ABSTRACT
BACKGROUND AND AIMS: Previously, we reported that granulocyte-colony stimulating factor (G-CSF) improves hepatic steatosis in experimental animals. It may also have preventive effects on the development of hepatic steatosis. Therefore, we investigated the preventive effects of G-CSF by using a high-fat diet (HFD) rat model. MATERIALS AND METHODS: Twelve rats were fed HFD and 6 rats were fed control diet from 10 weeks of age. Once little steatosis was confirmed in the liver (after 10 weeks of feeding the HFD; at 20 weeks of age), HFD rats were randomly divided into two groups and treated with either G-CSF (100 µg kg-1 day-1 for 5 consecutive days every other week; HFD/G-CSF rats) or saline (HFD/saline rats) for 10 weeks at 20 weeks of age. All rats were sacrificed at 30 weeks of age. Histology was examined by hematoxylin and eosin (H-E) and Oil Red O staining, and the expression levels of genes of associated with lipogenesis and ß-oxidation enzymes were determined by qRT-PCR. RESULTS: Histological examinations revealed that HFD/G-CSF rats had significantly lower lipid accumulation in their hepatocytes than did HFD/saline rats (p < 0.05). HFD/G-CSF rats also showed lower expression levels of genes associated with lipogenesis and higher expression levels of genes associated with ß-oxidation than HFD/saline rats (p < 0.05). CONCLUSION: In conclusion, we found that G-CSF prevented development of hepatic steatosis in an HFD rat model. The preventive effect may be associated with the regulation of gene expression involved in hepatic lipogenesis and ß-oxidation.
Subject(s)
Granulocyte Colony-Stimulating Factor/pharmacology , Lipid Metabolism/drug effects , Liver/drug effects , Non-alcoholic Fatty Liver Disease/prevention & control , Animals , Diet, High-Fat , Disease Models, Animal , Gene Expression Regulation , Humans , Lipid Metabolism/genetics , Lipogenesis/drug effects , Liver/metabolism , Liver/pathology , Male , Non-alcoholic Fatty Liver Disease/etiology , Non-alcoholic Fatty Liver Disease/genetics , Non-alcoholic Fatty Liver Disease/metabolism , Non-alcoholic Fatty Liver Disease/pathology , Oxidation-Reduction , Rats, Sprague-Dawley , Receptors, Granulocyte Colony-Stimulating Factor/agonists , Receptors, Granulocyte Colony-Stimulating Factor/metabolism , Recombinant Proteins/pharmacology , Time FactorsABSTRACT
OBJECTIVE: Because perivascular echo brightness (PEB) of coronary arteries has been proposed as a criterion for diagnosis of incomplete Kawasaki disease, we assessed the clinical importance of PEB during the acute phase of disease. STUDY DESIGN: We enrolled 58 patients with Kawasaki disease who underwent two-dimensional strain analysis of images of pericoronary tissue taken during the acute and the convalescent phases. Echogenicity of pericoronary tissue and of the blood pool was determined by speckle tracking in the respective areas of imaging as the averages of integrated backscatter over a single cardiac cycle. PEB was defined as echogenicity of pericoronary tissue minus blood pool. RESULTS: PEB did not differ in the acute phase in patients and control subjects (P = .10) and between phases of disease (P = .25). In comparison between patient groups, the presence of pericardial effusion was higher in patients with higher PEB during the acute phase (n = 30) than in the remaining patients (33% versus 4%, P < .01). CONCLUSIONS: PEB did not differ between patients and control subjects and is only associated with the presence of pericardial effusion during the acute phase of Kawasaki disease. Our data do not confirm the reliability of PEB as a useful diagnostic sign of incomplete Kawasaki disease.
Subject(s)
Coronary Vessels/diagnostic imaging , Mucocutaneous Lymph Node Syndrome/diagnostic imaging , Case-Control Studies , Child, Preschool , Echocardiography , Female , Humans , Image Processing, Computer-Assisted , Infant , Male , Pericardial Effusion/diagnostic imagingABSTRACT
OBJECTIVES: We have recently found a high prevalence of non-typeable pneumococcal isolates (NTPn) circulating in day-care centers in Central Brazil, besides serotype 14 isolates. We therefore examined the genetic relationship among NTPn and serotype 14 from carriage and invasive pneumococcal isolates obtained from children attending emergency rooms enrolled in a population-based surveillance. METHODS: The isolates were characterized by Quellung reaction serotyping, PCR for the presence of pneumolysin and the loci for a capsule gene (cpsA) and the type 14 gene (cps14H) in all NTPn, and by multilocus sequence typing and pulsed field gel electrophoresis. RESULTS: 87.2% of the isolates were clustered into nine clusters. The major cluster included 41 pneumococcal serotype 14 (28 carriage and 13 invasive isolates) and two NTPn related to the global pneumococcal clone Spain(9V)-3. Overall, 95.4% of the NTPn carriage strains were genetically related to carriage or invasive strains expressing serotype 14. A dominant NTPn lineage was found, that grouped 14 pneumococcal strains. Almost half of the multidrug-resistant isolates grouped into the NTPn cluster. CONCLUSION: These findings provide baseline data to assess the impact of the pneumococcal vaccination on the molecular epidemiology of Streptococcus pneumoniae. Changes in frequency of NTPn isolates and also genetic changes should be carefully monitored post vaccination, to detect potential vaccine-escape or replacement disease by capsule switched strains, especially in areas where colonization with NTPn has been frequently observed.
Subject(s)
Bacterial Typing Techniques , Carrier State/epidemiology , Carrier State/microbiology , Pneumococcal Infections/epidemiology , Pneumococcal Infections/microbiology , Streptococcus pneumoniae/classification , Streptococcus pneumoniae/genetics , Bacterial Proteins/genetics , Brazil/epidemiology , Child, Preschool , DNA Fingerprinting , Electrophoresis, Gel, Pulsed-Field , Female , Genotype , Humans , Infant , Infant, Newborn , Male , Molecular Epidemiology , Pneumococcal Vaccines/immunology , Polymerase Chain Reaction , Prevalence , Sequence Analysis, DNA , Serotyping , Streptococcus pneumoniae/isolation & purification , Streptolysins/genetics , Virulence Factors/geneticsABSTRACT
We cloned 2-keto-3-deoxy-gluconate kinase (KDGK), which catalyzes the phosphorylation of 2-keto-3-deoxygluconate (KDG) to 2-keto-3-deoxy-6-phophogluconate (KDPG) from Serratia marcescens KCTC 2172. The nucleotide sequence revealed a single open reading frame containing 1,208 bp and encoding for 309 amino acids, with a molecular weight of 33,993 Da. The enzyme was purified via GST affinity chromatography. The putative KdgT binding site was detected upstream of the initial codon. The KDG kinase utilized 2-ketogluconate (KG) and KDG as substrates. The optimal temperature and pH for KDGK activity were 50ºC and 8.0, respectively.