Synthesis of an analog of the thyroid hormone-binding protein transthyretin via regioselective chemical ligation

Transthyretin is an essential protein responsible for the transport of thyroid hormones and retinol in human serum and is also implicated in the amyloid diseases familial amyloidotic polyneuropathy and senile systemic amyloidosis. Its folding properties and stabilization by ligands are of current interest due to their importance in understanding and combating these diseases, Here we report the solid phase synthesis of the monomeric unit of a transthyretin analog (equivalent to 127 amino acids) using t-Boc chemistry and peptide ligation and its folding to form a functional 54-kDa tetramer, The monomeric unit of the protein was chemically synthesized in three parts (positions 1-51, 54-99, and 102-127) and ligated using a chemoselective thioether ligation chemistry. The synthetic protein was folded and assembled to a tetrameric structure in the presence of transthyretin's native ligand, thyroxine, as shown by gel filtration chromatography, native gel electrophoresis, transthyretin antibody recognition, and thyroid hormone binding. Other folding products included a high molecular weight aggregate as well as a transient dimeric species. This represents one of the largest macromolecules chemically synthesized to date and demonstrates the potential of protein chemical synthesis for investigations of protein-ligand interactions.

Resolving the puzzle of productive and destructive conflict in culturally heterogeneous workgroups: A communication accommodation theory approach

This study presents an investigation of the communicative behaviors and strategies employed in the stimulation and management of productive and destructive conflict in culturally heterogeneous workgroups. Using communication accommodation theory (CAT), we argue that the type and course of conflict in culturally heterogeneous workgroups is impacted by the communicative behaviors and strategies employed by group members during interactions. Analysis of data from participant observations, non-participant observations, semi-structured interviews, and self-report questionnaires support CA T-based predictions and provide fresh insights into the triggers and management strategies associated with conflict in culturally heterogeneous workgroups. In particular, results indicated that the more groups used discourse management Strategies, the more they experienced productive conflict. In addition, the use of explanation and checking of own and others' understanding was a major feature of productive conflict, while speech interruptions emerged as a strategy leading to potential destructive conflict. Groups where leaders emerged and assisted in reversing communication breakdowns were better able to manage their discourse, and achieved consensus On task processes. Contributions to the understanding of the triggers and the management of productive conflict in culturally heterogeneous workgroups are discussed.

Polarization squeezing and continuous-variable polarization entanglement

A concept of polarization entanglement for continuous variables is introduced. For this purpose the Stokes-parameter operators and the associated Poincare sphere, which describe the quantum-optical polarization properties of light, are defined and their basic properties are reviewed. The general features of the Stokes operators are illustrated by evaluation of their means and variances for a range of simple polarization states. Some of the examples show polarization squeezing, in which the variances of one or more Stokes parameters are smaller than the coherent-state value. The main object of the paper is the application of these concepts to bright squeezed light. It is shown that a light beam formed by interference of two orthogonally polarized quadrature-squeezed beams exhibits squeezing in some of the Stokes parameters. Passage of such a primary polarization-squeezed beam through suitable optical components generates a pair of polarization-entangled light beams with the nature of a two-mode squeezed state. Implementation of these schemes using the double-fiber Sagnac interferometer provides an efficient method for the generation of bright nonclassical polarization states. The important advantage of these nonclassical polarization states for quantum communication is the possibility of experimentally determining all of the relevant conjugate variables of both squeezed and entangled fields using only linear optical elements followed by direct detection.

Short communication: Extended shifts in ambulance work: influences on health

Shiftwork is a major source of stress for many worker's. This study highlights the role that organizational and psychosocial variables play in alleviating the negative health effects of 10 and 14-h shifts. It examines the direct and mediated effects of coping strategies, social support and control of shifts on work/non-work conflict and subjective health. Participants are 60 ambulance workers, aged 22 to SS years. A structural equation model with good fit demonstrates complex effects of social support from various sources (supervisors, co-workers and family), coping and control on work/non-work conflict and subjective health., Conceptually, the research contributes to the development of a theoretical framework that can assist in explaining how key psychosocial and organizational variables influence the psychological and physical symptoms experienced by shiftworkers. Copyright (C) 2002 John Wiley Sons, Ltd.

Air target detection via bistatic radar based on LEOS communication signals

The paper discusses the bistatic radar parameters for the case when the transmitter is a satellite emitting communication signals. The model utilises signals from an Iridium-like low earth orbiting satellite system. The maximum detection range, when thermal noise-limited, is discussed at the theoretical level and these results are compared with experimentation. Satellite-radar signal levels and the power of ground reflections are evaluated.

Combined chemical separation of Lu, Hf, Sm, Nd, and REEs from a single rock digest: Precise and accurate isotope Determinations of Lu-Hf and Sm-Nd using multicollector-ICPMS

A combined procedure for separating Lu, Hf, Sm, Nd, and rare earth elements (REEs) from a single sample digest is presented. The procedure consists of the following five steps: (1) sample dissolution via sodium peroxide sintering; (2) separation of the high field strength elements from the REEs and other matrix elements by a HF-free anion-exchange column procedure; (3) purification of Hf on a cation-exchange resin; (4) separation of REEs from other matrix elements by cation exchange; (5) Lu, Sm, and Nd separation from the other REEs by reversed-phase ion chromatography. Analytical reproducibilities of Sm-Nd and Lu-Hf isotope systematics are demonstrated for standard solutions and international rock reference materials. Results show overall good reproducibilities for Sm-Nd systematics independent of the rock type analyzed. For the Lu-Hf systematics, the reproducibility of the parent/daughter ratio is much better for JB-1 (basalt) than for two analyzed felsic crustal rocks (DR-N and an Archaean granitoid). It is demonstrated that this poorer reproducibility of the Lu/Hf ratio is truly caused by sample heterogeneity; thus, results are geologically reasonable.

A cassette ligation strategy with thioether replacement of three Gly-Gly peptide bonds: Total chemical synthesis of the 101 residue protein early pregnancy factor [psi(CH2S)(28-29,56-57,76-77)]

The 101 residue protein early pregnancy factor (EPF), also known as human chaperonin 10, was synthesized from four functionalized, but unprotected, peptide segments by a sequential thioether ligation strategy. The approach exploits the differential reactivity of a peptide-NHCH2CH2SH thiolate with XCH2CO-peptides, where X = Cl or I/Br. Initial model studies with short functionalized (but unprotected) peptides showed a significantly faster reaction of a peptide-NHCH2CH2SH thiolate with a BrCH2CO-peptide than with a CICH2CO-peptide, where thiolate displacement of the halide leads to chemoselective formation of a thioether surrogate for the Gly-Gly peptide bond. This rate difference was used as the basis of a novel sequential ligation approach to the synthesis of large polypeptide chains. Thus, ligation of a model bifunctional N-alpha-chloroacetyl, C-terminal thiolated peptide with a second N-alpha-bromoacetyl peptide demonstrated chemoselective bromide displacement by the thiol group. Further investigations showed that the relatively unreactive N-alpha-chloroacetyl peptides could be activated by halide exchange using saturated KI solutions to yield the highly reactive No-iodoacetyl peptides. These findings were used to formulate a sequential thioether ligation strategy for the synthesis of EPF, a 101 amino acid protein containing three Gly-Gly sites approximately equidistantly spaced within the peptide chain. Four peptide segments or cassettes comprising the EPF protein sequence (BrAc-[EPF 78-101] 12, ClAc-[EPF 58-75]-[NHCH2CH2SH] 13, ClAc-[EPF 30-55]-[NHCH2CH2SH] 14, and Ac-[EPF 1-27]-[NHCH2CH2SH] 15) of EPF were synthesized in high yield and purity using Boc SPPS chemistry. In the stepwise sequential ligation strategy, reaction of peptides 12 and 13 was followed by conversion of the N-terminal chloroacetyl functional group to an iodoacetyl, thus activating the product peptide for further ligation with peptide 14. The process of ligation followed by iodoacetyl activation was repeated to yield an analogue of EPF (EPF psi(CH2S)(28-29,56-57,76-77)) 19 in 19% overall yield.