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Biomedical subjects

Eric Carlson

Publications and source records attributed to Eric Carlson.

5 recordsLinked to original sources

CXCL1/KC and CXCL5/LIX are selectively produced by corneal fibroblasts and mediate neutrophil infiltration to the corneal stroma in LPS keratitis.

The severity of corneal inflammation depends on the activity of infiltrating neutrophils responding to chemotactic factors such as CXC chemokines. This study examines the relative contribution of CXCL1/keratinocyte-derived chemokine (KC), CXCL2/monocyte-inhibitory protein-2 (MIP-2), and CXCL5/LPS-induced chemokine (LIX) in neutrophil recruitment to the corneal stroma during LPS keratitis, where neutrophils infiltrate the corneal stroma at 6 h after LPS injection and peak at 24 h. Consistent with this timeframe, KC was detected after 3 h, reached peak levels at 24 h, and decreased thereafter. In contrast, LIX production was not detected until 8 h after injection and peaked at 24 h. MIP-2 was detected at 3 h but did not reach the levels of KC and LIX. Cell types associated with corneal inflammation produced markedly different chemokines in vitro: Murine corneal fibroblasts (MK/T-1) produced LIX and KC in response to LPS but did not produce MIP-2, whereas peritoneal macrophages and neutrophils produced MIP-2 and KC but did not produce LIX. To determine the role of these chemokines in neutrophil recruitment to the cornea, anti-LIX, anti-KC, or anti-MIP-2 was injected into the corneal stroma of enhanced GFP chimeric mice prior to LPS, and total cell and neutrophil infiltration was examined. Antibody to LIX and KC, injected individually or in combination, significantly inhibited neutrophil recruitment to the cornea, whereas anti-MIP-2 had no inhibitory effect. Together, these findings demonstrate cell-specific production of CXC chemokines and show that LIX and KC mediate neutrophil recruitment into the cornea during LPS keratitis.

Animals↗

Gaze following: why (not) learn it?

We propose a computational model of the emergence of gaze following skills in infant-caregiver interactions. The model is based on the idea that infants learn that monitoring their caregiver's direction of gaze allows them to predict the locations of interesting objects or events in their environment (Moore & Corkum, 1994). Elaborating on this theory, we demonstrate that a specific Basic Set of structures and mechanisms is sufficient for gaze following to emerge. This Basic Set includes the infant's perceptual skills and preferences, habituation and reward-driven learning, and a structured social environment featuring a caregiver who tends to look at things the infant will find interesting. We review evidence that all elements of the Basic Set are established well before the relevant gaze following skills emerge. We evaluate the model in a series of simulations and show that it can account for typical development. We also demonstrate that plausible alterations of model parameters, motivated by findings on two different developmental disorders - autism and Williams syndrome - produce delays or deficits in the emergence of gaze following. The model makes a number of testable predictions. In addition, it opens a new perspective for theorizing about cross-species differences in gaze following.

Animals↗

Reticulocyte lysate as a model system to study endoplasmic reticulum membrane protein degradation.

Recent studies have revealed that rabbit reticulocyte lysate (RRL) efficiently reconstitutes endoplasmic reticulum-associated degradation (ERAD) of mutant and misfolded membrane proteins. When supplemented with canine pancreas microsomal membranes, the RRL system faithfully carries out ER targeting, translocation, glycosylation, and membrane integration events and therefore provides a ready source of 35S-labeled protein with defined transmembrane topology. These substrates can be rapidly isolated in native ER membranes which, when incubated in RRL lacking exogenous hemin, are degraded in an ATP-dependent manner by the ubiquitin-proteasome pathway. Because the newly translated protein is the only source of radiolabel, degradation can be followed to its end state by conversion into trichloroacetic acid (TCA)-soluble peptide fragments. A particularly useful aspect of this system is that both membrane-associated and cytosolic components are amenable to biochemical and pharmacological manipulation. Here we describe techniques for preparing translation- and degradation-competent RRL, affinity depletion, identification of cytosolic factors involved in degrading the cystic fibrosis transmembrane conductance regulator (CFTR), and reconstitution of ERAD by add-back of purified recombinant proteins. These techniques provide a powerful tool for dissecting components involved in ubiquitination, degradation, and in particular, extraction of transmembrane ERAD substrates.

Animals↗

Molecular-dynamics simulation of the effect of ions on a liquid-liquid interface for a partially miscible mixture.

Molecular-dynamics simulations were performed to model the effect of added salt ions on the liquid-liquid interface in a partially miscible system. Simulations of the interface between saturated phases of a model 1-hexanol+water system show a bilayer structure of 1-hexanol molecules at the interface with -OH heads of the first layer directed into the water phase and the opposite orientation for the second layer. The alignment of the polar -OH groups at the interface stabilizes a charge separation of sodium and chloride ions when salt is introduced into the aqueous phase, producing an electrical double layer. Chloride ions aggregate nearer the interface and sodium ions move toward the bulk water phase, consistent with the explanation that the -OH alignment presents a region of partial positive charges to which the hydrated chloride atoms are attracted. Ions near the interface were found to be less solvated than those in the bulk phase. An electric field was also applied to drive ions through the interface. Ions crossing the interface tended to shed water molecules as they entered the hexanol bilayer, leaving a trail of water molecules. Stabilization and facilitated transport of the ion by interactions with the second layer of hexanol molecules appeared to be an important step in the mechanism of sodium ion transport.

Journal Article↗