Genetic screen for regulators of ind expression identifies shrew as encoding a novel twisted gastrulation-like protein involved in Dpp signaling.

Initiation and refinement of expression of the Ind homeodomain protein in the Drosophila embryo is coordinately regulated by global dorsoventral patterning pathways Dorsal, Egfr, and Dpp, and well as by Vnd, which positions the ventral boundary of Ind. Therefore, we set out to look for novel regulators of dorsoventral patterning by screening the Exelixis deficiency collection for modified expression of Ind. Indeed, we found deficiencies that remove components of the known signaling pathways had altered or lost ind expression. These findings included deficiencies that remove screw, dpp, and egfr as well as deficiencies that remove ind itself. In addition, we found several deficiencies that had altered or loss of ind expression. We also observed phenotypes suggestive of dorsoventral patterning defects such as twisting during gastrulation, and defects associated with loss of dorsal specification. These include a pair of overlapping deficiencies that produced ventralized embryos. We find that transheterozygotes of these two deficiencies are also ventralized. There are seven genes common to both deficiencies, including CG11582, which encodes a twisted gastrulation-like protein. These two deficiencies are also allelic with shrew mutations. Here, we present data supporting the conclusion that CG11582 is the gene affected in shrew mutants.

Population size estimation in Yellowstone wolves with error-prone noninvasive microsatellite genotypes.

Determining population sizes can be difficult, but is essential for conservation. By counting distinct microsatellite genotypes, DNA from noninvasive samples (hair, faeces) allows estimation of population size. Problems arise because genotypes from noninvasive samples are error-prone, but genotyping errors can be reduced by multiple polymerase chain reaction (PCR). For faecal genotypes from wolves in Yellowstone National Park, error rates varied substantially among samples, often above the 'worst-case threshold' suggested by simulation. Consequently, a substantial proportion of multilocus genotypes held one or more errors, despite multiple PCR. These genotyping errors created several genotypes per individual and caused overestimation (up to 5.5-fold) of population size. We propose a 'matching approach' to eliminate this overestimation bias.

Plicatamide, an antimicrobial octapeptide from Styela plicata hemocytes.

Plicatamide (Phe-Phe-His-Leu-His-Phe-His-dc Delta DOPA), where dc Delta DOPA represents decarboxy-(E)-alpha,beta-dehydro-3,4-dihydroxyphenylalanine, is a potently antimicrobial octapeptide from the blood cells of the solitary tunicate, Styela plicata. Wild type and methicillin-resistant Staphylococcus aureus (MRSA) responded to plicatamide exposure with a massive potassium efflux that began within seconds. Soon thereafter, treated bacteria largely ceased consuming oxygen, and most became nonviable. Native plicatamide also formed cation-selective channels in model lipid bilayers composed of bacterial lipids. Methicillin-resistant S. aureus treated with plicatamide for 5 min contained prominent mesosomes as well as multiple, small dome-shaped surface protrusions that suggested the involvement of osmotic forces in its antimicrobial effects. To ascertain the contribution of the C-terminal dc Delta DOPA residue to antimicrobial activity, we synthesized several analogues of plicatamide that lacked it. One of these peptides, PL-101 (Phe-Phe-His-Leu-His-Phe-His-Tyr-amide), closely resembled native plicatamide in its antimicrobial activity and its ability to induce potassium efflux. Plicatamide was potently hemolytic for human red blood cells but did not lyse ovine erythrocytes. The small size, rapid action, and potent anti-staphylococcal activity of plicatamide and PL-101 make them intriguing subjects for future antimicrobial peptide design.