Ligand binding to cytochrome c peroxidase from Pseudomonas aeruginosa

The high potential heme site of Pseudomonas cytochrome c peroxidase has His and Met as ligands. On reduction, the Fe-met bond becomes photosensitive. Following photolysis, the bond reforms with a half-time of 35 ps. The low potential heme peroxidatic site of the fully reduced enzyme has been shown to bind to a range of ligands. The compounds with carbon monoxide, methyl, ethyl, n-butyl, and t-butyl isonitriles have been investigated by laser flash photolysis. All are photosensitive and show different degrees of geminate recombination of ligand in the picosecond and nanosecond time ranges. Carbon monoxide shows the least effect. The three straight-chain isonitriles show about 50% geminate recombination with half-times of the order of 10 ns. t-Butyl isonitrile shows more and faster recombination. These results imply considerable freedom of movement within the active site for the smaller ligands.     

What it Takes to Get Passed On: Message Content,Style, and Structure as Predictors of Retransmission in the Boston Marathon Bombing Response

Message retransmission is a central aspect of information diffusion. In a disaster context, the passing on of official warning messages by members of the public also serves as a behavioral indicator of message salience, suggesting that particular messages are (or are not) perceived by the public to be both noteworthy and valuable enough to share with others. This study provides the first examination of terse message retransmission of official warning messages in response to a domestic terrorist attack, the Boston Marathon Bombing in 2013. Using messages posted from public officials’ Twitter accounts that were active during the period of the Boston Marathon bombing and manhunt, we examine the features of messages that are associated with their retransmission. We focus on message content, style, and structure, as well as the networked relationships of message senders to answer the question: what characteristics of a terse message sent under conditions of imminent threat predict its retransmission among members of the public? We employ a negative binomial model to examine how message characteristics affect message retransmission. We find that, rather than any single effect dominating the process, retransmission of official Tweets during the Boston bombing response was jointly influenced by various message content, style, and sender characteristics. These findings suggest the need for more work that investigates impact of multiple factors on the allocation of attention and on message retransmission during hazard events.     

Standardizing Immunohistochemistry: A New Reference Control for Detecting Staining Problems

A new standardized immunohistochemistry (IHC) control for breast cancer testing comprises formalin-fixed human epidermal growth factor receptor 2, estrogen receptor, or progesterone receptor peptide antigens covalently attached to 8-µm glass beads. The antigen-coated beads are suspended in a liquid matrix that hardens upon pipetting onto a glass microscope slide. The antigen-coated beads remain in place through deparaffinization, antigen retrieval, and immunostaining. The intensity of the beads’ stain provides feedback regarding the efficacy of both antigen retrieval and immunostaining. As a first report, we tested the sensitivity and specificity of the new IHC controls (“IHControls”). To evaluate sensitivity, various staining problems were simulated. IHControls detected primary and secondary reagent degradation similarly to tissue controls. This first group of IHControls behaved similarly to tissue controls expressing high concentrations of the antigen. The IHControls were also able to detect aberrations in antigen retrieval, as simulated by sub-optimal times or temperatures. Specificity testing revealed that each antigen-coated bead was specific for its cognate IHC test antibody. The data support the conclusion that, like tissue controls, IHControls are capable of verifying the analytic components of an immunohistochemical stain. Unlike tissue controls, IHControls are prepared in large bulk lots, fostering day-to-day reproducibility that can be standardized across laboratories.     

Genome amplification and gene expression in the ciliate macronucleus

The focus of this review is on the micronucleus and macronucleus in the ciliated protozoa and the organization and function of the DNA molecules within them. We present (1) some of the structural and functional differences which are known, (2) the genetic evidence for macronuclear units, (3) two hypotheses for the organization of the DNA molecules in the macronucleus to explain these units, and (4) experiments designed to discriminate between these hypotheses. We conclude that the size of the genome is not reduced in the macronucleus and that there are 45 copies of the haploid genome present in the macronucleus of normal strains of Tetrahymena pyriformis and 800 copies in the macronucleus of Paramecium aurelia. The ciliate genome is relatively simple in terms of repeated sequences. However, not all copies of the genes present in the macronucleus may be identical since fractions of differing thermal stability appear after renaturation.The work reported in this paper has been supported by a research grant, GM 15879, from the National Institute of General Medical Sciences, U.S. Public Health Service, and by grants from the Science and Medical Research Councils of Great Britain. We are also grateful to the Center for Human Growth and Development, University of Michigan, for a summer fellowship facilitating collaboration.