The immune-response of macrophages is an important area of investigation since it represents the major pathway by which early-stage defense barriers are established in skin, lungs, and mucosal systems to counteract foreign objects. In this study, we have examined the size-dependent inflammatory and toxicological effects of nanostructured silver particles (nano-Ag) on macrophage immune cells.
ObjectivesThe present study was designed to systematically characterize the denaturation and the renaturation of double stranded DNA (dsDNA), which is suitable for DNA hybridization.MethodsA series of physical and chemical denaturation methods were implemented on well-defined 86-bp dsDNA fragment. The degree of each denaturation was measured and the most suitable denaturation method was determined. DNA renaturation tendency was also investigated for the suggested denaturation method.ResultsHeating, beads mill, and sonication bath did not show any denaturation for 30 minutes. However probe sonication fully denatured DNA in 5 minutes. 1 mol/L sodium hydroxide (alkaline treatment) and 60% dimethyl sulfoxide (DMSO) treatment fully denatured DNA in 2-5 minutes.ConclusionsAmong all the physical methods applied, the direct probe sonication was the most effective way to denature the DNA fragments. Among chemical methods, 60% DMSO was the most adequate denaturation method since it does not cause full renaturation during DNA hybridization.
The portable lysis apparatus provides a standalone, rapid, low cost and power efficient way of obtaining genomic constituents prior to a variety of bioassays used in the field of environmental, biomedical and other applied microbiology.
Inhibitors to interfere protein-protein interactions (PPI) between programmed cell death 1 (PD-1) and programmed death ligand-1 (PD-L1) block evasion of cancers from immune surveillance. Analyzing hot spot residues in PPI is important for small-molecule drug development. In order to find out hot spots on PPI interface in PD-1/PD-L1 complex, we analyzed PPI in PD-1/PD-L1 with a new analysis method, 3-dimensional scattered pair interactions energies (3D-SPIEs), which assorts significant interactions with fragment molecular orbital (FMO) method. By additionally analyzing PPI in PD-1/antibody and PD-L1/antibody complexes, and small-ligand interactions in PD-L1/peptide and PD-L1/small-molecule complexes, we narrowed down the hot spot region with 3D-SPIEs-based interaction map, which integrates PPI and small-ligand interactions. Based on the map, there are two hot spot regions in PPI of PD-1/PD-L1 and the first hot spot region is important for inhibitors. In particular, LY56, LE58, and LN66 in the first hot spot of PD-L1 are important for PD-L1-antibodies and small-inhibitors in common, while LM115 is important for small-inhibitors. Therefore, the 3D-SPIEs-based map would provide valuable information for designing new small-molecule inhibitors to inhibit PPI of PD-1/PD-L1 and the FMO/3D-SPIEs method provides an effectual tool to understand PPI and integrate PPI and small-ligand interactions at a quantum mechanical level.
For the sensitive quantification of bisphenol A (BPA), we have developed NanoAptamer assay, which employs aptamer and complementary signaling DNA, a set of quantum dots (QD), and magnetic beads (MBs). Signaling DNA-QD was tethered to MB-QD via the aptamer. The affinity of the aptamer to BPA resulted in the release of the signaling DNA-QD from the complex and hence the corresponding decrease in the QD fluorescence measurement signal. Three new aptamers (23, 58, and 24-mer) were designed via truncation of the reference aptamer (73-mer). The sensitivity and selectivity of each aptamer for BPA detection via NanoAptamer assay were investigated. One of the truncated aptamers (24-mer) has shown a significantly better performance (limit of detection, LOD, 0.17 pg/mL) than the reference 73-mer aptamer (LOD, 570 pg/mL). It has also shown the best selectivity for BPA detection over BPA analogues (i.e., bisphenol B, bisphenol C, and diethylstilbestrol). It corresponded to a normalized fluorescence change of 33.7% at the environmentally relevant concentration of 1 ng/mL (1 ppb) BPA; however, the analogues remained unchanged (2.3-3.9%).
The prevalence of a novel β-coronavirus (SARS-CoV-2) was declared as a public health emergency of international concern on 30 January 2020 and a global pandemic on 11 March 2020 by WHO. The spike glycoprotein of SARS-CoV-2 is regarded as a key target for the development of vaccines and therapeutic antibodies. In order to develop anti-viral therapeutics for SARS-CoV-2, it is crucial to find amino acid pairs that strongly attract each other at the interface of the spike glycoprotein and the human angiotensin-converting enzyme 2 (hACE2) complex. In order to find hot spot residues, the strongly attracting amino acid pairs at the protein–protein interaction (PPI) interface, we introduce a reliable inter-residue interaction energy calculation method, FMO-DFTB3/D/PCM/3D-SPIEs. In addition to the SARS-CoV-2 spike glycoprotein/hACE2 complex, the hot spot residues of SARS-CoV-1 spike glycoprotein/hACE2 complex, SARS-CoV-1 spike glycoprotein/antibody complex, and HCoV-NL63 spike glycoprotein/hACE2 complex were obtained using the same FMO method. Following this, a 3D-SPIEs-based interaction map was constructed with hot spot residues for the hACE2/SARS-CoV-1 spike glycoprotein, hACE2/HCoV-NL63 spike glycoprotein, and hACE2/SARS-CoV-2 spike glycoprotein complexes. Finally, the three 3D-SPIEs-based interaction maps were combined and analyzed to find the consensus hot spots among the three complexes. As a result of the analysis, two hot spots were identified between hACE2 and the three spike proteins. In particular, E37, K353, G354, and D355 of the hACE2 receptor strongly interact with the spike proteins of coronaviruses. The 3D-SPIEs-based map would provide valuable information to develop anti-viral therapeutics that inhibit PPIs between the spike protein of SARS-CoV-2 and hACE2.
BACKGROUND/OBJECTIVESUse of nutrition labels in food selection is recommended for consumers. The aim of this study is to examine factors, mainly beliefs explaining nutrition label use in female college students based on the Theory of Planned Behavior (TPB).SUBJECTS/METHODSThe subjects were female college students from a university in Seoul, Korea. The survey questionnaire was composed of items examining general characteristics, nutrition label use, behavioral beliefs, normative beliefs, corresponding motivation to comply, and control beliefs. The subjects (n = 300) responded to the questionnaire by self-report, and data from 275 students were analyzed using t-test or χ2-test.RESULTSThe results showed that 37.8% of subjects were nutrition label users. Three out of 15 behavioral beliefs differed significantly by nutrition label use. Nutrition label users agreed more strongly on the benefits of using nutrition labels including 'comparing and selecting better foods' (P < 0.001), 'selecting healthy foods' (P < 0.05). The negative belief of 'annoying' was stronger in non-users than in users (P < 0.001). Three out of 7 sources (parents, siblings, best friend) were important in nutrition label use. Twelve out of 15 control beliefs differed significantly by nutrition label use. These included beliefs regarding constraints of using nutrition labels (e.g., time, spending money for healthy foods) and lack of nutrition knowledge (P < 0.001). Perceived confidence in understanding and applying the specifics of nutrition labels in food selection was also significantly related to nutrition label use (P < 0.001).CONCLUSIONSThis study found that the beliefs, especially control beliefs, suggested in the TPB were important in explaining nutrition label use. To promote nutrition label use, nutrition education might focus on increasing perceived control over constraints of using nutrition labels, acquiring skills for checking nutrition labels, as well as the benefits of using nutrition labels and receiving support from significant others for nutrition label use.
The Hippo pathway is an important signaling pathway modulating growth control and cancer cell proliferation. Dysregulation of the Hippo pathway is a common feature of several types of cancer cells. The modulation of the interaction between yes-associated protein (YAP) and transcriptional enhancer associated domain (TEAD) in the Hippo pathway is considered an attractive target for cancer therapeutic development, although the inhibition of PPI is a challenging task. In order to investigate the hot spots of the YAP and TEAD interacting complex, an ab initio Fragment Molecular Orbital (FMO) method was introduced. With the hot spots, pharmacophores for the inhibitor design were constructed, then virtual screening was performed to an in-house library. Next, we performed molecular docking simulations and FMO calculations for screening results to study the binding modes and affinities between PPI inhibitors and TEAD. As a result of the virtual screening, three compounds were selected as virtual hit compounds. In order to confirm their biological activities, cellular (luciferase activity, proximity ligation assay and wound healing assay in A375 cells, qRT-PCR in HEK 293T cells) and biophysical assays (surface plasmon resonance assays) were performed. Based on the findings of the study, we propose a novel PPI inhibitor BY03 and demonstrate a profitable strategy to analyze YAP–TEAD PPI and discover novel PPI inhibitors.
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