Tissue holder for experimental and Demonstration Surgery

Development of device for holding anatomical tissues during operations is discussed. Device consists of plastic dish with hemispherical recess in center to hold excised eyes. Low vacuum applied to underside of recess insures holding of part

Do genetic structure and landscape heterogeneity impact color morph frequency in a polymorphic salamander?

Landscape heterogeneity plays an important role in population structure and divergence, particularly for species with limited vagility. Here, we used a landscape genetic approach to identify how landscape and environmental variables affect genetic structure and color morph frequency in a polymorphic salamander. The eastern red- backed salamander, Plethodon cinereus, is widely distributed in northeastern North America and contains two common color morphs, striped and unstriped, that are divergent in ecology, behavior, and physiology. To quantify population structure, rates of gene flow, and genetic drift, we amplified 10 microsatellite loci from 648 individuals across 28 sampling localities. This study was conducted in northern Ohio, where populations of P. cinereus exhibit an unusually wide range of morph frequency variation. To test whether genetic distance was more correlated with morph frequency, elevation, canopy cover, waterways, ecological niche or geographic distance, we used resistance distance and least cost path analyses. We then examined whether landscape and environmental variables, genetic distance or geographic distance were correlated with variation in morph frequency. Tests for population structure revealed three genetic clusters across our sampling range, with one cluster monomorphic for the striped morph. Rates of gene flow and genetic drift were low to moderate across sites. Genetic distance was most correlated with ecological niche, elevation and a combination of landscape and environmental variables. In contrast, morph frequency variation was correlated with waterways and geographic distance. Thus, our results suggest that selection is also an important evolutionary force across our sites, and a balance between gene flow, genetic drift and selection interact to maintain the two color morphs

Exploration of factors driving incorporation of unnatural dNTPS into DNA by Klenow fragment (DNA polymerase I) and DNA polymerase α

In order to further understand how DNA polymerases discriminate against incorrect dNTPs, we synthesized two sets of dNTP analogues and tested them as substrates for DNA polymerase α (pol α) and Klenow fragment (exo(−)) of DNA polymerase I (Escherichia coli). One set of analogues was designed to test the importance of the electronic nature of the base. The bases consisted of a benzimidazole ring with one or two exocyclic substituent(s) that are either electron-donating (methyl and methoxy) or electron-withdrawing (trifluoromethyl and dinitro). Both pol α and Klenow fragment exhibit a remarkable inability to discriminate against these analogues as compared to their ability to discriminate against incorrect natural dNTPs. Neither polymerase shows any distinct electronic or steric preferences for analogue incorporation. The other set of analogues, designed to examine the importance of hydrophobicity in dNTP incorporation, consists of a set of four regioisomers of trifluoromethyl benzimidazole. Whereas pol α and Klenow fragment exhibited minimal discrimination against the 5- and 6-regioisomers, they discriminated much more effectively against the 4- and 7-regioisomers. Since all four of these analogues will have similar hydrophobicity and stacking ability, these data indicate that hydrophobicity and stacking ability alone cannot account for the inability of pol α and Klenow fragment to discriminate against unnatural bases. After incorporation, however, both sets of analogues were not efficiently elongated. These results suggest that factors other than hydrophobicity, sterics and electronics govern the incorporation of dNTPs into DNA by pol α and Klenow fragment

Genetic Analysis of a Cryptic Contact Zone between Mitochondrial Clades of the Eastern Red-Backed Salamander, Plethodon cinereus

When evolutionarily divergent lineages adjoin their geographic ranges after a period of isolation, myriad outcomes can occur, from population anastomosis to the evolution of reproductive isolation by way of reinforcement. Hybrid zones represent natural experiments that may indicate whether lineages will maintain their evolutionary independence. Here, we report on a hybrid zone in the Eastern Red-Backed Salamander, Plethodon cinereus, a highly abundant and wide-ranging terrestrial salamander found in the northeastern United States and in southeastern Canada. An earlier study identified six distinct mitochondrial clades across the range of P. cinereus. Populations of two of these clades were as close as 9.6 km apart in Lorain County, Ohio, USA. To investigate the nature of this contact zone, we sampled 316 individuals from 16 sites along a 53-km transect, and analyzed 10 microsatellite loci and one mitochondrial locus. We found a clinal transition for mtDNA haplotypes. In contrast, most studies of terrestrial plethodontid salamanders commonly exhibit sharp boundaries between mtDNA clades. Microsatellite markers, however, revealed little differentiation and weak population structure, suggesting the nuclear cline, if it exists, lies outside of our sampling region. Explanations for the discordance between the mitochondrial DNA and our microsatellite data include lineage sorting, male-biased dispersal, or historical introgression of mtDNA, among other possibilities. We compare our results with other studies of introgression in terrestrial salamanders, and discuss the causes of mitonuclear discordance

On the R-Matrix Formulation of Deformed Algebras and Generalized Jordan-Wigner Transformations

The deformed algebra $\cal{A(R)}$, depending upon a Yang-Baxter R- matrix, is considered. The conditions under which the algebra is associative are discussed for a general number of oscillators. Four types of solutions satisfying these conditions are constructed and two of them can be represented by generalized Jordan-Wigner transformations.Our analysis is in some sense an extension of the boson realization of fermions from single-mode to multimode oscillators.Comment: 10+ii pages,preprint RBI-TH-10/93,Latex,(to appear in Europhys.Lett.

Hermitian boson mapping and finite truncation

Starting from a general, microscopic fermion-to-boson mapping that preserves Hermitian conjugation, we discuss truncations of the boson Fock space basis. We give conditions under which the exact boson images of finite fermion operators are also finite (e.g., a 1+2-body fermion Hamiltonian is mapped to a 1+2-body boson Hamiltonian) in the truncated basis. For the most general case, where the image is not necessarily exactly finite, we discuss how to make practical and controlled approximations.Comment: 12 pages in RevTex with no figures, Los Alamos preprint # LA-UR-94-146

Largazole and Its Derivatives Selectively Inhibit Ubiquitin Activating Enzyme (E1)

Protein ubiquitination plays an important role in the regulation of almost every aspect of eukaryotic cellular function; therefore, its destabilization is often observed in most human diseases and cancers. Consequently, developing inhibitors of the ubiquitination system for the treatment of cancer has been a recent area of interest. Currently, only a few classes of compounds have been discovered to inhibit the ubiquitin-activating enzyme (E1) and only one class is relatively selective in E1 inhibition in cells. We now report that Largazole and its ester and ketone analogs selectively inhibit ubiquitin conjugation to p27Kip1 and TRF1 in vitro. The inhibitory activity of these small molecules on ubiquitin conjugation has been traced to their inhibition of the ubiquitin E1 enzyme. To further dissect the mechanism of E1 inhibition, we analyzed the effects of these inhibitors on each of the two steps of E1 activation. We show that Largazole and its derivatives specifically inhibit the adenylation step of the E1 reaction while having no effect on thioester bond formation between ubiquitin and E1. E1 inhibition appears to be specific to human E1 as Largazole ketone fails to inhibit the activation of Uba1p, a homolog of E1 in Schizosaccharomyces pombe. Moreover, Largazole analogs do not significantly inhibit SUMO E1. Thus, Largazole and select analogs are a novel class of ubiquitin E1 inhibitors and valuable tools for studying ubiquitination in vitro. This class of compounds could be further developed and potentially be a useful tool in cells

New Role for L-Arginine in Regulation of Inducible Nitric-Oxide-Synthase-Derived Superoxide Anion Production in Raw 264.7 Macrophages

Dietary supplementation with L-arginine was shown to improve immune responses in various inflammatory models. However, the molecular mechanisms underlying L-arginine effects on immune cells remain unrecognized. Herein, we tested the hypothesis that a limitation of L-arginine could lead to the uncoupled state of murine macrophage inducible nitric oxide synthase and, therefore, increase inducible nitric-oxide-synthase-derived superoxide anion formation. Importantly, we demonstrated that L-arginine dose- and time dependently potentiated superoxide anion production in bacterial endotoxin-stimulated macrophages, although it did not influence NADPH oxidase expression and activity. Detailed analysis of macrophage activation showed the time dependence between LPS-induced iNOS expression and increased O2∙− formation. Moreover, downregulation of macrophage iNOS expression, as well as the inhibition of iNOS activity by NOS inhibitors, unveiled an important role of this enzyme in controlling O2∙− and peroxynitrite formation during macrophage stimulation. In conclusion, our data demonstrated that simultaneous induction of NADPH oxidase, together with the iNOS enzyme, can result in the uncoupled state of iNOS resulting in the production of functionally important levels of O2∙− soon after macrophage activation with LPS. Moreover, we demonstrated, for the first time that increased concentrations of L-arginine further potentiate iNOS-dependent O2∙− formation in inflammatory macrophages

Elizabeth Cary and Intersections of Catholicism and Gender in Early Modern England

Historians have analyzed the life of Elizabeth Cary, Lady Falkland, primarily in the context of her highly publicized conversion to Catholicism and her equally public separation from her Protestant husband, Henry Cary. Through this scrutiny, she has become one among many English Catholic recusant heroines. Literary critics, in contrast, have celebrated Cary\u27s literary corpus both for its challenge to traditional ideals of early modern women as chaste, silent, and obedient and for its reevaluation of women\u27s roles within marriage.1 To circumscribe our understandings of Cary in such ways obscures one of her greatest contributions. Elizabeth Cary, albeit unintentionally, provided an alternative model of Catholic woman hood that sought to negotiate a new balance between religion and gender, thus challenging assumptions about women\u27s roles in English Catholic communities and about the rigid character of Catholicism in the Reformation era