Bitopic Ligands Support the Presence of a Metastable Binding Site at the β Adrenergic Receptor.

Metastable binding sites (MBS) have been observed in a multitude of molecular dynamics simulations and can be considered low affinity allosteric binding sites (ABS) that function as stepping stones as the ligand moves toward the orthosteric binding site (OBS). Herein, we show that MBS can be utilized as ABS in ligand design, resulting in ligands with improved binding kinetics. Four homobivalent bitopic ligands (-) were designed by molecular docking of ()-alprenolol (()-ALP) in the cocrystal structure of the β adrenergic receptor (βAR) bound to the antagonist ALP.

Fortuitous Compound Degradation Produces a Tractable Hit against Dethiobiotin Synthetase: A Cautionary Tale of What Goes In Does Not Always Come Out.

We previously reported potent ligands and inhibitors of dethiobiotin synthetase (DTBS), a promising target for antituberculosis drug development (Schumann et al., . 2021, , 2339-2347); here, the unconventional origin of the fragment compound they were derived from is described for the first time.

Inhibition of Dethiobiotin Synthase (DTBS): Toward Next-Generation Antituberculosis Agents.

dethiobiotin synthase (DTBS) is a crucial enzyme involved in the biosynthesis of biotin in the causative agent of tuberculosis, . Here, we report a binder of DTBS, cyclopentylacetic acid ( = 3.4 ± 0.

Probing the Existence of a Metastable Binding Site at the β-Adrenergic Receptor with Homobivalent Bitopic Ligands.

Herein, we report the development of bitopic ligands aimed at targeting the orthosteric binding site (OBS) and a metastable binding site (MBS) within the same receptor unit. Previous molecular dynamics studies on ligand binding to the β-adrenergic receptor (βAR) suggested that ligands pause at transient, less-conserved MBSs. We envisioned that MBSs can be regarded as allosteric binding sites and targeted by homobivalent bitopic ligands linking two identical pharmacophores.

Bitopic Ligands and Metastable Binding Sites: Opportunities for G Protein-Coupled Receptor (GPCR) Medicinal Chemistry.

G protein-coupled receptors (GPCRs) belong to a large superfamily of membrane receptors mediating a variety of physiological functions. As such they are attractive targets for drug therapy. However, it remains a challenge to develop subtype selective GPCR ligands due to the high conservation of orthosteric binding sites.

Silver Nanoparticles Decrease the Viability of Cryptosporidium parvum Oocysts.

Oocysts of the waterborne protozoan parasite Cryptosporidium parvum are highly resistant to chlorine disinfection. We show here that both silver nanoparticles (AgNPs) and silver ions significantly decrease oocyst viability, in a dose-dependent manner, between concentrations of 0.005 and 500 μg/ml, as assessed by an excystation assay and the shell/sporozoite ratio.

Exploitation of Nanotechnology for the Monitoring of Waterborne Pathogens: State-of-the-Art and Future Research Priorities.

Contaminated drinking water is one of the most important environmental contributors to the human disease burden. Monitoring of water for the presence of pathogens is an essential part of ensuring drinking water safety. In order to assess water quality it is essential to have methods available to sample and detect the type, level and viability of pathogens in water which are effective, cheap, quick, sensitive, and where possible high throughput.

Mechanism of neutrophil activation and toxicity elicited by engineered nanomaterials.

The effects of nanomaterials (NMs) on biological systems, especially their ability to stimulate inflammatory responses requires urgent investigation. We evaluated the response of the human differentiated HL60 neutrophil-like cell line to NMs. It was hypothesised that NM physico-chemical characteristics would influence cell responsiveness by altering intracellular Ca2+ concentration [Ca2+]i and reactive oxygen species production.

Inflammation and gene expression in the rat lung after instillation of silica nanoparticles: effect of size, dispersion medium and particle surface charge.

We investigated the effects of silica particles and nanoparticles (NPs) (50 nm and 200 nm) with a neutral and positively charged surface when dispersed in saline, bovine serum albumin (BSA) or lung lining fluid (LLF) 24 h post instillation into the lungs of rats. There was a significant increase in the recruitment of neutrophils in animals instilled with 50 nm plain and aminated NPs compared with 200 nm particles when dispersed in saline or BSA, but not when dispersed in LLF. There was no evidence of toxicity or an increase in the albumin content of the bronchoalveolar lavage fluid.

In vitro toxicological screening of nanoparticles on primary human endothelial cells and the role of flow in modulating cell response.

After passage through biological barriers, nanomaterials inevitably end up in contact with the vascular endothelium and can induce cardiovascular damage. In this study the toxicity and sub-lethal effects of six types of nanoparticle, including four of industrial and biomedical importance, on human endothelial cells were investigated using different in vitro assays. The results show that all the particles investigated induce some level of damage to the cells and that silver particles were most toxic, followed by titanium dioxide.

An in vitro assessment of panel of engineered nanomaterials using a human renal cell line: cytotoxicity, pro-inflammatory response, oxidative stress and genotoxicity.

Background: It has been shown that nanomaterials (NMs) are able to translocate to secondary tissues one of the important being the kidneys. Oxidative stress has been implicated as a possible mechanism for NM toxicity, hence effects on the human renal proximal tubule epithelial cells (HK-2) treated with a panel of engineered nanomaterials (NMs) consisting of two zinc oxide particles (ZnO - coated - NM 110 and uncoated - NM 111), two multi walled carbon nanotubes (MWCNT) (NM 400 and NM 402), one silver (NM 300) and five TiO2 NMs (NM 101, NRCWE 001, 002, 003 and 004) were evaluated.

Methods: In order to assess the toxicological impact of the engineered NMs on HK-2 cells - WST-1 cytotoxicity assay, FACSArray, HE oxidation and the comet assays were utilised.

Effects of silver nanoparticles on the liver and hepatocytes in vitro.

With the increasing use and incorporation of nanoparticles (NPs) into consumer products, screening for potential toxicity is necessary to ensure customer safety. NPs have been shown to translocate to the bloodstream following inhalation and ingestion, and such studies demonstrate that the liver is an important organ for accumulation. Silver (Ag) NPs are highly relevant for human exposure due to their use in food contact materials, dietary supplements, and antibacterial wound treatments.

Primary human hepatocytes versus hepatic cell line: assessing their suitability for in vitro nanotoxicology.

The use of hepatocyte cell lines as a replacement for animal models have been heavily criticised mainly due to low expression of metabolism enzymes. This study compares primary human hepatocytes with the C3A cell line and with respect to their response to a panel of nanomaterials (NMs; two ZnO, two MWCNTs, one Ag and one positively functionalised TiO₂). The cell line was very comparable with the primary hepatocytes with regards to their cytotoxic response to the NMs (Ag > uncoated ZnO > coated ZnO).

An in vitro liver model--assessing oxidative stress and genotoxicity following exposure of hepatocytes to a panel of engineered nanomaterials.

Background: Following exposure via inhalation, intratracheal instillation or ingestion some nanomaterials (NM) have been shown to translocate to the liver. Since oxidative stress has been implicated as a possible mechanism for NM toxicity this study aimed to investigate the effects of various materials (five titanium dioxide (TiO2), two zinc oxide (ZnO), two multi-walled carbon nanotubes (MWCNT) and one silver (Ag) NM) on oxidative responses of C3A cell line as a model for potential detrimental properties of nanomaterials on the liver.

Results: We noted a dose dependant decrease in the cellular glutathione content following exposure of the C3A cells to Ag, the ZnO and the MWCNTs.

Characterization of cerium oxide nanoparticles-part 1: size measurements.

The present study gives an overview of some of the major aspects for consideration in the characterization of nanomaterials (NMs). Part 1 focuses on the measurement of particle size and size-related parameters using several analytical techniques such as transmission electron microscopy, atomic force microscopy, dynamic light scattering, X-ray diffraction, and Brunauer, Emmett, and Teller surface area measurements as applied to commercially available cerium oxide nanoparticles (NPs) and microparticles (MPs). Part 2 (see companion paper) considers nonsize-related characterization and analysis.

In vitro assessment of engineered nanomaterials using a hepatocyte cell line: cytotoxicity, pro-inflammatory cytokines and functional markers.

Effects on the liver C3A cell line treated with a panel of engineered nanomaterials (NMs) consisting of two zinc oxide particles (ZnO; coated 100 nm and uncoated 130 nm), two multi-walled carbon nanotubes (MWCNTs), one silver (Ag < 20 nm), one 7 nm anatase, two rutile TiO2 nanoparticles (10 and 94 nm) and two derivatives with positive and negative covalent functionalisation of the 10 nm rutile were evaluated. The silver particles elicited the greatest level of cytotoxicity (24 h LC50 - 2 µg/cm(2)). The silver was followed by the uncoated ZnO (24 h LC50 - 7.

Interspecies comparisons on the uptake and toxicity of silver and cerium dioxide nanoparticles.

An increasing number and quantity of manufactured nanoparticles are entering the environment as the diversity of their applications increases, and this will lead to the exposure of both humans and wildlife. However, little is known regarding their potential health effects. We compared the potential biological effects of silver (Ag; nominally 35 and 600-1,600 nm) and cerium dioxide (CeO(2;) nominally <25 nm and 1-5 µm) particles in a range of cell (human hepatocyte and intestinal and fish hepatocyte) and animal (Daphnia magna, Cyprinus carpio) models to assess possible commonalities in toxicity across taxa.

Cytotoxicity and induction of inflammation by pepsin in Acid in bronchial epithelial cells.

Introduction. Gastroesophageal reflux has been associated with chronic inflammatory diseases and may be a cause of airway remodelling. Aspiration of gastric fluids may cause damage to airway epithelial cells, not only because acidity is toxic to bronchial epithelial cells, but also since it contains digestive enzymes, such as pepsin.

Effects of silver and cerium dioxide micro- and nano-sized particles on Daphnia magna.

Acute (96 h) and chronic (21 d) exposures of Daphnia magna neonates were carried out with nano- and micro-sized Ag and CeO(2) particles to assess the influence of both material and size of particles on mortality and moulting. Mortality rates for silver in the acute exposures were: AgNP, 56.7 ± 23.

Effects of aqueous exposure to silver nanoparticles of different sizes in rainbow trout.

Despite increasing application of silver nanoparticles (NPs) in industry and consumer products, there is still little known about their potential toxicity, particularly to organisms in aquatic environments. To investigate the fate and effects of silver NPs in fish, rainbow trout (Oncorhynchus mykiss) were exposed via the water to commercial silver particles of three nominal sizes: 10 nm (N(10)), 35 nm (N(35)), and 600-1600 nm (N(Bulk)), and to silver nitrate for 10 days. Uptake into the gills, liver, and kidneys was quantified by inductively coupled plasma-optical emission spectrometry, and levels of lipid peroxidation in gills, liver, and blood were determined by measurements of thiobarbituric acid reactive substances.

Assessing exposure, uptake and toxicity of silver and cerium dioxide nanoparticles from contaminated environments.

The aim of this project was to compare cerium oxide and silver particles of different sizes for their potential for uptake by aquatic species, human exposure via ingestion of contaminated food sources and to assess their resultant toxicity. The results demonstrate the potential for uptake of nano and larger particles by fish via the gastrointestinal tract, and by human intestinal epithelial cells, therefore suggesting that ingestion is a viable route of uptake into different organism types. A consistency was also shown in the sensitivity of aquatic, fish cell and human cell models to Ag and CeO2 particles of different sizes; with the observed sensitivity sequence from highest to lowest as: nano-Ag > micro Ag > nano CeO2 = micro CeO2.