The Experts below are selected from a list of 52245 Experts worldwide ranked by ideXlab platform
Matthias Griese - One of the best experts on this subject based on the ideXlab platform.
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Quantitation of surfactant protein B by HPLC in Bronchoalveolar Lavage Fluid
Journal of Chromatography B, 2005Co-Authors: Christian Paschen, Matthias GrieseAbstract:Abstract A sensitive reversed phase HPLC method with evaporative light scattering detection (ELSD) was developed for the determination of the hydrophobic surfactant protein B (SP-B) in human Bronchoalveolar Lavage Fluid. Samples were extracted two times with CHCl3:MeOH:HCl (2:3:0.005N) solution in a ratio of 1:2 by volume. The extract of the lower phase was separated on a C4 butyl silica gel column with an isocratic elution using a mobile phase, consisting of 97% methanol, 2.75% chloroform and 0.25% 0.1 M trifluoroacetic acid (by volume), at a flow rate of 1 ml/min. SP-B was detected by ELSD and quantified by comparison to an external standard. The duration of a run was 7 min, the quantification limit 30 ng and the limit of detection was at about 15 ng of SP-B. This method is suitable for the rapid routine quantification of SP-B in human Bronchoalveolar Lavage Fluid samples.
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Surfactant protein A and other Bronchoalveolar Lavage Fluid proteins are altered in cystic fibrosis
The European respiratory journal, 2001Co-Authors: C. Von Bredow, Peter Birrer, Matthias GrieseAbstract:Inflammation and proteolytic processes play an important role in the progression of cystic fibrosis (CF) lung disease. The goal of this study was to describe Bronchoalveolar Lavage Fluid (BALF) protein pattern of CF patients in comparison to controls and to assess if there is proteolytic degradation of surfactant protein A (SP-A), an important innate host defence component of the lungs. BALFs from 17 clinically stable CF patients and from eight healthy children were separated by two-dimensional gel electrophoresis. Silver staining was used to show BALF proteins and Western blotting to detect SP-A isoforms. In CF, BALF proteins of a low molecular weight ≤20 kD were more abundant than in controls. Various proteins were seen in CF which were not present in controls and vice versa . Degradation of SP-A was present in 15 of 17 CF BALFs but in none of the controls, in contrast polymeric isoforms were seen in all controls and in four of 17 CF patients. Proteolytic damage to surfactant protein A and significant changes of normal Bronchoalveolar Lavage Fluid proteins occur in lungs of cystic fibrosis patients. Identification of altered Bronchoalveolar Lavage Fluid proteins may give new insights into pathogenic mechanisms and provide new targets for therapy. This project was supported by a grant from the Wilhelm Sander Stiftung (Gr 93.002.1/2).
Nancy L Wengenack - One of the best experts on this subject based on the ideXlab platform.
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a real time polymerase chain reaction assay for detection of pneumocystis from Bronchoalveolar Lavage Fluid
Diagnostic Microbiology and Infectious Disease, 2006Co-Authors: Rodney C Arcenas, Seanne P Buckwalter, Andrew H Limper, Dana Crino, Glenn D Roberts, Nancy L WengenackAbstract:Pneumocystis jiroveci is an important cause of pneumonia in immunocompromised individuals. This organism cannot be cultured, and therefore, diagnosis relies on microscopic identification of the organism using stains or antibodies. Although simple, these tests are insensitive and require expertise for accurate interpretation. We developed a real-time polymerase chain reaction (PCR) assay that provides sensitive and objective detection of Pneumocystis from Bronchoalveolar Lavage Fluid. Primers and fluorescence resonance energy transfer probes were developed that target the cdc2 gene of P. jiroveci. Assay sensitivity is 6 copies of target per microliter of sample. No cross-reactivity occurs with other pathogens, and the PCR assay has a 21% increase in clinical sensitivity as compared with Calcofluor white staining. The real-time PCR assay provides a sensitive, rapid, and objective method for the detection of Pneumocystis from Bronchoalveolar Lavage Fluid.
Fariba Sabounchi-schütt - One of the best experts on this subject based on the ideXlab platform.
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Changes in Bronchoalveolar Lavage Fluid proteins in sarcoidosis: a proteomics approach
The European respiratory journal, 2003Co-Authors: Fariba Sabounchi-schütt, Jonas Åström, Ulf Hellman, Anders Eklund, Johan GrunewaldAbstract:In sarcoidosis, an inflammatory lung disease, the protein profile of Bronchoalveolar Lavage Fluid (BALF) is altered. To study the BALF protein pattern changes in sarcoidosis, samples from six patients and four healthy individuals were analysed by two-dimensional polyacrylamide gel electrophoresis. A comparison of the protein-spot patterns showed a significantly higher number of protein spots in the pH range 5.5-6.7 in patients compared to controls (472 versus 384). Furthermore, the number of protein spots in the patients were significantly decreased in the acidic pH range 4.5-5.5 (399 versus 518). Measurement of the optical density in the gels showed varying expression levels for several protein spots. Seventeen of the altered protein spots were identified, of which seven have previously not been reported for BALF. Many of these are nonplasma proteins involved in the inflammatory and oxidant-antioxidant processes. In conclusion, the Bronchoalveolar Lavage Fluid protein content is altered in sarcoidosis patients, especially for proteins that are not derived from plasma. The described proteomics approach will in the future be used to asses overall changes in the protein content associated with sarcoidosis and may offer the possibility of identifying disease-specific proteins.
Shi-chuan Chang - One of the best experts on this subject based on the ideXlab platform.
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Diagnosis of pulmonary alveolar proteinosis: usefulness of papanicolaou-stained smears of Bronchoalveolar Lavage Fluid.
Archives of internal medicine, 2001Co-Authors: Chung-wei Chou, Fang-chi Lin, Su-mei Tung, Rong-dih Liou, Shi-chuan ChangAbstract:Background The globules (stained green, orange, or orange in the center coated with a green rim) seen in Papanicolaou-stained smears of Bronchoalveolar Lavage Fluid are suggested to be characteristic of pulmonary alveolar proteinosis (PAP). Objective To evaluate the usefulness of Papanicolaou-stained smears of Bronchoalveolar Lavage Fluid in aiding a diagnosis of PAP. Methods Papanicolaou-stained smears of Bronchoalveolar Lavage Fluid obtained from 7 patients (5 idiopathic, 2 secondary) with PAP were evaluated. To serve as controls, the smears of 11 normal subjects and 128 patients with other pulmonary disorders were also examined. The findings on the presence and number of globules were recorded. To differentiate PAP from other pulmonary disorders, the highest globule value obtained from the control group was chosen as the cutoff point. Results The characteristic globules were not found in normal subjects and only found in 6 of 128 patients with other pulmonary disorders. Their clinical diagnoses were Sjogren syndrome in 2 cases; polymyositis, idiopathic pulmonary fibrosis, asbestosis, and hypersensitivity pneumonitis in 1 case each. The numbers of globules in these 6 patients were 1, 3, 17, 7, 3, and 2. In contrast, more than 100 globules were found in all patients with PAP. The number of globules was highly sensitive and specific in aiding a diagnosis of PAP when the cutoff value was set at 18. Conclusion The globules seen in Papanicolaou-stained smears of Bronchoalveolar Lavage Fluid may be valuable in aiding a diagnosis of PAP, especially when the number of globules is more than 18.
Johan Grunewald - One of the best experts on this subject based on the ideXlab platform.
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Bronchoalveolar Lavage Fluid exosomes contribute to cytokine and leukotriene production in allergic asthma
Allergy, 2012Co-Authors: Torregrosa P Paredes, Johan Grunewald, Julia Esser, Charlotte Admyre, Magnus Nord, Q K Rahman, Ana Lukic, Olof Radmark, R Gronneberg, Anders EklundAbstract:Background Leukotrienes (LTs) are potent pro-inflammatory mediators involved in asthma. Exosomes, nanosized vesicles released from various cells, can stimulate or down-regulate immune responses, depending on the state and nature of the originating cell. We have recently shown an altered exosome profile in Bronchoalveolar Lavage Fluid (BALF) of patients with sarcoidosis, but their role in asthma is unknown. Our aims were to investigate whether exosomes from BALF have LT biosynthetic capacity and to explore phenotypic and functional characteristics of BALF exosomes in asthma. Methods Bronchoalveolar Lavage Fluid exosomes were collected from healthy individuals (n = 13) and patients with mild allergic asthma to birch pollen (n = 12) before and after birch allergen provocation. Exosomes were characterized by flow cytometry and Western blot. Their capacity to induce IL-8 and LT production in the human bronchial epithelial cell (BEC) line 16HB14o- was measured by ELISA and reverse-phase HPLC, respectively. Results Compared to BALF exosomes from healthy individuals, BALF exosomes from asthmatics displayed higher levels of exosome-associated markers, such as the tetraspanins CD63 and CD81 and the scavenger receptor CD36. No major differences were observed between BALF exosomes from before and after allergen provocation. Furthermore, we show that BALF exosomes contain enzymes for LT biosynthesis. The effect of exosomes to promote LTC4 and IL-8 release in BEC was significantly increased for exosomes from asthmatics, and the CysLT1 receptor antagonist Montelukast reduced exosome-induced IL-8 secretion. Conclusions Bronchoalveolar Lavage Fluid exosomes from asthmatic and healthy individuals exhibit distinct phenotypes and functions. BALF exosomes from asthmatics might contribute to subclinical inflammation by increasing cytokine and LTC4 generation in airway epithelium.
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Changes in Bronchoalveolar Lavage Fluid proteins in sarcoidosis: a proteomics approach
The European respiratory journal, 2003Co-Authors: Fariba Sabounchi-schütt, Jonas Åström, Ulf Hellman, Anders Eklund, Johan GrunewaldAbstract:In sarcoidosis, an inflammatory lung disease, the protein profile of Bronchoalveolar Lavage Fluid (BALF) is altered. To study the BALF protein pattern changes in sarcoidosis, samples from six patients and four healthy individuals were analysed by two-dimensional polyacrylamide gel electrophoresis. A comparison of the protein-spot patterns showed a significantly higher number of protein spots in the pH range 5.5-6.7 in patients compared to controls (472 versus 384). Furthermore, the number of protein spots in the patients were significantly decreased in the acidic pH range 4.5-5.5 (399 versus 518). Measurement of the optical density in the gels showed varying expression levels for several protein spots. Seventeen of the altered protein spots were identified, of which seven have previously not been reported for BALF. Many of these are nonplasma proteins involved in the inflammatory and oxidant-antioxidant processes. In conclusion, the Bronchoalveolar Lavage Fluid protein content is altered in sarcoidosis patients, especially for proteins that are not derived from plasma. The described proteomics approach will in the future be used to asses overall changes in the protein content associated with sarcoidosis and may offer the possibility of identifying disease-specific proteins.
