Background: Trichomoniasis resulting from Trichomonas vaginalis (T. vaginalis) has been considered as a commonly seen disease with the transmission way of sex. At present, the detection methods of T. vaginalis mainly include wet mount microscopy, culture, PCR, immunofluorescence and ELISA. However, all of these detection methods exist shortcomings. Methods: In this study, a loop-mediated isothermal amplification (LAMP) assay that targeted the species-specific sequence of adhesion protein 65 (AP65) gene had been conducted to detect T. vaginalis. The optimum reaction system and conditions were optimized in this rapid detection method. Results: The results of sensitivity analysis showed that the LAMP assay targeting the AP65 gene was 1000 times more sensitive than the nested PCR targeting the actin gene commonly used for detection of T. vaginalis, and the detecting limitation of the former was 10 trichomonad. Moreover, the amplification of the target gene AP65 by LAMP assay exhibited high specificity and the product was exclusively from T. vaginalis. The detection technique of LAMP did not exhibit cross-reactivity with the common pathogens of Trichinella spiralis, Toxoplasma gondii, Escherichia coli, Candida albicans, Staphylococcus aureus, Haemophilus. Conclusions: According to the present study, the LAMP assay with the target of AP65 gene, was suitable for the early diagnosis of T. vaginalis infections. Consequently, the LAMP assay was proposed by the current study as a point-of-care examination and an alternative molecular tool which exhibited the potential value in the treatment, control and prevention of trichomoniasis transmission and relevant complication.
Fusarium crown rot (FCR), which is caused by Fusarium pseudograminearum, is one of the most important diseases affecting wheat production in the Huanghuai wheat-growing region of China. Although the phenylpyrrole fungicide fludioxonil is known to have a broad-spectrum activity against a wide range of plant pathogens, including F. pseudograminearum, it has not yet been registered for the control of FCR in China, and further research is required to assess the biological characteristics and molecular mechanisms associated with fludioxonil-resistance, and especially the potential for highly resistant isolates to emerge. The current study demonstrated that the baseline fludioxonil sensitivity of 61 F. pseudograminearum isolates collected from the Henan province of China during the summers of 2019-2021 conformed to a unimodal distribution with a mean effective concentration for 50% inhibition (EC50) value of 0.021 ± 0.003 μg/mL, which indicated that none of the isolates exhibited natural resistance to fludioxonil. Nevertheless, four fludioxonil-resistant mutants were attained after repeated exposure to fludioxonil under laboratory conditions. All the resistant mutants exhibited significantly reduced growth rates on potato dextrose agar (PDA), as well as reduced levels of sporulation and pathogenicity in wheat seedlings. In addition, the resistant mutants also exhibited reduced growth on PDA amended with either 0.5 M Mannitol, 0.5 M Glucose, 0.5 M MgCl2, or 0.5 M NaCl, which indicated that they had increased sensitivity to osmotic stress. Molecular analysis of the proposed fludioxonil target protein FpOs1 indicated that the predicted sequences of the resistant mutants contained none of the characteristic amino acid changes previously associated with fludioxonil resistance in other species. Further investigation using qPCR analysis revealed that the expression of FpOs1 gene was significantly altered in the resistant mutants in both the absence and presence of fludioxonil. Meanwhile, plate assays found evidence of cross resistance between fludioxonil and cyprodinil, as well as with the triazole fungicides tebuconazole and difenoconazole, but not with other commonly used fungicides including prochloraz, fluazinam, and carbendazim. Taken together these results provide new insights into the mechanism and biological characteristics associated with fludioxonil resistance in F. pseudograminearum, and indicate that fludioxonil could provide effective and sustained control of FCR during wheat production.
This article presents the influence of capping ligand and surface interaction types on the coarsening or reshaping behavior of surface-immobilized gold nanoparticles with different core size and shape. The morphological transformation of gold nanoparticles and nanorods on graphene oxide upon heating at temperatures ranging from 50 to 200 °C was investigated. The aggregation and coarsening behaviors of spherical nanoparticles on graphene oxide were slightly affected by the core size of nanoparticles (~ 1, 3, and 10 nm). The comparison of two different surface ligands revealed that glutathione ligands provide much better protection than cetyltrimethylammonium bromide ligands against the morphological transformation of nanoparticles. In addition, the evaluation of surface binding interactions indicated that the attachment of nanoparticles and nanorods onto graphene oxide with additional thiol functional groups could improve the immobilization of particles and therefore decelerate coarsening and reshaping of nanoparticle and nanorods.Electronic supplementary materialThe online version of this article (10.1186/s40580-018-0171-0) contains supplementary material, which is available to authorized users.
Dual-level electronic structure calculation has been performed to investigate the mechanism and all possible channels of OH radical reaction with CH 3 CH 2 F. Geometries and frequencies are computed at the B3LYP/6-311G(d,p) level of theory for all stationary points and complexes and transition states are located. Potential energy surfaces are constructed at the PMP2/cc-pVTZ//B3LYP/6-311G(d,p) level + ZPE correction. Four types of reaction channels are identified: hydrogen abstraction, fluorine abstraction and attack on carbon atom along or perpendicular to the C -C bond axis. Hydrogen abstraction channels have lower barriers and are more exothermic, while out-of-plane β -H abstraction with the lowest barrier is competitive with α -H abstraction. Due to the high energy barrier, contributions of non-H abstraction channels are excluded. The influence of hydrogen bonding interaction is clearly observed in the barrier heights.
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