Studies of Verapamil Binding to Human Serum Albumin By High-Performance Affinity Chromatography

The binding of verapamil to the protein human serum albumin (HSA) was examined by using high performance affinity chromatography. Many previous reports have investigated the binding of verapamil with HSA, but the exact strength and nature of this interaction (e.g., the number and location of binding sites) is still unclear. In this study, frontal analysis indicated that at least one major binding site was present for R- and S-verapamil on HSA, with estimated association equilibrium constants on the order of 104 M−1 and a 1.4-fold difference in these values for the verapamil enantiomers at pH 7.4 and 37°C. The presence of a second, weaker group of binding sites on HSA was also suggested by these results. Competitive binding studies using zonal elution were carried out between verapamil and various probe compounds that have known interactions with several major and minor sites on HSA. R/S-Verapamil was found to have direct competition with S-warfarin, indicating that verapamil was binding to Sudlow site I (i.e., the warfarin-azapropazone site of HSA). The average association equilibrium constant for R- and S-verapamil at this site was 1.4 (±0.1) × 104 M−1. Verapamil did not have any notable binding to Sudlow site II of HSA but did appear to have some weak allosteric interactions with L-tryptophan, a probe for this site. An allosteric interaction between verapamil and tamoxifen (a probe for the tamoxifen site) was also noted, which was consistent with the binding of verapamil at Sudlow site I. No interaction was seen between verapamil and digitoxin, a probe for the digitoxin site of HSA. These results gave good agreement with previous observations made in the literature and help provide a more detailed description of how verapamil is transported in blood and of how it may interact with other drugs in the body

Studies of Verapamil Binding to Human Serum Albumin By High-Performance Affinity Chromatography

The binding of verapamil to the protein human serum albumin (HSA) was examined by using high performance affinity chromatography. Many previous reports have investigated the binding of verapamil with HSA, but the exact strength and nature of this interaction (e.g., the number and location of binding sites) is still unclear. In this study, frontal analysis indicated that at least one major binding site was present for R- and S-verapamil on HSA, with estimated association equilibrium constants on the order of 104 M−1 and a 1.4-fold difference in these values for the verapamil enantiomers at pH 7.4 and 37°C. The presence of a second, weaker group of binding sites on HSA was also suggested by these results. Competitive binding studies using zonal elution were carried out between verapamil and various probe compounds that have known interactions with several major and minor sites on HSA. R/S-Verapamil was found to have direct competition with S-warfarin, indicating that verapamil was binding to Sudlow site I (i.e., the warfarin-azapropazone site of HSA). The average association equilibrium constant for R- and S-verapamil at this site was 1.4 (±0.1) × 104 M−1. Verapamil did not have any notable binding to Sudlow site II of HSA but did appear to have some weak allosteric interactions with L-tryptophan, a probe for this site. An allosteric interaction between verapamil and tamoxifen (a probe for the tamoxifen site) was also noted, which was consistent with the binding of verapamil at Sudlow site I. No interaction was seen between verapamil and digitoxin, a probe for the digitoxin site of HSA. These results gave good agreement with previous observations made in the literature and help provide a more detailed description of how verapamil is transported in blood and of how it may interact with other drugs in the body

Analysis Of Drug Interactions With High Density Lipoprotein By High-Performance Affinity Chromatography

Columns containing immobilized lipoproteins were prepared for the analysis of drug interactions with these particles by high-performance affinity chromatography. This approach was evaluated by using it to examine the binding of high density lipoprotein (HDL) to the drugs propranolol or verapamil. HDL was immobilized by the Schiff base method onto silica and gave HPLC columns with reproducible binding to propranolol over four to five days of continuous operation at pH 7.4. Frontal analysis experiments indicated that two types of interactions were occurring between R/S-propranolol and HDL at 37°C: saturable binding with an association equilibrium constant (Ka) of 1.1–1.9 × 105 M−1, and non-saturable binding with an overall affinity constant (n Ka) of 3.7–4.1 × 104 M−1. Similar results were found at 4 and 27°C. Verapamil also gave similar behavior, with a Ka of 6.0 × 104 M−1 at 37°C for the saturable sites and a n Ka value for the non-saturable sites of 2.5 × 104 M−1. These measured affinities gave good agreement with solution-phase values. The results indicated HPAC can be used to study drug interactions with HDL, providing information that should be valuable in obtaining a better description of how drugs are transported within the body

Additional file 1 of Validation of the Meet-URO score in metastatic clear cell renal cell carcinoma patients receiving second or third-line tyrosine kinase inhibitors-immune checkpoint inhibitors combination therapy

Additional file 1: Table S1. Summary of application of Meet-URO score in mRCC patients receiving systemic therapies

Design and evaluation of an innovative composite silicate-based surface treatment agent of concrete

Silicate-based surface treatment agent is a sustainable material to provide a sealing coat on the surface of Portland cement concrete to improve its hardness, impermeability, chemical durability, and abrasion resistance. A type of composite surface treatment agent of concrete is introduced in the paper, using potassium silicate and lithium silicate as main agents and different contents of lithium sulfate, urea, triethanolamine, organosilicon polyether surfactant, and fluorocarbon surfactant as additives. An orthogonal test was conducted to evaluate the effects of different proportions of the two main agents and five additives. The gelation time and viscosity of the agent and the rebound value and penetration depth of the tested concrete samples were tested to evaluate the performance of the designed composite material. The results showed that the two main agents had significant influences on the viscosity, while potassium silicate had the greatest influence on the rebound value. For penetration depth, urea, triethanolamine, organosilicon polyether nonionic surfactants, and fluorocarbon surfactants all had significant effects. The proper ingredients were selected based on the test results, and the optimum proportion of the designed surface treatment agent was determined based on variance analysis of the test results. Two representative commercial inorganic surface treatment agents were selected to compare with the optimum proportion of the designed material, and the effects of the material on the microstructures of the concrete specimens were analyzed by scanning electron microscope (SEM). The mechanism is that Calcium Silicate Hydrate (CSH) gel and ettringite generated can effectively improve surface properties of concrete. The results showed that the optimum proportion of the designed material can effectively block micropores in the surface layer, thus significantly decreasing the concrete porosity, which is conducive to the rapid improvement of concrete strength and compactness

Fused 3-Stage Image Segmentation for Pleural Effusion Cell Clusters

The appearance of tumor cell clusters in pleural effusion is usually a vital sign of cancer metastasis. Segmentation, as an indispensable basis, is of crucial importance for diagnosing, chemical treatment, and prognosis in patients. However, accurate segmentation of unstained cell clusters containing more detailed features than the fluorescent staining images remains to be a challenging problem due to the complex background and the unclear boundary. Therefore, in this paper, we propose a fused 3-stage image segmentation algorithm, namely Coarse segmentation-Mapping-Fine segmentation (CMF) to achieve unstained cell clusters from whole slide images. Firstly, we establish a tumor cell cluster dataset consisting of 107 sets of images, with each set containing one unstained image, one stained image, and one ground-truth image. Then, according to the features of the unstained and stained cell clusters, we propose a three-stage segmentation method: 1) Coarse segmentation on stained images to extract suspicious cell regions-Region of Interest (ROI); 2) Mapping this ROI to the corresponding unstained image to get the ROI of the unstained image (UI-ROI); 3) Fine Segmentation using improved automatic fuzzy clustering framework (AFCF) on the UI-ROI to get precise cell cluster boundaries. Experimental results on 107 sets of images demonstrate that the proposed algorithm can achieve better performance on unstained cell clusters with an F1 score of 90.40%