bluuebunny/crossref_metadata_2025 huggingface dataset:
Enrichment of Escherichia coli proteins by column chromatography on reactive dye columns
Authors: Birch Rosslyn M. , O'Byrne Conor , Booth Ian R. , Cash Phillip
Published: May 2003

The reliable identification and analysis of the low abundance proteins expressed by a cell remains a key challenge in the study of cellular proteomes. The analysis of low abundance proteins is a particular problem when using two‐dimensional gel electrophoresis (2‐DE) to resolve the cellular proteins since the technology is unable to display the wide dynamic range of protein levels typically synthesized by cells. We have investigated the use of reactive dye compounds for the enrichment of low abundance cellular proteins prior to analysis by 2‐DE. The capacity of reactive dye compounds to bind specific protein species was used as the basis for a general chromatographic tool for protein enrichment. Six reactive dye compounds were investigated in detail for the analysis of Escherichia coli proteins. Whole bacterial cell lysates were passed down columns prepared with the reactive dye compounds. The bound proteins were eluted with 1.5 M NaCl and analyzed by 2‐DE. Distinctive protein profiles were observed for the bound proteins recovered from the different reactive dye compounds. Selected proteins enriched by these methods were identified by peptide mass mapping. The enrichment procedure developed using reactive dye compounds were used to investigate acid‐induced changes in the proteome of E. coli grown at either pH 7.0 or pH 5.8. Increased levels of expression were observed for a number of proteins (for example, GdhA, PanC, ProC, TkrA, EF‐TS and YodA) were observed for E. coli grown at pH 5.8. Five identified proteins (AroG, FabI, GlyA, PurA and EF‐Tu) showed reduced levels of synthesis for bacteria grown at pH 5.8 compared to pH 7.0. In the case of PanC and FabI the altered expression profiles were only reliably demonstrated using the enrichment protocols. One theme emerging from these data was that the expression of proteins concerned with one‐carbon metabolism was perturbed at pH 5.8, which may point to a previously unrecognized affect of low pH stress on the physiology of E. coli cells. We conclude that the prefractionation of cell lysates on reactive dye columns will serve as a valuable generic tool for the analysis of low abundance proteins expressed by both prokaryotic and eukaryotic cells.
Navigated laser capture microdissection as an alternative to direct histological staining for proteomic analysis of brain samples
Authors: Moulédous Lionel , Hunt Sybille , Harcourt Rebecca , Harry Jenny , Williams Keith L. , Gutstein Howard B.
Published: May 2003

Proteomic analysis of the brain is complicated by the need to obtain cells from specific anatomical regions, or nuclei. Laser capture microdissection (LCM) is a technique that is precise enough to dissect single cells within a tissue section, and thus could be useful for isolating specific brain nuclei for analysis. However, we and others have previously demonstrated that histological staining protocols used to guide LCM have detrimental effects on protein separation by two‐dimensional electrophoresis (2‐DE). Here we describe a new LCM method called navigated LCM. This microdissection method uses fixed but unstained tissue as starting material and thus enables us to avoid artifacts induced by tissue staining. By comparing 2‐DE results obtained from fixed, unstained LCM brain tissue samples to those obtained from manually dissected samples, we demonstrated that this microdissection process gave similar protein recovery rates and similar resolution of protein spots on 2‐DE gels. Moreover, matrix‐assisted laser desorption/ionization‐time of flight mass spectrometry analysis of selected spots from gels derived from control and fixed, LCM samples revealed that the fixation‐LCM process had no effect on protein identification. Navigated LCM of tissue sections is therefore a practical and powerful method for performing proteomic studies in specifically defined brain regions.
A strategy for the comparative analysis of serum proteomes for the discovery of biomarkers for hepatocellular carcinoma
Authors: Steel Laura F. , Shumpert Donna , Trotter Michael , Seeholzer Steven H. , Evans Alison A. , London W. Thomas , Dwek Raymond , Block Timothy M.
Published: May 2003

Many of the emerging technologies for the global evaluation of gene expression, at both the RNA and protein level, are being applied to the problem of finding biomarkers for human disease progression. These analyses can be made difficult, however, by variation between samples that arises from both technical and nondisease related physiological or genetic causes. In an effort to identify serum polypeptides whose presence or absence correlates with the clinical status of patients at high risk for hepatocellular carcinoma (HCC), we have developed a strategy that helps to focus the analysis on meaningful changes in protein levels above the background of variation. For the current study we divided the patient population into four clinically defined diagnostic groups that represent a generally increasing risk for HCC. Chronic infection with hepatitis B virus (HBV) is a major risk factor for HCC and our groups included patients with no indication of liver disease (healthy), those with inactive chronic HBV, those with active chronic HBV, and patients with a diagnosis of HCC and history of chronic HBV infection. Serum polypeptides from these patients were first analyzed in two‐dimensional gels by combining the serum from patients in each of the four groups to generate composite gel profiles. Analysis of these composite gels allowed us to identify two relatively abundant features that were reduced in the HCC group as compared to the healthy group. Tryptic fragment mass fingerprinting identified the features as a carboxy terminal fragment of complement C3 and an isoform of apolipoprotein A1. These two features were examined by two‐dimensional gel electrophoresis of serum from each individual in the four groups in order to verify that the inter‐group differences seen in composite gels reported changes in abundance for most members of the group, rather than extreme changes for a small fraction of the group. These preliminary studies suggest that a proteomic methodology can be used for the identification of serum biomarkers for HCC and other liver disease.
Analysis of the Shewanella oneidensis proteome by two‐dimensional gel electrophoresis under nondenaturing conditions
Authors: Giometti Carol S. , Khare Tripti , Tollaksen Sandra L. , Tsapin Alexandre , Zhu Wenhong , Yates John R. , Nealson Kenneth H.
Published: May 2003

Proteomes are dynamic, i.e., the protein components of living cells change in response to various stimuli. Protein changes can involve shifts in the abundance of protein components, in the interactions of protein components, and in the activity of protein components. Two‐dimensional gel electrophoresis (2‐DE) coupled with peptide mass spectrometry is useful for the analysis of relative protein abundance, but the denaturing conditions of classical 2‐DE do not allow analysis of protein interactions or protein function. We have developed a nondenaturing 2‐DE method that allows analysis of protein interactions and protein functions, as demonstrated in our analysis of the cytosol and crude membrane fractions of the facultative anaerobe Shewanella oneidensis MR‐1. Our experiments demonstrate that enzymatic activity is retained under the sample and protein separation methods described, as shown by positive malate dehydrogenase activity results. We have also found protein interactions within both the soluble and membrane fractions. The method described will be useful for the characterization of the functional proteomes of microbial systems.
A reference map of a human pituitary adenoma proteome
Authors: Zhan Xianquan , Desiderio Dominic M.
Published: May 2003

In order to compare the proteomes from different cell types of pituitary adenomas for our long‐term goal to clarify the molecular mechanisms that participate in the formation of pituitary adenoma, and to detect any tumor‐related marker for an “early‐stage” diagnosis, the two‐dimensional gel electrophoresis (2‐DE) reference map of a pituitary adenoma tissue proteome is described here. A vertical, two‐dimensional (2‐D) polyacrylamide gel electrophoresis system and PDQuest image analysis software have been used to provide a high level of between‐gel reproducibility and to accurately array each protein expressed in a pituitary adenoma tissue. Mass spectrometry (matrix‐assisted laser desorption/ionization‐time of flight MALDI‐TOF and liquid chromatography‐electrospray ionization‐quadrupole‐ion trap LC‐ESI‐Q‐IT) and protein databases were used to characterize each protein in the 2‐D gel. The results demonstrate that a good reproducibility of the 2‐D gel pattern was attained. The position deviation of matched spots among four 2‐D gels was 1.95 ± 0.45 mm in the isoelectric focusing direction, and 1.70 ± 0.53 mm in the sodium dodecyl sulfate‐polyacrylamide gel electrophoresis direction. A total of ca. 1000 protein spots were separated by 2‐DE, and 135 protein spots that represent 111 proteins were characterized with mass spectrometry (96 spots for MALDI‐TOF, 39 spots for LC‐ESI‐Q‐IT). The characterized proteins include pituitary hormones, cellular signals, enzymes, cellular‐defense proteins, cell‐structure proteins, transport proteins, etc. Those proteins were located in the cytoplasmic, cellular membrane, mitochondrial, endoplasmic reticulum, nuclear, ribonucleosome, extracellular fractions, or were secreted in plasma, etc. Those identified proteins contribute to a functional profile of the pituitary adenoma proteome. These data will be used to expand the proteome database of the human pituitary, which can be accessed in the website http://www.utmem.edu /proteomics.
Proteomic study of human umbilical vein endothelial cells in culture
Authors: Bruneel Arnaud , Labas Valérie , Mailloux Agnès , Sharma Sanjiv , Vinh Joelle , Vaubourdolle Michel , Baudin Bruno
Published: May 2003

The endothelium is a single layer of cells lining the inside face of all blood vessels. It constitutes a major metabolic organ which is critically involved in the generation and the regulation of multiple physiological and pathological processes such as coagulation, hemostasis, inflammation, atherosclerosis, angiogenesis and cancerous metastasis dissemination. In order to increase our knowledge about the protein content and the main biological pathways of human vascular endothelial cells, we have undertaken the proteomic analysis of the most explored present endothelial cell model, i.e. primocultures of human umbilical vein endothelial cells (HUVECs). Using low levels of protein loads (~ 30 νg), the association of two‐dimensional electrophoresis with matrix‐assisted laser desorption/ionization‐time of flight mass spectrometry, liquid chromatography‐tandem mass spectrometry and database interrogations allowed us to identify 53 proteins of suspected endothelial origin in quiescent HUVECs. Beside cytoskeletal proteins such as actin, tubulin, tropomyosin and vimentin, we identified various proteins more especially implicated in cellular motility and plasticity (e.g. cofilin, F‐actin capping protein and prefoldin), in regulation of apoptosis and senescence (protease inhibitor 9, glucose related proteins, heat shock proteins, thioredoxin peroxidase, nucleophosmin) as well as other proteins implicated in coagulation (annexin V, high mobility group protein), antigen presentation (valosin containing protein and ubiquitin carboxyl terminal hydrolase isozyme L1) and enzymatic capabilities (glutathione‐S‐transferase, protein disulfide isomerases, lactate deshydrogenase). The presented annotated 2‐D maps of HUVECs will be soon available on the web at http://www. huvec.com.
Proteome analysis of secreted proteins during osteoclast differentiation using two different methods: Two‐dimensional electrophoresis and isotope‐coded affinity tags analysis with two‐dimensional chromatography
Authors: Kubota Kazuishi , Wakabayashi Kenji , Matsuoka Tatsuji
Published: May 2003

Bone is maintained by two cell types, bone‐forming osteoblasts and bone‐resorbing osteoclasts. Osteoblasts express two factors, osteoprotegerin and receptor activator of NF‐κB ligand (RANKL), inhibiting and promoting osteoclast differentiation, respectively. In contrast, modulators of bone resorption expressed by osteoclasts have not been so well studied enough. In the present study, we demonstrate proteome analysis of secreted proteins during osteoclast differentiation to elucidate the molecular mechanism of bone resorption and bone remodeling. To achieve this objective, we chose RAW264.7 cells with RANKL as a homogeneous osteoclast differentiation model and used two methods, two‐dimensional gel electrophoresis (2‐DE) and isotope‐coded affinity tags (ICAT) analysis with two‐dimensional liquid chromatography. We found 23 spots in 2‐DE and 19 proteins in ICAT analysis which were expressed differently during osteoclast differentiation. These two methods gave us closely related but different information about proteins, suggesting they are complementary or at least supplementary methods at present. Cathepsins, osteopontin, legumain, macrophage inflammatory protein‐1α, and other proteins were observed as up‐ or down‐regulated proteins and are discussed in the context of osteoclast differentiation and bone resorption. In addition to confirming previous observations, this study indicates novel proteins related to osteoclast differentiation which are potential therapeutic targets for the treatment of bone diseases, such as osteoporosis.
Identification of metastasis‐associated proteins by proteomic analysis and functional exploration of interleukin‐18 in metastasis
Authors: Jiang Daifeng , Ying Wantao , Lu Yinglin , Wan Jinghong , Zhai Yun , Liu Wanli , Zhu Yunping , Qiu Zongyin , Qian Xiaohong , He Fuchu
Published: May 2003

Very little is currently known about mechanisms underlying cancer metastasis. In the present study, metastasis‐associated proteomes were separated and identified by comparative proteomic analysis, and the metastasis‐related function of candidate protein interleukin‐18 (IL‐18) was further elucidated. First, a pair of highly and poorly metastatic sublines (termed PLA801D and PLA801C, respectively), originating from the same parental PLA801 cell line, was identified by spontaneous tumorigenicity and metastasis in vivo and characterized by metastatic phenotypes analysis in vitro. Subsequently, a proteomic approach was used to compare the protein expression profiles between PLA801C and PLA801D sublines. Eleven proteins were identified and further verified by one‐dimensional Western blotting, Northern blot and/or semiquantitative reverse transciptase polymerase chain reaction analysis. Compared with those in poorly metastatic PLA801C subline, cytokeratin 18, tissue transglutaminase, Rho GDP‐dissociation inhibitor 1, tropomyosin, fibroblast type, IL‐18 and annexin I were significantly up‐regulated, while protein disulfide isomerase, heat shock protein 60, peroxiredoxin 1, chlorine intracellular channel protein 1 (CLI1) and creatine kinase, B chain were significantly down‐regulated in the highly metastatic PLA801D subline. Intriguingly, all the identified candidate proteins except for CLI1 have been shown to be somehow associated with distinct aspects of tumor metastasis such as cell growth, motility, invasion, adhesion, apoptosis and tumor immunity, etc. Considering that IL‐18 was present in highly metastatic PLA801D but absent in poorly metastatic PLA801C, the association of IL‐18 with metastasis was further elucidated by introducing IL‐18 sense/IL‐18 antisense into PLA801C/PLA801D sublines simultaneously. The results demonstrated that ectopically expressed IL‐18 promoted cell motility in vitro and down‐regulated E‐cadherin expression of PLA801C transfectants, while IL‐18 antisense remarkably decreased cell invasion potency in vitro and notably increased E‐cadherin expression of PLA801D transfectants, indicating that IL‐18 might play a role in metastasis by inhibiting E‐cadherin expression.
Susceptibility to oxidative stress: proteomic analysis of bronchoalveolar lavage from ozone‐sensitive and ozone‐resistant strains of mice
Authors: Wattiez Ruddy , Noël‐Georis Isabelle , Cruyt Christine , Broeckaert Fabrice , Bernard Alfred , Falmagne Paul
Published: May 2003

Previous studies have shown that the pulmonary response to ozone (O3) varies greatly among strains of mice, but the factor(s) and the mechanism(s) that are responsible for this differential susceptibility have not yet been clearly identified. The present study explores the molecular bases for this differential O3 susceptibility by studying the expression of proteins associated to the epithelial lining fluid (ELF) from two strains of mice, C57BL/6J and the C3H/HeJ, respectively described as O3‐sensitive and O3‐resistant. The ELF proteins of these two strains were displayed by two‐dimensional gel electrophoresis (2‐DE) of bronchoalveolar lavage fluids (BALFs) and the protein patterns obtained with BALF samples of both strains were compared. Two major differences were observed between the BALF 2‐DE protein maps obtained from C57BL/6J and C3H/HeJ strains. First, two isoforms of the antioxidant protein 2 (AOP2) were detected in a strain‐dependent manner: C3J/HeJ possesses only AOP2a (isoelectric point 5.7) and C57BL/6J exhibits only AOP2b (isoelectric point 6.0). Second, the levels of anti‐inflammatory and immunosuppressive Clara cell protein‐16 (CC16) were 1.3 times higher in the BALF from resistant C3H/HeJ than from sensitive C57BL/6 mice. Moreover, two 6 kDa isoforms of CC16 with isoelectric points of 4.9 (CC16a) and 5.2 (CC16b) are detected in both strains. Interestingly, the C57BL/6J strain had a twice decreased level of the acidic isoform of CC16 compared to C3H/HeJ. Our results suggest that AOP2 and CC16 might participate in the protection of the pulmonary tract to O3‐induced lung injury. The possible differential contribution of specific protein isoforms in the differential susceptibility to oxidative stress is discussed.
Identification of novel proteins in culture filtrates of Mycobacterium bovis bacillus Calmette‐Guérin in the isoelectric point range 6–11
Authors: Florio Walter , Batoni Giovanna , Esin Semih , Bottai Daria , Maisetta Giuseppantonio , Pardini Manuela , Campa Mario
Published: May 2003

Two‐dimensional gel electrophoresis and mass spectrometry were used to identify proteins in the isoelectric point range 6–11 in culture filtrates of Mycobacterium bovis bacillus Calmette‐Guérin (BCG). Twelve proteins were identified, three of which had not been described previously. The expression of the identified proteins was comparatively analyzed in culture filtrates of BCG in different growth phases and culture conditions. For some of these proteins, the relative protein abundance in the different culture filtrate preparations was significantly different. The differential expression of the identified proteins is discussed in relation to their putative localization and/or biological function.
“Two‐in‐one” gel for spot matching after two‐dimensional electrophoresis
Authors: Wang Yuen‐Yuen , Cheung Pik‐Yuen , Wong Man‐Sau , Lo Samuel Chun‐Lap
Published: May 2003

Two‐dimensional electrophoresis (2‐DE) is one of the most commonly used techniques in proteomic investigations. However, due to the complex interplay of incidence including significant biological sample variations, lengthy steps involved in performing 2‐DE as well as exposure time with silver staining, it is sometimes difficult to differentiate authentic differences caused by drug treatment with those artifacts caused by sample variations, running conditions of 2‐DE as well as treatment time in silver staining etc. If we can compare pooled samples of control and treatment groups run in a single gel and stained together, we would be more comfortable with our findings. We propose here a low cost and highly effective method for locating differentially expressed proteins before and after drug treatment. This “two‐in‐one gel” technique might partially solve the problems mentioned above.
Proteome analysis of male gametophyte development in rice anthers
Authors: Kerim Tursun , Imin Nijat , Weinman Jeremy J. , Rolfe Barry G.
Published: May 2003

We used proteomic analysis to investigate the changing patterns of protein synthesis during pollen development in anthers from rice plants grown under strictly controlled growth conditions. Cytological analysis and external growth measurements such as anther length, auricle distances and days before flowering were used to determine pollen developmental stages. This allowed the collection of synchronous anther materials representing six discrete pollen developmental stages. Proteins were extracted from the anther samples and separated by two‐dimensional gel electrophoresis to produce proteome maps. The anther proteome maps of different developmental stages were compared and 150 protein spots, which were changed consistently during development, were analysed by matrix‐assisted laser desorption/ionization‐time of flight mass spectrometry to produce peptide mass fingerprint (PMF) data. Database searches using these PMF data revealed the identities of 40 of the protein spots analyzed. These 40 proteins represent 33 unique gene products. Four protein spots that could not be identified by PMF analysis were analysed by N‐terminal microsequencing. Multiple charge‐isoforms of vacuolar acid invertase, fructokinase, β‐expansin and profilin were identified. These proteins are closely associated with sugar metabolism, cell elongation and cell expansion, all of which are cell activities that are essential to pollen germination. The existence of multiple isoforms of the same proteins suggests that during the process of pollen development some kind of post‐translational modification of these proteins occurs.
Prevalence of symptoms of asthma and allergies in schoolchildren in Gondar town and its vicinity, northwest Ethiopia
Authors: Hailu Sirak , Tessema Tesfaye , Silverman Michael
Published: May 2003

A low level of allergic disorders has been generally reported in children from developing countries. A survey was conducted to determine the prevalence of symptoms of asthma and allergies among schoolchildren in Gondar, northwest Ethiopia. A standardized self‐administered questionnaire developed by the International Study of Asthma and Allergies in Children (ISAAC) was used. In total, 3,365 children aged 13–14 years were surveyed at school, with a response rate of 98.4%. The female to male ratio was 1.5:1. The 12‐month‐period prevalences of wheeze (16.2%), rhino‐conjunctivitis (14.5%), and eczema (10.9%) were comparable to findings from Addis Ababa, but significantly higher than reported from Jimma. A striking seasonal pattern of rhinitis symptoms is observed during the months of September–December with a peak in November, corresponding with the main pollen season of the area. Overall, about 36% of the children reported symptoms of atopic disorders. However, the prevalence of diagnosed atopic diseases (8.6%) is less than a quarter of the symptom prevalence rate. This may reflect a generally poor awareness and low health service utilization, or underdiagnosis by health facilities. In conclusion, it appears that the prevalence of atopic diseases in Gondar area is high, and there is also wide variation within the country. This calls for further epidemiological studies with more objective assessments of atopy, such as allergen and bronchial hyperresponsiveness tests and ecological analysis. Pediatr Pulmonol. 2003; 35:427–432. © 2003 Wiley‐Liss, Inc.
Simple and specific test for the diagnosis of aspiration into the airways using a corn flour‐milk mixture in a hamster model
Authors: Springer Chaim , Avital Avraham
Published: May 2003

Aspiration into the airways is one of the major causes of lung disease in infants and young children. Unfortunately, the diagnosis of aspiration is often delayed due to unawareness and low sensitivity and specificity of existing diagnostic tests. We tested the efficacy of a new method, using a mixture of corn flour in milk instilled into the trachea, to diagnose aspiration in a hamster model. Forty‐five female hamsters were used for the experiments. Twenty hamsters underwent tracheal instillation of 0.1 ml of a mixture of 7.5% corn flour (containing starch granules with a diameter of 2–30 μm) in milk. Twenty control animals underwent tracheal instillation of saline. Five hamsters served as naive controls. Five animals from the corn flour and saline groups underwent bronchoalveolar lavage on days 1, 3, 7, and 14 after tracheal instillation. Starch granules were identified under light microscopy, using both iodine and Diff‐Quik staining in all the corn flour‐instilled animals and during all days after tracheal instillation. No starch granules were detected in naive or saline‐treated animals. Starch granules were detected over 14 days following tracheal instillation, with a half‐life rate of disappearance of about 6 days. Corn flour aspiration in hamsters can be easily identified by light microscopy, using simple laboratory techniques. Pediatr Pulmonol. 2003; 35:400–404. © 2003 Wiley‐Liss, Inc.
Pulmonary manifestations and function tests in children genetically diagnosed with FMF
Authors: Brik Riva , Gershoni‐Baruch Ruth , Shinawi Marwan , Barak Larissa , Bentur Lea
Published: May 2003

Familial Mediterranean fever (FMF) is characterized by recurrent episodes of peritonitis, pleuritis, and synovitis. Among the pulmonary manifestations of FMF, pleuritis is the most common. Long‐term sequelae of the respiratory system have not been described in FMF patients. We describe the pulmonary manifestations and function tests in a group of children who were found by genetic screening to be homozygous for the FMF gene. We surveyed 48 patients of Mediterranean extraction (aged 6–18 years) who were evaluated for a variety of pulmonary symptoms, and in whom clinical and genetic studies confirmed a diagnosis of FMF. All patients underwent complete pulmonary function tests, which included spirometry, body plethysmography, and single‐breath carbon monoxide diffusion (Dlco). Forty percent of the Jewish patients, but only 8% of the Arab patients (P < 0.001), suffered from pulmonary manifestations during an attack of FMF. Jewish patients who were homozygotes for the M694V mutation suffered significantly more from episodes of pleuritis, cough, and rapid, shallow breathing than Arab patients, who were either homozygotes for the V726A mutation or bore any other combination of mutations. Three patients (6%) had mild restrictive lung disease, all of them homozygotes for the M694V mutation. In 3 further patients, obstructive lung impairment was found. Pulmonary manifestations during FMF attacks are significantly more common in the Jewish population bearing the M694V mutation. Restrictive lung impairment was found in a small number of these patients with a severe course of the disease; however, the series is too small to draw conclusions about long‐term sequelae of the respiratory system in FMF patients. Pediatr Pulmonol. 2003; 35:452–455. © 2003 Wiley‐Liss, Inc.
Iatrogenic pleuropulmonary charcoal instillation in a teenager
Authors: Godambe Sandip A. , Mack Jennifer W. , Chung David S. , Lindeman Robert , Lillehei Craig W. , Colin Andrew A.
Published: May 2003

Activated charcoal given through a nasogastric tube is a standard intervention for many types of toxic ingestions in the emergency department. This case study describes a teenage girl whose multidrug overdose was complicated by accidental charcoal instillation into her left lung and pleural space through a misplaced nasogastric tube. The ensuing empyema did not respond to antibiotic therapy alone, probably due to the inherent properties of charcoal, and required a chest tube placement with continuous irrigation. Unlike previously reported cases, this patient did well clinically, without long‐term morbidity. Pediatr Pulmonol. 2003; 35:490–493. © 2003 Wiley‐Liss, Inc.
Correlation of six different cystic fibrosis chest radiograph scoring systems with clinical parameters
Authors: Terheggen‐Lagro Suzanne , Truijens Neeltje , van Poppel Noor , Gulmans Vincent , van der Laag Johan , van der Ent Cornelis
Published: May 2003

In past decades, several chest radiograph scoring systems for cystic fibrosis were developed. This study was performed to compare interobserver variability of six different radiograph scores and to correlate them with clinical parameters. Thirty chest radiographs of 30 patients with cystic fibrosis were scored according to Shwachman‐Kulczycki scoring, Chrispin‐Norman scoring, adjusted Chrispin‐Norman scoring, Brasfield scoring, Wisconsin scoring, and the Northern scoring system by two independent observers. Data on clinical parameters such as lung function, nutritional status, and infectious exacerbation rate, obtained simultaneously with the chest radiograph, were reviewed. Interobserver variability was low (Pearson's correlation coefficients, 0.76–0.84; all P < 0.01), and scores had good limits of agreement (Bland and Altman). Correlation of radiograph score with clinical parameters was good for most pulmonary function test data (correlation coefficients from 0.72–0.78 for percent of forced expired volume in 1 sec (FEV1%) predicted and from 0.69–0.74 for FVC% predicted) and for infectious exacerbation rate (correlation coefficients from 0.68–0.73). All six radiograph scoring systems, especially the Chrispin‐Norman score, showed a low interobserver variability and correlated well with lung function tests, especially FEV1% predicted and infectious exacerbation rate, and moderately with maximum work capacity and thoracic mobility. Pediatr Pulmonol. 2003; 35:441–445. © 2003 Wiley‐Liss, Inc.
Noninvasive ventilation in the pediatric intensive care unit for children with acute respiratory failure
Authors: Teague W. Gerald
Published: May 2003

Noninvasive ventilation, a novel treatment to increase alveolar ventilation, is accomplished with either subatmospheric or positive pressure administered via an external interface. In adults with acute respiratory failure, noninvasive positive pressure ventilation (NPPV) is superior to standard therapy in preventing intubation and reducing mortality. The role of NPPV in pediatric‐age patients with acute respiratory distress is not as well established. Early case reports showed that NPPV treatment does acutely improve both the clinical manifestations of respiratory distress and respiratory gas exchange in children with respiratory distress. However, it is not clear whether NPPV in this setting can prevent vs. delay endotracheal intubation. Other uses of NPPV in the pediatric intensive care unit include the treatment of upper airway obstruction, atelectasis, and exacerbations of neuromuscular disorders, and to facilitate weaning from invasive mechanical ventilation. Successful use of NPPV in young infants with respiratory distress is impeded by the lack of suitable size interfaces, and the response characteristics of commercially available bilevel ventilators. Despite these challenges, NPPV is a promising alternate to standard therapies in the treatment of acute respiratory distress in the pediatric‐age patient. Pediatr Pulmonol. 2003; 35:418–426. © 2003 Wiley‐Liss, Inc.
Rapid progression of intrapulmonary arteriovenous shunting in polysplenia syndrome associated with biliary atresia
Authors: Kimura Takuya , Hasegawa Toshimichi , Sasaki Takashi , Okada Akira , Mushiake Sotarou
Published: May 2003

This report describes a patient with biliary atresia (BA) associated with polysplenia syndrome who showed a rapid progression of intrapulmonary arteriovenous shunting (IPS), resulting in a fatal outcome. Intrauterine ultrasonography at 36 weeks of gestation revealed fetal abnormalities, including situs inversus, absent retrohepatic inferior vena cava, and azygous connection. She was diagnosed postnatally as BA because of persistent acholic stool and neonatal jaundice. She underwent hepatic portoenterostomy at age 158 days. The gallbladder and the hepatic ducts were hypoplastic, and the common bile duct was absent. Magnetic resonance image and operative findings also identified polysplenia and an absent portal trunk. Liver histology showed cirrhotic changes and bile duct proliferation. Postoperatively, she achieved good bile secretion, with gradual decrease of total bilirubin. However, she had repeated febrile episodes, and computerized tomography at age 7 months showed multiple liver cysts. Thereafter, she presented with exertional dyspnea. Contrast‐enhanced echocardiography showed IPS with a degree of 2/III at age 8 months and 3/III at 10 months. 99mTechnetium‐labeled macroaggregated albumin (99mTc‐MAA) scintigraphy revealed a shunt ratio of 25.5% at 9 months and 39.7% at 10 months. Percutaneous transhepatic drainage of the bile cysts was performed without success. Sludged bile was obtained. However, respiratory distress rapidly progressed, and she died at age 11 months. In the present patient, the association of polysplenia syndrome and absent portal vein with BA, as well as liver cirrhosis, seemed to be contributing factors to rapid progression of IPS in early life. Pediatr Pulmonol. 2003; 35:494–498. © 2003 Wiley‐Liss, Inc.
Non‐CF bronchiectasis: Clinical and HRCT evaluation
Authors: Chang A.B. , Masel J.P. , Boyce N.C. , Wheaton G. , Torzillo P.J.
Published: May 2003

Childhood bronchiectasis not related to underlying disease is still common in some populations in affluent countries. The aims of the study were to: 1) describe demographics, 2) evaluate the effectiveness of routine investigations, and 3) determine the relationship between spirometry and radiology scoring systems, in children with chronic suppurative lung disease (CSLD) living in Central Australia. Data of children living in Central Australia aged ≤15 years with noncystic fibrosis CSLD were identified over 2 years. Chest high‐resolution computed tomography (HRCT) scans were interpreted by a pediatric radiologist and scored according to three systems. Sixty‐five children were identified, resulting in a prevalence of at least 14.7/1,000 aboriginal children aged ≤15 years. Comorbidities (most common: suppurative otitis media in >70%) and early hospitalisation for pneumonia were common (median age, 0.5 years). Previous admissions for pneumonia were almost universally present and significantly more common than bronchiolitis (95% CI for proportional difference, 0.4–0.51). Although the majority did not have a treatable underlying cause, investigations had significant impact on management in 12.3% of children. None of the chest HRCT scores related to any spirometry data. In conclusion, CSLD is unacceptably common in indigenous children of this region, commences early in life, and is associated with significant comorbidities. Spirometry data do not reflect the severity of lung disease in HRCT scans. While improvement in the living standards is of utmost importance, the medical management that includes thorough investigations of these children should not be neglected. Pediatr Pulmonol. 2003; 35:477–483. © 2003 Wiley‐Liss, Inc.
Oral administration of specific yolk antibodies (IgY) may prevent Pseudomonas aeruginosa infections in patients with cystic fibrosis: A phase I feasibility study
Authors: Kollberg Hans , Carlander David , Olesen Hanne , Wejåker Per‐Erik , Johannesson Marie , Larsson Anders
Published: May 2003

Respiratory infection is the major cause of morbidity and mortality in cystic fibrosis (CF) patients. Chronic Pseudomonas aeruginosa (PA) infections ultimately occur in virtually all patients. It is impossible to eradicate PA when a patient has been chronically colonized. Immunotherapy with specific egg‐yolk antibodies (IgY) may be an alternative to antibiotics for the prevention of PA infections. We wanted to determine if treatment with specific IgY can prolong the period between the first and the second PA colonization? And long‐term, can the treatment diminish the number of positive PA cultures and postpone the onset of chronic colonization? CF patients gargled daily with an IgY‐antibody preparation, purified from eggs of hens immunized with PA bacteria. They were compared to a group of patients who did not gargle with the preparation. Both groups had their first colonization with PA eradicated by antibiotics. The basic treatment was essentially the same in both groups. In the initial study, the period between the first and second colonization with PA was significantly prolonged for the treated vs. the control group (Kaplan‐Meier P = 0.015, Breslow test). In the prolonged study, the treated group had only 2.5 sputum cultures positive for PA per 100 months of observation, and none of these patients became chronically colonized with PA. No adverse events were reported. In the control group, 13.7 cultures per 100 months of observation were positive for PA, and 5 (24%) patients became chronically colonized with PA. This feasibility study shows that antipseudomonal IgY has the potential to effectively prevent PA colonization without any severe adverse effects. A phase III study should be initiated. Pediatr Pulmonol. 2003; 35:433–440. © 2003 Wiley‐Liss, Inc.
Early postnatal dexamethasone influences matrix metalloproteinase‐2 and ‐9, and their tissue inhibitors in the developing rat lung
Authors: Valencia Arwin M. , Beharry Kay D. , Ang Jorge G. , Devarajan Kamakshi , Van Woerkom Richard , Abrantes Maria , Nishihara Kenji , Chang Eileen , Waltzman Joshua , Modanlou Houchang D.
Published: May 2003

In order to test the hypothesis that early postnatal exposure to dexamethasone (Dex) influences matrix metalloproteinases (MMP)‐2 and ‐9, as well as their tissue inhibitors (TIMP‐1 and ‐2) in the developing rat lung, newborn rats (3 litters/group) were treated with low Dex (0.1 mg/kg/day, IM), high Dex (0.5 mg/kg/day), or equivalent volumes of saline at 5 days postnatal age (P5), P6, and P7. Lung weight and lung MMP and TIMP levels were determined at sacrifice (7 days postinjection, P14; at weaning, P21; and at adolescence, P45, n = 10/group and time). Dex did not adversely affect lung weight or lung MMP‐2 levels, which peaked in all groups at P21 and then fell by P45. In contrast, Dex decreased TIMP‐2 at all time intervals, but achieved statistical significance only at P45. An imbalance in MMP‐2/TIMP‐2 ratio was noted at P21, with elevations occurring in the low and high Dex‐treated groups. Lung MMP‐9 levels remained comparable with controls during low Dex treatment. However, high Dex exposure resulted in elevated lung MMP‐9 levels at P21 and P45. Lung TIMP‐1 levels increased only with high Dex exposure at P14 and P21, whereas the lung MMP‐9/TIMP‐1 ratio was elevated at P21 in the high Dex group, and at P45 in both Dex‐treated groups. These data provide evidence that early postnatal dexamethasone results in an imbalance between gelatinase‐A and ‐B, and their tissue inhibitors in the developing rat lung. These changes may be responsible, in part, for some of the known maturational effects of steroids on lung structure in the newborn. Pediatr Pulmonol. 2003; 35:456–462. © 2003 Wiley‐Liss, Inc.
Teaching of pulmonary auscultation in pediatrics: A nationwide survey of all U.S. accredited residencies
Authors: Mangione Salvatore , Torre Dario M.
Published: May 2003

Structured teaching of pulmonary auscultation is greatly underrepresented during internal medicine (IM) or family practice (FP) training. It is not known, however, whether this underrepresentation applies to the other major field of primary care, pediatrics. To answer this question, we surveyed all accredited U.S. residencies in pediatrics by mailing a 1‐page questionnaire to 174 pediatrics program directors, and by comparing results to those previously gathered from internal medicine and family practice residencies. Pediatrics directors' response rate was 62%. More pediatrics than family practice residencies offered structured teaching of pulmonary auscultation (21.5% vs. 9.7%, P < 0.004). When compared to internal medicine programs, this difference showed a trend toward significance (21.5% for pediatrics and 14.1%, for internal medicine, P = 0.08). Teaching modalities included: lectures (91.2%); audiotapes (13%); seminars (8.3%); and miscellaneous (21.7%). University‐affiliated residencies taught auscultation significantly more frequently than nonuniversity‐affiliated programs (25.4% vs.10.5%, P = 0.07). Pediatrics directors gave great importance to pulmonary auscultation, and wished for more time devoted to its teaching (5.52 ± 0.84 and 5.01 ± 1.07, respectively, on a 1–6 scale, with 6 indicating the highest value). They also attributed great clinical importance to 13 commonly encountered pulmonary auscultatory events (all rated, on average, between 4–5.8 on a 1–6 scale, with 6 indicating highest importance). In summary, training programs in pediatrics offered significantly more structured teaching of pulmonary auscultation than IM or FP residencies. Whether this difference in teaching may have a beneficial impact on the auscultatory proficiency of pediatric residents, as compared to internal medicine and family practice trainees, needs to be determined. Pediatr Pulmonol. 2003; 35:472–476. © 2003 Wiley‐Liss, Inc.
Montelukast does not protect against hyperoxia‐induced inhibition of alveolarization in newborn rats
Authors: Jouvencel Philippe , Fayon Michael , Choukroun Marie‐Luce , Carles Dominique , Montaudon Danielle , Dumas Eric , Begueret Hughes , Marthan Roger
Published: May 2003

Impaired lung development has been demonstrated in neonatal animals exposed to hyperoxia. High lung cys‐leukotriene levels may be a contributing factor towards the increase in oxygen toxicity. We investigated the effect of cysteinyl‐leukotriene inhibition using the receptor antagonist, montelukast (MK, Singulair®), on hyperoxia‐induced changes in lung parenchymal structure in neonatal rat pups. Rat pups were exposed to 21% O2 (air) or 50% O2 (moderate hyperoxia) from days 1–14 after birth, and were administered the cys‐leukotriene receptor antagonist MK (1 mg/kg/day) or normal saline from days 4–14. Somatic growth and morphometric measurements were done on day 15. There was a significant increase in bronchoalveolar lavage fluid cysteinyl‐leukotriene levels (+61.9%) when animals were exposed to hyperoxia. O2 exposure significantly decreased the specific internal surface area by 13%. There was a nonsignificant 5.8% and 19.6% increase in mean chord length and mean alveolar diameter, respectively, as well as an 8.6% decrease in lung volume to body weight ratio. Inhibition of only one arm of the arachidonic‐acid cascade by MK was not sufficient to prevent these oxygen‐induced changes. Pediatr Pulmonol. 2003; 35:446–451. © 2003 Wiley‐Liss, Inc.
Determination of maximal voluntary ventilation in children with cystic fibrosis
Authors: Stein Richard , Selvadurai Hiran , Coates Allan , Wilkes Donna L. , Schneiderman‐Walker Jane , Corey Mary
Published: May 2003

Maximal voluntary ventilation (MVV) may be determined directly by the sprint method or calculated from pulmonary function data, using the functions MVV = forced expired volume in 1 sec (FEV1) × 35 or MVV = FEV1 × 40. The purpose of this paper was to test the validity of the equation over a wide range of lung function in children. Cystic fibrosis (CF), a chronic lung disease where children typically have a wide range of pulmonary function, was chosen as the study requirement. Spirometric data from 332 children with CF who underwent pulmonary function testing between 1987–2000 were stratified according to disease severity, and box‐plots comparing the ratio of MVV to FEV1 for each category were generated. As results indicated that the equation underestimates true MVV proportionally to the degree of airflow limitation, a new function to predict MVV for this population was derived and tested. The new equation was derived using data from patients who were tested on odd‐numbered days (group A). The validity of the new equation was then tested on the patients tested on even‐numbered days (group B). To test its validity, the results were compared to the “gold standard” sprint values using a Bland and Altman plot. MVV was expressed as a function of FEV1 and predicted FEV1: MVV = 27.7(FEV1) + 8.8(PredFEV1) (R2 = 0.98, P < 0.05). In this way, the accuracy of the new equation was confirmed. Whenever possible, we recommend MVV be determined by the sprint method in accordance with ATS guidelines. If this is not feasible, we recommend considering the new prediction equation. Pediatr Pulmonol. 2003; 35:467–471. © 2003 Wiley‐Liss, Inc.
PRAC2: A new gene expressed in human prostate and prostate cancer
Authors: Olsson Pär , Motegi Akira , Bera Tapan K. , Lee Byungkook , Pastan Ira
Published: May 2003

BACKGROUND

The database of human Expressed Sequence Tags was previously used to identify PRAC (Prostate 47:125–131, 2001), a novel gene specifically expressed in human prostate, prostate cancer, rectum, and distal colon. In this report, we have identified PRAC2, another gene with a similar expression pattern that is located adjacent to the original PRAC gene on chromosome 17q21.3.

METHODS

Using a computer‐based analysis, a cluster of sequence homologous ESTs was identified that is mainly derived from human prostate cDNA libraries. The tissue specificity was examined by multiple tissue RNA dot blots and RT‐PCR. The PRAC2 transcript and protein were identified using Northern blot analysis, RACE‐PCR, primer extension, and Western blots.

RESULTS

PRAC2 encodes a 564 nucleotide RNA found in prostate, rectum, distal colon, and testis. Weak expression was also found in placenta, peripheral blood leukocytes, skin, and in two prostate cancer cell lines: LNCaP and PC‐3. The transcript seems to encode a 10.5‐kDa nuclear protein. The PRAC2 gene is located on chromosome 17 at position 17q21, between the Hoxb‐13 gene and the recently discovered PRAC gene.

CONCLUSIONS

Because of the higher expression of PRAC2 in prostate and its proximity to Hoxb‐13, PRAC2 may have a function in prostate growth and development. Prostate 56: 123–130, 2003. © 2003 Wiley‐Liss, Inc.
Anti‐proliferative and apoptotic effects of anandamide in human prostatic cancer cell lines: Implication of epidermal growth factor receptor down‐regulation and ceramide production
Authors: Mimeault Murielle , Pommery Nicole , Wattez Nicole , Bailly Christian , Hénichart Jean‐Pierre
Published: May 2003

BACKGROUND

Anandamide (ANA) is an endogenous lipid which acts as a cannabinoid receptor ligand and with potent anticarcinogenic activity in several cancer cell types.

METHODS

The inhibitory effect of ANA on the epidermal growth factor receptor (EGFR) levels expressed on the EGF‐stimulated prostatic cancer cells LNCaP, DU145, and PC3 was estimated by ELISA tests. The anti‐proliferative and cytotoxic effects of ANA were also evaluated on these human prostatic cancer cell lines by growth tests, flow cytometric analyses, trypan blue dye exclusion assays combined with the Papanicolaou cytological staining method.

RESULTS

ANA induced a decrease of EGFR levels on LNCaP, DU145, and PC3 prostatic cancer cells by acting through cannabinoid CB1 receptor subtype and this leaded to an inhibition of the EGF‐stimulated growth of these cells. Moreover, the G1 arrest of metastatic DU145 and PC3 growth was accompanied by a massive cell death by apoptosis and/or necrosis while LNCaP cells were less sensitive to cytotoxic effects of ANA. The apoptotic/necrotic responses induced by ANA on these prostatic cancer cells were also potentiated by the acidic ceramidase inhibitor, N‐oleoylethanolamine and partially inhibited by the specific ceramide synthetase inhibitor, fumonisin B1 indicating that these cytotoxic actions of ANA might be induced via the cellular ceramide production.

CONCLUSIONS

The potent anti‐proliferative and cytotoxic effects of ANA on metastatic prostatic cancer cells might provide basis for the design of new therapeutic agents for effective treatment of recurrent and invasive prostatic cancers. Prostate 56: 1–12, 2003. © 2003 Wiley‐Liss, Inc.