α1A-Adrenergic Receptor Induces Activation of Extracellular Signal-Regulated Kinase 1/2 through Endocytic Pathway

G protein-coupled receptors (GPCRs) activate mitogen-activated protein kinases through a number of distinct pathways in cells. Increasing evidence has suggested that endosomal signaling has an important role in receptor signal transduction. Here we investigated the involvement of endocytosis in α1A-adrenergic receptor (α1A-AR)-induced activation of extracellular signal-regulated kinase 1/2 (ERK1/2). Agonist-mediated endocytic traffic of α1A-AR was assessed by real-time imaging of living, stably transfected human embryonic kidney 293A cells (HEK-293A). α1A-AR was internalized dynamically in cells with agonist stimulation, and actin filaments regulated the initial trafficking of α1A-AR. α1A-AR-induced activation of ERK1/2 but not p38 MAPK was sensitive to disruption of endocytosis, as demonstrated by 4°C chilling, dynamin mutation and treatment with cytochalasin D (actin depolymerizing agent). Activation of protein kinase C (PKC) and C-Raf by α1A-AR was not affected by 4°C chilling or cytochalasin D treatment. U73122 (a phospholipase C [PLC] inhibitor) and Ro 31–8220 (a PKC inhibitor) inhibited α1B-AR- but not α1A-AR-induced ERK1/2 activation. These data suggest that the endocytic pathway is involved in α1A-AR-induced ERK1/2 activation, which is independent of Gq/PLC/PKC signaling

In Situ Expression of CD40, CD40L (CD154), IL-12, TNF-a, IFN-g and TGF-b1 in Murine Lungs during Slowly Progressive Primary Tuberculosis

The distribution and expression of CD40, its ligand CD40L (154) and related cytokines interleukin-12 (IL-12), tumour necrosis factor-α (TNF-α), interferon-γ (IFN-γ) and transforming growth factor-β1 (TGF-β1) were studied in the lungs of B6D2F1 hybrid mice during slowly progressive primary tuberculosis (TB) by immunohistochemistry. CD40 and CD40L are implicated in cell-mediated immunity (CMI) causing activation or apoptosis of infected cells. The phenomenon of apoptosis is associated with Mycobacterium tuberculosis survival. In this study, using frozen lung sections (n = 33), our results showed increased CD40, IL-12 and TGF-β1 expression in macrophages with progression of disease. High percentages of mycobacterial antigens (M.Ags), CD40L and IFN-γ expression were maintained throughout infection, and TNF-α-expressing cells were decreased. In lymphocytes, the percentage of IFN-γ-positive cells was increased, but CD40L and IL-12 were maintained with the progression of disease. M.Ags, CD40 and CD40L were expressed in the same areas of the lesions. We conclude that changes in the expression of CD40–CD40L and cytokines associated with M. tuberculosis infection favour the hypothesis that M. tuberculosis causes resistance of host cells to apoptosis causing perpetuation of infection

A Balanced Approach to Health Information Evaluation: A Vocabulary-based Naïve Bayes Classifier and Readability Formulas

Since millions seek health information online, it is vital for this information to be comprehensible. Most studies use readability formulas, which ignore vocabulary, and conclude that online health information is too difficult. We developed a vocabularly-based, naïve Bayes classifier to distinguish between three difficulty levels in text. It proved 98% accurate in a 250-document evaluation. We compared our classifier with readability formulas for 90 new documents with different origins and asked representative human evaluators, an expert and a consumer, to judge each document. Average readability grade levels for educational and commercial pages was 10th grade or higher, too difficult according to current literature. In contrast, the classifier showed that 70-90% of these pages were written at an intermediate, appropriate level indicating that vocabulary usage is frequently appropriate in text considered too difficult by readability formula evaluations. The expert considered the pages more difficult for a consumer than the consumer did