Investigating the pharmacodynamic durability of GalNAc–siRNA conjugates

One hallmark of trivalent N-acetylgalactosamine (GalNAc)-conjugated siRNAs is the remarkable durability of silencing that can persist for months in preclinical species and humans. Here, we investigated the underlying biology supporting this extended duration of pharmacological activity. We found that siRNA accumulation and stability in acidic intracellular compartments is critical for long-term activity. We show that functional siRNA can be liberated from these compartments and loaded into newly generated Argonaute 2 protein complexes weeks after dosing, enabling continuous RNAi activity over time. Identical siRNAs delivered in lipid nanoparticles or as GalNAc conjugates were dose-adjusted to achieve similar knockdown, but only GalNAc–siRNAs supported an extended duration of activity, illustrating the importance of receptor-mediated siRNA trafficking in the process. Taken together, we provide several lines of evidence that acidic intracellular compartments serve as a long-term depot for GalNAc–siRNA conjugates and are the major contributor to the extended duration of activity observed in vivo.     

Genetic control of tolerance induction to human gamma-globulin in mice.

Susceptibility to tolerance induction with monomeric human gamma-globulin (HGG) was tested in different inbred strains of mice. The results indicated a differential tolerance susceptibility among the strains and that the basis for the variation is genetic in nature. By using a protocol that permits genetic analysis, F1, F2, and backcross generations of the parental strains SJL/J and C3H/Bi were examined. A multigenic control model by H-2-linked and non-H-2-linked genes showing Mendelian autosomal inheritance is proposed.     

A Linear City Model with Asymmetric Consumer Distribution

The article analyzes a linear-city model where the consumer distribution can be asymmetric, which is important because in real markets this distribution is often asymmetric. The model yields equilibrium price differences, even though the firms’ costs are equal and their locations are symmetric (at the two endpoints of the city). The equilibrium price difference is proportional to the transportation cost parameter and does not depend on the good''s cost. The firms'' markups are also proportional to the transportation cost. The two firms’ prices will be equal in equilibrium if and only if half of the consumers are located to the left of the city’s midpoint, even if other characteristics of the consumer distribution are highly asymmetric. An extension analyzes what happens when the firms have different costs and how the two sources of asymmetry – the consumer distribution and the cost per unit – interact together. The model can be useful as a tool for further development by other researchers interested in applying this simple yet flexible framework for the analysis of various topics.     

Heterologous Expression and Metal-Binding Characterization of a Type 1 Metallothionein Isoform (OsMTI-1b) from Rice (Oryza sativa)

Metallothioneins (MTs) are ubiquitous, low molecular mass and cysteine-rich proteins that play important roles in maintaining intracellular metal homeostasis, eliminating metal toxification and protecting the cells against oxidative damages. MTs are able to bind metal ions through the thiol groups of their cysteine residues. Plants have several MT isoforms which are classified into four types based on the arrangement of cysteine residues. In the present study, a rice (Oryza sativa) gene encoding type 1 MT isoform, OsMTI-1b, was inserted in vector pET41a and overexpressed in Escherichia coli as carboxy-terminal extensions of glutathione-S-transferase (GST). The recombinant protein GST-OsMTI-1b was purified using affinity chromatography and its ability to bind with Ni²⁺, Cd²⁺, Zn²⁺ and Cu²⁺ ions was analyzed. The results demonstrated that this isoform has ability to bind Ni²⁺, Cd²⁺ and Zn²⁺ ions in vitro, whereas it has no substantial ability to bind Cu²⁺ ions. From competitive reaction with 5,5′-dithiobis(2-nitrobenzoic acid), DTNB, the affinity of metal ions for recombinant form of GST-OsMTI-1b was as follows: Ni²⁺/Cd²⁺ > Zn²⁺ > Cu²⁺     

Terbium‐sensitized fluorescence method for the determination of deferasirox in biological fluids and tablet formulation

A novel, rapid and sensitive spectroflurimetric method was developed and validated for the determination of deferasirox in urine, serum and tablet samples based on sensitization of terbium fluorescence. The excitation and emission wavelengths were 328 and 545 nm, respectively. The optimum conditions for the determination of deferasirox were investigated considering the effects of various parameters. The method was quantitatively evaluated in terms of linearity, recovery, reproducibility and limit of detection. Under the optimal conditions, the fluorescence intensities were linear with the concentration of deferasirox in the range of 5 × 10^?9 to 5×10^?6 mol L^?1, with a detection limit of 1.5 × 10^?9 mol L^?1 and a relative standard deviation of 1.1–2.3%. Linearity, reproducibility, recovery and limit of detection made the method suitable for determination of deferasirox in urine, serum and tablets samples. Copyright © 2010 John Wiley & Sons, Ltd.     

The earliest fossil schizopterid bug (Insecta: Heteroptera) in the Lower Cretaceous amber of Lebanon

The schizopterid bug Libanohypselosoma popovi n. gen., n. sp. belonging to the subfamily Hypselosomatinae is described from the Lower Cretaceous amber of Lebanon. This fossil is the earliest record of the Schizopteridae. The species is distinguished from its related taxa, a discussion is given.     

Two new non-biting midges from the Early Cretaceous Lebanese amber (Diptera: Chironomidae)

Abstract

Two chironomid flies, Ziadeus kamili n. gen., n. sp. and Paicheleria magnifica n. gen., n. sp., respectively attributed to the recent subfamilies Tanypodinae and Prodiamesinae, are described from the Early Cretaceous Lebanese amber. Although very old, this non-biting midge fauna was very diverse with no less than 11 genera and species. However, it was also strongly different from the recent faunas for the complete absence of the Chironominae, that is today the dominant subfamily. The development of the modern chironomid fauna occurred during the Late Cretaceous and/or the Early Paleogene, but when and how?     

Two novel gene orders and the role of light-strand replication in rearrangement of the vertebrate mitochondrial genome

Two novel mitochondrial gene arrangements are identified in an agamid lizard and a ranid frog. Statistical tests incorporating phylogeny indicate a link between novel vertebrate mitochondrial gene orders and movement of the origin of light-strand replication. A mechanism involving errors in light-strand replication and tandem duplication of genes is proposed for rearrangement of vertebrate mitochondrial genes. A second mechanism involving small direct repeats also is identified. These mechanisms implicate gene order as a reliable phylogenetic character. Shifts in gene order define major lineages without evidence of parallelism or reversal. The loss of the origin of light-strand replication from its typical vertebrate position evolves in parallel and, therefore, is a less reliable phylogenetic character. Gene junctions also evolve in parallel. Sequencing across multigenic regions, in particular transfer RNA genes, should be a major focus of future systematic studies to locate novel gene orders and to provide a better understanding of the evolution of the vertebrate mitochondrial genome.