Validation of Luminex immunological and competitive Luminex immunological assays for clinical immunogenicity assessment of a 14-valent recombinant human papillomavirus vaccine.

A novel virus-like particle (VLP)-based multivalent recombinant human papillomavirus (HPV) vaccine was developed and evaluated in human, including 14 HPV-type specific VLP antigens (HPV6, 11, 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, and 59). The pseudovirus-based neutralizing assay (PBNA) method is widely used for immunogenicity assessment of HPV vaccine in clinical trials. However, as many as 14 antigen-specific antibody levels need be determined, PBNA is, for many reasons, challenging and time-consuming. In this study, we developed a Luminex immunological assay (LIA) and a competitive Luminex immunological assay (cLIA). These methods increase the throughput, reproducibility and precision, as well as reduce the complexity. All assay parameters showed good characteristics in the validation of both methods, benefiting from highly purified and structurally correct VLPs, high specific antibodies, standard VLP-microspheres and PE-mAbs conjugating process, adequate assay development and stable system. Validation data support the use of both methods for immunogenicity assessment in clinical trials. LIA showed higher sensitivity than cLIA, and due to limited epitopes of mAb, cLIA detected lower antibody responses, and therefore, fewer antibodies. This work not only supports clinical trials of 14-valent HPV vaccines more efficiently and reliably, but also provides a set of validation strategies and usable standards for general vaccine immunogenicity testing.

Virtual screening for potential discoidin domain receptor 1 (DDR1) inhibitors based on structural assessment.

Discoidin domain receptor 1 (DDR1) (EC Number 2.7.10.1) has recently been considered as a promising therapeutic target for idiopathic pulmonary fibrosis (IPF). However, none of the currently discovered DDR1 inhibitors have been included in clinical studies due to low target specificity or druggability limitations, necessitating various approaches to develop novel DDR1 inhibitors. In this study, to assure target specificity, a docking assessment of the DDR1 crystal structures was undertaken to find the well-differentiated crystal structure, and 4CKR was identified among many crystal structures. Then, using the best pharmacophore model and molecular docking, virtual screening of the ChEMBL database was done, and five potential molecules were identified as promising inhibitors of DDR1. Subsequently, all hit compound complex systems were validated using molecular dynamics simulations and MM/PBSA methods to assess the stability of the system after ligand binding to DDR1. Based on molecular dynamics simulations and hydrogen-bonding occupancy analysis, the DDR1-Cpd2, DDR1-Cpd17, and DDR1-Cpd18 complex systems exhibited superior stability compared to the DDR1-Cpd1 and DDR-Cpd33 complex systems. Meanwhile, when targeting DDR1, the descending order of the five hit molecules' binding free energies was Cpd17 (- 145.820 kJ/mol) > Cpd2 (- 131.818 kJ/mol) > Cpd18 (- 130.692 kJ/mol) > Cpd33 (- 129.175 kJ/mol) > Cpd1 (- 126.103 kJ/mol). Among them, Cpd2, Cpd17, and Cpd18 showed improved binding characteristics, indicating that they may be potential DDR1 inhibitors. In this research, we developed a high-hit rate, effective screening method that serves as a theoretical guide for finding DDR1 inhibitors for the development of IPF therapeutics.

[Spatiotemporal Change and Source Apportionment of Non-point Source Nitrogen and Phosphorus Pollution Loads in the Three Gorges Reservoir Area].

The Three Gorges Reservoir area (TGRA) is a critical water source protection area in China and one of the regions with rapid economic development in the Yangtze River basin. Non-point source pollution is the leading cause of the deterioration of the water environment in the TGRA; therefore, studying the non-point source pollution status in the TGRA is of great significance to the regional ecological security and sustainable development. The improved export coefficient model was used to estimate the total non-point source nitrogen and phosphorus pollution loads in the TGRA from 1990 to 2015, the spatial and temporal characteristics of the non-point source nitrogen and phosphorus pollution were analyzed, and the primary sources of pollution were determined by calculating the contribution rate of each pollution source. The results concluded that the nitrogen and phosphorus pollution loads were highest in the hinterland of the reservoir, followed by the end of the reservoir, with the lowest in the head of the reservoir, showing significant spatial heterogeneity in the TGRA. The total loads of nitrogen and phosphorus pollution increased firstly and then decreased, which reached the highest value in 2000 and the lowest value in 2015. The contribution rate of each pollution source to the nitrogen and phosphorus pollution loads, from highest to lowest, were land use, rural life, livestock, and poultry farming. Among them, the land use type of dry land was the predominant source of non-point source nitrogen and phosphorus pollution.

Comprehensive assessment of the coupling coordination degree between urbanization and ecological environment in the Siberian and Far East Federal Districts, Russia from 2005 to 2017.

The urbanization growth in the 20th and 21st centuries has led to a series of unprecedented problems in the ecological environment. Based on constructing an integrated urbanization-ecological environment index system, this article conducts a comprehensive evaluation of the coupling coordination degree between urbanization and the ecological environment and uncovers its spatiotemporal variation characteristics in the Siberian and Far East Federal Districts, Russia from 2005 to 2017. The coupling coordination of urbanization and the ecological environment in the Siberian and Far East Federal Districts improve from slightly unbalanced development stage to barely balanced development stage from 2005 to 2017. In 2017, more than half regions achieved the barely balanced development of urbanization and the ecological environment. However, the most desirable development stage, the superior balanced development stage, is never achieved in the Siberian and Far East Federal Districts during the study period. The spatial pattern of the coupling coordination degree of urbanization and the ecological environment in the Siberian and Far East Federal District gradually changes from "dumbbell" to "high-north low-south". The south part of the Siberian and Far East Federal Districts should be paid more attention in the future urban development process. This research will provide support in the future coordination of urban development in the Siberian and Far East Federal Districts.