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51.
We recently described our finding that recombinant baculovirus-produced virus-like particles (VLPs) can induce cell-cell fusion similar to that induced by intact rotavirus in our assay for viral entry into tissue culture cells (J. M. Gilbert and H. B. Greenberg, J. Virol. 71:4555-4563, 1997). The conditions required for syncytium formation are similar to those for viral penetration of the plasma membrane during the course of viral infection. This VLP-mediated fusion activity was dependent on the presence of the outer-layer proteins, viral protein 4 (VP4) and VP7, and on the trypsinization of VP4. Fusion activity occurred only with cells that are permissive for rotavirus infection. Here we begin to dissect the role of VP4 in rotavirus entry by examining the importance of the precise trypsin cleavage of VP4 and the activation of VP4 function related to viral entry. We present evidence that the elimination of the three trypsin-susceptible arginine residues of VP4 by specific site-directed mutagenesis prevents syncytium formation. Two of the three arginine residues in VP4 are dispensable for syncytium formation, and only the arginine residue at site 247 appears to be required for activation of VP4 functions and cell-cell fusion. Using the recombinant VLPs in our syncytium assay will aid in understanding the conformational changes that occur in VP4 involved in rotavirus penetration into host cells. 相似文献
52.
The radiotoxicity of radium isotopes (especially the long-half-life 226Ra) requires their monitoring in drinking waters or nuclear wastes. We studied the applicability of the PERALS method of detection (photon electron rejecting alpha liquid scintillation) for radium measurement. This method combines alpha liquid scintillation with pulse shape analysis for beta rejection and specific chemical extractants included in the scintillating cocktail. Radium is separated by an extractive-scintillator cocktail called RADAEX containing 2-methyl-2-heptylnonanoic acid (HMHN) and dicyclohexano-21-crown-7 (Cy(2)21C7) as extractant molecules. The variation of the radium extraction has been studied relative to pH, salt concentrations, anion and cation effects, and the volume ratio between aqueous and organic phases. The main parameter affecting the radium extraction in mineral drinking water is its complexation by inorganic anions, especially sulfate. Due to the lack of thermodynamic data, some complexation constants had to be determined. For instance, the value reported in this paper for radium sulfate (log beta = 2.58 +/- 0.22) is in good agreement with that from the literature. The knowledge of complexation constants allows the determination of radium extraction recovery for any solution when the inorganic anion concentrations had been measured by capillary zone electrophoresis. The detection limit for this technique is found to be equal to 0.006 Bq.L-1 using only 6 mL of sample solution for analysis. Several French mineral waters have been studied and the results compared with determinations of uranium and thorium concentrations by ICPMS and time-resolved laser induced fluorescence (TRLIF). 相似文献
53.
Ana Ribeiro John Rasmussen Paulo Flores Luís F. Silva 《Multibody System Dynamics》2012,28(1-2):181-197
The development of a computational multibody knee model able to capture some of the fundamental properties of the human knee articulation is presented. This desideratum is reached by including the kinetics of the real knee articulation. The research question is whether an accurate modeling of the condyle contact in the knee will lead to reproduction of the complex combination of flexion/extension, abduction/adduction, and tibial rotation observed in the real knee. The model is composed by two anatomic segments, the tibia and the femur, whose characteristics are functions of the geometric and anatomic properties of the real bones. The biomechanical model characterization is developed under the framework of multibody systems methodologies using Cartesian coordinates. The type of approach used in the proposed knee model is the joint surface contact conditions between ellipsoids, representing the two femoral condyles, and points, representing the tibial plateau and the menisci. These elements are closely fitted to the actual knee geometry. This task is undertaken by considering a parameter optimization process to replicate experimental data published in the literature, namely that by Lafortune and his coworkers in 1992. Then kinematic data in the form of flexion/extension patterns are imposed on the model corresponding to the stance phase of the human gait. From the results obtained, by performing several computational simulations, it can be observed that the knee model approximates the average secondary motion patterns observed in the literature. Because the literature reports considerable inter-individual differences in the secondary motion patterns, the knee model presented here is also used to check whether it is possible to reproduce the observed differences with reasonable variations of bone shape parameters. This task is accomplished by a parameter study, in which the main variables that define the geometry of condyles are taken into account. It was observed that the data reveal a difference in secondary kinematics of the knee in flexion versus extension. The likely explanation for this fact is the elastic component of the secondary motions created by the combination of joint forces and soft tissue deformations. The proposed knee model is, therefore, used to investigate whether this observed behavior can be explained by reasonable elastic deformations of the points representing the menisci in the model. 相似文献
54.
Edwin Ramaroson Gilbert Blanchard Michel Che Jean-Michel Tatibouët 《Catalysis Letters》1992,15(4):393-400
The oxidative dimerisation of propene to 1,5-hexadiene has been investigated on Bi-Zn-O catalysts. The Bi48ZnO73 phase, observed in the catalysts calcined at 700 ° C is an active and selective catalyst for the formation of 1,5-hexadiene. The best catalytic performance (1,5-hexadiene selectivity 64%) has been obtained at 525 ° C, with a propene to oxygen ratio of 26, on a catalyst formed by Bi48ZnO73 with a small excess of ZnO. 相似文献
55.
T. D. Dudderar C. P. Burger J. A. Gilbert J. A. Smith B. R. Peters 《Journal of Nondestructive Evaluation》1987,6(3):135-146
An innovative approach to nondestructive evaluation (NDE) using noncontacting optical sensors has demonstrated. In this effort a single mode optical fiber interferometer (OFI) was used to sense the presence and form of Rayleigh waves traveling along the surface of a steel test bar at a velocity of nearly 3mm/µs. Acousto-optic time-domain data was successfully used to detect the presence and locate the position of a test flaw (a machined slot) in the bar, and spectrum analysis was used to estimate its geometry and size. This approach has many potential applications in the ultrasonic evaluation of real flaws in structures with complex geometries. Coupled with the authors' earlier work demonstrating the feasibility of generating acoustic waves in metals using laser light pulses transmitted through the fiber optic probes, this latest achievement points to the development of a fully noncontacting, fiber optic based thermal-acousto-photonic (TAP) NDE system, with potential applications to the reliability testing of many important structures where composition, scale, geometry, or restricted access preclude the use of conventional NDE techniques. 相似文献
56.
Marta Teixeira Pinto Ana Sofia Ribeiro Inês Conde Rita Carvalho Joana Paredes 《International journal of molecular sciences》2021,22(1)
The high plasticity of cancer stem-like cells (CSCs) allows them to differentiate and proliferate, specifically when xenotransplanted subcutaneously into immunocompromised mice. CSCs are highly tumorigenic, even when inoculated in small numbers. Thus, in vivo limiting dilution assays (LDA) in mice are the current gold standard method to evaluate CSC enrichment and activity. The chick embryo chorioallantoic membrane (CAM) is a low cost, naturally immune-incompetent and reproducible model widely used to evaluate the spontaneous growth of human tumor cells. Here, we established a CAM-LDA assay able to rapidly reproduce tumor specificities—in particular, the ability of the small population of CSCs to form tumors. We used a panel of organotropic metastatic breast cancer cells, which show an enrichment in a stem cell gene signature, enhanced CD44+/CD24−/low cell surface expression and increased mammosphere-forming efficiency (MFE). The size of CAM-xenografted tumors correlate with the number of inoculated cancer cells, following mice xenograft growth pattern. CAM and mice tumors are histologically comparable, displaying both breast CSC markers CD44 and CD49f. Therefore, we propose a new tool for studying CSC prevalence and function—the chick CAM-LDA—a model with easy handling, accessibility, rapid growth and the absence of ethical and regulatory constraints. 相似文献
57.
Jos L. Neira Ana Cmara-Artigas Jos Gins Hernndez-Cifre María Grazia Ortore 《International journal of molecular sciences》2021,22(6)
The histidine phosphocarrier protein (HPr) kinase/phosphorylase (HPrK/P) modulates the phosphorylation state of the HPr protein, and it is involved in the use of carbon sources by Gram-positive bacteria. Its X-ray structure, as concluded from crystals of proteins from several species, is a hexamer; however, there are no studies about its conformational stability, and how its structure is modified by the pH. We have embarked on the conformational characterization of HPrK/P of Bacillus subtilis (bsHPrK/P) in solution by using several spectroscopic (namely, fluorescence and circular dichroism (CD)) and biophysical techniques (namely, small-angle X-ray-scattering (SAXS) and dynamic light-scattering (DLS)). bsHPrK/P was mainly a hexamer in solution at pH 7.0, in the presence of phosphate. The protein had a high conformational stability, with an apparent thermal denaturation midpoint of ~70 °C, at pH 7.0, as monitored by fluorescence and CD. The protein was very pH-sensitive, precipitated between pH 3.5 and 6.5; below pH 3.5, it had a molten-globule-like conformation; and it acquired a native-like structure in a narrow pH range (between pH 7.0 and 8.0). Guanidinium hydrochloride (GdmCl) denaturation occurred through an oligomeric intermediate. On the other hand, urea denaturation occurred as a single transition, in the range of concentrations between 1.8 and 18 µM, as detected by far-UV CD and fluorescence. 相似文献
58.
Maria Buuales Maria Cristina Ballesteros-Briones Manuela Gonzalez-Aparicio Sandra Hervas-Stubbs Eva Martisova Uxua Mancheo Ana Ricobaraza Sara Lumbreras Cristian Smerdou Ruben Hernandez-Alcoceba 《International journal of molecular sciences》2021,22(8)
Immune checkpoint inhibitors (ICIs) have demonstrated remarkable efficacy in a growing number of malignancies. However, overcoming primary or secondary resistances is difficult due to pharmacokinetics issues and side effects associated with high systemic exposure. Local or regional expression of monoclonal antibodies (mAbs) using gene therapy vectors can alleviate this problem. In this work, we describe a high-capacity adenoviral vector (HCA-EFZP-aPDL1) equipped with a mifepristone-inducible system for the controlled expression of an anti-programmed death ligand 1 (PD-L1) blocking antibody. The vector was tested in an immune-competent mouse model of colorectal cancer based on implantation of MC38 cells. A single local administration of HCA-EFZP-aPDL1 in subcutaneous lesions led to a significant reduction in tumor growth with minimal release of the antibody in the circulation. When the vector was tested in a more stringent setting (rapidly progressing peritoneal carcinomatosis), the antitumor effect was marginal even in combination with other immune-stimulatory agents such as polyinosinic-polycytidylic acid (pI:C), blocking mAbs for T cell immunoglobulin, mucin-domain containing-3 (TIM-3) or agonistic mAbs for 4-1BB (CD137). In contrast, macrophage depletion by clodronate liposomes enhanced the efficacy of HCA-EFZP-aPDL1. These results highlight the importance of addressing macrophage-associated immunoregulatory mechanisms to overcome resistance to ICIs in the context of colorectal cancer. 相似文献
59.
Gilbert Audira Petrus Siregar Kelvin H.-C. Chen Marri Jmelou M. Roldan Jong-Chin Huang Hong-Thih Lai Chung-Der Hsiao 《International journal of molecular sciences》2021,22(11)
Recently, medaka has been used as a model organism in various research fields. However, even though it possesses several advantages over zebrafish, fewer studies were done in medaka compared to zebrafish, especially with regard to its behavior. Thus, to provide more information regarding its behavior and to demonstrate the behavioral differences between several species of medaka, we compared the behavioral performance and biomarker expression in the brain between four medaka fishes, Oryzias latipes, Oryzias dancena, Oryzias woworae, and Oryzias sinensis. We found that each medaka species explicitly exhibited different behaviors to each other, which might be related to the different basal levels of several biomarkers. Furthermore, by phenomics and genomic-based clustering, the differences between these medaka fishes were further investigated. Here, the phenomic-based clustering was based on the behavior results, while the genomic-based clustering was based on the sequence of the nd2 gene. As we expected, both clusterings showed some resemblances to each other in terms of the interspecies relationship between medaka and zebrafish. However, this similarity was not displayed by both clusterings in the medaka interspecies comparisons. Therefore, these results suggest a re-interpretation of several prior studies in comparative biology. We hope that these results contribute to the growing database of medaka fish phenotypes and provide one of the foundations for future phenomics studies of medaka fish. 相似文献
60.
Ivn Benito Ignacio J. Encío Fermín I. Milagro María Alfaro Ana Martínez-Peuela Miguel Barajas Florencio Marzo 《International journal of molecular sciences》2021,22(9)
Recent studies have suggested that flavonoids such as quercetin and probiotics such as Bifidobacterium bifidum (Bf) and Lactobacillus gasseri (Lg) could play a relevant role in inhibiting colon cancer cell growth. Our study investigated the role of dietary supplementation with microencapsulated probiotics (Bf and Lg) along with quercetin in the development of mouse colorectal cancer (CRC). Methods: Adenomatous polyposis coli/multiple intestinal neoplasia (ApcMin/+) mice were fed a standard diet or the same diet supplemented with microencapsulated probiotics (Bf and Lg strains, 107 CFU/100 g food) or both probiotics strains plus microencapsulated quercetin (15 mg/100 g food) for 73 days. Changes in body and organ weights, energy metabolism, intestinal microbiota, and colon tissue were determined. The expression of genes related to the Wnt pathway was also analyzed in colon samples. Results: Dietary supplementation with microencapsulated probiotics or microencapsulated probiotics plus quercetin reduced body weight loss and intestinal bleeding in ApcMin/+ mice. An improvement in energy expenditure was observed after 8 weeks but not after 10 weeks of treatment. A supplemented diet with microencapsulated Bf and Lg reduced the number of aberrant crypt foci (ACF) and adenomas by 45% and 60%, respectively, whereas the supplementation with Bf, Lg and quercetin decreased the number of ACF and adenomas by 57% and 80%, respectively. Microencapsulated Bf and Lg in combination with quercetin could exert inhibition of the canonical Wnt/β-catenin signaling pathway in the colon of ApcMin/+ mice Conclusions: The administration of microencapsulated Bf and Lg, individually or in combination with quercetin, inhibits the CRC development in ApcMin/+ mice. 相似文献