Neutrophil traction stresses are concentrated in the uropod during migration

We find that in contrast to strongly adherent, slow moving cells such as fibroblasts, neutrophils exert contractile stresses largely in the rear of the cell (uropod) relative to the direction of motion. Rather than the leading edge pulling the cell, the rear is both anchoring the cell and the area in which the contractile forces are concentrated. These tractions rapidly reorient themselves during a turn, on a timescale of seconds to minutes, and their repositioning precedes and sets the direction of motion during a turn. We find the total average root mean-squared traction force to be 28+/-10 nN during chemokinesis, and 67+/-10 nN during chemotaxis. We hypothesize that the contraction forces in the back of the neutrophil not only break uropodial adhesive contacts but also create a rearward squeezing contractility, as seen in amoeboid or amoeboidlike cells and the formation of blebs in cells, causing a flow of intracellular material to the fluidlike lamellipod. Our findings suggest an entirely new model of neutrophil locomotion.     

A comparative evaluation of aflatoxin B1 genotoxicity in fish models using the Comet assay

Aflatoxin B₁ (AFB₁) is classified as a Group I hepatocarcinogen in humans by the International Agency for Research on Cancer (IARC). The alkaline Comet assay is a simple and rapid method by which DNA damage can be demonstrated as a function of tail moment. The present work is the first to evaluate the genotoxicity of AFB₁ in fish using the Comet assay. Two different species of fish were selected as models due to previously established sensitivity to AFB₁: rainbow trout (sensitive) and channel catfish (resistant). Fish were i.p. injected with 0.5 mg AFB₁/1 ml DMSO/1 kg body weight. The Comet assay was performed after 4 and 24 h on whole blood, liver, and kidney cells of both species. Trout blood and kidney tissue tested displayed significant (p<0.05) and extensive DNA damage (shown by increased tail moment) after 4 h which then decreased by 24 h. In liver cells, damage progressively increased over time. Conversely, similarly treated catfish showed no elevation in DNA damage over controls at the same doses. These results suggest that the Comet assay is a useful tool for monitoring the genotoxicity of mycotoxins such as AFB₁ and for evaluating organ specific effects of these agents in different species.     

<Emphasis Type="Italic">Empruthotrema longipenis</Emphasis> n. sp. (Monogenoidea: Monocotylidae: Merizocotylinae) from the olfactory sacs of the smooth butterfly ray <Emphasis Type="Italic">Gymnura micrura</Emphasis> (Bloch &amp; Schneider) (Myliobatiformes: Gymnuridae) in the Gulf of Mexico

A new species of Empruthotrema Johnston & Tiegs, 1922 is described based on specimens collected from the olfactory sacs of smooth butterfly rays Gymnura micrura (Bloch & Schneider) captured in Mobile Bay (northcentral Gulf of Mexico), Alabama, USA. Empruthotrema longipenis n. sp. is most similar to the type-species Empruthotrema raiae (MacCallum, 1916) Johnston & Tiegs, 1922 by having 12 marginal and two interhamular loculi with members of haptoral hook pair 1 located midway along the periphery of each interhamular loculus and those of hook pair 2 located at the marginal termini of the bilateral septa flanking the interhamular loculi. Empruthotrema longipenis n. sp. differs from E. raiae by having a much longer male copulatory organ and from its remaining congeners by the sinistral and extracecal ejaculatory bulb flanking the pharynx, the number of interhamular and marginal septa, and the distribution of hook pairs 1 and 2 along the haptoral margin. This is the first report of a monocotylid from the smooth butterfly ray and from Mobile Bay. The diversity of haptoral morphotypes among the currently accepted species of Empruthotrema is detailed and discussed in the context of monophyly of the genus.     

Human stem cells for modeling neurological disorders: accelerating the drug discovery pipeline

The availability of human stem cells heralds a new era for modeling normal and pathologic tissues and developing therapeutics. For example, the in vitro recapitulation of normal and aberrant neurogenesis holds significant promise as a tool for de novo modeling of neurodevelopmental and neurodegenerative diseases. Translational applications include deciphering brain development, function, pathologies, traditional medications, and drug discovery for novel pharmacotherapeutics. For the latter, human stem cell-based assays represent a physiologically relevant and high-throughput means to assess toxicity and other undesirable effects early in the drug development pipeline, avoiding late-stage attrition whilst expediting proof-of-concept of genuine drug candidates. Here we consider the potential of human embryonic, adult, and induced pluripotent stem cells for studying neurological disorders and preclinical drug development.     

Alterations in peroxisome proliferator-activated receptor mRNA expression in skeletal muscle after acute and repeated bouts of exercise

Molecular and Cellular Biochemistry - Reticulon3 (RTN3), as a member of the reticulon family, is generally regarded as a novel human apoptosis-inducing protein. But the extensional role of RTN3...     

Foliar anatomy and fiber motifs in Mammea (Clusiaceae, Kielmeyeroideae)

Anatomical observations of the petiole and lamina of Mammea L. (Clusiaceae, Kielmeyeroideae) show extensive variation. All species have a complex folding pattern of the vascular bundle within the petiole, differing from most of the patterns found in the rest of the family. Except for the species found in the Neotropics and Africa, the laminas of almost all species contain fibers not immediately associated with the vascular tissue—a unique feature in the Clusiaceae. Fiber motifs range from bundles originating within the petiole, sheets forming an almost contiguous layer, to fibers with various patterns of organization derived from the secondary veins. Species groups based on fiber motif are recognized, and these correlate with other anatomical and morphological characters.     

Affinity maturation and characterization of a human monoclonal antibody against HIV-1 gp41

The human D5 monoclonal antibody binds to the highly conserved hydrophobic pocket on the N-terminal heptad repeat (NHR) trimer of HIV-1 gp41 and exhibits modest yet relatively broad neutralization activity. Both binding and neutralization depend on residues in the complementarity determining regions (CDRs) of the D5 IgG variable domains on heavy chain (VH) and light chain (VL). In an effort to increase neutralization activity to a wider range of HIV-1 strains, we have affinity matured the parental D5 scFv by randomizing selected residues in 5 of its 6 CDRs. The resulting scFv variants derived from four different CDR changes showed enhanced binding affinities to gp41 NHR mimetic (5-helix) which correlated to improved neutralization potencies by up to 8-fold. However, when converted to IgG1s, these D5 variants had up to a 12-fold reduction in neutralization potency over their corresponding scFvs despite their slightly enhanced in vitro binding affinities. Remarkably, D5 variant IgG1s bearing residue changes in CDRs that interact with epitope residues N-terminal to the hydrophobic pocket (such as VH CDR3 and VL CDR3) retained more neutralization potency than those containing residue changes in pocket-interacting CDRs (such as VH CDR2). These results provide compelling evidence for the existence of a steric block to an IgG that extends to the gp41 NHR hydrophobic pocket region, and can be a useful guide for developing therapeutic antibodies and vaccines circumventing this block.