ABSTRACT
BACKGROUND: Sepsis is a medical emergency during which early detection is closely associated with mortality. In sepsis, red blood cell (RBC) abnormalities have been reported. However, it is not known how early RBC abnormalities are expressed compared with various clinical manifestations used in sepsis-related organ failure assessment (SOFA). OBJECTIVE: Therefore, using a lipopolysaccharide (LPS)-induced sepsis model we investigated the clinical significance of RBC abnormalities as an early indicator in the detection of septic injury compared with clinical variables. METHODS: Sprague-Dawley rats received LPS (20âmg/kg) intraperitoneally. Aggregation indices (AIs) and aggregation half-time (T1/2), and elongation indices (EI max) were measured. Clinical data-related SOFA and lactate were measured at 2âh, 4âh, 8âh and 12âh after LPS injection. RESULTS: AIs increased at 4âh, and T1/2 decreased at 2âh after LPS injection. Platelet counts decreased at 4âh, and lactate increased at 2âh after LPS injection. AIs showed strong correlations with T1/2 and platelets, EI max increased at 2âh after LPS injection, while EI max had a positive correlation with lactate. CONCLUSIONS: RBC aggregation appears to be an early indicator of clinical deterioration in sepsis and may represent a diagnostic indicator in sepsis.
Subject(s)
Erythrocyte Aggregation/drug effects , Erythrocytes/drug effects , Sepsis/blood , Animals , Male , Rats , Rats, Sprague-Dawley , Sepsis/diagnosisABSTRACT
Glucagon-like peptide-1 and its potent agonist exendin-4 induce several immediate early response genes (IEGs) that code for transcription factors implicated in cell proliferation, differentiation, and apoptosis. We recently observed that early growth response factor-1 (EGR-1), an IEG product, was required for transcriptional activation of Ccnd1 (cyclin D1) gene by exendin-4. Herein, the regulatory mechanism whereby exendin-4 activates the transcription of EGR-1 gene was investigated in the pancreatic beta-cell line INS-1. Deletion analysis of rat EGR-1 promoter identified a critical region between -73 and -46 for the activation of EGR-1 in response to exendin-4. Mutation of the proximal putative cAMP response element (CRE, 5'-GTACGTCA-3') located at -69 resulted in a significant decrease in the EGR-1 transcription, whereas the mutation of the distal putative CRE at -139 was without such an effect. In immune supershift assays using exendin-4-treated cells, binding of cAMP response element-binding protein (CREB) phosphorylated on Ser(133) to the proximal CRE was increased. Employment of a CREB mutant containing Ala substitution at Ser(133) or a dominant negative CREB mutant that inhibits the binding of endogenous CREB to DNA significantly decreased the exendin-4-induced EGR-1 transcription. In experiments using specific protein kinase inhibitors, the effect of H-89 was more prominent than PD-98059, indicating the predominance of the PKA signaling over the MEK/ERK in induction of EGR-1. Therefore, it appears that the proximal CRE site is critical and the binding with CREB phosphorylated on Ser(133) is necessary for induction of the EGR-1 transcription by exendin-4.
Subject(s)
Cyclic AMP Response Element-Binding Protein/metabolism , Early Growth Response Protein 1/genetics , Peptides/pharmacology , Promoter Regions, Genetic/drug effects , Transcriptional Activation/drug effects , Venoms/pharmacology , Animals , Cells, Cultured , Cyclic AMP-Dependent Protein Kinases/metabolism , Early Growth Response Protein 1/metabolism , Exenatide , Extracellular Signal-Regulated MAP Kinases/metabolism , Mutation , Phosphorylation/drug effects , Rats , Signal Transduction/drug effectsABSTRACT
The volatile extract from dried pericarp of Zanthoxylum schinifolium that was obtained by simultaneous distillation with dichloromethane and water was composed of 29.9% geranyl acetate, 15.8% citronella, 15.4% sabinene and the minor volatile components included beta-myrcene, linalool, (-)-isopulegol, citronellyl acetate, 1,4-dimethyl pyrazole, alpha-terpinene, 3-methyl-6-(1-methylethyl)-2-cyclo-hexene-1-o1 and trans-geraniol. The volatile extract decreased the cell viability and induced apoptotic death in HepG2 human hepatoma cells in a concentration- and time-related manner. In addition, the volatile extract increased the production of reactive oxygen species in a dose-dependent manner. Pretreatment of the cells with Trolox, a well-known antioxidant, significantly suppressed the generation of reactive oxygen species and cell death induced by the extract. However, caspase-3 activity was not changed in the extract-treated cells, suggesting that the extract-induced apoptosis of HepG2 cells is caspase-3 independent. Furthermore, in nude mice inoculated with Huh-7 human hepatoma cells, the extract significantly inhibited tumor development. These results suggest that the volatile extract from Zanthoxylum schinifolium pericarpium is a good candidate for hepatocellular carcinoma (HCC) therapy and that reactive oxygen species are the key signaling molecules in the volatile extract-induced cell death in HepG2 cells.
Subject(s)
Apoptosis/drug effects , Carcinoma, Hepatocellular/metabolism , Oils, Volatile/pharmacology , Reactive Oxygen Species/metabolism , Zanthoxylum , Animals , Apoptosis/physiology , Carcinoma, Hepatocellular/drug therapy , Dose-Response Relationship, Drug , Female , Humans , Mice , Mice, Inbred BALB C , Mice, Nude , Oils, Volatile/isolation & purification , Oils, Volatile/therapeutic use , Plant Extracts/isolation & purification , Plant Extracts/pharmacology , Plant Extracts/therapeutic use , Xenograft Model Antitumor AssaysABSTRACT
The possibility of using waste Chinese cabbage (Brassica campestris) as a substrate for microbial biomass production was investigated. The juice from waste Chinese cabbage contains relatively high amounts of reducing sugars suitable for yeast culture. The cell mass and protein content of four species of yeast, Candida utilis, Pichia stipitis, Kluyveromyces marxianus, and Saccharomyces cerevisiae, were determined when cultured in juice extracted from cabbage waste. Compared to YM broth containing the same level of sugar, all the strains except C. utilis showed higher total protein production in cabbage juice medium (CJM).