To clone the first anion channel from Xenopus laevis (X. laevis), we isolated a calcium-activated chloride channel (CLCA)-like membrane protein 6 gene (CMP6) in X. laevis. As a first step in gene isolation, an expressed sequence tags database was screened to find the partial cDNA fragment. A putative partial cDNA sequence was obtained by comparison with rat CLCAs identified in our laboratory. First stranded cDNA was synthesized by reverse transcription polymerase-chain reaction (RT-PCR) using a specific primer designed for the target cDNA. Repeating the 5' and 3' rapid amplification of cDNA ends, full-length cDNA was constructed from the cDNA pool. The full-length CMP6 cDNA completed via 5'- and 3'-RACE was 2,940 bp long and had an open reading frame (ORF) of 940 amino acids. The predicted 940 polypeptides have four major transmembrane domains and showed about 50% identity with that of rat brain CLCAs in our previously published data. Semi-quantification analysis revealed that CMP6 was most abundantly expressed in small intestine, colon and liver. However, all tissues except small intestine, colon and liver had undetectable levels. This result became more credible after we did real-time PCR quantification for the target gene. In view of all CLCA studies focused on human or murine channels, this finding suggests a hypothetical protein as an ion channel, an X. laevis CLCA.
Amino Acids ; Animals ; Brain ; Chloride Channels ; Clone Cells ; Colon ; DNA, Complementary ; Expressed Sequence Tags ; Humans ; Intestine, Small ; Ion Channels ; Liver ; Membrane Proteins ; Membranes ; Open Reading Frames ; Peptides ; Rats ; Real-Time Polymerase Chain Reaction ; Resin Cements ; Reverse Transcription ; Staphylococcal Protein A ; Tissue Distribution ; Xenopus ; Xenopus laevis

Oroxylum indicum has long been used in Asian traditional medicine to prevent and treat respiratory diseases, diabetes, diarrhea and other conditions. Oroxylin A is a flavone that is present in Oroxylum indicum and in Scutellaria baicalensis. Because the root extracts of both plants have been shown to have anti-allergic effects, the authors investigated whether oroxylin A is likely to have beneficial effects on allergic asthma using female Balb/c mice and rat RBL-2H3 mast cells. Antigen-induced degranulation was measured in vitro by measuring β-hexosaminidase activity. A murine ovalbumin-induced allergic asthma model was used to test the in vivo efficacy of oroxylin A. Sensitization and challenge of ovalbumin induced allergic asthma responses, the accumulations of eosinophils and Th2 cytokine levels in bronchoalveolar lavage fluid and lung tissues. Oroxylin A administration decreased numbers of inflammatory cells, especially eosinophils, and reduced the expression and secretion of Th2 cytokines, including IL-4 and IL-13, in lung tissues and bronchoalveolar lavage fluid. Histologic studies showed oroxylin A reduced inflammatory signs and mucin production in lungs. These findings provide evidence that oroxylin A has potential use as an anti-allergic therapeutic.
Animals ; Asian Continental Ancestry Group ; Asthma ; Bronchoalveolar Lavage Fluid ; Cytokines ; Diarrhea ; Eosinophils ; Female ; Humans ; In Vitro Techniques* ; Interleukin-13 ; Interleukin-4 ; Lung ; Mast Cells ; Medicine, Traditional ; Mice ; Mucins ; Ovalbumin ; Rats ; Scutellaria baicalensis

BACKGROUND/OBJECTIVES: Diet plays an important role in growth and development of children. However, dietary intakes of children living in either rural or urban areas can be influenced by household income. This cross-sectional study examined energy, nutrient and food group intakes of 749 urban children (1-10 years old) by household income status. SUBJECTS/METHODS: Children's dietary intakes were obtained using food recall and record for two days. Diet adequacy was assessed based on recommended intakes of energy and nutrients and food group servings. RESULTS: For toddlers, all nutrients except dietary fiber (5.5 g) exceeded recommended intakes. Among older children (preschoolers and school children), calcium (548 mg, 435 mg) and dietary fiber (7.4 g, 9.4 g) did not meet recommendations while percentage of energy from total fat and saturated fats exceeded 30% and 10%, respectively. The mean sodium intakes of preschoolers (1,684 mg) and school children (2,000 mg) were relatively high. Toddlers in all income groups had similar energy and nutrient intakes and percentages meeting the recommended intakes. However, low income older children had lowest intakes of energy (P < 0.05) and most nutrients (P < 0.05) and highest proportions that did not meet recommended energy and nutrient intakes. For all food groups, except milk and dairy products, all age groups had mean intakes below the recommended servings. Compared to middle and high income groups, low income preschoolers had the lowest mean intake of fruits (0.07 serving), meat/poultry (0.78 serving) and milk/dairy products (1.14 serving) while low income toddlers and school children had the least mean intake of fruits (0.09 serving) and milk/dairy products (0.54 serving), respectively. CONCLUSION: Low socioeconomic status, as indicated by low household income, could limit access to adequate diets, particularly for older children. Parents and caregivers may need dietary guidance to ensure adequate quantity and quality of home food supply and foster healthy eating habits in children.
Calcium ; Caregivers ; Child ; Cross-Sectional Studies ; Dairy Products ; Diet ; Dietary Fiber ; Eating ; Family Characteristics* ; Fats ; Food Supply ; Fruit ; Growth and Development ; Humans ; Milk ; Parents ; Social Class ; Sodium

A sensitive and simple liquid chromatography-tandem mass spectrometry method was developed and validated for the simultaneous quantification of ticagrelor and its active metabolite, AR-C124910XX from 50 µL human plasma using tolbutamide as an internal standard as per regulatory guidelines. Analytes in plasma were extracted by simple protein precipitation using acetonitrile, followed by chromatographic separation with an Acclaim™ RSLC 120 C₁₈ column (2.2 µm, 2.1 × 100 mm) and a gradient acetonitrile-water mobile phase containing 0.1% formic acid within 8 min. Mass spectrometric detection and quantitation were conducted by selected reaction-monitoring on a negative electrospray ionization mode with the following transitions: m/z 521.11 → 361.10, 477.03 → 361.10, and 269.00 → 169.60 for ticagrelor, AR-C124910XX, and tolbutamide, respectively. The lower limit of quantifications was 0.2 ng/mL with linear ranges of 0.2–2,500 ng/mL (r² ≥ 0.9949) for both analytes. All validation data, including selectivity, cross-talk, precision, accuracy, matrix effect, recovery, dilution integrity, stability, and incurred sample reanalysis, were well within acceptable limits. This assay method was validated using K₂-EDTA as the specific anticoagulant. Also, the anticoagulant effect was tested by lithium heparin, sodium heparin, and K₃-EDTA. No relevant anticoagulant effect was observed. This validated method was effectively used in the determination of ticagrelor and its active metabolite, AR-C124910XX, in plasma samples from patients with myocardial infarction.
Heparin ; Humans ; Lithium ; Mass Spectrometry ; Methods ; Myocardial Infarction ; Pharmacokinetics ; Plasma ; Tolbutamide