The American Recovery and Reinvestment Act of 2009: An Investigation into the Determinants of Funds Awarded to the States

The American Recovery and Reinvestment Act of 2009 is one of the largest government responses to an economic crisis in the history of the United States. The purpose of this paper is to examine the determinants of the total funds awarded to states by federal agencies in the Recovery Act. A review of budgetary theory, distributive politics, and electoral vote maximization theory provides context of historical determinants of resource allocation by the federal government. Then from this literature I develop a model including economic and political variables. The dependent variable in my model is the total funds awarded to the fifty states expressed per capita for thirty-four federal agencies between February 17, 2009 and December 31, 2011. I organize the data into a panel for 1700 observations and use regression with agency fixed-effects to control for the average differences across agencies in observable and unobservable ways. I also cluster by state because the variances vary systematically based on unobserved, correlated state characteristics. The analysis provides strong evidence that four of my chosen independent variables affected the funds awarded in the American Recovery and Reinvestment Act. Economically, the total revenue growth of a state between 2007 and 2008 and the amount of federal aid received per capita have positive and statistically significant relationships with the amount of funds awarded. Politically, the presidential election competitiveness of states and the number of Representatives serving on the House Full Committee on Appropriations have negative and statistically significant relationships with the dependent variable. The results suggest that the awarding of funds in the American Recovery and Reinvestment Act are consistent with the literature indicating that a combination of politics and economics matter in allocating scarce resources among alternative uses

Characterization and optimization of a novel bacterial strain for the production of succinic acid

Bernsteinsäure (BS) ist ein wichtiger Grundbaustein der Chemie und findet als biobasierte Chemikalie weitreichende Anwendung. Aufgrund ihrer Eigenschaft als vielversprechende Plattformchemikalie dient sie als Rohstoff für verschiedene industriell produzierte Chemikalien und Polymere, vor allem als Basis für die Herstellung von Polyestern und Polyamiden. In den letzten Jahren wurden bereits viele Studien zur biotechnischen Produktion von BS hervorgebracht, in denen hohe Endkonzentrationen und Ertragskoeffizienten erreicht wurden. Ein entscheidender Vorteil, der sich aus der Verwendung von Wildtyp-Stämmen für diesen Prozess ergibt, ist, dass nachwachsende Rohstoffe zumeist die natürlichen Substrate der Mikroorganismen darstellen und so hohe Umwandlungsraten erzielt werden können. In dieser Arbeit wird ein bisher unbekannter Bodenorganismus für die biotechnische Produktion von BS verwendet. Relevant für die Bernsteinsäurebildung der aus Wildtyp-Stämmen bestehenden Mischkultur DSM 32268 ist ein neuartiger Organismus, der der Klasse Actinobacteria zuzuordnen ist. Infolgedessen wurde eine Isolierung des bernsteinsäurebildenden Organismus aus der Mischkultur DSM 32268 angestrebt. Dazu werden zahlreiche mikrobiologische Analyseverfahren angewandt. Hierbei wurde herausgestellt, dass es sich bei der Mischkultur DSM 32268 um eine mikrobielle Gemeinschaft handelt, wobei die Actinobacteria-Spezies auf die Interaktion des zweiten Stammes angewiesen ist. Unter mikroaerophilen Bedingungen und bei Verwendung von Glycerin und MgCO3 bildet die Mischkultur im Satzbetrieb als Hauptprodukt BS mit 85,4 g/L, bei einem Ertragskoeffizienten von 0,9 g/g und einer Gesamt-Produktivität von 0,25 g/(L*h). Ein mikroaerophiles Milieu wird dabei durch das Kultivierungssystem Laborflasche mit Kanüle gewährleistet. Daneben sind aber auch die Probenahmehäufigkeit, das Verhältnis Arbeits- zu Nennvolumen der Laborflaschen sowie die Schüttelfrequenz entscheidend für die Effizienz des Prozesses. Ebenso von Bedeutung für die Bernsteinsäurebildung der Mischkultur ist der Komplexmedienbestandteil Hefeextrakt. Die darin enthaltenen Aminosäuren und B-Vitamine wurden analysiert. Durch deren Supplementierung wird die Produktion zum Teil begünstigt. Eine Kostenreduktion des Prozesses wird durch die Verwendung von Rohglycerin der geringsten Aufarbeitungsstufe erzielt. Hiermit kann eine annähernd genauso effiziente Bernsteinsäureproduktion wie mit Pharmaglycerin realisiert werden.Succinic acid (SA) is an important building block chemical and is widely used as a bio-based chemical. Due to its properties as a promising platform chemical, it serves as a raw material for various industrially produced chemicals and polymers, especially as the basis for the production of polyesters and polyamides. In recent years, many studies on the biotechnical production of SA have already been produced, in which high final concentrations and yield coefficients have been achieved. A decisive advantage resulting from the use of wild-type strains for this process is that renewable raw materials usually represent the natural substrates of the microorganisms and thus high conversion rates can be achieved. In this work, a previously unknown soil organism is used for the biotechnical production of SA. Relevant for the formation of succinic acid in the mixed culture DSM 32268, consisting of wild-type strains, is a novel organism, which can be assigned to the Actinobacteria class. As a result, an isolation of the succinic acid-forming organism from the mixed culture DSM 32268 was aimed at. Numerous microbiological analysis methods are used for this purpose. It was found that the mixed culture DSM 32268 is a microbial community, with the Actinobacteria species relying on the interaction of the second strain. Under microaerophilic conditions and with the use of glycerol and MgCO3, the mixed culture forms in batch fermentation SA with 85.4 g/L as the main product with a yield coefficient of 0.9 g/g and a total productivity of 0.25 g/(L*h). A microaerophilic environment is ensured by the cultivation system laboratory bottle with cannula. In addition, the sampling frequency, the ratio of working to nominal volume of the laboratory bottles as well as the shaking frequency are decisive for the efficiency of the process. The complex media component yeast extract is also important for the formation of succinic acid of the mixed culture. The amino acids and B-vitamins contained therein were analyzed. Supplementing them partially favors production. The process is reduced in cost by using crude glycerol at the lowest processing stage. Thus, a succinic acid production can be realized almost as efficient as with pharmaceutical glycerol

The interplay between Src family kinases and receptor tyrosine kinases.

Src family tyrosine kinases (SFKs) are involved in a diverse array of physiological processes, as highlighted in this review. An overview of how SFKs interact with, and participate in signaling from, receptor tyrosine kinases (RTKs) is discussed. And also, how SFKs are activated by RTKs, and how SFKs, in turn, can activate RTKs, as well as how SFKs can promote signaling from growth factor receptors in a number of ways including participation in signaling pathways required for DNA synthesis, control of receptor turnover, actin cytoskeleton rearrangements and motility, and survival are discussed.Peer reviewe

Kinase- and rapsyn-independent activities of the muscle-specific kinase (MuSK).

The muscle-specific receptor tyrosine kinase (MuSK) is co-localized with nicotinic acetylcholine receptors (AChRs) in the postsynaptic membrane of the skeletal neuromuscular junction, and is required for all known aspects of postsynaptic differentiation. Studies in vitro have shown that Z(+)-agrin, a nerve-derived proteoglycan, activates MuSK's kinase activity to promote clustering of AChRs and MuSK itself with a cytoplasmic, receptor-associated protein, rapsyn. These studies, however, have used soluble forms of agrin, whereas agrin is cell- or matrix-attached in vivo. We show here that immobilized (particle- or cell-attached) agrin but not soluble agrin is able to aggregate MuSK in the absence of rapsyn and that this aggregation does not require MuSK's kinase activity but does require MuSK's cytoplasmic domain. Moreover, immobilized agrin can promote clustering of AChRs by a mechanism that requires MuSK and rapsyn but does not require MuSK's kinase activity. These results imply that rapsyn and signaling components activated by MuSK kinase may be dispensable for some early aspects of postsynaptic differentiation.Peer reviewe

A single amino acid substitution in the pleckstrin homology domain of phospholipase C delta1 enhances the rate of substrate hydrolysis.

The pleckstrin homology (PH) domain has been postulated to serve as an anchor for enzymes that operate at a lipid/water interface. To understand further the relationship between the PH domain and enzyme activity, a phospholipase C (PLC) delta1/PH domain enhancement-of-activity mutant was generated. A lysine residue was substituted for glutamic acid in the PH domain of PLC delta1 at position 54 (E54K). Purified native and mutant enzymes were characterized using a phosphatidylinositol 4,5-bisphosphate (PI(4, 5)P2)/dodecyl maltoside mixed micelle assay and kinetics measured according to the dual phospholipid model of Dennis and co-workers (Hendrickson, H. S., and Dennis, E. A. (1984) J. Biol. Chem. 259, 5734-5739; Carmen, G. M., Deems, R. A., and Dennis, E. A. (1995) J. Biol. Chem. 270, 18711-18714). Our results show that both PLC delta1 and E54K bind phosphatidylinositol bisphosphate cooperatively (Hill coefficients, n = 2.2 +/- 0.2 and 2.0 +/- 0.1, respectively). However, E54K shows a dramatically increased rate of (PI(4, 5)P2)-stimulated PI(4,5)P2 hydrolysis (interfacial Vmax for PLC delta1 = 4.9 +/- 0.3 micromol/min/mg and for E54K = 31 +/- 3 micromol/min/mg) as well as PI hydrolysis (Vmax for PLC delta1 = 27 +/- 3.4 nmol/min/mg and for E54K = 95 +/- 12 nmol/min/mg). In the absence of PI(4,5)P2 both native and mutant enzyme hydrolyze PI at similar rates. E54K also has a higher affinity for micellar substrate (equilibrium dissociation constant, Ks = 85 +/- 36 microM for E54K and 210 +/- 48 microM for PLC delta1). Centrifugation binding assays using large unilamelar phospholipid vesicles confirm that E54K binds PI(4,5)P2 with higher affinity than native enzyme. E54K is more active even though the interfacial Michaelis constant (Km) for E54K (0.034 +/- 0.01 mol fraction PI(4,5)P2) is higher than the Km for native enzyme (0.012 +/- 0.002 mol fraction PI(4,5)P2). D-Inositol trisphosphate is less potent at inhibiting E54K PI(4,5)P2 hydrolysis compared with native enzyme. These results demonstrate that a single amino acid substitution in the PH domain of PLC delta1 can dramatically enhance enzyme activity. Additionally, the marked increase in Vmax for E54K argues for a direct role of PH domains in regulating catalysis by allosteric modulation of enzyme structure.Peer reviewe

Narratives of thriving : Black lesbian and queer women negotiating racism, sexism, and heterosexism

This qualitative exploratory study explores the narratives that Black lesbian and queer women age 21 to 35 tell about their lived experience by addressing racism, sexism, and heterosexism and how Black lesbian and queer women live and negotiate in the world. In exploring these narratives, the research focused on the following questions: What are the ways in which Black Lesbian and Queer Women create their own story as they negotiate at the margins of society? How do Black lesbian women create meaning out of their experiences in the face of racism, sexism, and heterosexism? The study found that these 12 self-identified Black lesbian and queer Women were proactive and intentional about creating public and private spaces where they and other Black lesbian queer women could feel safe, comfortable, and free in being their full complex selves. The major findings included each participant exercised resistance strategies to maintain the integrity and expression of their identities, including engaging in practices of renaming to allow space for an intersectional and complex understanding of their identities. They were proactive in finding and building homeplaces to help them manage their complex and individually unique experiences of racism, sexism, and heterosexism. Their narratives revealed important themes regarding coming out and negotiating their identities within their family, faith communities, work, and other social groups. This study revealed Black lesbian and queer women are not simply surviving they are thriving in their communities and in their lives. I conclude with a recommendation that clinicians develop a sense of how their own identities interact and intersect within systems of oppression, and of how Black lesbian and queer women might be impacted by racism, sexism, heterosexism, and other oppressions

Identification and Characterization of a Novel Diterpene Gene Cluster in Aspergillus nidulans

Fungal secondary metabolites are a rich source of medically useful compounds due to their pharmaceutical and toxic properties. Sequencing of fungal genomes has revealed numerous secondary metabolite gene clusters, yet products of many of these biosynthetic pathways are unknown since the expression of the clustered genes usually remains silent in normal laboratory conditions. Therefore, to discover new metabolites, it is important to find ways to induce the expression of genes in these otherwise silent biosynthetic clusters. We discovered a novel secondary metabolite in Aspergillus nidulans by predicting a biosynthetic gene cluster with genomic mining. A Zn(II)2Cys6–type transcription factor, PbcR, was identified, and its role as a pathway-specific activator for the predicted gene cluster was demonstrated. Overexpression of pbcR upregulated the transcription of seven genes in the identified cluster and led to the production of a diterpene compound, which was characterized with GC/MS as ent-pimara-8(14),15-diene. A change in morphology was also observed in the strains overexpressing pbcR. The activation of a cryptic gene cluster by overexpression of its putative Zn(II)2Cys6–type transcription factor led to discovery of a novel secondary metabolite in Aspergillus nidulans. Quantitative real-time PCR and DNA array analysis allowed us to predict the borders of the biosynthetic gene cluster. Furthermore, we identified a novel fungal pimaradiene cyclase gene as well as genes encoding 3-hydroxy-3-methyl-glutaryl-coenzyme A (HMG-CoA) reductase and a geranylgeranyl pyrophosphate (GGPP) synthase. None of these genes have been previously implicated in the biosynthesis of terpenes in Aspergillus nidulans. These results identify the first Aspergillus nidulans diterpene gene cluster and suggest a biosynthetic pathway for ent-pimara-8(14),15-diene

The novel adaptor protein Tks4 (SH3PXD2B) is required for functional podosome formation.

Metastatic cancer cells have the ability to both degrade and migrate through the extracellular matrix (ECM). Invasiveness can be correlated with the presence of dynamic actin-rich membrane structures called podosomes or invadopodia. We showed previously that the adaptor protein tyrosine kinase substrate with five Src homology 3 domains (Tks5)/Fish is required for podosome/invadopodia formation, degradation of ECM, and cancer cell invasion in vivo and in vitro. Here, we describe Tks4, a novel protein that is closely related to Tks5. This protein contains an amino-terminal Phox homology domain, four SH3 domains, and several proline-rich motifs. In Src-transformed fibroblasts, Tks4 is tyrosine phosphorylated and predominantly localized to rosettes of podosomes. We used both short hairpin RNA knockdown and mouse embryo fibroblasts lacking Tks4 to investigate its role in podosome formation. We found that lack of Tks4 resulted in incomplete podosome formation and inhibited ECM degradation. Both phenotypes were rescued by reintroduction of Tks4, whereas only podosome formation, but not ECM degradation, was rescued by overexpression of Tks5. The tyrosine phosphorylation sites of Tks4 were required for efficient rescue. Furthermore, in the absence of Tks4, membrane type-1 matrix metalloproteinase (MT1-MMP) was not recruited to the incomplete podosomes. These findings suggest that Tks4 and Tks5 have overlapping, but not identical, functions, and implicate Tks4 in MT1-MMP recruitment and ECM degradation.Peer reviewe

A putative ariadne-like E3 ubiquitin ligase (PAUL) that interacts with the muscle-specific kinase (MuSK).

Formation of the postsynaptic membrane at the skeletal neuromuscular junction (NMJ) requires activation of the muscle-specific receptor tyrosine kinase (MuSK). Few intracellular mediators or modulators of MuSK actions are known. E3 ubiquitin ligases may serve this role, because activities of several receptor tyrosine kinases, G-protein-coupled receptors and channels are modulated by ubiquitination. Here, we report identification of a putative Ariadne-like ubiquitin ligase (PAUL) that binds to the cytoplasmic domain of MuSK. PAUL is expressed in numerous tissues of developing and adult mice, and is present at NMJs in muscle fibers but is not confined to them.Peer reviewe