Implementing Marriage: The Issuance of Marriage Licenses in Missouri after Obergefell

The Obergefell v. Hodges decision legalized same-sex marriage throughout the country. Some Missouri counties began issuing marriage licenses to same-sex couples immediately, while others did not until two weeks following the decision. The study examines attempts to explain the differences in the amount of time that counties in Missouri took to implement the Obergefell ruling. Factors such as partisanship, education, religion, and the service industry in each county are examined. The findings indicate that only educational levels in each county are a statistically significant predictor of when a county began issuing marriage licenses to same-sex couples

Breeding Systems in Spiranthes magnicamporum Sheviak

A study of the ecology, breeding systems, and pollinators of Spiranthes magnicamporum Sheviak was undertaken in fall 1992. An initial survey of three glacial drift hill prairies revealed sexual activity in this species via removed pollinia. Two hill prairie populations were used for pollination study. In each population, five individuals were randomly selected and crossed using autogamic, geitonogamic, and xenogamic methods. In addition, ten individuals were fitted with pollinator-exclusion bags. After capsule formation, all plants were harvested. They were examined in the lab for general morphology, the presence or absence of embryo formation, and the prescence of sexual or asexual characteristics. Samples were stained in order to determine the condition of the embryo(s) present. Asexual taxa of S. magnicamporum display adventitious polyembryony (agamospermy) while sexual taxa (autogamy) produce monoembryonic seeds. It was determined in both populations that for the most part, the breeding system exhibited was autogamy. Although no direct observations were made, it must be assumed that some pollination must be taking place due to the presence of removed pollinia

Breeding Systems in Spiranthes magnicamporum Sheviak

A study of the ecology, breeding systems, and pollinators of Spiranthes magnicamporum Sheviak was undertaken in fall 1992. An initial survey of three glacial drift hill prairies revealed sexual activity in this species via removed pollinia. Two hill prairie populations were used for pollination study. In each population, five individuals were randomly selected and crossed using autogamic, geitonogamic, and xenogamic methods. In addition, ten individuals were fitted with pollinator-exclusion bags. After capsule formation, all plants were harvested. They were examined in the lab for general morphology, the presence or absence of embryo formation, and the prescence of sexual or asexual characteristics. Samples were stained in order to determine the condition of the embryo(s) present. Asexual taxa of S. magnicamporum display adventitious polyembryony (agamospermy) while sexual taxa (autogamy) produce monoembryonic seeds. It was determined in both populations that for the most part, the breeding system exhibited was autogamy. Although no direct observations were made, it must be assumed that some pollination must be taking place due to the presence of removed pollinia

The Role of Money and Endorsements in Same-Sex Marriage Ballot Initiatives

This research examines how well certain theories of direct democracy explain outcomes of ballot measures on gay marriage bans. Most theories of direct democracy focus on the types of issues that are quite dissimilar from gay marriage bans. Therefore these theories will likely not do a good job of explaining the results of these elections. In particular I examine the role of campaign spending and elite endorsements in campaigns for gay marriage ban ballot initiatives. In candidate elections, voters commonly use heuristics such as party labels and past performance to help them decide. In most ballot initiative contests voters rely on the information provided by campaigns in lieu of these heuristics, since these are removed in ballot initiative contests. Greater campaign expenditures allow each side to get out more information regarding the proposition, which could be vital in swaying the minds of less informed voters. Elite endorsements can provide voters with partisan signals that may aid in their decision-making. Campaign expenditures and elite endorsements have been found to be important factors in determining which side wins in some ballot initiative elections. However, social issues such as gay marriage are quite different from the areas normally covered by ballot initiatives. As a moral issue, gay marriage has low information needs. Voters do not need to conduct a lot of research to be able to decide their opinion on gay marriage. Therefore, the effect of heuristics such as campaign expenditures and elite endorsements may be lower than it normally is in ballot initiative campaigns. My findings support these hypotheses. I find that campaign expenditures are not significantly correlated with the vote outcomes of same-sex marriage bans. Survey experiments also found that being primed with President Obama’s view on same-sex marriage did not affect most respondents’ opinion on marriage equality

Presenting Concerns and Treatment Issues of Children from Intact, Single-Parent and Stepfamilies

Applied Behavioral Studie

Targeted online liquid chromatography electron capture dissociation mass spectrometry for the localization of sites of in vivo phosphorylation in human Sprouty2

We demonstrate a strategy employing collision-induced dissociation for phosphopeptide discovery, followed by targeted electron capture dissociation (ECD) for site localization. The high mass accuracy and low background noise of the ECD mass spectra allow facile sequencing of coeluting isobaric phosphopeptides, with up to two isobaric phosphopeptides sequenced from a single mass spectrum. In contrast to the previously described neutral loss of dependent ECD method, targeted ECD allows analysis of both phosphotyrosine peptides and lower abundance phosphopeptides. The approach was applied to phosphorylation analysis of human Sprouty2, a regulator of receptor tyrosine kinase signaling. Fifteen sites of phosphorylation were identified, 11 of which are novel

Protein engineering to increase the potential of a therapeutic antibody Fab for long-acting delivery to the eye

To date, ocular antibody therapies for the treatment of retinal diseases rely on injection of the drug into the vitreous chamber of the eye. Given the burden for patients undergoing this procedure, less frequent dosing through the use of long-acting delivery (LAD) technologies is highly desirable. These technologies usually require a highly concentrated formulation and the antibody must be stable against extended exposure to physiological conditions. Here we have increased the potential of a therapeutic antibody antigen-binding fragment (Fab) for LAD by using protein engineering to enhance the chemical and physical stability of the molecule. Structure-guided amino acid substitutions in a negatively charged complementarity determining region (CDR-L1) of an anti-factor D (AFD) Fab resulted in increased chemical stability and solubility. A variant of AFD (AFD.v8), which combines light chain substitutions (VL-D28S:D30E:D31S) with a substitution (VH-D61E) to stabilize a heavy chain isomerization site, retained complement factor D binding and inhibition potency and has properties suitable for LAD. This variant was amenable to high protein concentration (>250 mg/mL), low ionic strength formulation suitable for intravitreal injection. AFD.v8 had acceptable pharmacokinetic (PK) properties upon intravitreal injection in rabbits, and improved stability under both formulation and physiological conditions. Simulations of expected human PK behavior indicated greater exposure with a 25-mg dose enabled by the increased solubility of AFD.v8

Combining SPR with atomic-force microscopy enables single-molecule insights into activation and suppression of the complement cascade

This work was supported by Leverhulme Trust Grant RPG-2015-109.Activation and suppression of the complement system compete on every serum-exposed surface, host or foreign. Potentially harmful outcomes of this competition depend on surface molecules through mechanisms that remain incompletely understood. Combining surface plasmon resonance (SPR) with atomic force microscopy (AFM), here we studied two complement system proteins at the single-molecule level: C3b, the proteolytically activated form of C3, and factor H (FH), the surface-sensing C3b-binding complement regulator. We used SPR to monitor complement initiation occurring through a positive-feedback loop wherein surface-deposited C3b participates in convertases that cleave C3, thereby depositing more C3b. Over multiple cycles of flowing factor B, factor D, and C3 over the SPR chip, we amplified C3b from ∼20 to ∼220 molecules·μm−2. AFM revealed C3b clusters of up to 20 molecules and solitary C3b molecules deposited up to 200 nm away from the clusters. A force of 0.17 ± 0.02 nanonewtons was needed to pull a single FH molecule, anchored to the AFM probe, from its complex with surface-attached C3b. The extent to which FH molecules stretched before detachment varied widely among complexes. Performing force-distance measurements with FH(D1119G), a variant lacking one of the C3b-binding sites and causing atypical hemolytic uremic syndrome, we found that it detached more uniformly and easily. In further SPR experiments, KD values between FH and C3b on a custom-made chip surface were 5-fold tighter than on commercial chips and similar to those on erythrocytes. These results suggest that the chemistry at the surface on which FH acts drives conformational adjustments that are functionally critical.Publisher PDFPeer reviewe

Identification of New Hematopoietic Cell Subsets with a Polyclonal Antibody Library Specific for Neglected Proteins

The identification of new markers, the expression of which defines new phenotipically and functionally distinct cell subsets, is a main objective in cell biology. We have addressed the issue of identifying new cell specific markers with a reverse proteomic approach whereby approximately 1700 human open reading frames encoding proteins predicted to be transmembrane or secreted have been selected in silico for being poorly known, cloned and expressed in bacteria. These proteins have been purified and used to immunize mice with the aim of obtaining polyclonal antisera mostly specific for linear epitopes. Such a library, made of about 1600 different polyclonal antisera, has been obtained and screened by flow cytometry on cord blood derived CD34+CD45dim cells and on peripheral blood derived mature lymphocytes (PBLs). We identified three new proteins expressed by fractions of CD34+CD45dim cells and eight new proteins expressed by fractions of PBLs. Remarkably, we identified proteins the presence of which had not been demonstrated previously by transcriptomic analysis. From the functional point of view, looking at new proteins expressed on CD34+CD45dim cells, we identified one cell surface protein (MOSC-1) the expression of which on a minority of CD34+ progenitors marks those CD34+CD45dim cells that will go toward monocyte/granulocyte differentiation. In conclusion, we show a new way of looking at the membranome by assessing expression of generally neglected proteins with a library of polyclonal antisera, and in so doing we have identified new potential subsets of hematopoietic progenitors and of mature PBLs