A regulator of G protein signaling interaction surface linked to effector specificity

Proteins of the regulator of G protein signaling (RGS) family accelerate GTP hydrolysis by the α subunits (Gα) of G proteins, leading to rapid recovery of signaling cascades. Many different RGS proteins can accelerate GTP hydrolysis by an individual Gα, and GTP hydrolysis rates of different Gαs can be enhanced by the same RGS protein. Consequently, the mechanisms for specificity in RGS regulation and the residues involved remain unclear. Using the evolutionary trace (ET) method, we have identified a cluster of residues in the RGS domain that includes the RGS-Gα binding interface and extends to include additional functionally important residues on the surface. One of these is within helix α3, two are in α5, and three are in the loop connecting α5 and α6. A cluster of surface residues on Gα previously identified by ET, and composed predominantly of residues from the switch III region and helix α3, is spatially contiguous with the ET-identified residues in the RGS domain. This cluster includes residues proposed to interact with the γ subunit of Gtα's effector, cGMP phosphodiesterase (PDEγ). The proximity of these clusters suggests that they form part of an interface between the effector and the RGS-Gα complex. Sequence variations in these residues correlate with PDEγ effects on GTPase acceleration. Because ET identifies residues important for all members of a protein family, these residues likely form a general site for regulation of G protein-coupled signaling cascades, possibly by means of effector interactions.

Cure of human carcinoma xenografts by a single dose of pretargeted yttrium-90 with negligible toxicity

A covalent conjugate (NR-LU-10/SA) was prepared between streptavidin (SA) and NR-LU-10, a mAb that binds an antigen expressed on the surface of most human carcinomas. NR-LU-10/SA was injected into nude mice bearing human tumor xenografts. Injection of biotinylated galactosyl-human serum albumin reduced the circulating levels of conjugate by 95%. Subsequent administration of 90Y-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid-biotin achieved peak uptake at the tumor within 2 hr while >80% of the radioactivity was eliminated in the urine. A single dose of 600–800 μCi of 90Y-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid-biotin produced cures in 10/10 mice with established (>200 mm3) s.c. human small cell lung or colon cancer xenografts and 8/10 cures in mice with human breast cancer xenografts without significant toxicity.

PCR candidate region mismatch scanning: adaptation to quantitative, high-throughput genotyping

Linkage and association analyses were performed to identify loci affecting disease susceptibility by scoring previously characterized sequence variations such as microsatellites and single nucleotide polymorphisms. Lack of markers in regions of interest, as well as difficulty in adapting various methods to high-throughput settings, often limits the effectiveness of the analyses. We have adapted the Escherichia coli mismatch detection system, employing the factors MutS, MutL and MutH, for use in PCR-based, automated, high-throughput genotyping and mutation detection of genomic DNA. Optimal sensitivity and signal-to-noise ratios were obtained in a straightforward fashion because the detection reaction proved to be principally dependent upon monovalent cation concentration and MutL concentration. Quantitative relationships of the optimal values of these parameters with length of the DNA test fragment were demonstrated, in support of the translocation model for the mechanism of action of these enzymes, rather than the molecular switch model. Thus, rapid, sequence-independent optimization was possible for each new genomic target region. Other factors potentially limiting the flexibility of mismatch scanning, such as positioning of dam recognition sites within the target fragment, have also been investigated. We developed several strategies, which can be easily adapted to automation, for limiting the analysis to intersample heteroduplexes. Thus, the principal barriers to the use of this methodology, which we have designated PCR candidate region mismatch scanning, in cost-effective, high-throughput settings have been removed.

Signals and signs in the nervous system: The dynamic anatomy of electrical activity is probably information-rich

The dichotomy between two groups of workers on neuroelectrical activity is retarding progress. To study the interrelations between neuronal unit spike activity and compound field potentials of cell populations is both unfashionable and technically challenging. Neither of the mutual disparagements is justified: that spikes are to higher functions as the alphabet is to Shakespeare and that slow field potentials are irrelevant epiphenomena. Spikes are not the basis of the neural code but of multiple codes that coexist with nonspike codes. Field potentials are mainly information-rich signs of underlying processes, but sometimes they are also signals for neighboring cells, that is, they exert influence. This paper concerns opportunities for new research with many channels of wide-band (spike and slow wave) recording. A wealth of structure in time and three-dimensional space is different at each scale—micro-, meso-, and macroactivity. The depth of our ignorance is emphasized to underline the opportunities for uncovering new principles. We cannot currently estimate the relative importance of spikes and synaptic communication vs. extrasynaptic graded signals. In spite of a preponderance of literature on the former, we must consider the latter as probably important. We are in a primitive stage of looking at the time series of wide-band voltages in the compound, local field, potentials and of choosing descriptors that discriminate appropriately among brain loci, states (functions), stages (ontogeny, senescence), and taxa (evolution). This is not surprising, since the brains in higher species are surely the most complex systems known. They must be the greatest reservoir of new discoveries in nature. The complexity should not deter us, but a dose of humility can stimulate the flow of imaginative juices.

Partial molar volume, surface area, and hydration changes for equilibrium unfolding and formation of aggregation transition state: High-pressure and cosolute studies on recombinant human IFN-γ

The equilibrium dissociation of recombinant human IFN-γ was monitored as a function of pressure and sucrose concentration. The partial molar volume change for dissociation was −209 ± 13 ml/mol of dimer. The specific molar surface area change for dissociation was 12.7 ± 1.6 nm2/molecule of dimer. The first-order aggregation rate of recombinant human IFN-γ in 0.45 M guanidine hydrochloride was studied as a function of sucrose concentration and pressure. Aggregation proceeded through a transition-state species, N*. Sucrose reduced aggregation rate by shifting the equilibrium between native state (N) and N* toward the more compact N. Pressure increased aggregation rate through increased solvation of the protein, which exposes more surface area, thus shifting the equilibrium away from N toward N*. The changes in partial molar volume and specific molar surface area between the N* and N were −41 ± 9 ml/mol of dimer and 3.5 ± 0.2 nm2/molecule, respectively. Thus, the structural change required for the formation of the transition state for aggregation is small relative to the difference between N and the dissociated state. Changes in waters of hydration were estimated from both specific molar surface area and partial molar volume data. From partial molar volume data, estimates were 25 and 128 mol H2O/mol dimer for formation of the aggregation transition state and for dissociation, respectively. From surface area data, estimates were 27 and 98 mol H2O/mol dimer. Osmotic stress theory yielded values ≈4-fold larger for both transitions.

Changes in Hexokinase Activity in Echinochloa phyllopogon and Echinochloa crus-pavonis in Response to Abiotic Stress1

Hexokinase (HXK; EC 2.7.1.1) regulates carbohydrate entry into glycolysis and is known to be a sensor for sugar-responsive gene expression. The effect of abiotic stresses on HXK activity was determined in seedlings of the flood-tolerant plant Echinochloa phyllopogon (Stev.) Koss and the flood-intolerant plant Echinochloa crus-pavonis (H.B.K.) Schult grown aerobically for 5 d before being subjected to anaerobic, chilling, heat, or salt stress. HXK activity was stimulated in shoots of E. phyllopogon only by anaerobic stress. HXK activity was only transiently elevated in E. crus-pavonis shoots during anaerobiosis. In roots of both species, anoxia and chilling stimulated HXK activity. Thus, HXK is not a general stress protein but is specifically induced by anoxia and chilling in E. phyllopogon and E. crus-pavonis. In both species HXK exhibited an optimum pH between 8.5 and 9.0, but the range was extended to pH 7.0 in air-grown E. phyllopogon to 6.5 in N2-grown E. phyllopogon. At physiologically relevant pHs (6.8 and 7.3, N2 and O2 conditions, respectively), N2-grown seedlings retained greater HXK activity at the lower pH. The pH response suggests that in N2-grown seedlings HXK can function in a more acidic environment and that a specific isozyme may be important for regulating glycolytic activity during anaerobic metabolism in E. phyllopogon.

Schleiermacher's Doctrine of Biblical Authority: An Alternative to Content-Based/Supernaturalist and Function-Based Rationalist Models

Author: Kerry W. Holton Title: SCHLEIERMACHER’S DOCTRINE OF BIBLICAL AUTHORITY: AN ALTERNATIVE TO CONTENT-BASED/SUPERNATURALIST AND FUNCTION- BASED/RATIONALIST MODELS Advisor: Theodore M. Vial, Jr. Degree Date: August 2015 This dissertation examines Friedrich Schleiermacher’s understanding of biblical authority and argues that, as an alternative to strictly supernaturalistic and rationalistic models, his understanding allows the New Testament to speak authoritatively in Christian religion in an age of critical, historical awareness. After classifying Schleiermacher’s position in a typology of the doctrine of biblical authority, this dissertation explores his conception of divine revelation and inspiration vis-à-vis scripture. It demonstrates that although he did not believe there is warrant for the claim of a direct connection between divine revelation and scripture, or that scripture is the foundation of faith, he nonetheless asserted that the New Testament is authoritative. He asserted the normative authority of the New Testament on the basis that it is the first presentation of Christian faith. This dissertation examines Schleiermacher’s “canon within the canon,” as well as his denial that the Old Testament shares the same normative worth and inspiration of the New. Although this dissertation finds difficulty with some of Schleiermacher’s views regarding the Old Testament, it names two significant strengths of what is identified as his evangelical, content-based, and rationalist approach to biblical authority. First, it recognizes and values the co-presence and co-activity of the supernatural and the natural !ii in the production of the New Testament canon. This allows both scripture and the church to share religious authority. Second, it allows Christian faith and the historical-method to coexist, as it does not require people to contradict what they know to be the case about science, history, and philosophy. Thus, this dissertation asserts that Schleiermacher’s understanding of biblical authority is a robust one, since, for him, the authority of scripture does not lie in some property of the texts themselves that historians or unbelievers can take away.

The woman who toils: being the experiences of two gentlewomen as factory girls

by Mrs. John Van Vorst and Marie Van Vorst.

The herpetology of Cuba / by Thomas Barbour and Charles T. Ramsden.

v.47:no.2 (1919)

Differentiated Integration in the EU - From the inside looking out. CEPS Paperbacks. January 2014

Flexibility might be convenient when it comes to accommodating member states’ differing socio-economic and political interests in an expanding EU. Yet, opt-outs, enhanced cooperation and cooperation between member states outside the EU’s legal framework are calling into question the boundaries of this constitutional, institutional and legal differentiation in an EU that is founded on common principles, with a specific legal order and common institutions.

Europe on the Road to the Third Stage of EMU: Recent Evidence. CES Germany & Europe Working Papers, No. 07.2, 1996

The paper deals with Europe's effort to proceed to the thud stage of EMU and establish a common currency. It is argued that the success of the common currency experiment will greatly depend on the fulfillment of the Optimum Currency Area (OCA) criteria, on the adoption of the proper adjustment policies as well as on the political desirability of the project. The paper is organized as follows: Section 1 deals briefly with the index of criteria that define an OCA. Section 2 examines the extent to which Europe experiences common demand disturbances, while sections 3 and 4 focus on evidence about the mobility of factors of production across Europe, namely labor and capital. Section 5 examines the possibility of an increase in trade volume across the EU under fixed exchange rates or a common currency regime. Section 6 sheds light on the possibility of the EURO (the ex-ECU} to become a vehicle currency in the international financial system, and Section 7 is concerned with the benefits and costs of the establishment of a European Central Bank (ECB), paying special attention to seigniorage revenues. Section 8 deals with the necessity of establishing an EU federal mechanism facilitating adjustment. Section 9 sketches out a proper role for a hegemonic power in a common currency regime. Finally, section 10 examines EMU prospects during the transitional period. The paper closes with some concluding remarks, where the role of politics and coordination of economic policies are particularly emphasized as of cardinal importance on the road to the third stage of EMU.

Paixão empreendedora: neoliberalismo, capital humano e processos seletivos de trainees

O neoliberalismo, do ponto de vista econômico e social, pode ser entendido como a instauração, na sociedade, de relações estritamente mercantis, fazendo com que a lógica da maximização do ganho e do rendimento seja estendida a todos os campos, promovendo a racionalidade econômica como forma de racionalidade em geral. A forma de governamentalidade neoliberal norte-americana, com sua pretensão de transmutar os indivíduos em sujeitos-microempresas e as relações humanas em relações de tipo concorrencial, faz com que os indivíduos passem a ser vistos como “capital humano”. Originalmente, o termo “capital humano” remete a uma teoria que, desenvolvida sob influência do paradigma econômico neoclássico e liderança de Theodore Schultz, foi responsável por assimilar e transferir princípios econômicos para uma realidade anteriormente isenta de significados dessa natureza, fazendo emergir um discurso que associa o humano ao capital, transportando-o, dessa forma, para uma lógica onde ele deve gerir a si mesmo, tal como uma empresa. A empresa é, assim, promovida a modelo de subjetivação, sendo cada indivíduo um capital a ser gerenciado e valorizado conforme as demandas do mercado. É por isso que o modelo de conduta empreendedora, advindo do discurso do capital humano de inspiração neoliberal e de teorias clássicas propostas por Werner Sombart e Joseph A. Schumpeter, acomete os profissionais das organizações sediadas nos países capitalistas. Esse fato é bastante expressivo entre os jovens que procuram inserir-se no mercado de trabalho, principalmente em posições estratégicas valorizadas dentro das organizações, como as de trainee. No Brasil, os programas de trainee são considerados uma estratégia de busca de atração de jovens com perfil diferenciado, sendo uma resposta encontrada por muitas organizações desde 1970 para ganhar vantagem em um cenário econômico altamente competitivo. Esses profissionais são vistos como os “talentos” da organização, sendo treinados para ocuparem cargos estratégicos em um curto espaço tempo. A fim de esclarecer de que maneira o modelo de conduta empreendedora está presente nos processos seletivos de trainee, foi realizada uma análise dos textos que descrevem as competências exigidas na seleção desses jovens, a partir da Análise Crítica do Discurso (ACD) de Fairclough (2001, 2003), a partir das categorias analíticas “modalidade” e “avaliação”, e reflexões acerca da ideologia neoliberal. Chegou-se à conclusão de que o modelo de conduta empreendedora que está presente nos processos seletivos de trainee é marcada pela expressão de um comportamento apaixonado, que, no campo do management, é entendido a partir do conceito de “paixão empreendedora”. A pesquisa desenvolvida é relevante para o campo da Administração, tanto para o campo acadêmico (uma vez que há poucos estudos que têm como objeto de pesquisa a seleção de trainees e que procuram entendê-lo a partir de um viés crítico utilizando-se da análise do discurso do capital humano), como para quem está inserido nas organizações e convive com as dificuldades e desafios de selecionar jovens para programas de trainees, já que levanta questões importantes sobre os impactos dessas iniciativas tanto para os jovens, como para as organizações que os contratam.