FRET probe for detecting two mutations in one EGFR mRNA

Technologies for visualizing and tracking RNA are essential in molecular biology, including in disease-related fields. In this study, we propose a novel probe set (DAt-probe and T-probe) that simultaneously detects two mutations in the same RNA using fluorescence resonance energy transfer (FRET). The DAt-probe carrying the fluorophore Atto488 and the quencher Dabcyl were used to detect a cancer mutation (exon19del), and the T-probe carrying the fluorophore Tamra was used to detect drug resistance mutations (T790M) in epidermal growth factor receptor (EGFR) mRNA. These probes were designed to induce FRET when both mutations were present in the mRNA. Gel electrophoresis confirmed that the two probes could efficiently bind to the mutant mRNA. We measured the FRET ratios using wild-type and double-mutant RNAs and found a significant difference between them. Even in living cells, the FRET probe could visualize mutant RNA. As a result, we conclude that this probe set provides a method for detecting two mutations in the single EGFR mRNA via FRET

Long-Term Prognosis of Patients with Major Depression and Silent Cerebral Infarction

.OBJECTIVE: Many studies have examined the effects of cerebrovascular change on treatment response in geriatric depression. However, few such studies have examined the relationship between cerebrovascular changes and long term prognosis. We examined the effects of cerebrovascular change on the course of geriatric depressive symptoms, dementia rates, and mortality over a follow-up period of approximately 10 years. METHOD: Participants were 84 patients with major depression (age of onset greater than 50 years old); patients suffering from strokes, neurological disorders, and other psychiatric disorders were excluded. MRI findings were used to classify all patients into silent cerebral infarction (SCI) positive (n = 37) or negative groups (n = 47). Patient prognoses were ascertained using a review of clinical charts and over the mail. RESULTS: Only 5% of patients with SCI were able to maintain remission, whereas 36% of those patients without SCI were able to do so. Total duration of depressive episodes was significantly longer in the SCI positive group than in the SCI negative group. In addition, SCI was associated with a higher risk of the onset of dementia. CONCLUSION: The results of this long-term follow-up study demonstrate that the presence of SCI is associated with a relatively poor prognosis for geriatric depressio

Effect of Paddy-Upland Rotation System on the Net Greenhouse Gas Balance as the Sum of Methane and Nitrous Oxide Emissions and Soil Carbon Storage: A Case in Western Japan

To investigate the effect of paddy-upland (PU) rotation system on greenhouse gas emissions, methane (CH4) and nitrous oxide (N2O) emissions were monitored for three years for a PU rotation field (four cultivations (wheat-soybean-rice-rice) over three years) and continuous paddy (CP) field on alluvial soil in western Japan. Soil carbon storage was also calculated using an improved Rothamsted Carbon (RothC) model. The net greenhouse gas balance was finally evaluated as the sum of CO2eq of the CH4, N2O and changes in soil carbon storage. The average CH4 emissions were significantly lower and the average N2O emissions were significantly higher in the PU field than those in the CP field (p 2 equivalent basis, CH4 emissions were much higher than N2O emission. In total, the average CO2eq emissions of CH4 plus N2O in the PU field (1.81 Mg CO2 ha−1 year−1) were significantly lower than those in the CP field (7.42 Mg CO2 ha−1 year−1) (p 2eq emissions of 0.57 and 0.09 Mg CO2 ha−1 year−1 in the both fields, respectively. Consequently, the net greenhouse gas balance in the PU and CP fields were estimated to be 2.38 and 7.51 Mg CO2 ha−1 year−1, respectively, suggesting a 68% reduction in the PU system. In conclusion, PU rotation system can be regarded as one type of the climate-smart soil management

Effect of Paddy-Upland Rotation System on the Net Greenhouse Gas Balance as the Sum of Methane and Nitrous Oxide Emissions and Soil Carbon Storage: A Case in Western Japan

To investigate the effect of paddy-upland (PU) rotation system on greenhouse gas emissions, methane (CH4) and nitrous oxide (N2O) emissions were monitored for three years for a PU rotation field (four cultivations (wheat-soybean-rice-rice) over three years) and continuous paddy (CP) field on alluvial soil in western Japan. Soil carbon storage was also calculated using an improved Rothamsted Carbon (RothC) model. The net greenhouse gas balance was finally evaluated as the sum of CO2eq of the CH4, N2O and changes in soil carbon storage. The average CH4 emissions were significantly lower and the average N2O emissions were significantly higher in the PU field than those in the CP field (p < 0.01). On CO2 equivalent basis, CH4 emissions were much higher than N2O emission. In total, the average CO2eq emissions of CH4 plus N2O in the PU field (1.81 Mg CO2 ha−1 year−1) were significantly lower than those in the CP field (7.42 Mg CO2 ha−1 year−1) (p < 0.01). The RothC model revealed that the changes in soil carbon storage corresponded to CO2eq emissions of 0.57 and 0.09 Mg CO2 ha−1 year−1 in the both fields, respectively. Consequently, the net greenhouse gas balance in the PU and CP fields were estimated to be 2.38 and 7.51 Mg CO2 ha−1 year−1, respectively, suggesting a 68% reduction in the PU system. In conclusion, PU rotation system can be regarded as one type of the climate-smart soil management