ABSTRACT:Prediction of human volume of distribution at steady state (V ss ) before first administration of a new drug candidate to humans has become an important part of the drug development process. This study examines the assumptions behind interspecies scaling techniques used to predict human V ss from preclinical data, namely the equivalency of V ss,u and/or f ut across species. In addition, several interspecies scaling techniques are evaluated side by side using a set of 67 reference compounds where observed V ss from rats, dogs, monkeys, and humans were compiled from the literature and where plasma protein binding was determined across species using an ultracentrifugation technique. Species similarity in V ss,u or f ut does not appear to be the norm among rats, dogs, monkeys, or humans. Despite this, interspecies scaling from rats, dogs, and monkeys is useful and can provide reasonably accurate predictions of human V ss , although some interspecies scaling approaches were better than others. For example, the performance of the common V ss,u or f ut equivalency approaches using average V ss,u or f ut across three preclinical species was superior to allometric scaling techniques. In addition, considering data from several preclinical species, using the equivalency approach, was superior to scaling from any single species. Although the mechanistic tissue composition equations available in the Simcyp populationbased pharmacokinetic simulator did not necessarily provide the most accurate predictions, and the equations used likely need refinement, they still provide the best opportunity for a mechanistic understanding and prediction of human V ss .

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Objective The CagA (cytotoxin-related gene A, CagA) protein is an important factor for the pathogenicity of Helicobacter pylori (H. pylori). Although H. pylori has previously been shown to activate the NLRP3 inflammasome, it remains unclear what role CagA plays in this process. In the current study, we aimed to investigate the effect of CagA on NLRP3 activation and how it is linked to gastric cancer cell migration and invasion. Methods CagA positive H. pylori strain (Hp/CagA + ) and CagA gene knockout mutant (Hp/ΔCagA) infected and the pcDNA3.1/CagA plasmid transfected gastric epithelial cell lines, respectively. The morphological alterations of cells under a microscope; the NLRP3 inflammasome-related markers: NLRP3, caspase-1, and ASC protein levels were detected by Western blot, IL-1β and IL-18 levels were determined by ELISA; cell migration and invasion were determined by transwell assay; and the pyroptosis levels and intracellular ROS were determined by flow cytometry analysis. Then, pretreated with 5 mM NAC for 2 h and subsequently transfected with the pcDNA3.1/CagA plasmid for 48 h, the effects of NAC pretreatment on CagA-induced NLRP3 inflammasome-related markers expression and cell pyroptosis were examined, finally assessed the effect of CagA on migration and invasion in NLRP3-silenced cells. Results We found that Hp/CagA + strain infection and pcDNA3.1/CagA vector transfection result in NLRP3 inflammasome activation, generation of intracellular ROS, and increased invasion and migration of gastric cancer cells. Moreover, we found that ROS inhibition via NAC effectively blocks NLRP3 activation and pyroptosis. Silencing of NLRP3 reduces the effects of CagA on gastric cancer cell migration and invasion. ConclusionOur study shows that CagA can promote the invasion and migration of gastric cancer cells by activating NLRP3 inflammasome pathway. These findings provide novel insights into the mechanism of gastric cancer induction by H. pylori.

It is generally agreed that early diversification of animals and significant rise of atmospheric and oceanic oxygen (O2) levels occurred in the Ediacaran (635–541 million years ago, Ma) and early Cambrian (ca. 541–509 Ma). The strength and nature of their relationship, however, remain unclear and debated. A recent wave of paleoredox research — with a particular focus on the fossiliferous sections in South China — demonstrates high spatial heterogeneity of oceanic O2 (redox) conditions and dynamic marine shelf oxygenation in a dominantly anoxic ocean during the Ediacaran and early Cambrian. This pattern shows a general spatiotemporal coupling to early animal evolution. We attribute dynamic shelf oxygenation to a complex interplay among the evolving atmosphere, continents, oceans, and biosphere during a critical period in Earth history. Our review supports the idea of a complex coevolution between increasing O2 levels and early diversification of animals, although additional work is required to fully delineate the timing and patterns of this coevolution and the mechanistic underpinnings.

Exclusive liquid repellency (ELR) describes an extreme wettability phenomenon in which a liquid phase droplet is completely repelled from a solid phase when exposed to a secondary immiscible liquid phase. Earlier, we developed a multi-liquid-phase open microfluidic (or underoil) system based on ELR to facilitate rare-cell culture and single-cell processing. The ELR system can allow for the handling of small volumes of liquid droplets with ultra-low sample loss and biofouling, which makes it an attractive platform for biological applications that require lossless manipulation of rare cellular samples (especially for a limited sample size in the range of a few hundred to a few thousand cells). Here, we report an automated platform using ELR microdrops for single-particle (or single-cell) isolation, identification, and retrieval. This was accomplished via the combined use of a robotic liquid handler, an automated microscopic imaging system, and real-time image-processing software for single-particle identification. The automated ELR technique enables rapid, hands-free, and robust isolation of microdrop-encapsulated rare cellular samples.