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Clara cell protein in nasal lavage fluid and nasal nitric oxide - biomarkers with anti-inflammatory properties in allergic rhinitis
Kristina Irander, J?rgen P Palm, Magnus P Borres, Bijar Ghafouri
Clinical and Molecular Allergy , 2012, DOI: 10.1186/1476-7961-10-4
Abstract: The levels of CC16 in nasal lavage fluids, achieved from subjects with persistent allergic rhinitis (n = 13), intermittent allergic rhinitis in an allergen free interval (n = 5) and healthy controls (n = 7), were analyzed by Western blot. The levels of nNO were measured by the subtraction method using NIOX?. The occurrences of effector cells in allergic inflammation, i.e. metachromatic cells (MC, mast cells and basophiles) and eosinophils (Eos) were analyzed by light microscopy in samples achieved by nasal brushing.The levels of CC16 correlated with nNO levels (r2 = 0.37; p = 0.02) in allergic subjects.The levels of both biomarkers showed inverse relationships with MC occurrence, as higher levels of CC16 (p = 0.03) and nNO (p = 0.05) were found in allergic subjects with no demonstrable MC compared to the levels in subjects with demonstrable MC. Similar relationships, but not reaching significance, were observed between the CC16 and nNO levels and Eos occurrence. The levels of CC16 and nNO did not differ between the allergic and the control groups.The correlation between nasal CC16 and nNO levels in patients with allergic rhinitis, along with an inverse relationship between their levels and the occurrences of MC in allergic inflammation, may indicate that both biomarkers have anti-inflammatory effects by suppression of cell recruitment. The mechanisms behind these observations warrant further analyses.Clara cell protein (CC16, identical to CC10 and uteroglobin) is a biomarker of high interest in airway diseases. The protein, initially described in the epithelium of the tracheobronchial tree as a secretory product from non-ciliated Clara cells, diffuses passively from the respiratory tract into serum and is excreted via the urinary tract [1]. CC16 is ascribed a protective role against oxidative stress and inflammation in the respiratory tract [2]. Due to a particular vulnerability of Clara cells to lung toxicants CC16 has also been evaluated as a useful biomarker of r
Effects of ultrafine particles-induced oxidative stress on Clara cells in allergic lung inflammation
Francesca Alessandrini, Ingrid Weichenmeier, Erik van Miert, Shinji Takenaka, Erwin Karg, Cornelia Blume, Martin Mempel, Holger Schulz, Alfred Bernard, Heidrun Behrendt
Particle and Fibre Toxicology , 2010, DOI: 10.1186/1743-8977-7-11
Abstract: Ovalbumin (OVA)-sensitized mice were exposed to EC-UFP (507 μg/m3 for 24 h) or filtered air immediately prior to allergen challenge and systemically treated with N-acetylcysteine (NAC) or vehicle prior and during EC-UFP inhalation. CC16 was measured up to one week after allergen challenge in bronchoalveolar lavage fluid (BALF) and in serum. The relative expression of CC16 and TNF-α mRNA were measured in lung homogenates. A morphometrical analysis of mucus hypersecretion and electron microscopy served to investigate goblet cell metaplasia and Clara cell morphological alterations.In non sensitized mice EC-UFP inhalation caused alterations in CC16 concentration, both at protein and mRNA level, and induced Clara cell hyperplasia. In sensitized mice, inhalation of EC-UFP prior to OVA challenge caused most significant alterations of BALF and serum CC16 concentration, BALF total protein and TNF-α relative expression compared to relevant controls; their Clara cells displayed the strongest morphological alterations and strongest goblet cell metaplasia occurred in the small airways. NAC strongly reduced both functional and morphological alterations of Clara cells.Our findings demonstrate that oxidative stress plays an important role in EC-UFP-induced augmentation of functional and morphological alterations of Clara cells in allergic lung inflammation.There is rapidly increasing epidemiological evidence associating respiratory health effects and exposures to ultrafine particles (UFP) in a susceptible population, such as elderly, young children, and people with pre-existing respiratory conditions [1]. For adult asthmatics, ambient levels of UFP (aerodynamic diameter <0.1 μm) were found to have a higher specific toxicological role compared to larger particles [2]. UFP, due to their large surface area, have enhanced capabilities to produce reactive oxygen species [3].Clara cells are non-ciliated secretory epithelial cells lining the pulmonary airways, distinct from mucous and ser
Novel Method for Isolation of Murine Clara Cell Secretory Protein-Expressing Cells with Traces of Stemness  [PDF]
Xiao-Yang Wang,Kathleen M. Keefe,Sandra M. Jensen-Taubman,Danlei Yang,Kai Yan,R. Ilona Linnoila
PLOS ONE , 2012, DOI: 10.1371/journal.pone.0043008
Abstract: Clara cells are non-ciliated, secretory bronchiolar epithelial cells that serve to detoxify harmful inhaled substances. Clara cells also function as stem/progenitor cells for repair in the bronchioles. Clara cell secretory protein (CCSP) is specifically expressed in pulmonary Clara cells and is widely used as a Clara cell marker. In addition CCSP promoter is commonly used to direct gene expression into the lung in transgenic models. The discovery of CCSP immunoreactivity in plasma membranes of airway lining cells prompted us to explore the possibility of enriching Clara cells by flow cytometry. We established a novel and simple method for the isolation of CCSP-expressing cell Clara cells using a combination of mechanical and enzymatic dissociation followed by flow cytometry sorting technology. We showed that ~25% of dissociated cells from whole lung expressed CCSP. In the resulting preparation, up to 98% of cells expressed CCSP. Notably, we found that several common stem cell markers including CD44, CD133, Sca-1 and Sox2 were expressed in CCSP+ cells. Moreover, CCSP+ cells were able to form spheroid colonies in vitro with 0.97‰ efficiency. Parallel studies in vivo confirmed that a small population of CCSP?expressing cells in mouse airways also demonstrates stem cell-like properties such as label retention and harboring rare bronchioalveolar stem cells (BASCs) in terminal bronchioles (TBs). We conclude that CCSP+ cells exhibit a number of stem cell-like features including stem cell marker expression, bronchosphere colony formation and self-renewal ability. Clara cell isolation by flow cytometry sorting is a useful method for investigating the function of primary Clara cells in stem cell research and mouse models.
Plasma CC16 levels are associated with development of ALI/ARDS in patients with ventilator-associated pneumonia: a retrospective observational study
Rogier M Determann, Julian L Millo, Sam Waddy, Rene Lutter, Chris S Garrard, Marcus J Schultz
BMC Pulmonary Medicine , 2009, DOI: 10.1186/1471-2466-9-49
Abstract: We performed serial measurements of Clara cell protein (CC16), soluble receptor for advanced glycation end products (sRAGE), surfactant protein D (SP-D) and Krebs von den Lungen (KL-6) in plasma of patients with VAP and mechanically ventilated control patients without VAP. ALI/ARDS was diagnosed using the criteria of the North-American European consensus conference.Thirty-seven patients were enrolled - 22 patients with VAP and 15 control patients. Ten patients with pneumonia met the ALI/ARDS consensus criteria. Control patients never met these criteria. Plasma CC16 had a good diagnostic capacity for ALI/ARDS as shown by the receiver operating characteristic curve with an area under the curve of 0.91 (95% confidence interval (CI) 0.79 - 1.00; p < 0.001). Identification of ALI/ARDS patients by sudden increases in plasma CC16 of 30% or more yielded a sensitivity of 90% and a specificity of 92%. Of note, levels of CC16 increased 2 days before ALI/ARDS diagnosis. A cut-off level of 50 ng/ml SP-D yielded a specificity of 100% while the sensitivity was 70%. The area under the curve for SP-D was 0.80 (95% CI 0.58 - 1.00; p = 0.02). The diagnostic accuracies of KL-6 and sRAGE were low.Plasma CC16 seems a potential biological marker for ALI/ARDS in patients with VAP. Plasma levels of sRAGE, SP-D and KL-6 have limited discriminative power for diagnosing ALI/ARDS in VAP.Intubated and mechanically ventilated patients are at risk for ventilator-associated pneumonia (VAP) [1]. Development of acute lung injury (ALI) or its more severe form, acute respiratory distress syndrome (ARDS) decreases the chance of survival [2]. Early and adequate recognition of ALI/ARDS is mandatory for intensive care physicians to take sufficient action at the right time (e.g., the use of so-called lung-protective mechanical ventilation [3], and refinement of fluid management [4]). In today's intensive care practice, ALI/ARDS is diagnosed by means of the North-American European consensus conference (NAECC
Clara cell adhesion and migration to extracellular matrix
Jeffrey J Atkinson, Tracy L Adair-Kirk, Diane G Kelley, Daphne deMello, Robert M Senior
Respiratory Research , 2008, DOI: 10.1186/1465-9921-9-1
Abstract: Using a modification of existing procedures for Clara cell isolation, we examined mouse Clara cells and a mouse Clara-like cell line (C22) for adhesion to and migration toward matrix substrate gradients, to establish the nature and integrin dependence of migration in Clara cells.We observed that Clara cells adhere preferentially to fibronectin (Fn) and type I collagen (Col I) similar to previous reports. Migration of Clara cells can be directed by a fixed gradient of matrix substrates (haptotaxis). Migration of the C22 cell line was similar to the Clara cells so integrin dependence of migration was evaluated with this cell line. As determined by competition with an RGD containing-peptide, migration of C22 cells toward Fn and laminin (Lm) 511 (formerly laminin 10) was significantly RGD integrin dependent, but migration toward Col I was RGD integrin independent, suggesting that Clara cells utilize different receptors for these different matrices.Thus, Clara cells resemble alveolar type II and bronchiolar ciliated epithelial cells by showing integrin mediated pro-migratory changes to extracellular matrix components that are present in tissues after injury.Clara cells are epithelial cells on the luminal surface of airways with a dome shaped cytoplasmic protrusion and no cilia [1,2]. In addition to their secretory and xenobiotic roles [3,4], Clara cells are the progenitor cell in small airways [5]. After airway injury, Clara cells in stem cell niches proliferate and migrate to replenish the injured terminally differentiated epithelial cells [6]. In fact, after alveolar injury, Clara cells can be seen in the alveolus (alveolar bronchiolization), suggesting the response of the terminal airway epithelium to alveolar injury exceeds the rate of alveolar epithelial cell repair [7,8].Epithelial repair requires a complex series of steps including cell spreading and/or migration over the exposed interstitial matrix and provisional matrix to form intact tight junctions (restitutio
Comparison of Compression Schemes for CLARA  [PDF]
Peter H. Williams,James K. Jones,Julian W. McKenzie
Physics , 2012,
Abstract: CLARA (Compact Linear Advanced Research Accelerator)at Daresbury Laboratory is proposed to be the UK's national FEL test facility. The accelerator will be a ~250 MeV electron linac capable of producing short, high brightness electron bunches. The machine comprises a 2.5cell RF photocathode gun, one 2 m and three 5 m normal conducting S-band (2998MHz) accelerating structures and a variable magnetic compression chicane. CLARA will be used as a test bed for novel FEL configurations. We present a comparison of acceleration and compression schemes for the candidate machine layout.
A clara e a gema
Márcia Denise de Oliveira Godoy
ETD : Educa??o Temática Digital , 2001,
Abstract: RONCA, Afonso Caruso; GON ALVES, Luiz Carlos . A clara e a gema. S o Paulo: Edesplan,1998.
Herbal treatment of secretory diarrhea  [cached]
H.M. Asif,Khan Usmanghni,Muhammad Akram,Naveed Akhtar
International Journal of Phytomedicine , 2011,
Abstract: The research study is conducted to understand interaction of illness, symptoms, context, patients response and the clinical skill vis a vis better management of secretory diarrhea. Specific aim of this study is to determine the impact of intensive medical intervention with herbal drug “Dirasif” (Test) and allopathic drug “Furoxone” (Control) to treat secretory diarrhea. This is randomized controlled clinical trial in primary care with an open intervention. All patients judged by the physician to need either herbal or allopathic medicine for secretory diarrhea are randomized in treatment therapy. Clinical trial was conducted on hundred patients from both groups i.e.50 patient from control and 50 from experimental group having age between 12-40 year.Comparison of data recorded by participants relating to these variables showed significant differences between test and control groups (p < 0.05) despite the fact that no side effects were recorded in test group. Overall clinical success was observed in both treatment groups however the efficacy of the test treated medication (Dirasif) was superior as p=0.03. Dirasif is more effective than the Furoxone in the treatment of secretory diarrhea in Gadap community Karachi, Pakistan. Keywords: Secretory diarrhea, efficacy, dirasif, furoxone
Cell proliferation and interferon-gamma response to recombinant MBP-3, NarL, MT-10.3, and 16kDa Mycobacterium tuberculosis antigens in Brazilian tuberculosis patients
Tavares, Ricardo Candido Oliveira;Salgado, Jorge;Moreira, Valéria Barbosa;Ferreira, M?nica Antonia S;Mello, Fernanda Carvalho Queoz;Leung, Janaína AW;Singh, Mahavir;Fonseca, Leila de Souza;Saad, Maria Helena Feres;
Memórias do Instituto Oswaldo Cruz , 2006, DOI: 10.1590/S0074-02762006000800006
Abstract: human pulmonary tuberculosis (tb) is a worldwide public health problem. in resistant individuals, control of the infection mainly requires development of a th1 cell immune response with production of cytokines, of which interferon-g (ifn-g)plays an important role. several antigens from mycobacterium tuberculosis complex has been described for use in vaccine development or for diagnostic purposes, however little evaluation has been done in endemic area for tb. the proliferative and ifn-g human t cell immune responses, to four recombinant proteins (mbp-3, narl, mt-10.3, 16 kda) and ppd, of 38 brazilian tb patients (6 untreated and 32 treated) and 67 controls (38 positive and 29 negative tuberculin skin test - tst) were compared. the highest reactivity mean rate was obtained with ppd followed by 16 kda in tb patients. while most of the patients (87%) and controls (> 64%) respond to the ppd, 16kda was more specifically recognized (> 21%) although less sensitive (54%). when tb patients were divided according to treatment status, opposite to ppd, higher average level of ifn-g was induced by 16kda in untreated (505 pg/ml) compared to treated tb patients and tst+ (269.8 pg/ml x 221.6pg/ml, respectively), although the difference was not significant. these data show that in contrast with the other recombinant proteins, the stimulatory potency of 16kda to induce proliferative and inf-g response was more effective and is more recognized by active tb untreated patients, eliciting in control individuals a more selective immune response than ppd.
Prevalence of Aeroallergens in Allergic Rhinitis in Shiraz
Sara Kashef,Mohammad Amin Kashef,Fardin Eghtedari
Iranian Journal Of Allergy, Asthma and Immunology , 2003,
Abstract: Allergic rhinitis is an extremely common disease worldwide. Aeroallergens are very often involved in allergic rhinitis and their prevalence may vary in differ ent regions. The causative allergens of allergic rhinitis in our area are unknown.The purpose of this study was to determine the prevalence of skin reactivity to different aeroallergens in patients with allergic rhinitis in the city of Shiraz, Iran.A total of 212 patients who were referred to Motahari Allergy Clinic with chronic rhinitis were subjected to skin prick test (SPT) with a series of common allergenic extracts including grasses, weeds, trees, house dust mites and moulds.One hundred and thirty two subjects (62.2%) had positive SPT to at least one aeroallergen. Male to female ratio was 1.2 and mean age was 18.2 years. The prevalence rates for allergen groups were: pollens (92.4%), mites (22.7%) and moulds (8.3%). Among 122 patients reactive to pollens, 92 (75.4%) showed skin reactivity to weeds, 78 (63.9%) to grasses and 68 (55.7%) to trees. Polysensitization was common, with 75.7% of all sensitized patients being posi tive to more than one aeroallergen.Pollens are the main sensitizing allergens among patients with allergic rhinitis in Shiraz. This pattern of prevalence was expected based on herbal geography, climate and also found to be compatible with the results from studies carried out in places with the same habitat.
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