Close association of the alpha subunits of Gq and G11 G proteins with actin filaments in WRK1 cells: relation to G protein-mediated phospholipase C activation.

A selective polyclonal antibody directed toward the C-terminal decapeptide common to the alpha subunits of Gq and G11 G proteins (G alpha q/G alpha 11) was prepared and used to investigate the subcellular distribution fo these proteins in WRK1 cells, a rat mammary tumor cell line. In immunoblots, the antibody recognized purified G alpha q and G alpha 11 proteins and labeled only two bands corresponding to these alpha subunits. Functional studies indicated that this antibody inhibited vasopressin- and guanosine 5'-[alpha-thio]triphosphate-sensitive phospholipase C activities. Immunofluorescence experiments done with this antibody revealed a filamentous labeling corresponding to intracytoplasmic and perimembranous actin-like filament structures. Colocalization of G alpha q/G alpha 11 and F-actin filaments (F-actin) was demonstrated by double-labeling experiments with anti-G alpha q/G alpha 11 and anti-actin antibodies. Immunoblot analysis of membrane, cytoskeletal, and F-actin-rich fractions confirmed the close association of G alpha q/G alpha 11 with actin. Large amounts of G alpha q/G alpha 11 were recovered in the desmin- and tubulin-free F-actin-rich fraction obtained by a double depolymerization-repolymerization cycle. Disorganization of F-actin filaments with cytochalasin D preserved G alpha q/G alpha 11 and F-actin colocalization but partially inhibited vasopressin- and fluoroaluminate-sensitive phospholipase C activity, suggesting that actin-associated G alpha q/G alpha 11 proteins play a role in signal transduction.

cAMP- and rapamycin-sensitive regulation of the association of eukaryotic initiation factor 4E and the translational regulator PHAS-I in aortic smooth muscle cells.

Incubating rat aortic smooth muscle cells with either platelet-derived growth factor BB (PDGF) or insulin-like growth factor I (IGF-I) increased the phosphorylation of PHAS-I, an inhibitor of the mRNA cap binding protein, eukaryotic initiation factor (eIF) 4E. Phosphorylation of PHAS-I promoted dissociation of the PHAS-I-eIF-4E complex, an effect that could partly explain the stimulation of protein synthesis by the two growth factors. Increasing cAMP with forskolin decreased PHAS-I phosphorylation and markedly increased the amount of eIF-4E bound to PHAS-I, effects consistent with an action of cAMP to inhibit protein synthesis. Both PDGF and IGF-I activated p70S6K, but only PDGF increased mitogen-activated protein kinase activity. Forskolin decreased by 50% the effect of PDGF on increasing p70S6K, and forskolin abolished the effect of IGF-I on the kinase. The effects of PDGF and IGF-I on increasing PHAS-I phosphorylation, on dissociating the PHAS-I-eIF-4E complex, and on increasing p70S6K were abolished by rapamycin. The results indicate that IGF-I and PDGF increase PHAS-I phosphorylation in smooth muscle cells by the same rapamycin-sensitive pathway that leads to activation of p70S6K.

Stimulation of human monocytes with macrophage colony-stimulating factor induces a Grb2-mediated association of the focal adhesion kinase pp125FAK and dynamin.

Macrophage colony-stimulating factor (M-CSF) is required for the growth and differentiation of mononuclear phagocytes. In the present studies using human monocytes, we show that M-CSF induces interaction of the Grb2 adaptor protein with the focal adhesion kinase pp125FAK. The results demonstrate that tyrosine-phosphorylated pp125FAK directly interacts with the SH2 domain of Grb2. The findings indicate that a pYENV site at Tyr-925 in pp125FAK is responsible for this interaction. We also demonstrate that the Grb2-FAK complex associates with the GTPase dynamin. Dynamin interacts with the SH3 domains of Grb2 and exhibits M-CSF-dependent tyrosine phosphorylation in association with pp125FAK. These findings suggest that M-CSF-induced signaling involves independent Grb2-mediated pathways, one leading to Ras activation and another involving pp125FAK and a GTPase implicated in receptor internalization.

Cdc37 is required for association of the protein kinase Cdc28 with G1 and mitotic cyclins.

Studies of the temperature-sensitive cdc37-1 mutant of Saccharomyces cerevisiae suggest that Cdc37 is required for passage through the G1 phase of the cell cycle, but its precise function is not known. We have investigated the role of Cdc37 in the regulation of the cyclin-dependent protein kinase Cdc28. We find that G1 arrest in the cdc37-1 mutant is accompanied by a decrease in the Cdc28 activity associated with the G1 cyclin Cln2. This defect appears to be caused by a decrease in the binding of Cdc28 and Cln2. cdc37-1 mutants also exhibit a defect in the binding and activation of Cdc28 by the mitotic cyclin Clb2. Thus Cdc37 may be a regulator that is required for the association of Cdc28 with multiple cyclins.

Intersubunit contacts made by tryptophan 120 with biotin are essential for both strong biotin binding and biotin-induced tighter subunit association of streptavidin.

In natural streptavidin, tryptophan 120 of each subunit makes contacts with the biotin bound by an adjacent subunit through the dimer-dimer interface. To understand quantitatively the role of tryptophan 120 and its intersubunit communication in the properties of streptavidin, a streptavidin mutant in which tryptophan 120 is converted to phenylalanine was produced and characterized. The streptavidin mutant forms a tetrameric molecule and binds one biotin per subunit, as does natural streptavidin, indicating that the mutation of tryptophan 120 to phenylalanine has no significant effect on the basic properties of streptavidin. However, its biotin-binding affinity was reduced substantially, to approximately 10(8) M-1, indicating that the contact made by tryptophan 120 to biotin has a considerable contribution to the extremely tight biotin binding by streptavidin. The mutant retained bound biotin over a wide pH range or with the addition of urea up to 6 M at neutral pH. However, bound biotin was efficiently released by the addition of excess free biotin due, presumably, to exchange reactions. Electrophoretic analysis revealed that the intersubunit contact made by tryptophan 120 to biotin through the dimer-dimer interface is the major interaction responsible for the biotin-induced, tighter subunit association of streptavidin. In addition, the mutant has weaker subunit association than natural streptavidin even in the absence of biotin, indicating that tryptophan 120 also contributes to the subunit association of tetramers in the absence of biotin.

Asperger's Disorder in College Students: Application of Acceptance and Commitment Therapy

College students with Asperger's Disorder (AD) are likely to face significant challenges as they encounter the unpredictable and socially demanding environment of their university setting. Acceptance and Commitment Therapy (ACT) can be a valuable approach to addressing many of these struggles. This paper will explore the application of ACT to problems that are commonly associated with individuals diagnosed with AD. Utilizing ACT and the Hexaflex Model as a guide for working with AD students can assist in establishing a greater understanding and acceptance of their unique internal experience, as well as their interaction with the external world. ACT can offer these students the ability to be more present in the moment, to recognize efforts to avoid negative experiences, and to move in directions in life that are personally meaningful.

Perceptions of Usability and Usefulness in Digital Libraries

This paper provides an overview of a case study research that investigated the use of Digital Library (DL) resources in two undergraduate classes and explored faculty and students’ perceptions of educational digital libraries. This study found that students and faculty use academic DLs primarily for textual resources, but turn to the open Web for visual and multimedia resources. The study participants did not perceive academic libraries as a useful source of digital images and used search engines when searching for visual resources. The limited use of digital library resources for teaching and learning is associated with perceptions of usefulness and ease of use, especially if considered in a broader information landscape, in conjunction with other library information systems, and in the context of Web resources. The limited use of digital libraries is related to the following perceptions: 1) Library systems are not viewed as user-friendly, which in turn discourages potential users from trying DLs provided by academic libraries; 2) Academic libraries are perceived as places of primarily textual resources; perceptions of usefulness, especially in regard to relevance of content, coverage, and currency, seem to have a negative effect on user intention to use DLs, especially when searching for visual materials.

Draft of the Rules, Directions, and Statutes of the Boylston Professorship of Rhetoric and Oratory in Harvard College, [ca. 1804]

This original draft was probably written by Eliphalet Pearson (1752-1826) as a member of the committee charged with the task of establishing the rules, directions, and statutes for the Boylston Professorship by the Harvard Corporation and the Board of Overseers. This draft is heavily edited and contains many cross outs through the text.

Plan of College buildings and lot from Mr. Bulfinch, [ca. 1814]

Known as the Bulfinch view, this proposed site plan of the College grounds by Charles Bulfinch depicts University Hall at the center of the drawing surrounded by Massachusetts, Harvard, Hollis, Stoughton, and Holworthy Halls. Several unlabeled buildings are displayed in the plan.

A statement of the bills due to Thomas Adams, late Butler of Harvard College and the amount of their appropriations to discharge his debts, 1794

Statement with list of creditors owed funds. Verso is dated 1797, and includes list of cash paid out to creditors.

Rules and orders relating to a Professor of the Mathematicks, of Natural and Experimental Philosophy, in Harvard College, 18 January 1726

This folder contains a single document describing the "rules and orders" of the Hollis Professor of Mathematics and Natural Philosophy. The document begins by defining the subjects to be taught by the Hollis Professor including natural and experimental philosophy, elements of geometry, and the principles of astronomy and geography. It then outlines the number of public and private lectures to be given to students, how much extra time the professor should spend with students reviewing any difficulties they may encounter understanding class subject matter discussed, and stipulates that the professor's duties shall be restricted solely to his teaching activities and not involve him in any religious activities at the College or oblige him to teach any additional studies other than those specified for the Hollis Professor of Mathematics and Natural Philosophy. Furthermore, the rules establish the professor's salary at £80 per year and allow the professor to receive from students, except those students studying theology under the Hollis Professor of Divinity, an additional fee as determined by the Corporation and Board of Overseers, to supplement his income. Moreover, the rules assert that all professorship candidates selected by the Harvard Corporation must be approved by Thomas Hollis during his lifetime or by his executor after his death. Finally, the rules state that the Hollis professor take an oath to the civil government and declare himself a member of the Protestant reformed religion. This document is signed by Thomas Hollis and four witnesses, John Hollis, Joshua Hollis, Richard Solly, and John Williams.

Inventory of Silver Plate, Belonging to Harvard College, and in the Keeping of the President, 1886 December 8

Black and white composition book sent to the Harvard College Library containing a typed "copy of notes made in the spring of 1886" by John H. Buck. Includes historical information, and physical descriptions and valuations of the Great Salt, the Stoughton Cup, the Browne Cup, and the christening basin acquired with the donation of Oliver Wendell, as well as notes on other gifts of silver.

Annotated bibliography of research on operational characteristics and geometric implications of longer and wider trucks. Interim report.

Texas State Department of Highways and Public Transportation, Transportation Planning Division, Austin