ABSTRACT
Obesity and type 2 diabetes (T2D) are characterized by decreased insulin sensitivity and higher concentrations of free fatty acids (FFAs) in plasma. Among FFAs, saturated fatty acids (SFAs), such as palmitate, have been proposed to promote inflammatory responses. Primary Sjögren's syndrome (SS) is an autoimmune disease characterized by inflammatory mononuclear cell infiltration and destruction of epithelial cells in the salivary and lacrimal glands. IL-6 production and α-fodrin degradation are increased in salivary gland epithelial cells of patients with primary SS. Although previous studies have shown a link between SS and either dyslipidemia or T2D, little is known about the clinical significance of FFAs in primary SS. Here we report that SFAs, but not unsaturated fatty acids, induced IL-6 production via NF-κB and p38 MAPK activation in human salivary gland epithelial cells. Moreover, palmitate induced apoptosis and α-fodrin degradation by caspase-3 activation. Unlike salivary gland epithelial cells, induction of IL-6 production and the degradation of α-fodrin in response to palmitate were undetectable in squamous carcinoma cells and keratinocytes. Taken together, SFAs induced IL-6 production and α-fodrin degradation in salivary gland epithelial cells, implicating a potential link between the pathogenesis of primary SS and SFAs level in plasma.
Subject(s)
Fatty Acids, Nonesterified/pharmacology , Parotid Gland/drug effects , Sjogren's Syndrome/pathology , Submandibular Gland/drug effects , Apoptosis/drug effects , Carcinoma, Squamous Cell/pathology , Carrier Proteins/drug effects , Caspase 3/drug effects , Cell Line , Cell Line, Tumor , Cell Survival/drug effects , Enzyme Inhibitors/pharmacology , Epithelial Cells/drug effects , Fatty Acids, Unsaturated/pharmacology , Humans , Inflammation Mediators/pharmacology , Interleukin-6/analysis , Keratinocytes/drug effects , Membrane Proteins/drug effects , Microfilament Proteins/drug effects , Mouth Neoplasms/pathology , NF-kappa B/drug effects , Palmitates/pharmacology , Parotid Gland/cytology , Phosphorylation , Stearates/pharmacology , Submandibular Gland/cytology , p38 Mitogen-Activated Protein Kinases/drug effectsABSTRACT
Cancer-specific gene promoter methylation has been described in many types of cancers, and various semi-quantified results have shown their usefulness. Here, we show a more sensitive and specific second-generation system for profiling the DNA methylation status. This method is based on bisulfite reaction of DNA and real-time PCR using two TaqMan MGB probes labeled with different fluorescence, followed by clustering analysis. Primers were designed with CpG-less sequences, and TaqMan MGB probes were designed to contain three or four CpG sites and to be shorter than conventional TaqMan probes. We have added new criteria for primer and probe design for further specificity. We confirmed the reliability of this system and applied it to analysis of lung cancers. Using 10 promoters, 90 primary lung cancers were clustered into six groups consisting of cases having similar smoking status and pathological findings. EGFR mutation and p16 promoter DNA methylation were exclusive, as previously reported; however, DNA methylation in other genes was unrelated to EGFR mutation. This system was also useful to distinguish double primary lung cancers from a single cancer with intrapulmonary metastasis. As above, our system has widespread availability in clinical use and biological research.
Subject(s)
DNA Methylation , Lung Neoplasms/genetics , Promoter Regions, Genetic , Reverse Transcriptase Polymerase Chain Reaction/methods , Cluster Analysis , CpG Islands , DNA Probes , Genes, erbB-1 , Humans , Molecular Probe Techniques , MutationABSTRACT
Dyspepsia is a common complaint, but its course and associated resource utilization have not been well described. In this study, 288 adult, primary care patients with dyspepsia treated at ambulatory clinics were followed prospectively for one year. Medical chart, utilization, and baseline and one-year follow-up survey data were collected. These patients had 13.3 medical visits (sex- and age-standardized) during the follow-up period, 55% above standardized mean visits for a comparison group of nondyspepsia patients. Standardized mean charges of $3542 for dyspeptics was 126% above nondyspepsia patient charges. Over half had gastrointestinal-related follow-up visits; 61% used gastrointestinal drugs; and 43% had gastrointestinal procedures. NSAID users had higher gastrointestinal-related utilization than did nonusers, recording an additional gastrointestinal visit (P < 0.001) and $678 more in charges (P = 0.03). Eighty-six percent of the 189 follow-up survey respondents experienced gastrointestinal symptoms at some time during the follow-up year. This study showed that most primary care dyspepsia patients remained symptomatic after one year and were intensive users of medical care.
