Due to the development of acaricide resistance in Tetranychus urticae, a rapid and accurate monitoring method for acaricide resistance gene frequency is required for resistance management.In this study, we developed a new diagnostic gene frequency prediction method, using quantitative real-time PCR with allele-specific primer sets, for mutation of I1017F in chitin synthase I, G126S in mitochondrial cytochrome b, and H110R in the PSST subunit of mitochondrial electron transport complex I, which are involved in etoxazole, bifenazate, and pyridaben resistance in T. urticae, respectively. Resistance allele frequencies computed using the 2 −ΔΔCq method, in mixtures of resistance and wild-type (susceptible) alleles, were strongly correlated with the actual frequencies of the resistance alleles in DNA samples for all three acaricides (regression slopes of 0.945-0.996; R 2 > 0.99). This result strongly indicates that resistance allele frequency in a population can be accurately predicted using a diagnostic quantitative real-time PCR method with a resistance allele-specific primer set. Finally, we applied this method to nine field populations and successfully determined resistance allele frequencies for the three acaricides. Overall, this diagnostic prediction method may contribute to the development of acaricide resistance management strategies, via monitoring resistance allele frequencies for a wide range of acaricides.
To develop a novel type of cheese using Aspergillus oryzae, four strains (AHU 7139, AHU 7140, AHU 7141 and AHU 7146) were tested with respect to their extracellular protease production on solid-state media containing heatdenatured whey protein under various temperatures and pH levels. Proteolytic activity of the crude enzyme extracts was evaluated under acidic condition considering the circumstance of cheese ripening. Zymographic analysis revealed that several kinds of proteases were secreted, which were dependent on the strain, incubation temperature and initial pH value. Using three strains, six types of culture products were selected as adjunctive materials in combination with an incubation temperature of 15 or 20ºC and an initial pH condition of 4.0 or 6.5. Gouda-type cheese curd was prepared, and the curd particle received 1% (w/w) of the culture product, followed by pressing, cooling, salting and ripening for 3 months at 11.5ºC. An increase in water extractable nitrogen and free fatty acid was observed in some experimental cheeses, depending on the strain and culture history. In particular, incorporation of culture products from A. oryzae AHU 7139, incubated at pH 6.5 and 20ºC affected these parameters. As a novel dairy technology, this study propose new insights into the potential use of A. oryzae for providing an enzyme cocktail to develop palatable cheese flavor when suitable strains and culture conditions offer products with an optimum balance of proteases and lipases.
[Purpose] The purpose of this study was to examine the relationship between the degree
of thoracic deformity (TD) and the angle formed by a line drawn on transverse plane
computed tomography (CT) images, connecting the sternum and the spinous process of the
vertebrae at the level of the xiphisternum, and the perpendicular line from the floor
(ANGLE), in individuals with severe motor and intellectual disorders (SMID). [Subjects]
Twenty seven individuals with SMID were examined. [Methods] CT transverse images were
acquired at the level of the xiphisternum of each patient. Two protocols were used to
measure the anteroposterior (AP) and laterolateral (LL) diameters. The largest AP
diameters were measured along a perpendicular line from the floor (protocol 1) and the
line from the midline of the sternum to the spinous process of the vertebrae (protocol 2).
The largest LL diameters were measured along the lines perpendicular to the AP diameters
in each protocol. The ratios of the AP to LL diameters and the difference between the
ratios of protocols 1 and 2 (DIFFERENCE) were calculated. [Results] Moderate to good
correlation between DIFFERENCE and ANGLE was observed, and DIFFERENCE became larger with
increasing ANGLE. [Conclusions] These results show that ANGLE indicates the degree of
TD.
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