Appressorium formation is an important event in establishing a successful interaction between the rice blast fungus, Magnaporthe oryzae, and its host plant, rice. An understanding of molecular events occurring in appressorium differentiation will give new strategies to control rice blast. A quick and reliable method to extract total RNA from appressorium is essential for studying gene expression during appressorium formation and its mechanism. We found that duplicate film is an efficient substratum for appressorium formation, even when inoculated with high density conidia. When inoculated with conidia at 1×10 6 ml −1 , the percentages of conidium germination and appressorium formation were (97.98±0.67)% and (97.88±0.45)%, respectively. We applied Trizol before appressorium collection for total RNA isolation, and as much as 113.6 μg total RNA was isolated from the mature appressoria at 24 h after inoculation. Functional analysis of two genes, MNH6 and MgATG1, isolated from the cDNA subtractive library, revealed that the quantity of RNA was good enough to construct a cDNA (complementary DNA) library or a cDNA subtractive library. This method may be also applicable for the appressorium RNA isolation of other pathogenic fungi in which conidia differentiate into appressoria in the early stages of host infection.
Introduction. The relationship between ventricular pre-excitation and left ventricular dysfunction has been described in the absence of sustained supraventricular tachycardia in a series of case reports. However, there have been no systematic studies about the effect of ventricular pre-excitation on cardiac function in adult patients with different accessory pathway locations. Methods and Results. Patients were divided into four groups based on the type and location of their accessory pathway: septal, right free wall, left free wall, and concealed. N-terminal pro-B-type natriuretic peptide (NT-proBNP) levels, electrocardiogram recordings, electrophysiological properties, and transthoracic echocardiographic data (septal-to-posterior wall motion delay (SPWMD) and interventricular mechanical delay (IVMD) indicating intraventricular and interventricular dyssynchrony) were compared before and after successful ablation. Before radiofrequency catheter ablation, left ventricular ejection fraction (LVEF) was significantly lower in patients with septal and right free wall accessory pathways. Within three months after radiofrequency catheter ablation, NT-proBNP levels decreased, left ventricular function improved, and intraventricular left ventricular dyssynchrony disappeared. There was a negative correlation between initial LVEF with initial QRS duration and initial SPWMD. Notably, SPWMD had a stronger correlation with LVEF than initial QRS duration. Conclusions. Anterograde conduction with a septal or right free wall accessory pathway may cause left ventricular dyssynchrony and impair left ventricular function. Intraventricular left ventricular dyssynchrony seems to be responsible for the pathogenesis of left ventricular dysfunction. Radiofrequency catheter ablation results in decreased NT-proBNP levels, normalized QRS duration, mechanical resynchronization, and improved left ventricular function.
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