muramidase and Pseudomonas-Infections

Topics: alpha 1-Antitrypsin; Blood Proteins; Bronchi; Humans; Immunoglobulin A, Secretory; Immunoglobulin E; Immunoglobulin G; Immunoglobulin M; L-Lactate Dehydrogenase; Lipids; Lung Diseases, Obstructive; Malate Dehydrogenase; Mucins; Muramidase; Peptide Hydrolases; Protease Inhibitors; Pseudomonas Infections; Serotonin

Lysozyme, a 14-kDa protein, is one of the most abundant antimicrobials in the lungs. Its concentration in airway surface sufficient to kill several bacterial pathogens in vitro. The purpose of this study was to determine if administration of exogenous lysozyme would further enhance bacterial killing in vivo.. To assess the effect of acute lung infection on endogenous lysozyme protein levels, mice were infected by intratracheal instillation of Pseudomonas aeruginosa and bronchoalveolar (BAL) fluid assessed for lysozyme concentration and for muramidase activity. In order to inform in vivo testing, species-specific bacterial killing efficacy was determined by incubating mucoid P. aeruginosa with 2×10. These results indicate that endogenous lysozyme is increased during acute lung infection and that early administration of exogenous lysozyme further enhances bacterial killing in vivo.

Topics: Animals; Anti-Infective Agents; Bronchoalveolar Lavage Fluid; Drug Synergism; Drug Therapy, Combination; Female; Lung; Male; Mice; Microbial Viability; Muramidase; Pneumonia, Bacterial; Pseudomonas aeruginosa; Pseudomonas Infections; Treatment Outcome

Pseudomonas aeruginosa colonises the upper airway of cystic fibrosis (CF) patients, providing a reservoir of host-adapted genotypes that subsequently establish chronic lung infection. We previously experimentally-evolved P. aeruginosa in a murine model of respiratory tract infection and observed early-acquired mutations in pmrB, encoding the sensor kinase of a two-component system that promoted establishment and persistence of infection. Here, using proteomics, we show downregulation of proteins involved in LPS biosynthesis, antimicrobial resistance and phenazine production in pmrB mutants, and upregulation of proteins involved in adherence, lysozyme resistance and inhibition of the chloride ion channel CFTR, relative to wild-type strain LESB65. Accordingly, pmrB mutants are susceptible to antibiotic treatment but show enhanced adherence to airway epithelial cells, resistance to lysozyme treatment, and downregulate host CFTR expression. We propose that P. aeruginosa pmrB mutations in CF patients are subject to an evolutionary trade-off, leading to enhanced colonisation potential, CFTR inhibition, and resistance to host defences, but also to increased susceptibility to antibiotics.

Topics: A549 Cells; Adaptation, Physiological; Animals; Anti-Infective Agents; Bacterial Adhesion; Bacterial Proteins; Biological Evolution; Colony Count, Microbial; Cystic Fibrosis Transmembrane Conductance Regulator; Down-Regulation; Epithelial Cells; Fimbriae, Bacterial; Host-Pathogen Interactions; Humans; Lung; Mice; Microbial Sensitivity Tests; Models, Biological; Movement; Muramidase; Mutation; Principal Component Analysis; Proteomics; Pseudomonas aeruginosa; Pseudomonas Infections; Transcription Factors

Topics: Animals; Anti-Bacterial Agents; beta-Lactams; Caenorhabditis elegans; Cell Wall; Disease Models, Animal; DNA Transposable Elements; Gene Knockout Techniques; Genetic Complementation Test; Genetic Testing; Mice, Inbred C57BL; Microbial Viability; Muramidase; Mutagenesis, Insertional; Pseudomonas aeruginosa; Pseudomonas Infections; Vancomycin; Virulence

Graphene oxide (GO)-modified sinapinic acid (3,5-dimethoxy-4-hydroxycinnamic acid, SA) (SA@GO) was synthesized and characterized; it was then investigated as a new surface assisted laser desorption/ionization mass spectrometry (SALDI-MS) for proteomics and pathogenic bacteria biosensing. SA@GO could effectively decrease the time necessary for sweet spotting searching, reducing the amount of organic matrix and solvent and enhance the sensitivity. SA@GO shows high performance as a matrix alone without the need to add trifluoroacetic acid (TFA). However, the analysis of the intact bacteria cells shows improvement in the signal intensity (2-5 fold) and offers a low limit of detection. All these analyses could be performed with low concentrations (1-10 fmol) and tiny volumes (0.5-1 μL). This study demonstrated that the exploration of new hybrid materials is pivotal to achieve high performance and high ionization. Because of the plane of GO, it assists protein-protein interactions that make it undergo softer ionization.

Topics: Biosensing Techniques; Cellulase; Coumaric Acids; Fluorescence; Graphite; Humans; Lactalbumin; Lasers; Muramidase; Nanocomposites; Proteomics; Pseudomonas aeruginosa; Pseudomonas Infections; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Spectrophotometry, Ultraviolet; Spectroscopy, Fourier Transform Infrared; Staphylococcal Infections; Staphylococcus aureus; Trypsin

Type I interferon (IFN-I) predisposes to bacterial superinfections, an important problem during viral infection or treatment with interferon-alpha (IFN-α). IFN-I-induced neutropenia is one reason for the impaired bacterial control; however there is evidence that more frequent bacterial infections during IFN-α-treatment occur independently of neutropenia.. We analyzed in a mouse model, whether Pseudomonas aeruginosa control is influenced by co-infection with the lymphocytic choriomeningitis virus (LCMV). Bacterial titers, numbers of neutrophils and the gene-expression of liver-lysozyme-2 were determined during a 24 hours systemic infection with P. aeruginosa in wild-type and Ifnar(-/-) mice under the influence of LCMV or poly(I:C).. Virus-induced IFN-I impaired the control of Pseudomonas aeruginosa. This was associated with neutropenia and loss of lysozyme-2-expression in the liver, which had captured P. aeruginosa. A lower release of IFN-I by poly(I:C)-injection also impaired the bacterial control in the liver and reduced the expression of liver-lysozyme-2. Low concentration of IFN-I after infection with a virulent strain of P. aeruginosa alone impaired the bacterial control and reduced lysozyme-2-expression in the liver as well.. We found that during systemic infection with P. aeruginosa Kupffer cells quickly controlled the bacteria in cooperation with neutrophils. Upon LCMV-infection this cooperation was disturbed.

Topics: Animals; Granulocytes; Immunity, Innate; Interferon Type I; Kupffer Cells; Liver; Lymphocytic choriomeningitis virus; Mice, Inbred C57BL; Muramidase; Neutropenia; Poly I-C; Pseudomonas aeruginosa; Pseudomonas Infections; Spleen; Virulence

In septic shock (SS), dysfunction of many organ systems develops during the course of the illness, although the mechanisms are not clear. In earlier studies, we reported that lysozyme-c (Lzm-S), a protein that is released from leukocytes and macrophages, was a mediator of the myocardial depression and vasodilation that develop in a canine model of Pseudomonas aeruginosa SS. Whereas both of these effects of Lzm-S are dependent on its ability to intrinsically generate hydrogen peroxide, we subsequently showed that Lzm-S can also deposit within the vascular smooth muscle layer of the systemic arteries in this model. In the present study, we extend our previous findings. We used a canine carotid artery organ bath preparation to study the time course and dose dependence of Lzm-S deposition within the vascular smooth muscle layer. We used a human aortic vascular smooth muscle cell preparation to determine whether Lzm-S can persistently inhibit contraction in this preparation. We also used a canine P. aeruginosa model to determine whether Lzm-S deposition might occur in other organs such as the kidney, liver, and small intestine. The results showed that, in the carotid artery organ bath preparation, Lzm-S deposition occurred within minutes of instillation and there was a dose-response effect. In the human aortic vascular smooth muscle cell preparation, Lzm-S inhibited contraction during a 4-day period. In the in vivo model, Lzm-S accumulated in the kidney and the superior mesenteric artery. In a canine renal epithelial preparation, we further showed that Lzm-S can be taken up by the renal tubules to activate inflammatory pathways. We conclude that Lzm-S can deposit in the systemic vasculature and kidneys in SS, where this deposition could lead to acute organ dysfunction.

Topics: Animals; Aorta; Carotid Arteries; Cells, Cultured; Disease Models, Animal; Dogs; Humans; Intestine, Small; Kidney Tubules, Distal; Macrophages; Mesenteric Arteries; Muramidase; Muscle, Smooth, Vascular; Myocytes, Smooth Muscle; Organ Culture Techniques; Pseudomonas aeruginosa; Pseudomonas Infections; Sepsis

The spread of drug-resistant bacterial pathogens is a growing global concern and has prompted an effort to explore potential adjuvant and alternative therapies derived from nature's repertoire of bactericidal proteins and peptides. In humans, the airway surface liquid layer is a rich source of antibiotics, and lysozyme represents one of the most abundant and effective antimicrobial components of airway secretions. Human lysozyme is active against both Gram-positive and Gram-negative bacteria, acting through several mechanisms, including catalytic degradation of cell wall peptidoglycan and subsequent bacterial lysis. In the infected lung, however, lysozyme's dense cationic character can result in sequestration and inhibition by polyanions associated with airway inflammation. As a result, the efficacy of the native enzyme may be compromised in the infected and inflamed lung. To address this limitation, we previously constructed a charge-engineered variant of human lysozyme that was less prone to electrostatic-mediated inhibition in vitro. Here, we employ a murine model to show that this engineered enzyme is superior to wild-type human lysozyme as a treatment for mucoid Pseudomonas aeruginosa lung infections. The engineered enzyme effectively decreases the bacterial burden and reduces markers of inflammation and lung injury. Importantly, we found no evidence of acute toxicity or allergic hypersensitivity upon repeated administration of the engineered biotherapeutic. Thus, the charge-engineered lysozyme represents an interesting therapeutic candidate for P. aeruginosa lung infections.

Topics: Animals; Anti-Bacterial Agents; Bronchoalveolar Lavage Fluid; Colony Count, Microbial; Cytokines; Glycosaminoglycans; Humans; Inflammation; Lung; Male; Mice; Mice, Inbred C57BL; Muramidase; Protein Engineering; Pseudomonas aeruginosa; Pseudomonas Infections; Static Electricity

Vitamin A deficiency (VAD) alters the phenotype of airway epithelium and attenuates the epithelial defense system, and many studies have reported the association of VAD with respiratory disease. In this study, we investigated changes in submucosal glands (SMG) in a mouse model of VAD. C57BL/6 mice were fed a vitamin A-devoid diet and the others were fed a control diet (1.2 mg retinol/kg). The areas of serous and mucous cells of SMG were measured in 4-, 8-, and 20-wk-old male mice. The volume and lysozyme concentration of glandular secretions were also measured. The 2 groups did not differ in body weight or general morbidity at 3-10 wk of age, although serum retinol concentrations were greater in the control mice than in the VAD mice after 4 wk. Upon histological evaluation, we found that the areal ratio of serous cells:total SMG cells was significantly lower after 8 wk in the VAD mice compared with the control mice, although the total area of SMG did not differ between groups throughout the 20-wk experiment. The number of secretory bubbles did not differ between the groups, but total secretion volume was reduced by 35% in 8-wk-old VAD mice compared with controls. Furthermore, the concentration of lysozyme in secretions from 8-wk-old VAD mice was also less than in controls, compounding the effect of diminished secretion volume. In this study, we found serous cell hypotrophy/hypoplasia and dysfunction in VAD mice, which may contribute to the susceptibility to airway infection linked to VAD.

Topics: Animals; Bodily Secretions; Cell Count; Chemokine CXCL2; Disease Resistance; Down-Regulation; ErbB Receptors; Male; Mice; Mice, Inbred C57BL; Mucus; Muramidase; Phosphorylation; Protein Processing, Post-Translational; Pseudomonas aeruginosa; Pseudomonas Infections; Respiratory Mucosa; Respiratory Tract Infections; RNA, Messenger; Severity of Illness Index; Up-Regulation; Vitamin A Deficiency

Previous studies from this laboratory have shown that aerosolized recombinant human lysozyme (rhLZ) mitigates Pseudomonas aeruginosa (PA)-induced pneumonia. In the current investigation, our laboratory tested the hypothesis that aerosolized rhLZ can potentiate the effects of tobramycin (TBMN), thereby reducing the effective dose of this agent in the treatment of PA-induced pneumonia. Syrian hamsters were instilled intratracheally with PA, then exposed to an aerosol containing either 1% rhLZ, 3 μg TBMN, or a combination of both agents. In contrast to the initial studies with rhLZ, which involved 3 separate aerosol exposures, only a single treatment was used in the current investigation. Twenty-four hours after completion of the aerosol regimen, the following parameters were measured: (1) whole-lung colony-forming units (CFU), (2) total bronchoalveolar lavage fluid (BALF) CFU, (3) lung histopathology, and (4) total BALF neutrophils. The combination of rhLZ and TBMN significantly reduced whole-lung and BALF CFU, as well as the inflammatory index, compared to TBMN alone. Similar results were seen in vitro with regard to bactericidal activity. These findings provide a rationale for clinical testing of rhLZ as an adjunct to commercial antibiotic treatment.

Topics: Administration, Inhalation; Animals; Anti-Bacterial Agents; Bronchoalveolar Lavage Fluid; Cricetinae; Disease Models, Animal; Drug Synergism; Female; Humans; Lung; Mesocricetus; Muramidase; Neutrophils; Pneumonia, Bacterial; Pseudomonas aeruginosa; Pseudomonas Infections; Recombinant Proteins; Tobramycin

Nutritional fatty acids are known to have an impact on membrane lipid composition of body cells, including cells of the immune system, thus providing a link between dietary fatty acid uptake, inflammation and immunity. In this study we reveal the significance of macrophage membrane lipid composition on gene expression and cytokine synthesis thereby highlighting signal transduction processes, macrophage activation as well as macrophage defense mechanisms. Using RAW264.7 macrophages as a model system, we identified polyunsaturated fatty acids (PUFA) of both the n-3 and the n-6 family to down-regulate the synthesis of: (i) the pro-inflammatory cytokines IL-1β, IL-6 and TNF-α; (ii) the co-stimulatory molecule CD86; as well as (iii) the antimicrobial polypeptide lysozyme. The action of the fatty acids partially depended on the activation status of the macrophages. It is particularly important to note that the anti-inflammatory action of the PUFA could also be seen in case of infection of RAW264.7 with viable microorganisms of the genera R. equi and P. aeruginosa. In summary, our data provide strong evidence that PUFA from both the n-3 and the n-6 family down-regulate inflammation processes in context of chronic infections caused by persistent pathogens.

Topics: Actinomycetales Infections; Animals; B7-2 Antigen; Cell Line; Dietary Fats; Down-Regulation; Fatty Acids, Unsaturated; Immunity, Innate; Inflammation; Interleukin-1beta; Interleukin-6; Macrophage Activation; Macrophages; Membrane Lipids; Mice; Muramidase; Myeloid Differentiation Factor 88; Pseudomonas aeruginosa; Pseudomonas Infections; Rhodococcus equi; Tumor Necrosis Factor-alpha

As an alternative to conventional antibiotics, aerosolized recombinant human lysozyme (rhLZ) was used to treat experimentally induced pneumonia. Syrian hamsters were inoculated intratracheally with a nonmucoid strain of Pseudomonas aeruginosa (PA), then exposed to a 1.0% solution of rhLZ in water for 2 hours per day for 3 consecutive days (controls were treated with aerosolized water alone). Compared to controls, the rhLZ-treated group showed statistically significant reductions in the following parameters: (1) lung histopathological changes, (2) bacterial colony-forming units in whole lung and bronchoalveolar lavage fluid (BALF), (3) total BALF leukocytes, (4) percent BALF neutrophils, and (5) alveolar septal apoptosis. Exposure to aerosolized rhLZ also resulted in a large increase in BALF lysozyme activity. These findings indicate that aerosolized rhLZ may be potentially useful in reducing the level of bacterial colonization and inflammation in the lungs of patients with PA pneumonia.

Topics: Administration, Inhalation; Animals; Anti-Infective Agents; Apoptosis; Bronchoalveolar Lavage Fluid; Cricetinae; Drug Evaluation, Preclinical; Female; Humans; Leukocyte Count; Lung; Mesocricetus; Muramidase; Pneumonia, Bacterial; Pseudomonas aeruginosa; Pseudomonas Infections; Recombinant Proteins

Lysozymes contain a disproportionately large fraction of cationic residues, and are thereby attracted toward the negatively charged surface of bacterial targets. Importantly, this conserved biophysical property may inhibit lysozyme antibacterial function during acute and chronic infections. A mouse model of acute pulmonary Pseudomonas aeruginosa infection demonstrated that anionic biopolymers accumulate to high concentrations in the infected lung, and the presence of these species correlates with decreased endogenous lysozyme activity. To develop antibacterial enzymes designed specifically to be used as antimicrobial agents in the infected airway, the electrostatic potential of human lysozyme (hLYS) was remodeled by protein engineering. A novel, high-throughput screen was implemented to functionally interrogate combinatorial libraries of charge-engineered hLYS proteins, and variants with improved bactericidal activity were isolated and characterized in detail. These studies illustrate a general mechanism by which polyanions inhibit lysozyme function, and they are the first direct demonstration that decreasing hLYS's net cationic character improves its antibacterial activity in the presence of disease-associated biopolymers. In addition to avoiding electrostatic sequestration, at least one charge-engineered variant also kills bacteria more rapidly in the absence of inhibitory biopolymers; this observation supports a novel hypothesis that tuning the cellular affinity of peptidoglycan hydrolases may be a general strategy for improving kinetics of bacterial killing.

Topics: Animals; Anti-Bacterial Agents; Bacteria; Gram-Negative Bacteria; Humans; Lung; Mice; Micrococcus luteus; Models, Molecular; Muramidase; Mutation; Protein Engineering; Pseudomonas aeruginosa; Pseudomonas Infections; Static Electricity

Lysozyme is an abundant, cationic antimicrobial protein that plays an important role in pulmonary host defense. Increased concentration of lysozyme in the airspaces of transgenic mice enhanced bacterial killing whereas lysozyme deficiency resulted in increased bacterial burden and morbidity. Lysozyme degrades peptidoglycan in the bacterial cell wall leading to rapid killing of Gram-positive organisms; however, this mechanism cannot account for the protective effect of lysozyme against Gram-negative bacteria. The current study was therefore designed to test the hypothesis that the catalytic activity (muramidase activity) of lysozyme is not required for bacterial killing in vivo. Substitution of serine for aspartic acid at position 53 (D53S) in mouse lysozyme M completely ablated muramidase activity. Muramidase-deficient recombinant lysozyme (LysM(D53S)) killed both Gram-positive and Gram-negative bacteria in vitro. Targeted expression of LysM(D53S) in the respiratory epithelium of wild-type (LysM(+/+)/LysM(D53S)) or lysozyme M(null) mice (LysM(-/-)/LysM(D53S)) resulted in significantly elevated lysozyme protein in the airspaces without any increase in muramidase activity. Intratracheal challenge of transgenic mice with Gram-positive or Gram-negative bacteria resulted in a significant increase in bacterial burden in LysM(-/-) mice that was completely reversed by targeted expression of LysM(D53S). These results indicate that the muramidase activity of lysozyme is not required for bacterial killing in vitro or in vivo.

Topics: Animals; Aspartic Acid; Blood Bactericidal Activity; Bronchoalveolar Lavage Fluid; Klebsiella Infections; Klebsiella pneumoniae; Mice; Mice, Knockout; Mice, Transgenic; Muramidase; Mutagenesis, Site-Directed; Peptidoglycan; Pseudomonas aeruginosa; Pseudomonas Infections; Recombinant Proteins; Respiratory Mucosa; Serine; Staphylococcal Infections; Staphylococcus aureus

Macrolide antibiotics have clinical benefits in patients with diffuse panbronchiolitis and in patients with cystic fibrosis. Although many mechanisms have been proposed, the precise mechanisms are still uncertain. We examined the effects of erythromycin on bactericidal activity of airway surface liquid secreted by cultured human tracheal epithelial cells. Airway surface liquid was collected by washing the surface of human tracheal epithelial cells with a sodium solution (40 meq/l). Methicillin-resistant Staphylococcus aureus and Pseudomonas aeruginosa were incubated with airway surface liquid, and the number of surviving bacteria was examined. The number of bacteria in airway surface liquid from the cells cultured in medium alone was significantly lower than that in the sodium solution. Furthermore, the number of bacteria in airway surface liquid from the cells treated with erythromycin was significantly lower than that in airway surface liquid from the cells treated with solvent alone. The production of mRNA and protein of human beta-defensin-1 and human beta-defensin-2 was significantly increased by erythromycin. Bactericidal activity of airway surface liquid was observed at low concentrations (40 meq/l) of sodium but not at higher concentrations (> or =80 meq/l). Airway surface liquid did not contain significant amounts of antibiotics supplemented in the culture medium. Erythromycin at the levels in airway surface liquid and in culture medium did not inhibit bacterial growth. These results suggest that erythromycin may increase bactericidal activity of airway surface liquid in human airway epithelial cells through human beta-defensins production and reduce susceptibility of the airway to bacterial infection.

Topics: Anti-Bacterial Agents; beta-Defensins; Cell Survival; Cells, Cultured; Erythromycin; Gene Expression; Humans; Lactoferrin; Methicillin Resistance; Muramidase; Pseudomonas Infections; Respiratory Mucosa; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sodium; Staphylococcal Infections; Trachea

Lysozyme is a ubiquitous and abundant, cationic, antimicrobial polypeptide of leukocytes and epithelia, but its biological function in host defense is largely unexplored. To ascertain the role of lysozyme during bacterial infection of murine airways, we exposed the airways of lysozyme M-deficient (lys M-/-) mice to the pulmonary pathogen Pseudomonas aeruginosa and examined the host's response to infection. Despite partial compensation as a result of the appearance of lysozyme P in the infected airways of lys M-/- mice, these lys M-/- mice showed decreased clearance of P. aeruginosa compared with their lys M+/- or lys M+/+ littermates. Lysozyme contributes to optimal clearance of P. aeruginosa from the murine airways.

Topics: Animals; Bronchoalveolar Lavage Fluid; Lung; Mice; Mice, Inbred C57BL; Mice, Transgenic; Mucociliary Clearance; Muramidase; Pseudomonas aeruginosa; Pseudomonas Infections

We established a mouse model in which fatal pneumonia was induced by pneumococcal superinfection following influenza virus infection in chronic Pseudomonas aeruginosa infected mice. In this mouse model, influenza virus infection caused a significant increase in inflammatory cells, cytokines and severe tissue damage in the lungs of these P. aeruginosa infected mice, before pneumococcal infection. Intrapulmonary virus titres were significantly increased in mice with chronic P. aeruginosa infection, compared with control mice. Neutrophil function analysis showed significant reduction of myeloperoxidase (MPO) activity and lysozyme secretion by influenza virus infection in these mice. Our results suggest that influenza virus infection may play an important role in inducing pneumococcal pneumonia in chronic P. aeruginosa infected mice. Our results suggested that our mouse model is useful for investigating the pathogenesis of influenza virus infection in patients with chronic lung infection.

Topics: Acute Disease; Animals; Chronic Disease; Colony Count, Microbial; Cytokines; Disease Models, Animal; Disease Susceptibility; Lung; Lung Diseases, Parasitic; Male; Mice; Mice, Inbred Strains; Muramidase; Orthomyxoviridae Infections; Peroxidase; Pneumococcal Infections; Pneumonia, Pneumococcal; Pseudomonas Infections; Superinfection

Nasal polyps frequently appear in patients with cystic fibrosis (CF). The aims of this study were to focus on what problems (symptoms, endoscopic findings, and laboratory correlates) nasal polyps cause the CF patient, and how these correlate to the total health situation of this patient group.. The clinical histories, endoscopic investigations of the nasal cavity, and analyses of nasal lavage fluid of 44 patients with CF complicated with nasal polyposis have been compared with those of 67 CF control subjects. The patients were examined at annual control examinations (with pulmonary tests, working capacity, liver tests, and bacterial and blood tests) from 1995 to 1996 at Stockholm Cystic Fibrosis Center, Huddinge University Hospital. All patients were > 2 years of age. The endoscopic findings were related to the actual pulmonary function, inflammatory blood parameters, colonizing pathogens, antibodies (Staphylococcus aureus and Pseudomonas aeruginosa), and genotype.. The patients with nasal polyps differed with respect to chronic colonization of P aeruginosa in sputum samples and had a higher occurrence of serum antibodies against the same species. The two groups did not differ in pulmonary functions, inflammatory parameters, or genotype. The polyps found were mainly small (within the meatus media) and gave no significant increase in ongoing clinical symptoms such as rhinorrhea, nasal obstruction, or hyposmia. Neither was any significantly marked finding concerning the nose (mucosal swellings, secretion, etc.) made in the polyp patients. The patients with CF scored slightly lower in a smell identification test in comparison with the healthy control group. The nasal lavage fluid was analyzed (in 93 of the 111 patients) for the occurrence of P aeruginosa (by polymerase-chain reaction [PCR]), interleukin [IL]-5, IL-8, and lysozyme. The lysozyme and IL-8 content was equal in the two CF groups but increased in comparison with the healthy control group. P aeruginosa was not detected with PCR in any nasal lavage fluid. No measurable levels of IL-5 in the nasal lavage were found.. There was a higher frequency of chronic colonization of P aeruginosa in the lower respiratory tract in patients with nasal polyps. Otherwise, nonsevere nasal polyposis was not an indicator of lower respiratory tract morbidity in CF patients.

Topics: Adolescent; Adult; Antibodies, Bacterial; Child; Child, Preschool; Cystic Fibrosis; Endoscopy; Female; Humans; Interleukin-5; Interleukin-8; Male; Muramidase; Nasal Lavage Fluid; Nasal Polyps; Pseudomonas aeruginosa; Pseudomonas Infections; Respiratory Tract Infections; Risk Factors; Staphylococcal Infections

To assess the role of lysozyme in pulmonary host defense in vivo, transgenic mice expressing rat lysozyme cDNA in distal respiratory epithelial cells were generated. Two transgenic mouse lines were established in which the level of lysozyme protein in bronchoalveolar (BAL) lavage fluid was increased 2- or 4-fold relative to that in WT mice. Lung structure and cellular composition of BAL were not altered by the expression of lysozyme. Lysozyme activity in BAL was significantly increased (6.6- and 17-fold) in 5-wk-old animals from each transgenic line. To determine whether killing of bacteria was enhanced by expression of rat lysozyme, 5-wk-old transgenic mice and WT littermates were infected with 10(6) CFU of group B streptococci or 10(7) CFU of a mucoid strain of Pseudomonas aeruginosa by intratracheal injection. Killing of group B streptococci was significantly enhanced (2- and 3-fold) in the mouse transgenic lines at 6 h postinfection and was accompanied by a decrease in systemic dissemination of pathogen. Killing of Pseudomonas aeruginosa was also enhanced in the transgenic lines (5- and 30-fold). Twenty-four hours after administration of Pseudomonas aeruginosa, all transgenic mice survived, whereas 20% of the WT mice died. Increased production of lysozyme in respiratory epithelial cells of transgenic mice enhanced bacterial killing in the lung in vivo, and was associated with decreased systemic dissemination of pathogen and increased survival following infection.

Topics: Adjuvants, Immunologic; Animals; Bronchoalveolar Lavage Fluid; Cell Movement; Gene Expression Regulation; Humans; Inflammation Mediators; Lung; Mice; Mice, Inbred Strains; Mice, Transgenic; Muramidase; Neutrophils; Pseudomonas aeruginosa; Pseudomonas Infections; Rats; Transgenes

The effect of trichlorphon, one of the most widely used organophosphorus insecticides, on the nonspecific immune response in carp (Cyprinus carpio) was studied. The effect of 20,000 ppm trichlorphon on the immune response was followed for 3 and 56 days after intoxication. The effect of 10,000 ppm trichlorphon on the nonspecific immune response of carp experimentally infected by Pseudomonas alcaligenes and Aeromonas punctata was also examined. Leucocyte number, phagocytic ability of neutrophils, percentage NBT-positive PMN cells, phagocytic index, lysozyme level in serum, and ceruloplasmin activity in plasma were examined on Days 2, 4, 6, 8, 10, 14, 18, 22, and 26 after carp were exposed. After intoxication leukopenia was observed as decreases in phagocytic ability of neutrophils and in phagocytic index. Lysozyme level in serum was also decreased compared to that of control. The percentage of NBT-positive PMN cells decreased when the ceruloplasmin activity in plasma increased in intoxicated fish.

Topics: Aeromonas; Animals; Bacterial Infections; Carps; Ceruloplasmin; Cyprinidae; Leukocyte Count; Muramidase; Neutrophils; Phagocytosis; Pseudomonas Infections; Trichlorfon

Experimental investigations indicate that localized forms of staphylococcal, Pseudomonas aeruginosa and associated (Pseudomonas aeruginosa-staphylococcal) infection inhibit the lysozyme activity, cause disorders in the metabolism of P-450 cytochrome, free-radical and iron-containing liver proteins.

Topics: Animals; Cytochrome P-450 Enzyme System; Free Radicals; Iron; Liver; Male; Metalloproteins; Mice; Muramidase; Pseudomonas Infections; Staphylococcal Infections; Time Factors

The oral administration of forphenicinol increased the survival rate of both normal and immunodepressed mice intraperitoneally or intratracheally infected with clinically isolated strains of Pseudomonas aeruginosa. The therapeutic effect of amikacin on intraperitoneal infection with P. aeruginosa was enhanced by combined use with forphenicinol. Forphenicinol did not enhance the bactericidal activity of polymorphonuclear cells (PMN) towards P. aeruginosa in vitro, but enhanced it in vivo. In vitro study indicated that the macrophages taken from mice treated with forphenicinol or the cultured supernatant of these macrophages enhanced the bactericidal activity of PMN. The protective effect of forphenicinol against P. aeruginosa infection was thus suggested to be due to macrophage activation followed by the enhancement of the bactericidal activity of PMN.

Topics: Adjuvants, Immunologic; Amikacin; Animals; Female; Glycine; Mice; Mice, Inbred ICR; Muramidase; Neutrophils; Pseudomonas aeruginosa; Pseudomonas Infections; Superoxides

Topics: Animals; Cystic Fibrosis; Fatty Acids, Essential; Lung; Macrophages; Microscopy, Electron; Muramidase; Pseudomonas Infections; Pulmonary Alveoli; Rabbits; Respiratory Tract Infections; Staphylococcal Infections

We prospectively evaluated concentrations of beta-D-galactosidase, alpha-L-fucosidase, beta-D-N-acetylglucosaminidase, and lysozyme in urine from normal subjects, ambulatory patients with cystic fibrosis (CF), and CF patients with previously normal renal function who were receiving intravenous aminoglycoside (AG) therapy. Enzyme activities were generally low or negligible in subjects not receiving AG. Enzymuria was documented during 12 of 13 AG treatment courses and most frequently involved beta-D-N-acetylglucosaminidase excretion. In nine courses, enzymuria occurred in the absence of proteinuria or elevations of blood urea nitrogen and serum creatinine. In three courses attended by enzymuria and evidence of nephrotoxicity, neither the time of appearance nor the magnitude of enzymuria was different from that of nonnephrotoxic patients. In two of these three treatment courses, enzymuria preceded clinical evidence of nephrotoxicity of 16 and 5 days, and in the third course enzymuria and elevation of blood urea nitrogen and serum creatinine occurred simultaneously. We conclude that enzymuria is not a reliable predictor of nephrotoxicity due to AG in CF patients and is not an indication of discontinue AG therapy.

Topics: Acetylglucosaminidase; Adolescent; Adult; alpha-L-Fucosidase; Aminoglycosides; beta-Galactosidase; Child; Child, Preschool; Cystic Fibrosis; Female; Gentamicins; Glycoside Hydrolases; Humans; Kidney; Lung Diseases; Male; Middle Aged; Muramidase; Pseudomonas Infections; Tobramycin

Topics: Acute Disease; Bacteriuria; Child; Child, Preschool; Enterobacteriaceae Infections; Humans; Muramidase; Pseudomonas Infections; Staphylococcal Infections

Topics: Animals; Cystitis; Dog Diseases; Dogs; Edetic Acid; Male; Muramidase; Otitis Externa; Pseudomonas Infections; Tromethamine

Topics: Animals; Bacteriuria; Cystitis; Dog Diseases; Dogs; Edetic Acid; Gentamicins; Injections, Subcutaneous; Male; Muramidase; Pseudomonas aeruginosa; Pseudomonas Infections; Therapeutic Irrigation; Tromethamine; Urinary Catheterization; Urinary Tract Infections

Topics: Animals; Dog Diseases; Dogs; Drug Therapy, Combination; Edetic Acid; Female; Male; Muramidase; Otitis Externa; Otitis Media; Pseudomonas aeruginosa; Pseudomonas Infections; Therapeutic Irrigation; Tromethamine

Topics: Edetic Acid; Enterobacteriaceae Infections; Humans; Muramidase; Pseudomonas Infections; Therapeutic Irrigation; Tromethamine; Urinary Bladder Diseases; Urinary Tract Infections

Topics: Adolescent; Adult; Age Factors; Aged; Antibodies; Antineoplastic Agents; Blood Bactericidal Activity; Burns; Child; Complement System Proteins; Drug Resistance, Microbial; Escherichia coli Infections; Female; Humans; Immunity; Middle Aged; Muramidase; Parotitis; Phagocytosis; Pneumonia; Polysaccharides, Bacterial; Pseudomonas Infections; Sepsis; Serratia marcescens; Staphylococcal Infections; Stimulation, Chemical

Topics: Animals; Burns; Injections, Intraperitoneal; Muramidase; Phagocytosis; Pseudomonas aeruginosa; Pseudomonas Infections; Rats; Time Factors

Topics: Cross Infection; Eye Diseases; Humans; Muramidase; Ophthalmic Solutions; Pseudomonas Infections; Staphylococcal Infections; Sterilization