Analysis of the effect of chronic morphine treatment on miRNA profile and introduction of the MAPK pathway as the target of differentially expressed miRNAS

Different signaling pathways have been identified that are involved in the cellular response to opiates. The mitogen-activated protein kinase pathway is one of the most important signaling pathways underlying the neuronal response to opiates. MicroRNAs [miRNAs] are considered to be post-transcriptional regulators of gene expression with paramount significance, which plays key roles in modulating cellular processes such as neuronal plasticity and synaptic consolidation. The purpose of this study is to identify miRNAs that are differentially expressed in response to chronic morphine treatment, and predict those genes that have a possible role in this process. Because the MAPK pathway in involved in morphine dependence and participates in hypersensitivity to pain, determining miRNAs that modulate this pathway could be insightful in morphine dependence treatment and pain control. In this study, the BE[2]-C neuroblastoma cell line was chronically treated with morphine sulphate and the changes in expression of 750 miRNAs were analyzed by real time PCR. Two up- and down- regulated groups of miRNAs were determined to be differentially expressed in response to morphine: i] has-mir-193a-3p, -212, -181c, -362-3p, -639, -646 and ii] has-mir-412, -937, -558, -552, -943, -628-5p, -593, -555, -636, -643, 566, -571, -642, -653, -611, -31, let7-g. The analysis of differentially expressed miRNAs showed that the MAPK signaling pathway could be regarded as a signaling pathway with utmost significance in chronic morphine response. Due to the role played by MAPK pathway in cellular response to morphine exposure, we can propose that protein phosphorylation has a presumable part in this response

Detection of types 40 and 41 adenoviruses in stool samples of diarrheal children by solid phase PCR

Adenoviruses [AdVs] types 40 and 41 are the causative agents of diarrhea in children. Hence, rapid sensitive and specific detection of these viruses are of clinical importance. The customary methods such as propagation of virus in cell culture suffer from limitations. Detection of immobilized amplified products in a one phase system [DIAPOPS] method has the potential to overcome these problems. A DIAPOPS method for detection of AdV types 40 and 41 was designed. Forward primers were covalently linked to the Nucleolink surface. After amplification of a 745 bp sequence of DNA binding protein gene, the amplified product was hybridized with the biotinylated probe. The hybrids were detected by the antibody-peroxidase conjugate. After optimization of the DIAPOPS conditions, 80 stool samples from children with clinical manifestation of viral diarrhea were tested. Their DIAPOPS results were compared with those of the conventional polymerase chain reaction [PCR] assay. Positive results were obtained in 11 samples. The comparison between conventional PCR and DIAPOPS showed a significant increase in sensitivity of the DIAPOPS test, 6 samples shown to be negative by conventional PCR, were demonstrated positive by DIAPOPS [p=0.00]. The DIAPOPS assay presented in this study can provide a rapid, sensitive, specific and economic method for detection of viral infections. The assay can be performed for numerous samples simultaneously in a day. This DIAPOPS method can provide a practical and reliable tool for diagnosis of enteric adenoviruses. In addition, the risk of contamination in this assay is low