The cytoplasmic polyadenylation element (CPE)-binding protein (CPEB) binds to CPE containing mRNAs on their 3' untranslated regions (3'UTRs). This RNA binding protein comes out many important tasks, especially in learning and memory, by modifying the translational efficiency of target mRNAs via poly (A) tailing. Overexpressed CPEB has been reported to induce the formation of stress granules (SGs), a sort of RNA granule in mammalian cell lines. RNA granule is considered to be a potentially important factor in learning and memory. However, there is no study about RNA granule in Aplysia. To examine whether an Aplysia CPEB, ApCPEB1, forms RNA granules, we overexpressed ApCPEB1-EGFP in Aplysia sensory neurons. Consistent with the localization of mammalian CPEB, overexpressed ApCPEB1 formed granular structures, and was colocalized with RNAs and another RNA binding protein, ApCPEB, showing that ApCPEB1 positive granules are RNA-protein complexes. In addition, ApCPEB1 has a high turnover rate in RNA granules which were mobile structures. Thus, our results indicate that overexpressed ApCPEB1 is incorporated into RNA granule which is a dynamic structure in Aplysia sensory neuron. We propose that ApCPEB1 granule might modulate translation, as other RNA granules do, and furthermore, influence memory.
Animals ; Aplysia/genetics/*metabolism ; Fluorescence Recovery After Photobleaching ; RNA/genetics/metabolism ; Sensory Receptor Cells/*metabolism ; mRNA Cleavage and Polyadenylation Factors/genetics/metabolism/*physiology

OBJECTIVETo analyze the cleavage activity of two deoxyribozymes targeting at hepatitis C virus (HCV) RNA in vitro and evaluate their prospects of antiviral therapy.

METHODSTwo specific sequences containing 5' ...A / U... 3' in HCV 5'-noncoding region and 5'-fragment of C region (5'-NCR-C) were selected as the target sites, and with the active region of 5'GGCTAGCTACAACGA3', two phosphorothioate deoxyribozymes (TDRz) named as TDRz-127 and TDRz1 were synthesized. HCV RNA 5'-NCR-C was transcribed in vitro from plasmid pHCV-neo which was completely linearized with restriction endonuclease Nar I, and its 5'-end phosphoric acid was deleted by calf intestinal alkaline phosphatase (CIP), then radiolabelled with T4 polynucleotide kinase and gamma-32P-ATP. Under the conditions such as pH 7.5 and a 10 mmol/L Mg2+ concentration, TDRz-127 and TDRz1 were separately (a 5 micromol/L final concentration) or combinedly (each 2.5 micromol/L) mixed with the substrate RNA (200 nmol/L). After denaturation and then renaturation, the reaction systems were incubated in 37 degrees C, and aliquots were removed to terminate the reaction at intended time points. The cleavage products were separated with 8% denaturated polyacrylamide gel electrophoresis and displayed by autoradiography. Finally, the optical density of each product band was measured with Gel Documentation-Analyzing Systems for calculating the percentages of cleaved HCV 5'-NCR-C.

RESULTSAfter reaction for 15, 30, 45, 60, 75 and 90 min under the adopted conditions, about 8.3%, 16.1%, 24.3%, 26.2%, 29.4% and 31.1% of HCV 5'-NCR-C was cleaved by TDRz-127 respectively; 7.4%, 13.0%, 15.6%, 18.7%, 19.4% and 20.3% by TDRz1; and 15.1%, 29.6%, 37.8%, 39.1%, 41.5%, 42.6% by combining the two TDRzs.

CONCLUSIONSCleavage percentage of both TDRz-127 and TDRz1 increases with the time, and the effect of combining the two TDRzs is better than that of anyone.

5' Untranslated Regions ; metabolism ; Base Sequence ; DNA, Catalytic ; genetics ; metabolism ; Hepacivirus ; enzymology ; genetics ; Humans ; Molecular Sequence Data ; RNA Processing, Post-Transcriptional ; RNA, Catalytic ; metabolism ; RNA, Viral ; metabolism ; mRNA Cleavage and Polyadenylation Factors ; genetics ; metabolism

BACKGROUND: Eosinophilia may be associated with various primary and reactive conditions. The incidence and the causes of eosinophilia might have been changed according to the changes in the incidence of diseases such as cancer, chronic degenerative diseases, etc. We have conducted a retrospective study to investigate the incidence and causes of eosinophilia. METHODS: Eosinophilia and hypereosinophilia were defined when absolute eosinophil count was greater than 500/microL and 1,500/microL, respectively. Patient's clinical records were reviewed to find out the underlying clinical conditions responsible for causes of hypereosinophilia. Conventional chromosomal analysis, reverse transcriptase PCR and FISH for gene rearrangement were performed to check the presence of clonal eosinophilia. RESULTS: Out of 41,137 patients who had a hematology profile performed, 5,019 (12.2%) and 373 patients (0.9%) were found to have eosinophilia and hypereosinophilia, respectively. Among patients with hypereosinophilia, 227 patients (60.9%) had identifiable and/or possible causes. The major causes of hypereosinophilia were malignancy (35.2%), allergy and skin diseases (18.1%), infectious diseases (15.4%), hepatobiliary diseases (7.5%), bone marrow clonal diseases (6.6%) and parasite infections (6.6%). We also found a rare case of FIP1L1-PDGFRalpha positive chronic eosinophilic leukemia combined with light chain multiple myeloma. CONCLUSIONS: We found a difference in the distribution of causes of hypereosinophilia in comparison with previous Korean studies, and the most common cause of hypereosinophilia in the current study was malignancy. A rare case of clonal eosinophilia (chronic eosinophilic leukemia) associated with multiple myeloma was confirmed using molecular studies.
Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Bone Marrow/pathology ; Child ; Child, Preschool ; Eosinophilia/epidemiology/*etiology/genetics ; Female ; Hospitals, University ; Humans ; Hypereosinophilic Syndrome/epidemiology/*etiology/genetics ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Receptor, Platelet-Derived Growth Factor alpha/genetics/metabolism ; Retrospective Studies ; Sex Factors ; Young Adult ; mRNA Cleavage and Polyadenylation Factors/genetics/metabolism