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1.
Eur J Clin Microbiol Infect Dis ; 34(7): 1403-13, 2015 Jul.
Article in English | MEDLINE | ID: mdl-25820290

ABSTRACT

Drug susceptibility testing (DST) of rapidly growing mycobacteria (RGM) are recommended for guiding the antimicrobial therapy. We have evaluated the use of resazurin in Mueller-Hinton medium (MHR) for MIC determination of RGM and compared the results with those obtained with the reference standard broth microdilution in Mueller-Hinton (MH) and with the resazurin microtiter assay (REMA) in 7H9 broth. The MIC of eight drugs: amikacin (AMI), cefoxitin (FOX), ciprofloxacin (CIP), clarithromycin (CLA), doxycycline (DOX), linezolid (LZD), moxifloxacin (MXF) and trimethoprim-sulfamethoxazole (TMP-SMX) were evaluated against 76 RGM (18 species) using three methods (MH, MHR, and REMA) in a 96-well plate format incubated at 37 °C over 3-5 days. Results obtained in the MH plates were interpreted by the appearance of turbidity at the bottom of the well before adding the resazurin. MHR and 7H9-REMA plates were read by visual observation for a change in color from blue to pink. The majority of results were obtained at day 5 for MH and 1 day after for MHR and 7H9-REMA. However, the preliminary experiment on time to positivity results using the reference strain showed that the resazurin can be added to the MH at day 2 to produce the results at day 3, but future studies with large sets of strains are required to confirm this suggestion. A high level of agreement (kappa 1.000-0.884) was obtained between the MH and the MHR. Comparison of results obtained with 7H9-REMA, on the other hand, revealed several discrepancies and a lower level of agreement (kappa 1.000-0.111). The majority of the strains were resistant to DOX and TMP-SMX, and the most active antimicrobials for RGM were AMI and FOX. In the present study, MHR represented an excellent alternative for MIC determination of RGM. The results could be read reliably, more easily, and more quickly than with the classical MH method.


Subject(s)
Anti-Bacterial Agents/pharmacology , Microbial Sensitivity Tests/methods , Mycobacterium/drug effects , Humans , Mycobacterium/growth & development , Mycobacterium/isolation & purification , Mycobacterium Infections/microbiology , Nontuberculous Mycobacteria/drug effects , Nontuberculous Mycobacteria/growth & development , Nontuberculous Mycobacteria/isolation & purification , Time Factors
2.
Clin Microbiol Infect ; 16(2): 152-6, 2010 Feb.
Article in English | MEDLINE | ID: mdl-19686282

ABSTRACT

Fifty-seven nosocomial Klebsiella pneumoniae isolates producing extended-spectrum beta-lactamases (ESBLs) were collected between February 2007 and November 2007 in different wards of the Sarajevo (Bosnia-Herzegovina) reference hospital. These isolates comprise two major epidemic pulsed-field electrophoresis-defined clones plus two minor clones. In addition to the ESBL-mediated resistance, all strains uniformly showed resistance to ciprofloxacin, gentamicin and tobramycin. The beta-lactamases involved in this resistance phenotype were TEM-1, SHV-1, and CTX-M-15, as demonstrated by isoelectric focusing, PCR amplification, and sequencing. TEM-1 and CTX-M-15 beta-lactamases, as well as the aminoglycoside resistance determinants, were encoded in plasmids that could be transferred to Escherichia coli by conjugation. In three of the infected patients with the predominant clone, cefoxitin resistance development (MICs >128 mg/L) was documented. The analysis of the outer membrane proteins of the cefoxitin-susceptible and cefoxitin-resistant isolates revealed that the former expressed only one of the two major porins, OmpK36, whereas in the latter, the expression of Ompk36 was altered or abolished. This is the first report of CTX-M-15-producing K. pneumoniae in Bosnia-Herzegovina. Furthermore, we document and characterize for the first time cefoxitin resistance development in CTX-M-15-producing K. pneumoniae.


Subject(s)
Bacterial Proteins/biosynthesis , Cross Infection/microbiology , Klebsiella Infections/microbiology , Klebsiella pneumoniae/enzymology , beta-Lactamases/biosynthesis , Adult , Anti-Bacterial Agents/pharmacology , Bacterial Outer Membrane Proteins/analysis , Bacterial Typing Techniques , Bosnia and Herzegovina , Cluster Analysis , DNA Fingerprinting , Electrophoresis, Gel, Pulsed-Field , Genotype , Hospitals , Humans , Isoelectric Focusing , Klebsiella pneumoniae/chemistry , Klebsiella pneumoniae/drug effects , Klebsiella pneumoniae/isolation & purification , Plasmids , Polymerase Chain Reaction , Young Adult , beta-Lactamases/chemistry , beta-Lactamases/classification , beta-Lactamases/genetics
3.
Hum Exp Toxicol ; 24(3): 101-8, 2005 Mar.
Article in English | MEDLINE | ID: mdl-15901049

ABSTRACT

Fluoride has been in focus as a possible causal agent for respiratory symptoms amongst aluminium potroom workers for several decades. Previously, using bronchoalveolar lavage (BAL), we demonstrated airway inflammation in healthy volunteers 24 hours after exposure to hydrogen fluoride (HF). The objective of the present study was to examine early lung responses to HF exposure. Bronchoscopy with BAL was performed 2 hours after the end of 1-hour exposure to HE Significant reductions in the total cell number and the number of neutrophils and lymphocytes were observed in bronchoalveolar portion (BAP), whereas there were no significant changes in the bronchial portion (BP). Significantly decreased concentrations of beta2-MG, IL-6 and total protein were found in both BAP and BP. Additionally, IL-8 was significantly reduced in BP, and ICAM-1 and albumin were present in lower concentrations in BAP. Lung function measurements were not affected by HF exposure. These reported effects are presumably transitory, as many were not present in the airways 24 hours after a similar HF exposure.


Subject(s)
Air Pollutants, Occupational/toxicity , Bronchoalveolar Lavage Fluid , Hydrofluoric Acid/toxicity , Pneumonia/immunology , Administration, Inhalation , Adult , Antioxidants/analysis , Biomarkers/analysis , Bronchoalveolar Lavage Fluid/chemistry , Bronchoalveolar Lavage Fluid/cytology , Bronchoalveolar Lavage Fluid/immunology , Cell Count , Humans , Male , Pneumonia/chemically induced
4.
Inhal Toxicol ; 14(2): 119-32, 2002 Feb.
Article in English | MEDLINE | ID: mdl-12122575

ABSTRACT

The development of asthmalike symptoms among aluminum potroom workers has been associated with exposure to fluorides. In the present study, the immediate nasal response in humans was examined subsequent to short-term hydrogen fluoride (HF) exposure. Ten healthy subjects were exposed to HF (3.3-3.9 mg/m(3)) for 1 h. Nasal lavage (NAL) was performed before, immediately after, and 1.5 h after the end of exposure. Control lavages were performed on the same subjects at the same time points but without HF exposure. At the end of HF exposure, 7 of 10 individuals reported upper airway symptoms. A significant increase was observed in the number of neutrophils and total cells, while there was a decrease in cell viability. The changes in neutrophil numbers correlated significantly with the reported airway symptoms. HF also induced a significant increase in tumor necrosis factor-alpha and the total protein content of NAL fluid. Among the eicosanoids, prostaglandin E(2), leukotriene B(4), and peptide leukotrienes were elevated after exposure. Of the antioxidants measured, the concentration of uric acid increased after exposure. In conclusion, exposure to HF induced immediate nasal inflammatory and antioxidant responses in healthy human volunteers. These findings may contribute to a further understanding of the way HF exerts damage to the airways and show that HF could represent an occupational hazard.


Subject(s)
Air Pollutants, Occupational/adverse effects , Antioxidants/analysis , Eicosanoids/biosynthesis , Hydrofluoric Acid/adverse effects , Nasal Lavage Fluid/chemistry , Neutrophils/cytology , Respiration Disorders/chemically induced , Acute Disease , Administration, Inhalation , Adult , Cell Survival/drug effects , Humans , Leukocyte Count , Male , Nasal Lavage Fluid/cytology , Respiration Disorders/metabolism
5.
Eur Respir J ; 15(3): 582-9, 2000 Mar.
Article in English | MEDLINE | ID: mdl-10759457

ABSTRACT

Respiratory epithelial cells are actively involved in the host defence and inflammatory reactions of the airways. Nuclear factor-kappaB (NF-kappaB) is a transcription factor that plays a pivotal role in many cellular responses to environmental changes. The inducible nitric oxide synthase (iNOS) isoform has been implicated in airway inflammation as well as in normal airway function. In this study, the hypothesis that NF-kappaB may be associated with iNOS expression in airway epithelium, not only in inflammatory processes but also under physiological conditions was examined. NF-kappaB deoxyribonucleic acid-binding activity was assayed by means of electrophoretic mobility shift assay (EMSA) and iNOS expression examined using immunohistochemical techniques in healthy nasal mucosa and chronically inflamed nasal polyps. Further NF-kappaB activity was assayed; by means of EMSA, in nasal epithelial cells isolated from both tissues. NF-kappaB was activated in nasal polyps, but also to the same extent in healthy nasal mucosa. Uniform iNOS expression was localized within the airway epithelium in both inflamed and noninflamed tissues. Along with iNOS expression, concomitant NF-kappaB activation was found in nasal epithelial cells obtained from both tissues and no differences were observed when nasal mucosa and nasal polyp were compared. These results suggest that constitutive nuclear factor-kappaB and concurrent inducible nitric oxide synthase expression in epithelial cells may play a physiological role in airway function.


Subject(s)
Epithelial Cells/physiology , NF-kappa B/physiology , Female , Humans , Male , Middle Aged , Nasal Mucosa/metabolism , Nasal Polyps/metabolism , Nitric Oxide/biosynthesis
6.
Acta Otorrinolaringol Esp ; 49(7): 503-7, 1998 Oct.
Article in Spanish | MEDLINE | ID: mdl-9866213

ABSTRACT

Nitric oxide (NO) plays an important regulatory role in airway function and seems to be implicated in the pathophysiology of several airway diseases. We studied the presence of NO synthase activity in human nasal mucosa and nasal polyp tissues obtained from patients undergoing septoplasty or polypectomy, respectively. NO synthase activity was quantified in tissue homogenates using citrulline release assay and was located in tissue sections using NADPH-diaphorase histochemistry. The results indicated that nasal polyps contain higher levels of total NO synthase activity than nasal mucosa tissue. In addition, nasal polyps contained mainly inducible NO synthase activity whereas all NO synthase activity detected in the nasal mucosa was in constitutive form. In both cases, NO synthase activity was localized in epithelial cells. In view of these results, we conclude that NO may be an important inflammatory mediator in the respiratory system and that the epithelium may be a source of NO production.


Subject(s)
Nasal Mucosa/enzymology , Nitric Oxide Synthase/metabolism , Adult , Aged , Female , Humans , Male , Middle Aged , Nasal Polyps/enzymology , Nasal Polyps/surgery
7.
Environ Res ; 75(1): 23-33, 1997 Oct.
Article in English | MEDLINE | ID: mdl-9356191

ABSTRACT

Groups of industrial workers exposed to heavy metals (cadmium, mercury, and lead) or solvents were studied together with corresponding control groups. The cohorts were collected from several European centers (countries). Eighty-one measurements were carried out on urine, blood, and serum samples and the results of these analyses together with questionnaire information on each individual were entered into a central database using the relational database package Rbase. After the completion of the database construction phase, the data were exported in a format suitable for analysis by the statistical package SAS. The potential value of each test as an indicator of nephrotoxicity was then assessed. Rigorous exclusion criteria were applied which resulted in the elimination of some tests and samples from the dataset. The measurable contributions of smoking, gender, metal exposure, and site were either singly or in combination assessed by biomarkers for nephrotoxicity. The parameters measured included three urinary enzymes, six specific proteins, total protein, two extracellular matrix markers, four prostaglandins and anti-GBM antibodies, and beta 2-microglobulin in serum. The most sensitive renal tests included the urinary enzymes N-acetyl-beta-D-glucosaminidase (NAG) and intestinal alkaline phosphatase (IAP), brush border antigens, and urinary low-molecular-weight proteins. Of the newer tests investigated the prostaglandins were the most promising. Different patterns of biomarker excretion were observed following exposure to lead, cadmium, or mercury. The dataset provides a unique repository of data which could provide the basis of an enlarging source of information on normal human reference ranges and on the effects of exposure to toxins and the use of biomarkers for monitoring nephrotoxicity.


Subject(s)
Database Management Systems , Hazardous Substances , Kidney/drug effects , Occupational Exposure , Biomarkers , Blood Chemical Analysis , Cohort Studies , Europe/epidemiology , Humans , Surveys and Questionnaires
8.
Hum Exp Toxicol ; 15 Suppl 1: S10-9, 1996 Mar.
Article in English | MEDLINE | ID: mdl-8882556

ABSTRACT

Within the framework of an European Commission-funded project, groups of industrial workers exposed to heavy metals (cadmium, mercury and lead) or solvents were studied together with corresponding control groups. Eighty-one measurements were carried out on urine and serum samples and the scientific results together with individual questionnaire information were entered into a central database. Data obtained was assessed centrally and individually in subsidiary studies. The measurable contributions were assessed either singly or in combination, of smoking, gender, metal exposure and site, to nephrotoxicity. The potential value of each test as an indicator of nephrotoxicity was then assessed on the basis of sensitivity and specificity. A number of new tests including prostaglandins and for extracellular matrix components were investigated as well as established tests for renal damage and dysfunction. The data obtained from this comprehensive study emphasises the value of noninvasive biomarkers for the early detection of nephrotoxicity due to environmental toxins. The urinary profile varied with the type of environmental/occupational toxin. By careful selection of a small panel of markers they can be used to indicate the presence of renal damage, the principal region affected, and to monitor the progress of disease and damage. Biomarkers were also used to confirm and tentatively establish safe exposure levels to nephrotoxins.


Subject(s)
Drug Evaluation, Preclinical , Environmental Pollutants/toxicity , Kidney/drug effects , Biomarkers , Drug Evaluation, Preclinical/methods , Drug Evaluation, Preclinical/standards , Drug Evaluation, Preclinical/trends , Humans
9.
Eur Respir J ; 9(2): 202-6, 1996 Feb.
Article in English | MEDLINE | ID: mdl-8777951

ABSTRACT

Nitric oxide (NO) plays an important regulatory role in airway function and seems to be implicated in the pathophysiology of several airway diseases. To better understand the involvement of NO in the upper airways, we examined the presence of nitric oxide synthase (NOS) activity in human nasal mucosa and nasal polyp tissues. Nasal mucosa was obtained from seven patients undergoing septoplasty, and nasal polyps came from nine patients following polypectomy. NOS activity was quantified in tissue homogenates using the citrulline release assay and localized in tissue sections using reduced nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase histochemistry. The results showed that nasal polyps (n = 9) contained higher levels of total NOS activity (mean +/- SD 5.94 +/- 5.71, range 1.29-18.0 pmol.min-1.mg protein) than nasal mucosa tissues (n = 7) (0.28 +/- 0.22, range 0.01-0.57 pmol.min-1.mg protein). In addition, nasal polyps mainly contained inducible NOS activity (4.67 +/- 4.57, range 1.23-15.5 pmol.min-1.mg protein) whereas in nasal mucosa all NOS activity detected was in constitutive form. In both cases, NOS activity was localized in the epithelial cells. Since NO synthase is induced in inflamed upper airways, we conclude that NO may be an important inflammatory mediator in the respiratory system and that the epithelium may be a source of NO production in the human upper airways.


Subject(s)
Nasal Mucosa/enzymology , Nasal Polyps/enzymology , Nitric Oxide Synthase/metabolism , Adult , Aged , Female , Histocytochemistry , Humans , Male , Middle Aged , NADPH Dehydrogenase , Nasal Mucosa/cytology , Nasal Polyps/pathology
10.
Am J Respir Crit Care Med ; 153(1): 350-5, 1996 Jan.
Article in English | MEDLINE | ID: mdl-8542142

ABSTRACT

Cytokines, such as interleukin-3 (IL-3), have been suggested to play an important role in mediating the increased number of airway eosinophils and metachromatic cells in patients with even mild asthma. We used immunohistochemistry to determine the presence of IL-3 protein in bronchial biopsies from nonasthmatics (n = 10) and subjects with mild (n = 8) and allergen-induced (n = 7) asthma. We also examined whether IL-3 was related to airway eosinophil number and activation, the number of airway metachromatic cells, or airway function. We found that the number and activation of eosinophils and the number of metachromatic cells were increased in the airways of asthmatics, compared with nonasthmatics, with further increases evident after allergen challenge. IL-3 protein was localized primarily to the epithelium in nonasthmatic and asthmatic subjects, with no difference apparent between groups or after allergen inhalation challenge. The extent of staining for IL-3 in the tissue was not correlated with eosinophil number or activity, metachromatic cell number, airway responsiveness, or the severity of the late asthmatic response. This study provides the first demonstration of IL-3 protein localization in bronchial tissue from human airways. The results suggest that the increases in eosinophils and metachromatic cells associated with mild and allergen-induced asthma occur independent of IL-3.


Subject(s)
Asthma/metabolism , Bronchi/chemistry , Interleukin-3/analysis , Adult , Allergens/administration & dosage , Asthma/diagnosis , Asthma/pathology , Biopsy , Bronchi/pathology , Bronchial Provocation Tests , Bronchoalveolar Lavage Fluid/cytology , Bronchoscopy , Coloring Agents , Data Interpretation, Statistical , Eosinophils , Humans , Immunohistochemistry , Middle Aged , Radioimmunoassay , Skin Tests
11.
Thorax ; 50(5): 490-6, 1995 May.
Article in English | MEDLINE | ID: mdl-7597660

ABSTRACT

BACKGROUND: An abnormal platelet release of oxygen-free radicals has been described in acetylsalicylic acid (aspirin)-induced asthma, a finding which might suggest the existence of an intrinsic, specific platelet abnormality of arachidonic acid metabolism in these patients. The objective of this study was to evaluate platelet arachidonic acid metabolism in asthmatic patients with or without intolerance to aspirin. METHODS: Thirty subjects distributed into three groups were studied: group 1, 10 healthy subjects; group 2, 10 asthmatic patients with aspirin tolerance; and group 3, 10 aspirin-intolerant asthmatics. Platelets were isolated from blood, preincubated with 3H-arachidonic acid for 30 minutes and then incubated for 10 minutes with platelet activating factor (PAF) and aspirin. Cyclo-oxygenase (thromboxane, PGE2, PGF2 alpha, and HHT) and lipoxygenase (12-HETE) arachidonic acid metabolites were measured by high pressure liquid chromatography. Release of oxygen free radicals after incubation with PAF and aspirin was measured by chemiluminescence. Platelet levels of glutathione peroxidase (GSH-Px) were also measured using spectrophotometry. RESULTS: Platelets from aspirin-intolerant asthmatic patients produced higher quantities of arachidonic acid metabolites than the control group at baseline conditions. This increase was significant only for lipoxygenase products. No differences were found amongst the three groups in the response of arachidonic acid metabolism to PAF and aspirin. Incubation with aspirin but not with PAF caused an increase in oxygen-free radical production in aspirin-intolerant patients whereas in aspirin-tolerant patients PAF, rather than aspirin, was the more potent stimulus for oxygen-free radical production. No differences in GSH-Px levels were found amongst the three groups. CONCLUSIONS: These results suggest that the platelet lipoxygenase pathway is activated in aspirin-intolerant patients and that the production of oxygen-free radicals may differentiate aspirin-tolerant from aspirin-intolerant asthmatic subjects. Our study, however, does not support the hypothesis that an increase in lipoxygenase products may be responsible for oxygen-free radical production. Moreover, a lowered platelet GSH-Px activity does not seem to be involved in this phenomenon.


Subject(s)
Arachidonic Acid/metabolism , Aspirin , Asthma/blood , Blood Platelets/metabolism , Drug Hypersensitivity/metabolism , Glutathione Peroxidase/metabolism , Superoxides/metabolism , Adult , Blood Platelets/enzymology , Chromatography, High Pressure Liquid , Female , Humans , In Vitro Techniques , Lipoxygenase/metabolism , Luminescent Measurements , Male
13.
Eur Respir J ; 7(12): 2212-22, 1994 Dec.
Article in English | MEDLINE | ID: mdl-7713206

ABSTRACT

Inflammation is a response that has evolved over millions of years to become an extremely complex process. This complexity reflects the host's need to deal effectively with a wide variety of potentially injurious agents, as well as the need to incorporate an adequate set of checks and balances. An inappropriately checked response, which occurs rarely, results in disease, either acute or chronic. However, in most instances, inflammation is a beneficial response, essential for survival. Inflammation comprises an extensive network of cellular interactions implemented by an overwhelming number of molecules. One category of signal includes soluble products, such as neuropeptide, lipid mediators, cytokines and growth factors, most of which can be produced by inflammatory/haemopoietic cells. However, resident structural cells can also produce many of these products and, on this basis only, fibroblasts, epithelial, endothelial and smooth muscle cells should be considered as active contributors to the regulation of the inflammatory response. Extracellular matrix (ECM) proteins comprise another category of signals. Whilst the most recognized activities of these proteins are those concerned with providing structural tissue integrity, it is clear that they also have powerful inductive effects. Indeed, ECM proteins can influence the shape, movement and state of activation of inflammatory cells in the tissue. Recent evidence indicates that these signals may also play substantial roles in homing of inflammatory cells to certain sites and in the handling of a number of cytokines and growth factors. In so far as fibroblasts are the main producers of ECM proteins, these new data establish an indirect but important role for fibroblasts in the regulation of the inflammatory response.


Subject(s)
Fibroblasts/immunology , Inflammation/immunology , Lung Diseases/immunology , Lung/immunology , Animals , Cytokines/physiology , Dinoprostone/physiology , Extracellular Matrix Proteins/physiology , Fibroblasts/physiology , Growth Substances/physiology , Humans , Lung/cytology
14.
J Pharm Biomed Anal ; 11(11-12): 1135-9, 1993.
Article in English | MEDLINE | ID: mdl-8123725

ABSTRACT

We have developed a fully automated on-line extraction-purification method for peptide leukotrienes in biological fluids using a column-switching technique. Individual leukotrienes in HPLC-collected fractions are determined by immunoassay techniques. Leukotrienes were extracted from nasal lavage samples and eluted from the solid-phase extraction cartridge into the HPLC column with methanol-ammonium acetate buffer (60:40, v/v) pH 5.4, as a mobile phase. This method provides good recoveries, excellent resolution of leukotrienes and is suitable to be combined with off-line radioimmunoassay (RIA) or enzymoimmunoassay (EIA). The most important losses are observed after evaporation-concentration and lyophilization. We also have compared RIA with EIA determination. Significant differences are found between RIA values and EIA values.


Subject(s)
Chromatography, High Pressure Liquid , Leukotriene C4/isolation & purification , Leukotriene D4/isolation & purification , Nasal Lavage Fluid/chemistry , Autoanalysis , Humans , Immunoenzyme Techniques , Radioimmunoassay
15.
Agents Actions ; 40(1-2): 72-7, 1993 Sep.
Article in English | MEDLINE | ID: mdl-8147272

ABSTRACT

The differentiation of HL-60 human promyelocytic leukaemia cells into specific monocytic or granulocytic lineage cells depending of the inductor agent is accompanied by selective regulation of several key enzymes involved in the synthesis of eicosanoids. In this communication we have investigated the changes in arachidonic acid metabolic profiles during phorbol 12-myristate 13-acetate (PMA)-induced monocytic differentiation of HL-60 cells. Our results show that HL-60 cells have spontaneous capacity to synthesize large amounts of LTB4, but PMA-differentiated cells lose the ability to release LTB4. Significant differences are found between HL-60 cells and PMA-treated cells in basal conditions and under ionophore stimulation. The addition of LTB4 at the time of PMA differentiation did not have effects on cell proliferation, but nordihydroguaiaretic acid (NDGA), a potent 5-lipoxygenase inhibitor, also inhibited HL-60 cell proliferation and did not have any effect on PMA-differentiated cell proliferation.


Subject(s)
Leukotriene B4/metabolism , Macrophages/physiology , Monocytes/physiology , Arachidonic Acids/metabolism , Cell Differentiation/drug effects , Cell Differentiation/physiology , Cell Division/drug effects , Cell Line , Dinoprostone/metabolism , Humans , Hydroxyeicosatetraenoic Acids/metabolism , Lipoxygenase/metabolism , Macrophages/enzymology , Macrophages/metabolism , Masoprocol/pharmacology , Monocytes/enzymology , Monocytes/metabolism , Prostaglandin-Endoperoxide Synthases/metabolism , Tetradecanoylphorbol Acetate/pharmacology , Thromboxane B2/metabolism
16.
J Allergy Clin Immunol ; 92(2): 248-54, 1993 Aug.
Article in English | MEDLINE | ID: mdl-8349935

ABSTRACT

BACKGROUND: We evaluated the effect of furosemide on allergen-induced rhinitis in a double-blind, crossover, placebo-controlled experiment. METHODS: Fourteen patients with rhinitis who were allergic to house dust were nebulized with an intranasal dose of 20 mg of furosemide or placebo before allergen challenge with an extract of Dermatophagoides pteronyssinus (100 BU). Clinical evaluation and nasal lavages with normal saline solution were performed at baseline; after placebo-furosemide nebulization, and at 10, 30, and 60 minutes after allergen challenge. Number of sneezes and a composite symptom score were recorded to evaluate clinical response. Prostaglandin E2 (PGE2), PGD2 peptide leukotrienes and 15-hydroxy, 5,8,11,13-eicosatetraenoic acid (15-HETE) were measured by radioimmunoassay in nasal lavages. Cells were counted and classified as epithelial cells, neutrophils, eosinophils, and others. RESULTS: No differences in either clinical symptoms or cell influx after allergen challenge were found between furosemide and placebo groups. PGE2 levels did not change after provocation, and furosemide had no effect on its production. Ten minutes after antigen challenge there was a marked increase of PGD2 (p < 0.01), peptide leukotrienes (p < 0.01), and 15-HETE (not significant) on both study days. However, no significant differences in the release of eicosanoids were found between furosemide and placebo groups. CONCLUSIONS: Our observations in the nasal mucosa suggest that furosemide has no effect on the release of proinflammatory and bronchoconstrictor metabolites (PGD2, peptide leukotrienes, and 15-HETE). In contrast to bronchial asthma, allergen-induced rhinitis was not effectively prevented by furosemide.


Subject(s)
Furosemide/therapeutic use , Rhinitis, Allergic, Perennial/drug therapy , Rhinitis, Allergic, Perennial/metabolism , Adult , Arachidonic Acid/antagonists & inhibitors , Arachidonic Acid/metabolism , Cell Count/drug effects , Cyclooxygenase Inhibitors/therapeutic use , Double-Blind Method , Eosinophils/pathology , Female , Humans , Lipoxygenase Inhibitors/therapeutic use , Male , Nasal Mucosa/drug effects , Nasal Mucosa/metabolism , Nasal Mucosa/pathology , Neutrophils/pathology , Rhinitis, Allergic, Perennial/pathology
17.
Thorax ; 48(3): 260-3, 1993 Mar.
Article in English | MEDLINE | ID: mdl-8497826

ABSTRACT

BACKGROUND: It has been suggested that inhaled frusemide protects subjects with asthma against bronchoconstriction by enhancing the synthesis of prostaglandin E2 (PGE2). To evaluate this hypothesis the effect of frusemide on PGE2 production from nasal mucosa was studied. METHODS: Two main arachidonic acid metabolites produced by epithelial cells, PGE2 and 15-hydroxy 5,8,11,13-eicosatetraenoic acid (15-HETE), were measured by radioimmunoassay in nasal secretions obtained by nasal lavages with saline. Eleven healthy volunteers were randomly assigned to two study days, one week apart, in a double blind crossover study. Nasal instillation with three increasing doses of frusemide (5, 10, and 20 mg) or placebo was carried out at intervals of 15 minutes. Nasal lavages were performed immediately before nasal instillations and 15, 30, and 60 minutes after the last instillation. RESULTS: Baseline concentrations of 15-HETE were at least six times higher than PGE2. No differences between frusemide and placebo were detected either on PGE2 or 15-HETE release. CONCLUSIONS: The findings do not support the hypothesis that the antiasthmatic effect of frusemide may be due to increased synthesis of PGE2 or release in the respiratory mucosa.


Subject(s)
Dinoprostone/biosynthesis , Furosemide/pharmacology , Nasal Mucosa/drug effects , Adult , Double-Blind Method , Female , Furosemide/administration & dosage , Humans , Hydroxyeicosatetraenoic Acids/biosynthesis , Instillation, Drug , Male , Nasal Mucosa/metabolism
18.
Br J Ind Med ; 50(1): 28-36, 1993 Jan.
Article in English | MEDLINE | ID: mdl-8431388

ABSTRACT

The present study has been carried out in the framework of a collaborative research project on the development of new markers of nephrotoxicity. A battery of more than 20 potential indicators of renal changes has been applied to 50 workers exposed to lead (Pb) and 50 control subjects. After application of selection criteria 41 exposed and 41 control workers were eventually retained for the final statistical analysis. The average blood Pb concentration of exposed workers was 480 micrograms/l and their mean duration of exposure was 14 years. The battery of tests included parameters capable of detecting functional deficits (for example, urinary proteins of low or high molecular weight), biochemical alterations (for example, urinary eicosanoids, glycosaminoglycans, sialic acid) or cell damage (for example, urinary tubular antigens or enzymes) at different sites of the nephron or the kidney. The most outstanding effect found in workers exposed to Pb was an interference with the renal synthesis of eicosanoids, resulting in lower urinary excretion of 6-keto-PGF1 alpha and an enhanced excretion of thromboxane (TXB2). The health significance of these biochemical alterations, detectable at low exposure to Pb is unknown. As they were not associated with any sign of renal dysfunction, they may represent reversible biochemical effects or only contribute to the degradation of the renal function from the onset of clinical Pb nephropathy. The urinary excretion of some tubular antigens was also positively associated with duration of exposure to Pb. Another effect of Pb that might deserve further study is a significant increase in urinary sialic acid concentration.


Subject(s)
Kidney Diseases/chemically induced , Kidney/drug effects , Lead Poisoning/complications , Occupational Diseases/chemically induced , Occupational Exposure , Adult , Biomarkers/blood , Biomarkers/urine , Humans , Kidney Diseases/blood , Kidney Diseases/urine , Lead Poisoning/blood , Lead Poisoning/urine , Occupational Diseases/blood , Occupational Diseases/urine
19.
Br J Ind Med ; 50(1): 37-48, 1993 Jan.
Article in English | MEDLINE | ID: mdl-8431389

ABSTRACT

Cadmium (Cd) was the third heavy metal investigated in the European collaborative research project on the development and validation of new markers of nephrotoxicity. Fifty workers exposed to Cd and 50 control workers were examined. After application of selection criteria 37 workers (mean age 43) exposed to Cd for an average of 11.3 years; and 43 age matched referents were retained for final analysis. The average concentrations of Cd in blood (Cd-B) and urine (Cd-U) of exposed workers were 5.5 micrograms Cd/l and 5.4 micrograms Cd/g creatinine respectively. By contrast with lead and mercury, Cd had a broad spectrum of effects on the kidney, producing significant alterations in amounts of almost all potential indicators of nephrotoxicity that were measured in urine--namely, low and high molecular weight proteins, kidney derived antigens or enzymes, prostanoids, and various other biochemical indices such as glycosaminoglycans and sialic acid. An increase in beta 2-microglobulin and a decrease of sialic acid concentration were found in serum. Dose-effect/response relations could be established between most of these markers and Cd-U or Cd-B. The thresholds of Cd-U associated with a significantly higher probability of change in these indicators were estimated by logistic regression analysis. Three main groups of thresholds could be identified: one around 2 micrograms Cd/g creatinine mainly associated with biochemical alterations, a second around 4 micrograms Cd/g creatinine for high molecular weight proteins and some tubular antigens or enzymes, and a third one around 10 micrograms Cd/g creatinine for low molecular weight proteins and other indicators. The recent recommendation by the American Conference of Governmental Industrial Hygienists (ACGIH) of 5 micrograms Cd/g creatinine in urine as the biological exposure limit for occupational exposure to Cd appears thus justified, although for most of the effects occurring around this threshold the link with the subsequent development of overt Cd nephropathy is not established. In that respect, the very early interference with production of some prostanoids (threshold 2 micrograms Cd/g creatinine) deserves further investigation; although this effect might contribute to protect the filtration capacity of the kidneys, it might also play a part in the toxicity of Cd on bone.


Subject(s)
Cadmium Poisoning/complications , Kidney Diseases/chemically induced , Kidney/drug effects , Occupational Diseases/chemically induced , Occupational Exposure , Adult , Biomarkers/blood , Biomarkers/urine , Cadmium Poisoning/blood , Cadmium Poisoning/urine , Humans , Kidney Diseases/blood , Kidney Diseases/urine , Occupational Diseases/blood , Occupational Diseases/urine
20.
Prostaglandins ; 44(6): 597-609, 1992 Dec.
Article in English | MEDLINE | ID: mdl-1475378

ABSTRACT

N-phenyllinoleamide (NPLA), the anilide of linoleic acid, has been associated with the epidemiology of Toxic Oil Syndrome, but no data are available on its metabolism. On account of the similarity in chemical structure between the linoleic acid and NPLA, the aim of this study has been to investigate the oxidative metabolism of this xenobiotic by the human nasal polyp, a tissue with elevated 15-lipoxygenase activity. For this purpose, tissue homogenates have been incubated for 2 h with NPLA (0.1 mM) spiked with either N-(ring G-3H)PLA (0.2 microCi/ml) or N-P(1-14C)LA (0.05 microCi/ml). Gas chromatographic/mass spectrometric analysis of the high performance liquid radiochromatographic fractions shows that the 9,12,13-trihydroxy, 12,13-epoxy-11-hydroxy and 13-hydroxy NPLA derivatives are the major metabolites. These results revealed that NPLA metabolites are chemical structures related to the linoleic acid derivatives, some of which may show biological activity.


Subject(s)
Anilides/metabolism , Linoleic Acids/metabolism , Nasal Polyps/metabolism , Chromatography, High Pressure Liquid , Gas Chromatography-Mass Spectrometry , Humans , In Vitro Techniques , Molecular Structure , Oxidation-Reduction
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