. Diffuse areas were seen in 72 cases (31 FL3a, 28 FL3b, and 13 FLC). There were no significant differences in the clinical characteristics, overall survival, or event-free survival between patients with grades FL3a, FL3b, or FLC. However, those cases with a predominant diffuse component (> 50% diffuse) had a significantly worse overall survival (P = .0037) and event-free survival (P = .012). Therefore, we conclude that the subdivision of FL3 into cytologic subtypes does not appear to be important clinically. However, patients with FL3 having a diffuse component of more than 50% have an inferior survival that is similar to the survival of those with DLBCL.
Increased inflammation in lysozyme M–deficient mice in response to Micrococcus luteus and its peptidoglycan
Authors: Ganz Tomas , Gabayan Victoria , Liao Hsiang-I , Liu Lide , Oren Ami , Graf Thomas , Cole Alexander M.
Published: February 2003

More than 70 years ago, Alexander Fleming discovered lysozyme and proposed that nonpathogenic bacteria fail to cause disease because they are very susceptible to destruction by lysozyme, an enzyme that is one of the principal proteins of phagocytes. Although much has been learned about the effects of lysozyme in vitro, its biological role in vivo has not been determined. We examined transgenic mice deficient in lysozyme M after challenge by the normally nonpathogenic and highly lysozyme-sensitive bacterium Micrococcus luteus. Despite partial compensation by newly expressed lysozyme P in macrophages, lysozyme M–deficient mice developed much more severe lesions than wild-type mice. The tissue injury was due to the failure of lysozyme M–deficient mice to inactivate peptidoglycan, resulting in an intense and prolonged inflammatory response. Our data indicate that tissue injury is normally limited by prompt degradation of bacterial macromolecules that trigger innate immunity and inflammation.
Anti-CD45–mediated cytoreduction to facilitate allogeneic stem cell transplantation
Authors: Wulf Gerald G. , Luo Kang-Li , Goodell Margaret A. , Brenner Malcolm K.
Published: February 2003

The CD45 antigen is present on all cells of the hematopoietic lineage. Using a murine model, we have determined whether a lytic CD45 monoclonal antibody can produce persistent aplasia and whether it could facilitate syngeneic or allogeneic stem cell engraftment. After its systemic administration, we found saturating quantities of the antibody on all cells expressing the CD45 antigen, both in marrow and in lymphoid organs. All leukocyte subsets in peripheral blood were markedly diminished during or soon after anti-CD45 treatment, but only the effect on the lymphoid compartment was sustained. In contrast to the prolonged depletion of T and B lymphocytes from the thymus and spleen, peripheral blood neutrophils began to recover within 24 hours after the first anti-CD45 injection and marrow progenitor cells were spared from destruction, despite being coated with saturating quantities of anti-CD45. Given the transient effects of the monoclonal antibody on myelopoiesis and the more persistent effects on lymphopoiesis, we asked whether this agent could contribute to donor hematopoietic engraftment following nonmyeloablative transplantation. Treatment with anti-CD45 alone did not enhance syngeneic engraftment, consistent with its inability to destroy progenitor cells and permit competitive repopulation with syngeneic donor stem cells. By contrast, the combination of anti-CD45 and an otherwise inactive dose of total-body irradiation allowed engraftment of H2 fully allogeneic donor stem cells. We attribute this result to the recipient immunosuppression produced by depletion of CD45+ lymphocytes. Monoclonal antibodies of this type may therefore have an adjunctive role in nonmyeloablative conditioning regimens for allogeneic stem cell transplantation.
Low-penetrance genetic susceptibility and resistance loci implicated in the relative risk for radiation-induced acute myeloid leukemia in mice
Authors: Boulton Emma , Cole Clare , Knight Abigail , Cleary Helen , Snowden Roger , Plumb Mark
Published: February 2003

Inbred CBA/H mice are susceptible to radiation-induced acute myeloid leukemia (r-AML), and C57BL/6 mice are resistant. A genome-wide screen for linkage between genotype and phenotype (r-AML) of 67 affected (CBA/H × C57BL/6)F1 × CBA/H backcross mice has revealed at least 2 suggestive loci that contribute to the overall lifetime risk for r-AML. Neither is necessary or sufficient for r-AML, but relative risk is the net effect of susceptibility (distal chromosome 1) and resistance (chromosome 6) loci. An excess of chromosome 6 aberrations in mouse r-AML and bone marrow cells up to 6 months after irradiation in vivo suggests the locus confers a proliferative advantage during the leukemogenic process. The stem cell frequency regulator 1 (Scfr1) locus maps to distal chromosome 1 and determines the frequency of hemopoietic stem cells (HSCs) in inbred mice, suggesting that target size may be one factor in determining the relative susceptibility of inbred mice to r-AML.
Retroviral transduction efficiency of G-CSF+SCF–mobilized peripheral blood CD34+ cells is superior to G-CSF or G-CSF+Flt3-L–mobilized cells in nonhuman primates
Authors: Hematti Peiman , Sellers Stephanie E. , Agricola Brian A. , Metzger Mark E. , Donahue Robert E. , Dunbar Cynthia E.
Published: February 2003

Gene transfer experiments in nonhuman primates have been shown to be predictive of success in human clinical gene therapy trials. In most nonhuman primate studies, hematopoietic stem cells (HSCs) collected from the peripheral blood or bone marrow after administration of granulocyte colony-stimulating factor (G-CSF) + stem cell factor (SCF) have been used as targets, but this cytokine combination is not generally available for clinical use, and the optimum target cell population has not been systematically studied. In our current study we tested the retroviral transduction efficiency of rhesus macaque peripheral blood CD34+ cells collected after administration of different cytokine mobilization regimens, directly comparing G-CSF+SCF versus G-CSF alone or G-CSF+Flt3-L in competitive repopulation assays. Vector supernatant was added daily for 96 hours in the presence of stimulatory cytokines. The transduction efficiency of HSCs as assessed by in vitro colony-forming assays was equivalent in all 5 animals tested, but the in vivo levels of mononuclear cell and granulocyte marking was higher at all time points derived from target CD34+ cells collected after G-CSF+SCF mobilization compared with target cells collected after G-CSF (n = 3) or G-CSF+Flt3-L (n = 2) mobilization. In 3 of the animals long-term marking levels of 5% to 25% were achieved, but originating only from the G-CSF+SCF–mobilized target cells. Transduction efficiency of HSCs collected by different mobilization regimens can vary significantly and is superior with G-CSF+SCF administration. The difference in transduction efficiency of HSCs collected from different sources should be considered whenever planning clinical gene therapy trials and should preferably be tested directly in comparative studies.
RANK ligand and osteoprotegerin in myeloma bone disease
Authors: Sezer Orhan , Heider Ulrike , Zavrski Ivana , Kühne Christian Alexander , Hofbauer Lorenz Christian
Published: February 2003

Myeloma bone disease is due to interactions of myeloma cells with the bone marrow microenvironment, and is associated with pathologic fractures, neurologic symptoms and hypercalcemia. Adjacent to myeloma cells, the formation and activation of osteoclasts is increased, which results in enhanced bone resorption. The recent characterization of the essential cytokine of osteoclast cell biology, receptor activator of NF-κB ligand (RANKL) and its antagonist osteoprotegerin (OPG), have led to a detailed molecular and cellular understanding of myeloma bone disease. Myeloma cells induce RANKL expression in bone marrow stromal cells, and direct RANKL expression by myeloma cells may contribute to enhanced osteoclastogenesis in the bone microenvironment in myeloma bone disease. Furthermore, myeloma cells inhibit production and induce degradation of OPG. These effects result in an increased RANKL-to-OPG ratio that favors the formation and activation of osteoclasts. Patients with myeloma bone disease have inappropriately low serum and bone marrow levels of OPG. Specific blockade of RANKL prevented the skeletal complications in various animal models of myeloma, and suppressed bone resorption in a preliminary study of patients with myeloma bone disease.
The molecular genetics of the human I locus and molecular background explain the partial association of the adult i phenotype with congenital cataracts
Authors: Yu Lung-Chih , Twu Yuh-Ching , Chou Ming-Lun , Reid Marion E. , Gray Alan R. , Moulds Joann M. , Chang Ching-Yi , Lin Marie
Published: February 2003

The human i and I antigens are characterized as linear and branched repeats of N-acetyllactosamine, respectively. Conversion of the i to the I structure requires I-branching β-1,6-N-acetylglucosaminyltransferase activity. It has been noted that the null phenotype of I, the adult i phenotype, is associated with congenital cataracts in Asians. Previously, the identification of molecular changes in the IGnT gene, associated with the adult i phenotype, has been reported. In the present study, we demonstrate that the human I locus expresses 3 IGnT forms, designated IGnTA, IGnTB, and IGnTC, which have different exon 1, but identical exons 2 and 3, coding regions. The molecular genetics proposed for the I locus offer a new perspective on the formation and expression of the I antigen in different cells and provide insight into the questions derived from investigation of the adult i phenotype. Molecular genetic analyses of the Iloci of the 2 adult i groups, with and without congenital cataracts, were performed, and enzyme function assays and expression patterns for the 3 IGnT transcripts in reticulocytes and lens-epithelium cells were analyzed. The results suggest a molecular genetic mechanism that may explain the partial association of the adult i phenotype with congenital cataracts and indicate that a defect in theI locus may lead directly to the development of congenital cataracts. The results also suggest that the human blood groupI gene should be reassigned to the IGnTC form, not the IGnTB form, as described previously.
Transcript profiling of human platelets using microarray and serial analysis of gene expression
Authors: Gnatenko Dmitri V. , Dunn John J. , McCorkle Sean R. , Weissmann David , Perrotta Peter L. , Bahou Wadie F.
Published: February 2003

Human platelets are anucleate blood cells that retain cytoplasmic mRNA and maintain functionally intact protein translational capabilities. We have adapted complementary techniques of microarray and serial analysis of gene expression (SAGE) for genetic profiling of highly purified human blood platelets. Microarray analysis using the Affymetrix HG-U95Av2 approximately 12 600-probe set maximally identified the expression of 2147 (range, 13%-17%) platelet-expressed transcripts, with approximately 22% collectively involved in metabolism and receptor/signaling, and an overrepresentation of genes with unassigned function (32%). In contrast, a modified SAGE protocol using the Type IIS restriction enzyme MmeI (generating 21–base pair [bp] or 22-bp tags) demonstrated that 89% of tags represented mitochondrial (mt) transcripts (enriched in 16S and 12S ribosomal RNAs), presumably related to persistent mt-transcription in the absence of nuclear-derived transcripts. The frequency of non-mt SAGE tags paralleled average difference values (relative expression) for the most “abundant” transcripts as determined by microarray analysis, establishing the concordance of both techniques for platelet profiling. Quantitative reverse transcription-polymerase chain reaction (PCR) confirmed the highest frequency of mt-derived transcripts, along with the mRNAs for neurogranin (NGN, a protein kinase C substrate) and the complement lysis inhibitor clusterin among the top 5 most abundant transcripts. For confirmatory characterization, immunoblots and flow cytometric analyses were performed, establishing abundant cell-surface expression of clusterin and intracellular expression of NGN. These observations demonstrate a strong correlation between high transcript abundance and protein expression, and they establish the validity of transcript analysis as a tool for identifying novel platelet proteins that may regulate normal and pathologic platelet (and/or megakaryocyte) functions.
Von Willebrand disease type 1: a diagnosis in search of a disease
Authors: Sadler J. Evan
Published: February 2003

Von Willebrand disease (VWD) type 1 is reported to be common but frequently is difficult to diagnose. Many people have nonspecific mild bleeding symptoms, von Willebrand factor (VWF) levels display low heritability, and low VWF levels (15% to 50% of normal) are weak risk factors for bleeding. Therefore, bleeding and low VWF levels often associate by chance. Even with stringent diagnostic criteria based on a triad of bleeding symptoms, a low VWF level, and a positive family history, the prevalence of “false-positive” VWD type 1 is comparable to the published prevalence of the disease. Consequently, many patients diagnosed with VWD type 1 do not have a specific hemorrhagic disease at all, which limits the utility of the diagnosis. This unfortunate reality is a consequence of trying to force patients into binary categories of “diseased” or “healthy” that are incompatible with the continuous biologic context in which VWF functions. The problem may be avoided by substituting an empirical epidemiologic approach like that applied to other modest risk factors for disease such as elevated cholesterol and high blood pressure. Such a risk management strategy could be generalized to include other hemorrhagic and thrombotic risk factors.
T(14;18)(q32;q21) involving IGH andMALT1 is a frequent chromosomal aberration in MALT lymphoma
Authors: Streubel Berthold , Lamprecht Andrea , Dierlamm Judith , Cerroni Lorenzo , Stolte Manfred , Ott German , Raderer Markus , Chott Andreas
Published: February 2003

T(11;18)(q21;q21) is the most common structural abnormality in extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma) leading to the fusion of the apoptosis inhibitor-2 (API2) gene and the MALT lymphoma-associated translocation (MALT1) gene. In 2 patients with MALT lymphoma of the liver and skin, respectively, t(14;18)(q32;q21) was observed by cytogenetic analysis. Subsequent fluorescence in situ hybridization (FISH) studies disclosed that the immunoglobulin heavy-chain locus (IGH) and the MALT1 gene were rearranged by this translocation. In order to screen a large series of MALT lymphomas for this aberration, a 2-color interphase FISH assay was established. Among a total of 66 cases, t(14;18)(q32;q21) involving IGH and MALT1 was detected in MALT lymphomas of the liver (4 of 4), skin (3 of 11), ocular adnexa (3 of 8), and salivary gland (2 of 11), but did not occur in MALT lymphomas of the stomach (n = 10), intestine (n = 9), lung (n = 7), thyroid (n = 4), or breast (n = 2). In total, 12 of 66 (18%) MALT lymphomas harbored t(14;18)(q32;q21); 7 additional cases of splenic marginal zone lymphoma tested negative. All of the 12 MALT lymphomas featuring the t(14;18)(q32;q21) were negative for t(11;18)(q21;q21) by reverse transcriptase–polymerase chain reaction (RT-PCR). However, trisomy 3 and/or 18 was found in 4 of 12 cases, suggesting that the t(14;18)(q32;q21) does not occur as the sole genetic abnormality. This study identifies IGH as a new translocation partner of MALT1 in MALT lymphomas, which tend to arise frequently at sites other than the gastrointestinal tract and lung. In contrast to t(11;18)(q21;q21)+ MALT lymphomas, those with t(14;18)(q32;q21) may harbor additional genetic abnormalities.
MDR1 gene overexpression confers resistance to imatinib mesylate in leukemia cell line models
Authors: Mahon François-Xavier , Belloc Francis , Lagarde Valérie , Chollet Claudine , Moreau-Gaudry François , Reiffers Josy , Goldman John M. , Melo Junia V.
Published: February 2003

Inappropriate expression of the multidrug resistance(MDR1) gene encoding the P-glycoprotein (Pgp) has been frequently implicated in resistance to different chemotherapeutic drugs. We have previously generated chronic myeloid leukemia (CML) cell lines resistant to the tyrosine kinase inhibitor imatinib mesylate (STI571), and one line (LAMA84-r) showed overexpression not only of the Bcr-Abl protein but also of Pgp. In the present study, we investigated this phenomenon in other cell lines overexpressing exclusively Pgp. Thus, cells from the K562/DOX line, described as resistant to doxorubicin due to MDR1 gene overexpression, grew continuously in the presence of 1 μM imatinib, but died in 4 to 5 days if the Pgp pump modulators verapamil or PSC833 were added to the imatinib-treated culture. Analysis of cell proliferation by the MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay confirmed the differential sensitivity of K562/DOX to imatinib, which was also reversed by verapamil or PSC833. Flow cytometric analysis of the total phosphotyrosine content by intracytoplasmic staining after a 2-hour incubation with escalating doses of imatinib showed that the inhibitory concentrations of 50% (IC50) for inhibition of cellular protein tyrosine phosphorylation were 15, 10, and 5 μM for K562/DOX, K562/DOX plus verapamil, and K562, respectively. Retroviral-mediated transfection of theBCR-ABL+ AR230 cell line with theMDR1 gene decreased its sensitivity to imatinib, an effect that was also reversed by verapamil. The possible role of MDR overexpression in clinical resistance to imatinib remains to be defined. We therefore confirm that imatinib should be added to the extensive list of drugs that can be affected by the MDR phenomenon.
The effect of a novel orally active selective PDE4 isoenzyme inhibitor (CDP840) on allergen-induced responses in asthmatic subjects
Authors: Harbinson PL , MacLeod D , Hawksworth R , O'Toole S , Sullivan PJ , Heath P , Kilfeather S , Page CP , Costello J , Holgate ST , Lee TH
Published: February 2003

Recent studies have suggested that theophylline, a nonspecific phospho-diesterase inhibitor, has useful anti-inflammatory actions in asthma. Phosphodiesterase 4 (PDE4) represents the predominant PDE isoenzyme present in inflammatory cells. PDE4 inhibitors might, therefore, have beneficial effects in asthma. Side-effects, specifically nausea, have limited the use of existing agents. CDP840 is an orally active, potent and selective PDE4 inhibitor. We have examined the effect of CDP840 on the allergen-induced asthmatic response, its possible modes of action, and its tolerability at therapeutic doses. A total of 54 patients were recruited to three double-blind, placebo-controlled studies. The first study examined the effect of CDP840 (15 mg b.i.d. for 9.5 days) on the allergen-induced asthmatic response in patients with known dual response to allergen. A second study examined the effect of CDP840 (15 mg b.i.d. for 9.5 days) on airway responsiveness to histamine. A third study examined whether single dose CDP840 (15 and 30 mg) had significant bronchodilatory effects. In all studies, CDP840 was well-tolerated, with no patients reporting nausea. CDP840 did not lead to changes in baseline forced expiratory volume in one second (FEV1) as compared to placebo. The late asthmatic response (LAR) to allergen, expressed as area under the curve at 3-8 h (AUC3-8h), was inhibited by 30% (p=0.016), an effect which persisted to the end of the observation period. The early asthmatic response (EAR) was unaffected, and there was no bronchodilatory effect at the doses used. Treatment with CDP840 did not affect bronchial hyperresponsiveness to histamine. In conclusion, CDP840 significantly attenuated the late asthmatic response to allergen challenge in the absence of any bronchodilatory or histamine antagonist effect. This suggests that CDP840 may exert its effects via an anti-inflammatory mechanism.
Airway endothelin levels in asthma: influence of endobronchial allergen challenge and maintenance corticosteroid therapy
Authors: Redington AE , Springall DR , Ghatei MA , Madden J , Bloom SR , Frew AJ , Polak JM , Holgate ST , Howarth PH
Published: February 2003

Endothelins (ETs) are 21 amino acid peptides which, in addition to their other properties, are potent bronchoconstrictors. Whilst there is evidence of the involvement of ET in the pathophysiology of chronic asthma, its contribution to the acute allergic response is undefined. To examine this, we have undertaken segmental bronchoprovocation with allergen and saline at separate sites in six atopic asthmatics receiving treatment with bronchodilators only and six atopic asthmatics additionally receiving treatment with inhaled corticosteroids. Each challenged segment was lavaged 10 min after bronchoprovocation and concentrations of immunoreactive ET were measured in bronchoalveolar lavage fluid. In the non-steroid-treated subjects, there were significantly lower ET levels at the allergen-challenged sites compared to the saline-challenged sites (p<0.05). In the steroid-treated subjects, on the other hand, there was no significant difference between the two sites. Levels of ET at the saline-challenged sites were significantly lower in the steroid-treated subjects compared to the non-steroid-treated subjects (p<0.04). These findings do not support the hypothesis that allergen exposure in asthma results in immediate release of endothelin. However, release at later time-points and a role for endothelin in late-phase bronchoconstriction are not excluded.
Changing prevalence of asthma in school children: evidence for diagnostic changes in asthma in two surveys 13 yrs apart
Authors: Nystad W , Magnus P , Gulsvik A , Skarpaas IJ , Carlsen KH
Published: February 2003

It is still unclear whether the reported increase in the prevalence of asthma is real or due to changes in diagnostic criteria. The objectives of this study were to compare the prevalence of diagnosed asthma with the prevalence of respiratory symptoms, and to compare the association between asthma and other atopic diseases in 1981 and 1994. The study populations comprised randomly selected school classes in Oslo in 1981 (n=1,772) and 1994 (n=2,577). The main outcomes in these comparable cross-sectional studies of children, 6-16 yrs of age, were parent-reported prevalence of diagnosed asthma, respiratory symptoms, eczema and hay fever. The questionnaire was identical in 1981 and 1994. The response rates were 94% (1,674 out of 1,772) in 1981 and 85% (2,188 out of 2,577) in 1994. The lifetime prevalence of asthma increased from 3.4% in 1981 to 9.3% in 1994; odds ratio (OR) 2.9 (95% confidence interval (95% CI) 2.1-4.0) comparing 1994 to 1981. The prevalence of occasional wheezing increased from 9.0 to 10.8%; OR 1.2 (95% CI 1.0-1.5), and attacks of wheezing from 3.7 to 6.8%; OR 1.8 (95% CI 1.3-2.5). Survival analyses for 3 year birth cohorts showed that asthma was more readily diagnosed in the latest birth cohort (1985-1988). The association between asthma and other atopic diseases decreased during the period under study. The increase in diagnosed asthma and respiratory symptoms supports a true increase in asthma. However, the larger increase in diagnosed asthma than wheezing and a reduced association between asthma and other atopic diseases suggest that the increase in asthma may be explained, in part, by changes in diagnostic criteria.
Disabling dyspnoea in patients with advanced disease: lack of effect of nebulized morphine
Authors: Noseda A , Carpiaux JP , Markstein C , Meyvaert A , de Maertelaer V
Published: February 2003

The purpose of this placebo-controlled, double-blind, randomized study was to assess the effect of nebulized morphine on dyspnoea perceived at rest by patients with advanced disease. Seventeen hospital in-patients with disabling dyspnoea received isotonic saline or morphine via nebulization for 10 min through a mouthpiece, combined with oxygen via nasal prongs. On four consecutive days, they were given one of the four following treatments in random order: saline with 2 L x min(-1) oxygen; 10 mg morphine with 2 L x min(-1) oxygen; 20 mg morphine with 2 L x min(-1) oxygen; and 10 mg morphine without oxygen (prongs fixed, no flow). Dyspnoea was assessed on a bipolar visual analogue scale (VAS) (-100% much more short of breath, +100% much less short of breath), and arterial oxygen saturation (Sa,O2) and respiratory frequency (fR) were recorded at the end of nebulization and 10 min later. In 14 subjects who completed the study, mean VAS ratings 10 min after the end of nebulization ranged +30 to +43%, with no significant difference between the four study days (VAS 20 mg morphine minus VAS saline, 95% confidence interval (95% CI) -6 to +8%). Sa,O2 significantly increased on the 3 days with supplemental oxygen, and remained stable on the zero flow day. Respiratory frequency significantly decreased on the 4 days, with a trend to correlation between VAS rating and parallel change in respiratory frequency (Spearman's rank correlation coefficient (r(s))=0.46; p=0.09). We conclude that the subjects benefited from saline or morphine via a placebo effect and/or a nonspecific effect, and that nebulized morphine had no specific effect on dyspnoea.
Serum and BAL Clara cell 10 kDa protein (CC10) levels and CC10-positive bronchiolar cells are decreased in smokers
Authors: Shijubo N , Itoh Y , Yamaguchi T , Shibuya Y , Morita Y , Hirasawa M , Okutani R , Kawai T , Abe S
Published: February 2003

Cigarette smoking has diverse effects on the structure and function of the lung. Smoking appears to reduce the levels of Clara cell 10 kDa protein (CC10) in the alveolar lining fluid, but the influence of smoking serum on CC10 levels is still debated, and it has not been clear whether smoking reduces the number of CC10-producing lung cells. The aims of this study were to clarify the influence of smoking on CC10 levels in the alveolar lining fluid and bloodstream, and on the number of CC10-producing lung cells. CC10 concentrations were measured in sera and bronchoalveolar lavage (BAL) fluids, by means of enzyme-linked immunosorbent assay using monoclonal and polyclonal antibody, and the immunohistochemical expression of CC10 was examined in the lungs of nonsmokers and smokers using the monoclonal antibody, TY-5, against CC10/human urinary protein-1. CC10 concentrations in sera and in BAL fluids from healthy smokers were significantly lower than in healthy nonsmokers. Immunohistochemical expression of CC10 was found exclusively in nonciliated bronchiolar epithelial cells. As compared to that of nonsmokers, the mean percentage of CC10-positive bronchiolar epithelial cells was significantly decreased in lung tissue specimens obtained from smokers who had normal results in pulmonary function tests. It was concluded that smoking reduces the proportion of Clara cell 10 kDa protein-producing bronchiolar epithelial cells, resulting in decreased levels of Clara cell 10 kDa protein in the lower respiratory tract and in the bloodstream. The protein is a new blood biochemical and immunohistochemical marker, reflecting structural changes in peripheral airways induced by cigarette smoking.
White blood cells, C-reactive protein and erythrocyte sedimentation rate in pneumococcal pneumonia in children
Authors: Korppi M , Heiskanen-Kosma T , Leinonen M
Published: February 2003

We evaluated the applicability of C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), white blood cell count (WBC), and absolute neutrophil count (ANC), in the screening of pneumococcal (PNC) pneumonia in children. In 1981-1982, 161 children were treated for radiologically verified community-acquired pneumonia in the hospital during a period of 12 months. The Streptococcus pneumoniae aetiology of infection was studied by antigen, antibody and immune complex assays in acute and convalescent sera. In acute blood samples, CRP was measured by the immunonephelometric method, ESR by the Westergren method, WBC using an automatic cell counter, and thereafter the ANC was calculated after microscopic examination of peripheral smears. CRP and ESR were significantly higher in patients with alveolar (n=53) than in those with interstitial (n=108) pneumonia. CRP, ESR and ANC were significantly higher in PNC (n=29) than in viral (n=23) pneumonia. The values in mixed PNC and viral infections (n=17) were approximately midway between PNC and viral cases. All cases with serologic evidence of S. pneumoniae aetiology were combined (n=46) for calculation of diagnostic parameters. When a cut-off limit of 60 mg x L(-1) was used, CRP had a sensitivity of 26% and a specificity of 83% in the screening of PNC pneumonia. We conclude that C-reactive protein and erythrocyte sedimentation rate have a limited capacity to differentiate between pneumococcal and nonpneumococcal pneumonia. C-reactive protein is recommended as the first-line method of screening, and the value of 60 mg x L(-1) as the cut-off limit.
Penetration of sparfloxacin and ciprofloxacin into alveolar macrophages, epithelial lining fluid, and polymorphonuclear leucocytes
Authors: Schuler P , Zemper K , Borner K , Koeppe P , Schaberg T , Lode H
Published: February 2003

Sparfloxacin (SPX), a novel fluoroquinolone with a broad spectrum of antibacterial activity, has been shown, in vitro, to be more effective against common pulmonary pathogens, particularly Streptococcus pneumoniae, Mycoplasma pneumoniae and some intracellular organisms, than ciprofloxacin (CPX). The objective of this open, comparative, randomized study in two parallel groups was to assess the serum concentrations and penetration of SPX and CPX into alveolar macrophages (AMs), epithelial lining fluid (ELF), and peripheral polymorphonuclear leucocytes (PMNs). Patients received either a single oral dose of SPX, 400 mg (16 patients), or CPX, 500 mg (15 patients), at various times before a routine diagnostic bronchoscopy was performed. Antibiotic concentrations were determined by using a microbiological assay. The median serum elimination half-lives were 17.3 h for SPX and 5.3 h for CPX. Peak levels (median+/-68% confidence range) of CPX were attained in AMs (7.6+/-1.7 mg x L(-1)) at 5 h after admission, and in both ELF (2.13+/-0.91 mg x L(-1)) and PMNs (9.1+/-6.2 mg x L(-1)) at 2.5 h after administration. SPX achieved comparable peak levels in PMNs (median+/-68% range, 9.4+/-1.1 mg x L(-1) at 5 h), but peak concentrations attained in AMs and ELF (35.1+/-15.4 and 32.2+/-28.2 mg x L(-1), respectively, at 24 h) were several times higher than those of CPX. SPX demonstrated substantially greater accumulation in all three sites than did CPX (peak site to serum ratios of 11.0+/-4.3, 65.5+/-28.4 and 63.0+/-66 versus 4.94+/-0.64, 10.6+/-3.7 and 10.6+/-3.7 for PMNs, AMs and ELF, respectively). The results indicate, that sparfloxacin has a long elimination half-life and that it achieves higher concentrations in alveolar macrophages and epithelial lining fluid than ciprofloxacin. Site concentrations of sparfloxacin greatly exceeded the minimal inhibitory concentrations of common respiratory pathogens.
Chylothorax and pseudochylothorax
Authors: Hillerdal G
Published: February 2003

Chylothorax is the occurrence of chylus (lymph) in the pleura due to damage to the thoracic duct. There is a high content of triglycerides, and chylomicrons can be seen. It is usually right-sided, since most of the duct is within the right hemithorax. With damage at the level of the aorta, the chyle will appear on the left. Traumatic rupture occurs after accidents or surgery. Of nontraumatic causes, the most common is a malignant lymphoma. Computed tomography (CT) scan of the thorax and upper abdomen should be performed. Lymphography can show where the leakage or blockage is situated. With repeated drains, large amounts of fat, proteins, and lymphocytes are lost. Treatment is with low-fat diet or parenteral nutrition to decrease the amount of chyle, but chemical pleurodesis or ligation of the thoracic duct, usually via thoracoscopy, is often necessary. Pseudochylothorax (cholesterol pleurisy) occurs with long-standing fluid in a fibrotic pleura. The fluid has a high content of cholesterol but no triglycerides or chylomicrons. In both conditions, the pleural fluid is thick, opalescent, whitish or the colour of cafe-au-lait.
Pneumocystis carinii pneumonia in a HIV-seronegative patient with untreated rheumatoid arthritis and CD4+ T-lymphocytopenia
Authors: Prekates A , Kyprianou T , Paniara O , Roussos C
Published: February 2003

Pneumocystis carinii pneumonia (PCP) usually occurs in immunocompromised patients, and it is a life-threatening infection. We report the case of a human immunodeficiency virus (HIV)-seronegative patient with untreated rheumatoid arthritis (RA), who developed fatal PCP related to uncommon CD4+ T-lymphocytopenia. Although extremely rare and of uncertain aetiology, suppression of cellular immunity and subsequent opportunistic infections should be suspected in such patients.
Pre-emptive control of moisture content in paper manufacturing using surrogate measurements
Authors: Li Perry Y. , Ramaswamy Shri , Bjegovic Petar
Published: February 2003

Paper manufacturing consists of the sequential removal of over 90% of the water from pulp through gravity, vacuum dewatering, pressing and thermal drying. Control of moisture content is important for paper quality and energy economy. Current strategy for the control of moisture content uses a feedback sensor at the end of the process to adjust the dryers. This introduces a long dead-time and causes excessive use of the dryers, which translate to limitations in performance, robustness and inefficient energy usage. In this paper, we investigate a new control approach in which in-process moisture contents are estimated using air-flow as surrogate measurements, and the pressure settings in the vacuum dewatering boxes are adjusted according to the surrogate measurements. A pre-emptive control algorithm is developed which has the ability to decouple and eliminate the effects of the disturbances that occur upstream in the process from the downstream. Robustness analysis and simulation studies suggest that as long as the surrogate measurements are accurate, the proposed control scheme will be robust and accurate.
Application of exponentially weighted principal component analysis for the monitoring of a polymer film manufacturing process
Authors: Lane S. , Martin E. B. , Morris A. J. , Gower P.
Published: February 2003

Multivariate statistical representations have been widely used in the process manufacturing industries for process performance monitoring, in particular for the detection of changes in current operation and the onset of process disturbances or faults. Applications of the technology have focused to a lesser extent on manufacturing processes where drift occurs over time as part of normal process operation, e.g., due to reactor fouling, machine wear, ramping of temperatures during process operation, and changes due to set-point adjustments. In this paper, an extension to the methodology based on the statistical projection technique of principal component analysis (PCA) is proposed for the monitoring of processes where drift and set-points changes are common place, i.e., exponentially weighted PCA. The technique is illustrated through its application to a polymer film manufacturing process where the representation is required to adapt quickly to changes in the process that are part of normal operating procedures, but remain sensitive to the detection of deviations from normal operation.
British Disarmament Policy and the Rome Naval Conference, 1924
Authors: Silverlock Gerard
Published: February 2003

This article provides the first detailed examination of an important but largely unnoticed aspect of British defence and foreign policy in the period after the Great War. It is based upon a sophisticated definition of disarmament which takes account of both aims and process in international relations. Following the Washington conference, 1921-22, the British put forward a plan which sought to extend the principles of the naval limitation treaty to nonsignatory powers. Although the initiative was ultimately unsuccessful, its history reveals that the Admiralty was prepared to use the League of Nations in pursuit of a disarmament agreement which, it believed, could benefit the country’s naval interests.
Prognostic Value of Circulating Melanoma Cells Detected by Reverse Transcriptase–Polymerase Chain Reaction
Authors: Palmieri Giuseppe , Ascierto Paolo A. , Perrone Francesco , Satriano Sabrina M.R. , Ottaiano Alessandro , Daponte Antonio , Napolitano Maria , Caracò Corrado , Mozzillo Nicola , Melucci Maria T. , Cossu Antonio , Tanda Francesco , Gallo Ciro , Satriano Rocco A. , Castello Giuseppe
Published: February 2003

Purpose: Factors that are predictive of prognosis in patients who are diagnosed with malignant melanoma (MM) are widely awaited. Detection of circulating melanoma cells (CMCs) by reverse transcriptase-polymerase chain reaction (RT-PCR) has recently been postulated as a possible negative prognostic factor. Two main questions were addressed: first, whether the presence of CMCs, defined as the patient being positive for any of the three markers, had a prognostic role; and second, what the predictive value of each individual marker was.

Patients and Methods: A consecutive series of 200 melanoma patients observed between January 1997 and December 1997, with stage of disease ranging from I to IV, was analyzed by semiquantitative RT-PCR. Tyrosinase, p97, and MelanA/MART1 were used as markers to CMCs on baseline peripheral blood samples. Progression-free survival (PFS) was used as a unique end point and was described by the product limit method. Multivariable analysis was applied to verify whether the auspicated prognostic value of these markers was independent of the stage of disease, and a subgroup analysis was performed that excluded patients with stage IV disease.

Results: Overall, 32% (64 of 200) of patients progressed, and a median PFS of 52 months in the whole series was observed. The presence of CMCs and the markers individually or combined was predictive of prognosis in the univariate analysis but did not provide additional prognostic information to the stage of disease in multivariable models. In the subgroup analysis of stage (ie, I–III subgroup), similar results were observed.

Conclusion: Detection of CMCs in peripheral blood samples at the time of MM diagnosis by semiquantitative RT-PCR does not add any significant predictive value to the stage of disease. Thus, this approach should not be used in clinical practice, and further studies are required to determine its usefulness.
Prognostic Value of Tumor Size, Metastases, Extension into Bone, and Increased Tumor Marker in Children With Malignant Sacrococcygeal Germ Cell Tumors: A Prospective Evaluation of 71 Patients Treated in the German Cooperative Protocols Maligne Keimzelltumoren (MAKEI) 83/86 and MAKEI 89
Authors: Calaminus G. , Schneider D.T. , Bökkerink J.P.M. , Gadner H. , Harms D. , Willers R. , Göbel U.
Published: February 2003

Purpose: To evaluate the prognostic value of metastases, extension into bone, and alpha-fetoprotein (AFP) elevation in children with malignant sacrococcygeal germ cell tumors (GCTs) prospectively collected in two cooperative Maligne Keimzelltumoren (MAKEI) protocols (83/86 and 89).

Patients and Methods: Between October 1983 and October 1995, 76 of 210 registered patients with sacrococcygeal primaries presented either with pure yolk sac tumor, embryonal carcinoma (EC), or yolk sac tumor and EC mixed with immature and mature teratoma elements. Stages T1 and T2 disease were diagnosed in 15 and 61 children, respectively, 41 patients had metastases, and 35 children presented with extension into bone. At diagnosis, 22 children had an AFP elevation of less than 10,000 ng/mL. Thirty-six children showed an AFP level between 10,000 and 100,000 ng/mL, and 12 patients had values of greater than 100,000 ng/mL. Five patients died of complication during treatment and were excluded from further evaluation. Seventy-one patients could be analyzed.

Results: The 5-year relapse-free survival rate (RFS, Kaplan-Meier) was 0.76 ± 0.03 (54 of 71 patients; median observation time, 54 months after diagnosis). The RFS of patients with and without metastases was different, but not significantly so (0.71 v 0.82). The outcome of patients with extension into bone (n = 31) and without this extension (n = 40) was 0.71 versus 0.80 (RFS, 5 years). Above-normal AFP level had no prognostic significance (P = .52).

Conclusion: In children with malignant sacrococcygeal GCTs treated with an intensive, short-interval, platinum-based regimen, the stage, extent of metastases, extension into bone, and AFP level had no prognostic significance.
Shared Treatment Decision Making: What Does It Mean to Physicians?
Authors: Charles Cathy A. , Whelan Tim , Gafni Amiram , Willan Andy , Farrell Sylvia
Published: February 2003

Purpose: Physicians are urged to practice shared treatment decision making (STDM), yet this concept is poorly understood. We developed a conceptual framework describing essential characteristics of a shared approach. This study assessed the degree of congruence in the meanings of STDM as described in the framework and as perceived by practicing physicians.

Methods: A cross-sectional survey questionnaire was mailed to eligible Ontario medical and radiation oncologists and surgeons treating women with early-stage breast cancer. Open-ended and structured questions elicited physicians’ perceptions of shared decision making.

Results: Most study physicians spontaneously described STDM using characteristics identified in the framework as essential to this concept. When presented with clinical examples in which the decision-making roles of physicians and patients were systematically varied, study physicians overwhelmingly identified example 4 as illustrating a shared approach. This example was deliberately constructed to depict STDM as defined in the framework. In addition, more than 85.0% of physicians identified as important to STDM specific patient and physician roles derived from the framework. These included the following: the physician gives information to the patient on treatment benefits and risks; the patient gives information to the physician about her values; the patient and physician discuss treatment options; both agree on the treatment to implement.

Conclusion: Substantial congruence was found between the meaning of STDM as described in the framework and as perceived by study physicians. This supports use of the framework as a conceptual tool to guide research, compare different treatment decision-making approaches, clarify the meaning of STDM, and enhance its translation into practice.
Final Results of a Randomized Phase III Trial Comparing Cyclophosphamide, Epirubicin, and Fluorouracil With a Dose-Intensified Epirubicin and Cyclophosphamide + Filgrastim as Neoadjuvant Treatment in Locally Advanced Breast Cancer: An EORTC-NCIC-SAKK Multicenter Study
Authors: Therasse P. , Mauriac L. , Welnicka-Jaskiewicz M. , Bruning P. , Cufer T. , Bonnefoi H. , Tomiak E. , Pritchard K. I. , Hamilton A. , Piccart M.J.
Published: February 2003

Purpose: To compare the efficacy of a standard anthracycline-based regimen to a dose-intensified anthracycline regimen in locally advanced breast cancer.

Patients and Methods: Locally advanced breast cancer patients were randomly assigned onto a study comparing cyclophosphamide (C; 75 mg/m2 orally days 1 to 14), epirubicin (E; 60 mg/m2 intravenously [IV] days 1, 8), and fluorouracil (F; 500 mg/m2 IV days 1, 8) six cycles every 28 days versus E (120 mg/m2 IV day 1), C (830 mg/m2 IV day 1), and granulocyte colony-stimulating factor (filgrastim; 5 μg/kg/d subcutaneously days 2 to 13) six cycles every 14 days. The study was designed to detect a 15% improvement; that is, from 50% to 65% in median progression-free survival (PFS) in favor of the dose-intensified regimen.

Results: A total of 448 patients were enrolled over a period of 3 years. The median dose intensity delivered for C and E reached, respectively, 85% and 87% of that planned in the CEF arm and 96% and 95% of that planned in the EC arm. The dose-intensified arm was slightly more emetogenic and generated more grade 3 to 4 anemia but less febrile neutropenia episodes. After a median follow-up of 5.5 years, 277 events have been reported. The median PFS was 34 and 33.7 months for CEF and EC, respectively (P = .68), and the 5-year survival rate was 53% and 51% for CEF and EC, respectively (P = .94).

Conclusion: Dose-intensified EC does not provide a measurable therapeutic benefit over CEF as neoadjuvant chemotherapy for unselected locally advanced breast cancer patients.