Inflammatory cells in the bronchial glands of smokers with chronic bronchitis

To characterize the inflammatory process in the bronchial glands of smokers with chronic sputum production, we examined lobar bronchi from 18 subjects undergoing lung resection for localized pulmonary lesions, all with a history of cigarette smoking. Nine of the subjects had symptoms of chronic bronchitis and chronic airflow obstruction, and nine were asymptomatic, with normal lung function. The number of neutrophils, eosinophils, mast cells, macrophages, CD4+ and CD8+ T-lymphocytes, and the ratio of CD4+ to CD8+ cells were assessed in the bronchial glands, epithelium, and submucosa. Cells were identified through immunohistochemistry. Smokers with symptoms of chronic bronchitis had an increased number of neutrophils (p = 0.01) and macrophages (p = 0.03) and a decreased CD4+/CD8+ ratio (p = 0.01) in the bronchial glands as compared with asymptomatic smokers. Chronic bronchitic smokers also had an increased number of epithelial neutrophils (p = 0.04), whereas the numbers of macrophages and CD4+ and CD8+ T-lymphocytes in the epithelium and submucosa were similar in the two groups of smokers. No differences in numbers of eosinophils or mast cells were observed between bronchitic and asymptomatic smokers in any of the compartments examined. In conclusion, smokers with chronic sputum production have an increased infiltration of neutrophils and macrophages and an increased proportion of CD8+ T-lymphocytes in their bronchial glands, supporting the important role of bronchial-gland inflammation in the pathogenesis of chronic bronchitis.     

Characterization of an N-acetylglucosamine-6-O-sulfotransferase from human respiratory mucosa active on mucin carbohydrate chains

A microsomal GlcNAc-6-O-sulfotransferase activity from human bronchial mucosa, able to transfer a sulfate group from adenosine 3'-phosphate 5'-phosphosulfate onto methyl-N-acetylglucosaminides or terminal N-acetylglucosamine residues of carbohydrate chains from human respiratory mucins, has been characterized. The reaction products containing a terminal HO3S-6GlcNAc were identified by high performance anion-exchange chromatography. Using methyl-beta-N-acetylglucosaminide as a substrate, the optimal activity was obtained with 0.1% Triton X-100, 30 mM NaF, 20 mM Mn2+, 5 mM AMP in a 30 mM MOPS (3-(N-morpholino) propanesulfonic acid) buffer at pH 6.7. The apparent Km values for adenosine 3'-phosphate 5'-phosphosulfate and methyl-beta-N-acetylglucosaminide were observed at 9.1 x 10(-6) M and 0.54 x 10(-3) M, respectively. The enzyme had more affinity for carbohydrate chains with a terminal GlcNAc residue than for methyl-beta-N-acetylglucosaminide; it was unable to catalyze the transfer of sulfate to position 6 of the GlcNAc residue contained in a terminal Galbeta1-4GlcNAc sequence. However, oligosaccharides with a nonreducing terminal HO3S-6GlcNAc were substrates for a beta1-4 galactosyltransferase from human bronchial mucosa. These data point out that GlcNAc-6-O-sulfotransferase must act before beta1-4 galactosylation in mucin-type oligosaccharide biosynthesis.     

The impact of bronchial colonization in the quality of life of patients with chronic, stable bronchitis

BACKGROUND: The aim of this study was to determine the impact of respiratory function and bacterial colonization of the lower airway on the quality of life of patients with chronic, stable bronchitis (CB). MATERIALS AND METHODS: A series of 41 patients with stable CB was studied (age: 63.8; standard deviation (SD) 9.1 years; FVC% 91.0 (18.9); FEV1% 74.6 (23.7); FEV1/FEC 62.8 (11.2) with normal thoracic radiography. Patients with previous diagnosis of bronchiectasia, bronchial asthma and/or positive bronchodilatory tests (> 15%) were not included in the study. Bacterial growth in a sputum sample of grade 4-5 of the Murray-Washington scale was considered diagnostic of bronchial colonization. Measurement of the quality of life was performed with the Nottingham Health Profile (NHP) and the St. George Respiratory Questionnaire (SGRQ). RESULTS: The patients presented a moderate alteration in their quality of life with scores over 25 in most of the dimensions of the NHP and the SGRQ. In 9 out of 41 cases (22%), the sputum cultures demonstrated bronchial colonization with the most frequently isolated bacterias being Haemophilus influenzae and Moraxella catarrhalis. Multivariate analysis performed with the quality of life as the dependent variable showed an association between FEV1/FEC1 and the SGRQ score (R2 = 0.18), and energy (R2 = 0.09) and physical mobility (R2 = 0.05) of NHP. CONCLUSIONS: Bronchial obstruction is the main determinant in the quality of life in patients with stable CB. Colonization of the lower airway is observed in 22% of the patients and also influences the quality of life of the patients but to a much lesser extent.     

Effects of seaprose on the rheology of bronchial mucus in patients with chronic bronchitis. A double-blind study vs placebo

Approximately 2000 children aged between 0 and 14 years die annually of accidents and their sequelae in Japan. Moreover, accidents and their sequelae are the leading cause of death in the 1-4 year, 5-9 year and 10-14 year age groups. Accidental drowning and traffic accidents account for 2/3 of all such deaths. Measures for preventing injury in these age groups are clearly necessary, since the mortality rate due to accidents and their sequelae in children 0 to 4 years old is higher in Japan than in European countries. It is considered that the lives of 824 infants aged between 0 and 4 years might be saved annually in Japan if the mortality rate could be reduced to a level comparable to that in Sweden by accident prevention and control. Therefore, a systematic approach for prevention of childhood injury is a high priority in Japan.     

Heterogeneity of plasma glucagon. Circulating components in normal subjects and patients with chronic renal failure

Plasma immunoreactive glucagon (IRG) concentrations were measured in 36 patients with chronic renal failure (CRF) and 32 normal subjects. In addition, the components of circulating IRG were analyzed by gel filtration in the fasting state and after physiological stimuli. Fasting IRG was elevated (P less than 0.001) in CRF patients (534 +/- 32 pg/ml) compared with the levels found in healthy subjects (113 +/- 9 pg/ml). Oral glucose suppressed plasma IRG in CRF patients from a basal level of 568 +/- 52 to a nadir of 354 +/- 57 pg/ml (120 min). This degree of suppression (38%) was comparable to that found in normal subjects (basal = 154 +/- 20 to 100 +/- 23 pg/ml) at 120 min (35%). Intravenous infusion of arginine (250 mg/kg) resulted in a 71% rise in IRG in CRF patients and a 166% increase in normal subjects. Gel filtration of fasting plasma from CRF patients showed three major peaks. The earliest (A) was found in the void volume (mol wt greater than 40,000) and constituted 16.5 +/- 4.7% of the elution profile. The middle peak (B) eluted just beyond the proinsulin marker (approximately 9,000 mol wt) and constituted the largest proportion of the elution profile (56.5 +/- 3.4%). The third peak (C) coincided with the standard glucagon and [125I]glucagon markers (3,485 mol wt) and comprised 27.0 +/- 4% of the IRG profile. In contrast, only peaks A and C were found in fasting plasma of normal subjects (53.6 +/- 10.4% in A and 46.4 +/- 10.4 in C). After oral glucose, glucagon immunoreactivity in the 3,500 mol wt peak (C) was markedly suppressed, while the B peak in patients with CRF declined to a lesser extent. The A peak in both groups was unchanged. After an arginine infusion only the C peak increased in both groups of subjects. Gel filtration of plasma in 3 M acetic acid gave similar profiles to those obtained in glycine albumin buffer. Exposure of serum to trypsin indicated that the B and C peaks were digestible, while the A peak was resistant to the action of the enzyme. In one sample, peak C increased after a 2-h exposure of serum to trypsin. We conclude that circulating IRG in normal subjects and patients with CRF is heterogenous. The hyperglucagonemia of renal failure is largely due to an increase in IRG material of approximately 9,000 mol wt, consistent with proglucagon, although the 3,500 mol wt component is also considerably elevated (threefold). The significance of circulating IRG levels should be interpreted with caution until the relative biological activity of the three components is established.     

Bronchial inflammation in chronic bronchitis assessed by measurement of cell products in bronchial lavage fluid

BACKGROUND: Bronchial inflammation in chronic bronchitis has not been characterised as well as in asthma. The present study was undertaken to assess whether a characteristic pattern of bronchial inflammatory markers could be found in patients with chronic bronchitis. METHODS: Bronchoscopy with bronchial lavage was performed in 42 patients with chronic bronchitis and in 13 healthy controls. Twenty three of the patients had non-obstructive chronic bronchitis and 19 had chronic bronchitis and chronic obstructive pulmonary disease (COPD). Eighteen of the patients with bronchitis had recurrent infective exacerbations and 24 did not. Intrabronchial bacterial cultures were taken with a protected specimen brush. RESULTS: Increased activity of neutrophils, fibroblasts, and eosinophils was found in the patients with chronic bronchitis as assessed by the levels of myeloperoxidase (MPO) and interleukin-8 (IL-8), hyaluronan, and eosinophil cationic protein (ECP), respectively. The levels of tryptase did not differ from the controls. High correlations were found between the levels of MPO and IL-8, as well as ECP and IL-8. No differences were found between the patients with COPD and those with non-obstructive chronic bronchitis. CONCLUSIONS: Recruitment and activation of both neutrophils and eosinophils seem to be a characteristic of chronic bronchitis. This activation is associated with IL-8. The patients with intrabronchial cultures of Streptococcus pneumoniae had the highest individual levels of MPO, ECP, and IL-8 of all subjects in the study, indicating that colonisation with S pneumoniae could promote bronchial inflammation.     

A comparative assessment of the clinical efficacy of standard and low-volume plasmapheresis in treating patients with bronchial asthma and chronic bronchitis

21 patients with bronchial asthma and chronic obstructive bronchitis resistant to standard therapy have been exposed to discrete plasmapheresis. Changes in the bronchial tree inflammation measured by fibre bronchoscopy and activity of neuron-specific enolase (NSE) in bronchial wash-offs made it clear that low-volume plasmapheresis is not inferior in efficacy to the standard method. The trend in the fall of NSE activity as a marker of diffuse neuroendocrine system objectively reflects attenuating inflammation in bronchial mucosa in response to treatment of chronic obstructive diseases of the lungs using different techniques of plasmapheresis.     

Heterogeneity of protein profiles of Helicobacter pylori isolated from individual patients

A standardized neuropsychological test battery was administered to 167 patients with different forms of mild-to-moderate dementia: probable Alzheimer dementia (AD: n = 49), multi-infarct dementia (n = 43), idiopathic Parkinson disease with dementia (n = 35), depressive pseudodementia (n = 26), and progressive supranuclear palsy (n = 14). Results obtained were used (a) to analyze the profiles of cognitive impairment shown by the different dementia groups; (b) to assess the incidence of some neuropsychological patterns that we hypothesized to be more characteristic of AD, in the various groups; and hence (c) to evaluate the reliability of these patterns as diagnostic markers of AD. Four of the patterns investigated were derived from a verbal learning task (Rey's Auditory Verbal Learning test): (1) absence of the primacy effect; (2) tendency to produce intrusion errors during free recall of a word list; (3) absolute decay of memory trace; and (4) tendency to produce false alarms during delayed recognition of the same word list. Two additional patterns were derived from visual-spatial tasks (copying drawings and Raven's Coloured Progressive Matrices): (5) occurrence of the closing-in phenomenon in copying drawings; and (6) tendency to choose globalistic or odd responses in Raven's matrices. Though all the six patterns were somewhat useful for identifying AD patients, no pattern met the criteria of being both highly sensitive and highly specific, which should characterize an ideal marker. In fact, intrusions and false alarms were observed in many AD patients, but also in patients affected by other forms of dementia. The absence of the primacy effect, the closing-in phenomenon, and the absolute decay of memory trace were more specific, but could be observed in only one-third of AD patients. We also computed the number of positive patterns shown by each patient and assumed the presence of two or more patterns as a global index suggestive of a dementia of the Alzheimer type. With this cumulative method, a higher level of sensitivity and specificity was achieved in the identification of AD patients.     

Heterogeneity of staphylococci and other bacteria isolated from six-week-old broiler chickens

In broiler operations, various health problems develop during the final 2 wk of the growing period, resulting in increased mortality and condemnation losses. At this stage, sickly birds were found to be systemically infected by various bacteria regardless of varied clinical signs, and the purpose of this study was to carry out thorough microbiological investigations on this problem. Thirty-one 6-wk-old broilers showing signs of illness were obtained from three farms, and bacterial isolations were carried out from the blood, liver, and hock joint. Bacteria were isolated from 87, 90, and 71% of the blood, liver, and hock joint samples, respectively. Mean bacterial counts in log10 of the blood (per milliliter) and liver (per gram) were 2.15 and 2.93, respectively. Among 132 bacterial isolates, major species were Staphylococcus (60%), Corynebacterium (18%), Escherichia coli (5%), and Stomatococcus (4%). Among 79 Staphylococcus isolates, 77 were coagulase-negative. Major species of staphylococci were S. lentus (19%), S. simulans (18%), S. cohnii (13%), S. gallinarum (10%), and S. captis (7%). In addition, six species of gram-positive and five species of gram-negative organisms were isolated. Thus, the apparent systemic infections were not caused by predominant pathogenic bacterial species, and adequately described as mixed infections. There were some significant relationships between isolated bacterial species and sampling sites, suggesting that certain organisms were abundant in the environment of a particular poultry house. These results indicate that systemic infections in market age broilers are caused by mixed bacterial species and suggest that they are caused by suppressed host antibacterial systems rather than pathogenic factors of microorganisms.     

Increased iron and ferritin content of sputum from patients with cystic fibrosis or chronic bronchitis

PURPOSE: Extracellular free iron, or iron bound to ferritin, may promote oxidative injury and bacterial growth in airways of patients with chronic airway inflammation due to cystic fibrosis (CF) or chronic bronchitis (CB). In this study, we assessed sputum content of total iron, ferritin, and transferrin in patients with CF or CB as well as sputum from normal subjects with acute airway inflammation caused by viral upper respiratory tract infections (URTIs). METHODS: Spontaneously produced sputum was obtained from 33 subjects, including 10 subjects with CF, 18 subjects with CB (10 acute exacerbations, 8 with stable CB), and 5 subjects with URTIs (control subjects). After lysing and dilution, total iron concentrations were determined by controlled coulometry, ferritin was measured by radioimmunoassay, and transferrin was measured by enzyme-linked immunosorbent assay. RESULTS: Iron was not present in detectable amounts in control sputums, but ferritin was present (6+/-2 ng/mg protein, mean+/-SE), as was transferrin (2.37+/-0.44 microg/mg). Compared with control subjects, concentrations of iron in sputum were increased in patient groups with higher amounts in CF patients (242+/-47 ng/mg, p<0.01) than CB patients with acute exacerbations or patients with stable CB (98+/-50 and 42+/-12 ng/mg, p<0.05 for both). Ferritin content of sputum was also increased in each group, with CF patients (113+/-22 ng/mg, p<0.001) higher than CB patients (acute, 45+/-10 ng/mg; stable, 87+/-24 ng/mg; p<0.01 for both). Compared with control subjects, sputum transferrin was decreased in CF patients (1.09+/-0.40 microg/mg, p<0.05), but not CB patients. CONCLUSIONS: These findings indicate there are increased airway concentrations of total iron and ferritin-bound iron in patients with CB and, to a greater extent, in patients with CF. Particularly in CF patients who also demonstrated decreased airway concentrations of transferrin, ferritin-bound iron in airways may promote oxidative injury and enhance bacterial growth.     

Psychophysiologic predictors of weaning from mechanical ventilation in chronic bronchitis and emphysema

This study identified psychophysiologic variables related to successful weaning in 27 ventilator-dependent patients with chronic bronchitis and emphysema (CBE) from two long-term care pulmonary-specialty hospitals in South Central Florida. Subjects were studied from admission until weaning occurred (successful weaning without mechanical ventilation) or until they were transferred without being weaned or died (unsuccessful weaning). The study subjects, 15 males and 12 females, ranged in age from 56 to 89. Baseline data on the variables (age, mastery, hope, social support, dyspnea, and rapid shallow breathing index [RSBI]) were not statistically significant by gender. The logistic regression model identified mastery and RSBI to be the best predictors of successful weaning (model chi 2 = 16.33, df = 2, and p value = .0003; prediction rate 82%).     

The structure of the keratan sulphate chains attached to fibromodulin isolated from articular cartilage

Fibromodulin has been isolated from bovine and equine articular cartilage and the attached keratan sulphate chains subjected to digestion by keratanase II. The oligosaccharides generated have been reduced and subsequently isolated by strong anion-exchange chromatography. Their structures have been determined by high-field 1H-NMR spectroscopy and high-pH anion-exchange chromatography. Both alpha(2-6)- and alpha(2-3)-linked N-acetylneuraminic acid have been found in the capping oligosaccharides, and, fucose which is alpha(1-3)-linked to N-acetylglucosamine has been found as a branch in both repeat region and capping oligosaccharides. These data demonstrate that there are fundamental differences between the structures present in the N-linked keratan sulphate chains attached to fibromodulin from articular cartilage and those from tracheal cartilage, which lack both alpha(2-6)-linked N-acetylneuraminic acid and alpha(1-3)-linked fucose. It has been confirmed that the keratan sulphate chains are short, being only eight or nine disaccharides in length. Very significant differences in the levels of galactose sulphation have been identified at the non-reducing end of the chain. The galactose residue adjacent to the non-reducing cap is sulphated in only 1-3% of chains, compared with a sulphation level of over 40% closer to the reducing end. This highlights the difference between the chain termini and the repeat region in terms of structure and points to the potential for functional importance. The repeat region and capping fragments of the N-linked keratan sulphates from bovine and equine articular cartilage fibromodulin have been found to have the following general structure: NeuAc-(alpha 2-3/6)Gal[6SO3-](beta 1-4)GlcNAc6SO3-(beta 1-3)Gal[6SO3-] (beta 1-4)?[Fuc(alpha 1-3)]0-1GlcNAc6SO3-(beta 1-3)Gal-[6SO3-](beta 1-4)? 6-7GlcNAc6SO3-.     

Functional characteristics of bronchial epithelium obtained by brushing from asthmatic and normal subjects

Airways epithelial cells may be involved in the pathogenesis of asthma, but their role remains to be determined. Epithelial cells can release large amounts of 15-hydroxy-eicosatetranoic acid (15-HETE) and smaller amounts of prostaglandin E2 (PGE2) as well as fibronectin, a mediator involved in epithelial repair after injury. Epithelial cells obtained after bronchial brushing of 16 asthmatic (age 38 +/- 5 yr) and 11 normal subjects (age 36 +/- 5 yr) were studied. The percentage of epithelial cells was assessed by immunocytochemistry using an anti-cytokeratin antibody. The viability of the cells was assessed by trypan blue exclusion. The release of 15-HETE PGE2 and fibronectin was studied in resting cells and after A23187 calcium ionophore stimulation. Epithelial cells always comprised more than 86% of cells recovered, and the viability of epithelial cells was significantly (p < 0.001, Mann-Whitney U test) greater in normal subjects (54 +/- 5%) compared with asthmatic subjects (13 +/- 1%). The release of 15-HETE and fibronectin by resting epithelial cells was significantly greater in asthmatics (p < 0.05, Mann-Whitney U test) than in normal subjects. A23187 significantly (p < 0.05, Wilcoxon W test) increased the release of 15-HETE and fibronectin. There was no significant difference in the release of PGE2 by resting cells from either asthmatics or normal subjects, but challenge with A23187 induced a significant (p < 0.03, Wilcoxon W test) increase in PGE2 from cells of asthmatics but not from cells of normal subjects. This study shows that epithelial cells are activated and less viable in asthma and suggests a role for these cells in asthma.     

Synthesis of a fully protected glycooctaosyl serine isolated from blood group A human ovarian mucin

We have developed a competitive heparin binding assay employing protamine-coated magnetic beads for detection and measurement of heparin. The assay utilizes 125-iodine specifically bound to newly synthesized low-molecular-mass (LMM) heparin-tyramine. The tracer was stable over a period of 3 weeks, as demonstrated by gel filtration chromatography. The protamine-coated beads were found to be stable over at least two months. The heparin-tyramine bead assay had in buffer a lower detection limit of 0.04 microgram/ml and in plasma of 0.23 microgram heparin/ml. 50% binding was obtained at 0.7 microgram/ml and 20% binding at 4 micrograms/ml in plasma. The within assay coefficient of variation ranged from 9 to 28% for unfractionated, high molecular mass (HMM) heparin and from 12 to 15% for LMM-heparins in buffer system and in plasma. Various heparin fractions displaced the tracer from the protamine-coated magnetic beads to different extents. The validity of the assay was proven after intravenous administration of unfractionated and LMM-heparin in man. The elimination rate was similar using the heparin-tyramine bead assay compared with the anti-factor Xa coagulation assay. After intravenous dosing of LMM-heparin the maximal concentration was lower using the heparin-tyramine bead assay compared with the anti-factor Xa coagulation assay. The bead assay was found to be reproducible, valid, and rapid for measurement of the concentration of heparin preparations in purified systems and for HMM-heparin in plasma. Measurement of the concentration of LMM-heparin in plasma has a high coefficient of variation using the binding assay.     

Treatment of patients with pneumonia, chronic obstructive bronchitis and mucoviscidosis with cefpirome

Twenty two patients with inflammatory respiratory tract infection were treated with cefpirome. Among the patients 14 were with severe pneumonia, 4 with exacerbated obstructive chronic purulent bronchitis and 4 with mucoviscidosis. All the patients were subjected to clinical, laboratory and x-ray examinations, electrocardiography, estimation of the external respiration and sputum bacteriological tests. The cefpirome susceptibility was determined by the agar diffusion assay with standard disks from Roussel Uclaf. Cefpirome was administered by slow intravenous infusion in a daily dose of 2 to 4 g every 12 hours depending on the disease severity. After 2 or 3 days of the patient afebrile temperature and normal differential blood count the therapy was discontinued. The favourable time course of the disease was recorded in 12 out of the 14 patients with pneumonia. Recovery and clinical improvement were stated in 64.3 and 21.4 per cent of the cases respectively. In 2 patients the treatment failed. In all the patients with exacerbated severe chronic purulent bronchitis the cefpirome therapy resulted in the disease remission. The clinical effect of the mucoviscidosis treatment was observed in 3 out of the 4 patients. The drug tolerance in the doses used was good.