RESUMO
The ATP-binding cassette (ABC) transporters are a superfamily of membrane proteins. These active transporters are involved in the export of different substances such as xenobiotics. ABC transporters from subfamily C (ABCC) have also been described as functional receptors for different insecticidal proteins from Bacillus thuringiensis (Bt) in several lepidopteran species. Numerous studies have characterized the relationship between the ABCC2 transporter and Bt Cry1 proteins. Although other ABCC transporters sharing structural and functional similarities have been described, little is known of their role in the mode of action of Bt proteins. For Heliothis virescens, only the ABCC2 transporter and its interaction with Cry1A proteins have been studied to date. Here, we have searched for paralogs to the ABCC2 gene in H. virescens, and identified two new ABC transporter genes: HvABCC3 and HvABCC4. Furthermore, we have characterized their gene expression in the midgut and their protein topology, and compared them with that of ABCC2. Finally, we discuss their possible interaction with Bt proteins by performing protein docking analysis.
Assuntos
Toxinas de Bacillus thuringiensis , Proteínas de Bactérias , Endotoxinas , Proteínas Hemolisinas , Proteína 2 Associada à Farmacorresistência Múltipla , Proteínas Associadas à Resistência a Múltiplos Medicamentos , Proteínas de Bactérias/metabolismo , Proteínas de Bactérias/genética , Proteínas de Bactérias/química , Animais , Toxinas de Bacillus thuringiensis/metabolismo , Endotoxinas/metabolismo , Endotoxinas/genética , Endotoxinas/química , Proteínas Hemolisinas/metabolismo , Proteínas Hemolisinas/genética , Proteínas Hemolisinas/química , Proteínas Associadas à Resistência a Múltiplos Medicamentos/metabolismo , Proteínas Associadas à Resistência a Múltiplos Medicamentos/genética , Proteínas Associadas à Resistência a Múltiplos Medicamentos/química , Proteínas de Insetos/genética , Proteínas de Insetos/metabolismo , Proteínas de Insetos/química , Mariposas/metabolismo , Mariposas/genética , Bacillus thuringiensis/metabolismo , Bacillus thuringiensis/genética , Simulação de Acoplamento Molecular , Transportadores de Cassetes de Ligação de ATP/metabolismo , Transportadores de Cassetes de Ligação de ATP/genética , Transportadores de Cassetes de Ligação de ATP/químicaRESUMO
Background: On the basis of the hypothesis that virtual noniodine (VNI)-based coronary artery calcium scoring (CACS) is feasible at reduced radiation doses, this study assesses the impact of radiation dose reduction on the accuracy of this VNI algorithm on a photon-counting detector (PCD)-CT. Methods: In a systematic in vitro setting, a phantom for CACS simulating three chest sizes was scanned on a clinical PCD-CT. The standard radiation dose was chosen at volumetric CT dose indices (CTDIVol) of 1.5, 3.3, 7.0 mGy for small, medium-sized, and large phantoms, and was gradually reduced by adjusting the tube current resulting in 100, 75, 50, and 25%, respectively. VNI images were reconstructed at 55 keV, quantum iterative reconstruction (QIR)1, and at 60 keV/QIR4, and evaluated regarding image quality (image noise (IN), contrast-to-noise ratio (CNR)), and CACS. All VNI results were compared to true noncontrast (TNC)-based CACS at 70 keV and standard radiation dose (reference). Results: INTNC was significantly higher than INVNI, and INVNI at 55 keV/QIR1 higher than at 60 keV/QIR4 (100% dose: 16.7 ± 1.9 vs. 12.8 ± 1.7 vs. 7.7 ± 0.9; p < 0.001 for every radiation dose). CNRTNC was higher than CNRVNI, but it was better to use 60 keV/QIR4 (p < 0.001). CACSVNI showed strong correlation and agreement at every radiation dose (p < 0.001, r > 0.9, intraclass correlation coefficient > 0.9). The coefficients of the variation in root-mean squared error were less than 10% and thus clinically nonrelevant for the CACSVNI of every radiation dose. Conclusion: This phantom study suggests that CACSVNI is feasible on PCD-CT, even at reduced radiation dose while maintaining image quality and CACS accuracy.
RESUMO
The Vip3Aa insecticidal protein from Bacillus thuringiensis (Bt) is produced by specific transgenic corn and cotton varieties for efficient control of target lepidopteran pests. The main threat to this technology is the evolution of resistance in targeted insect pests and understanding the mechanistic basis of resistance is crucial to deploy the most appropriate strategies for resistance management. In this work, we tested whether alteration of membrane receptors in the insect midgut might explain the >2000-fold Vip3Aa resistance phenotype in a laboratory-selected colony of Heliothis virescens (Vip-Sel). Binding of 125I-labeled Vip3Aa to brush border membrane vesicles (BBMV) from 3rd instar larvae from Vip-Sel was not significantly different from binding in the reference susceptible colony. Interestingly, BBMV from Vip-Sel larvae showed dramatically reduced levels of membrane-bound alkaline phosphatase (mALP) activity, which was further confirmed by a strong downregulation of the membrane-bound alkaline phosphatase 1 (HvmALP1) gene. However, the involvement of HvmALP1 as a receptor for the Vip3Aa protein was not supported by results from ligand blotting and viability assays with insect cells expressing HvmALP1.