Poems : dedicated to the people of Branch County /

Mode of access: Internet.

Of Elephants and Toothaches : Ethics, Politics, and Religion in Krzysztof Kieslowski's 'Decalogue' /

This collection is the first to offer a genuinely interdisciplinary approach to Krzysztof Kie?lowski’s Decalogue, a ten-film cycle of modern tales that touch on the ethical dilemmas of the Ten Commandments. The cycle’s deft handling of moral ambiguity and inventive technique established Kie?lowski as a major international director. Kie?lowski once said, “Both the deep believer and the habitual skeptic experience toothaches in exactly the same way.” Of Elephants and Toothaches takes seriously the range of thought, from theological to skeptical, condensed in the cycle’s quite human tales. Bringing together scholars of film, philosophy, literature, and several religions, the volume ranges from individual responsibility, to religion in modernity, to familial bonds, to human desire and material greed. It explores Kie?lowski’s cycle as it relentlessly solicits an ethical response that stimulates both inner disquiet and interpersonal dialogue.

Performance of resurfacing of JRCP and CRCP on the Illinois interstate highway system /

"Project IHR-517, Illinois Cooperative Highway Research Program."

Library of mesmerism and psychology ...

Mode of access: Internet.

Metallurgical calculations /

"Published first as a serial in Electrochemical and metallurgical industry, from March, 1905, to July, 1908."--Pref.

UM Men's Swimming Team, 1955

Front Row: John Hubley, Jim P. Thurlow, Burwell O. Jones, Glennwood I. Miller, Tom O'Keiffe.

Middle Row: head Gus Stager, Ron Pudduck, Thomas W. Prunk, J. Michael Delaney, Peter A. Dow, Donald E. Potter, Joseph W. Haselby, Al Hangelman.

Back Row: Lee Gunn, David Fritz Meyers, Theodore C. Reissing, Harrison G. Wehner, Jr., Wally Goldsmith, Buz Gutowski, Robert Wardrop, John C. Wardrop.

UM Men's Swimming Team, 1956, diving team

Front Row: Lawrason D. Thomas, James M. Kruthers, John P. O'Reilly, J. Michael Delaney, Charles C. Bates.

Middle Row: Charles H. Carscallen, Harrison G. Wehner, Jr., Theodore C. Reising, John Narcy, David Fritz Myers, John W. Murphy, Dick Hausler.

Back Row: head coach Gus Stager, Joseph W. Haselby, Donald F. Adamski, Ed Dauw, James P. Thurlow, diving coach Bruce Harlan.

Sparks from the campfire /

Includes index.

Study of temporary 100 percent ratings for service-connected disabilities requiring inpatient treatment /

Item 985

A yellow fluorescent protein-based assay for high-throughput screening of glycine and GABA(A) receptor chloride channels

There is a significant clinical need to identify novel ligands with high selectivity and potency for GABA(A), GABA(C) and glycine receptor Cl- channels. Two recently developed, yellow fluorescent protein variants (YFP-I152L and YFP-V163S) are highly sensitive to quench by small anions and are thus suited to reporting anionic influx into cells. The aim of this study was to establish the optimal conditions for using these constructs for high-throughput screening of GABA(A), GABA(C) and glycine receptors transiently expressed in HEK293 cells. We found that a 70% fluorescence reduction was achieved by quenching YFP-I152L with a 10 s influx of I- ions, driven by an extemal I- concentration of at least 50 mM. The fluorescence quench was rapid, with a mean time constant of 3 s. These responses were similar for all anion receptor types studied. We also show the assay is sufficiently sensitive to measure agonist and antagonist concentration-responses using either imaging- or photomultiplier-based detection systems. The robustness, sensitivity and low cost of this assay render it suited for high-throughput screening of transiently expressed anionic ligand-gated channels. (c) 2005 Elsevier Ireland Ltd. All rights reserved.

A picrotoxin-specific conformational change in the glycine receptor M2-M3 loop.

The external loop linking the M2 and M3 transmembrane domains is crucial for coupling agonist binding to channel gating in the glycine receptor chloride channel (GlyR). A substituted cysteine accessibility scan previously showed that glycine activation increased the surface accessibility of 6 contiguous residues (Arg(271) Lys(276)) toward the N-terminal end of the homomeric alpha 1 GlyR M2 - M3 loop. In the present study we used a similar approach to determine whether the allosteric antagonist, picrotoxin, could impose conformational changes to this domain that cannot be induced by varying agonist concentrations alone. Picrotoxin slowed the reaction rate of a sulfhydryl-containing compound ( MTSET) with A272C, S273C, and L274C. Before interpreting this as a picrotoxin-specific conformational change, it was necessary to eliminate the possibility of steric competition between picrotoxin and MTSET. Accordingly, we showed that picrotoxin and the structurally unrelated blocker, bilobalide, were both trapped in the R271C GlyR in the closed state and that a point mutation to the pore-lining Thr(6') residue abolished inhibition by both compounds. We also demonstrated that the picrotoxin dissociation rate was linearly related to the channel open probability. These observations constitute a strong case for picrotoxin binding in the pore. We thus conclude that the picrotoxin-specific effects on the M2 - M3 loop are mediated allosterically. This suggests that the M2 - M3 loop responds differently to the occupation of different binding sites.

Payroll Taxes and Personal Liability

Many corporate officers and responsible employees may be exposing their personal assets to liability for unpaid corporate payroll taxes. The authors discuss where liability may arise and identify strategies to avoid personal liability.

Insights into Nonpilus Adhesin Functionality and the Molecular Determinants of Nontypeable Haemophilus influenzae Colonization

Bacterial colonization of the upper respiratory tract is the first step in the pathogenesis of nontypeable Haemophilus influenzae (NTHi) disease. Examination of the determinants of NTHi colonization process has been hampered by the lack of an appropriate animal model. To address this, we have developed a model of NTHi colonization in adult rhesus macaques that involves intranasal inoculation of 1x105 CFU and results in persistent colonization of the upper respiratory tract for at least three weeks with no signs of disease, mimicking asymptomatic colonization of humans. Using this model, we assessed the contributions to colonization of the HMW1 and HMW2 adhesive proteins. In competition experiments, the parent strain expressing both HMW1 and HMW2 was able to efficiently out-compete an isogenic mutant strain expressing neither HMW1 nor HMW2. In experiments involving inoculation of single isogenic derivatives of NTHi strain 12, the strains expressing HMW1 or HMW2 or both were able to colonize efficiently, while the strain expressing neither HMW1 nor HMW2 colonized inefficiently. Furthermore, colonization resulted in antibody production against HMW1 and HMW2 in one-third of the animals, demonstrating that colonization can be an immunizing event. In conclusion, we have established that NTHi is capable of colonizing the upper respiratory tract of rhesus macaques, in some cases associated with stimulation of an immune response. The HMW1 and HMW2 adhesive proteins play a major role in the process of colonization.

After establishing that the HMW1 and HMW2 proteins are colonization factors we further investigated the determinants of HMW1 function. HMW1 is encoded in the same genetic locus as two other proteins, HMW1B and HMW1C, with which HMW1 must interact in order to be functional. Interaction with HMW1C in the cytoplasm results in the glycosylation of HMW1. By employing homologues of HMW1C that glycosylate HMW1 in slightly different patterns we show that the pattern of modification is critical to HMW1 function. Structural analysis showed a change in protein structure when the pattern of HMW1 modification differed. We also identified two specific sites which must be glycosylated for HMW1 to function properly. These point mutations did not have a significant effect on protein structure, suggesting that glycosylation at those specific sites is instead necessary for interaction of HMW1 with its receptor. HMW1B is an outer membrane pore through which HMW1 is transported to reach the bacterial cell surface. We observed that HMW1 isolated from the cytoplasm has a different structure than HMW1 isolated from the bacterial cell surface. By forcing HMW1 to be secreted in a non-HMW1B dependent manner, we show that secretion alone is not sufficient for HMW1 to obtain a functional structure. This leads us to hypothesize that there is something specific in the interaction between HMW1 and HMW1B that aids in proper HMW1 folding.

The NTHi HMW1C glycosyltransferase mediates unconventional N-linked glycosylation of HMW1. In this system, HMW1 is modified in the cytoplasm by sequential transfer of hexose residues. To determine if this mechanism of N-linked glycosylation is employed by species other than NTHi, we examined Kingella kingae and Aggregatibacter aphrophilus homologues of HMW1C. We found both homologues to be functional glycosyltransferases and identified their substrates as the K. kingae Knh and the A. aphrophilus EmaA trimeric autotransporter proteins. LC-MS/MS analysis revealed multiple sites of N-linked glycosylation on Knh and EmaA. Without glycosylation, Knh and EmaA failed to facilitate wild type levels of bacterial autoaggregation or adherence to human epithelial cells, establishing that glycosylation is essential for proper protein function.