SFQ data processing with set/reset information

We propose a new class of SFQ logic circuits. In this new approach, an SFQ pulse represents the transition between "zero" and "one". By using these two bits of information, a one-to-one correspondence between input and output can be realized. Since the correspondence is then the same as in semiconductor circuits, this method permits logic design without clock elements. In order to carry out this logic, we propose the most fundamental element, a new DC/SFQ converter. A computer simulation and low-speed test were performed. Both results showed that this converter operates correctly with a wide margin. Moreover, this converter also pro, ides the basis for many other logic elements such as AND, OR, and XOR

β2 adrenergic agonist suppresses eosinophil-induced epithelial-to-mesenchymal transition of bronchial epithelial cells

Abstract Background Epithelial-mesenchymal transition is currently recognized as an important mechanism for the increased number of myofibroblasts in cancer and fibrotic diseases. We have already reported that epithelial-mesenchymal transition is involved in airway remodeling induced by eosinophils. Procaterol is a selective and full β2 adrenergic agonist that is used as a rescue of asthmatic attack inhaler form and orally as a controller. In this study, we evaluated whether procaterol can suppress epithelial-mesenchymal transition of airway epithelial cells induced by eosinophils. Methods Epithelial-mesenchymal transition was assessed using a co-culture system of human bronchial epithelial cells and primary human eosinophils or an eosinophilic leukemia cell line. Results Procaterol significantly inhibited co-culture associated morphological changes of bronchial epithelial cells, decreased the expression of vimentin, and increased the expression of E-cadherin compared to control. Butoxamine, a specific β2-adrenergic antagonist, significantly blocked changes induced by procaterol. In addition, procaterol inhibited the expression of adhesion molecules induced during the interaction between eosinophils and bronchial epithelial cells, suggesting the involvement of adhesion molecules in the process of epithelial-mesenchymal transition. Forskolin, a cyclic adenosine monophosphate-promoting agent, exhibits similar inhibitory activity of procaterol. Conclusions Overall, these observations support the beneficial effect of procaterol on airway remodeling frequently associated with chronic obstructive pulmonary diseases

Eosinophils Promote Epithelial to Mesenchymal Transition of Bronchial Epithelial Cells

<div><p>Eosinophilic inflammation and remodeling of the airways including subepithelial fibrosis and myofibroblast hyperplasia are characteristic pathological findings of bronchial asthma. Epithelial to mesenchymal transition (EMT) plays a critical role in airway remodelling. In this study, we hypothesized that infiltrating eosinophils promote airway remodelling in bronchial asthma. To demonstrate this hypothesis we evaluated the effect of eosinophils on EMT by <i>in vitro</i> and <i>in vivo</i> studies. EMT was assessed in mice that received intra-tracheal instillation of mouse bone marrow derived eosinophils and in human bronchial epithelial cells co-cultured with eosinophils freshly purified from healthy individuals or with eosinophilic leukemia cell lines. Intra-tracheal instillation of eosinophils was associated with enhanced bronchial inflammation and fibrosis and increased lung concentration of growth factors. Mice instilled with eosinophils pre-treated with transforming growth factor(TGF)-β1 siRNA had decreased bronchial wall fibrosis compared to controls. EMT was induced in bronchial epithelial cells co-cultured with human eosinophils and it was associated with increased expression of TGF-β1 and Smad3 phosphorylation in the bronchial epithelial cells. Treatment with anti-TGF-β1 antibody blocked EMT in bronchial epithelial cells. Eosinophils induced EMT in bronchial epithelial cells, suggesting their contribution to the pathogenesis of airway remodelling.</p></div

Eol-1 cells induce EMT in BEAS-2B cells via the JNK/Smad3 pathway.

<p>(A) The concentration of TGF-β1 in the supernatant of BEAS-2B in the presence of PI3 kinase (LY294002), JNK (SP600125), MEK1/2 (MUO126) or p38 kinase (SB203580) inhibitor. Inhibitors of both PI3 and JNK kinases suppress the morphological features (B, C, D, E) and molecular markers (F, G, H) of EMT. Data are expressed as means ± SEM. *P<0.05; **P<0.01.</p

Eol-1 cell lines induce EMT in BEAS-2B cells by increasing TGF-β1 expression.

<p>(A, B) Control and BEAS-2B cells co-cultured with EoL-1 cells. (C) BEAS-2B cells co-cultured with EoL-1 cells in the presence of anti-TGF-β1 neutralizing antibody. BEAS-2B cells lysates were subjected to RT-PCR analysis. The expression level of E-cadherin (D) and vimentin (E) mRNA was measured by RT-PCR and normalized to that of GAPDH mRNA. TGF-β1 (F) levels in the supernatant from each culture condition, and the ratio of p-Smad3 to Smad3 (G) in BEAS-2B cells as analyzed by densitometry. The scale bars indicate 50 µm. Data are expressed as means ± SEM. *P<0.05.</p

Relevant role of TGF-β1 in airway remodeling and the need of direct contact of eosinophils for TGF-β1 production.

<p>(A, B, C) Mice receiving instillation of eosinophils pre-treated with TGF-β1 siRNA have less peribronchial fibrosis than those receiving eosinophils pre-treated with scrambled RNA as assessed by Masson staining. (D) Quantification of Masson positive areas disclosed significant difference. (E, F) TGFβ1 levels in the supernatant in Boyden chamber assay and using 2% paraformaldehyde-fixed eosinophils. Culture supernatants were used to measure TGF-β1 by ELISA. Data are expressed as means ± SEM. **P<0.05. The scale bars indicate 100 µm. Data are expressed as means ± SEM. **P<0.05.</p

Eosinophils induced secretion of TGF-β1 from BEAS-2B cells.

<p>(A) TGF-β1 levels in the supernatant from each cell condition. (B) Representative Western blotting of Smad3 and p-Smad3. (C) The ratio of p-Smad3 to Smad3 analyzed by densitometry. (D, E, F) Morphological changes in each cell condition. (G, H, I, J) Effect of anti-TGF-β1 antibody on the protein level of TGF-β1 and RNA expression of E-cadherin and vimentin during each cell condition. BEAS-2B cells were stained with DIFF-QUICK. The scale bars indicate 50 µm. Data are expressed as means ± SEM. *P<0.05.</p

Inflammation and fibrosis of the airway walls induced by eosinophils instillation <i>in vivo</i>.

<p>(A–F) Representative lung sections from control (n = 3; A, C, E) and eosinophil-instilled mice (n = 4; B, D, F). Specimens were stained with Hematoxylin-Eosin (A, B), Masson trichrome (C, D) and immunostained for type I collagen (E, F) and examined by light microscopy. Mice receiving intra-tracheal eosinophils had increased infiltration of inflammatory cells around the bronchi (B), peribronchial fibrosis (D) and deposition of type I collagen (F) in lung tissue compared to controls (A, C and E). (G, H) Collagen deposition (G) and immunoreactive areas for type I collagen (H) were quantified using WinROOF software. Lung tissue of eosinophil-instilled mice showed significant deposition of collagen as compared to control. The data are expressed as the mean percentage of the total lung field area ± S.E.M. The scale bars indicate 50 µm. Data are expressed as means ± SEM. *P<0.05.</p

Human eosinophils induced EMT morphological changes in BEAS-2B cells.

<p>(A, D) control, (B, E) BEAS-2B cells co-cultured with eosinophils. (C, F) BEAS-2B cells cultured in the presence of TGF-β1. BEAS-2B cells were stained with Diff-Quick after washing out eosinophils (A, B, C). (D, E, F) Immunofluorescence staining was performed with phalloidin (red) and DAPI for nuclei staining (blue) after washing out eosinophils. The scale bars indicate 50 µm. (G, H, I) RNA expression of E-cadherin and vimentin in BEAS-2B cells co-cultured with or without eosinophils and in the presence of TGF-β1 as assessed by RT-PCR and densitometry analysis. (K, L, M) Protein expression of E-cadherin and αSMA in BEAS-2B cells co-cultured with or without eosinophils as assessed by Western blotting and densitometry analysis. Data are expressed as means ± SEM. **P<0.01; *P<0.05.</p