Participatory ergonomic processes to reduce musculoskeletal disorders: summary of a Québec experience.

This article critically reviews 11 participatory ergonomic interventions carried out in Québec by the Occupational Health and Safety Research Institute (IRSST). In the introduction, the characteristics of the approach used are situated in relation to the literature on this subject. Based on the "Ergo team" formula, the approach aims to provide company personnel with the skills to analyze and correct hazardous workstations in relation to musculoskeletal disorders (MSD), using an analysis process that the researchers developed. Although isolated workstations were corrected, the process aims for more general impacts on the company. In the 11 interventions, 40 work situations were analyzed, and in 31 cases, changes were implemented to reduce MSD risks. The most common changes dealt with the tools/equipment (77.4% and physical layouts (84%); changes involving work methods (29% and work organization (12.9%) were less common. The difficulties encountered in the interventions are summarized, and the possible impacts of the interventions on the organization and psychosocial factors are discussed. The authors then address the limitations of the paper and the factors that should be considered in evaluating such a participatory process. The authors conclude that the participatory process was successful in implementing changes in companies and that other studies are necessary for a better understanding of the process and its impacts.

Exploiting antibodies as catalysts: potential therapeutic applications.

In this review, we explore recent developments in the generation of catalytic antibodies and their potential in therapy.

Modulation of the regioselectivity of a Bacillus alpha-galactosidase by directed evolution.

Alpha-galactosidase AgaB of Bacillus stearothermophilus was subjected to directed evolution in an effort to modify its regioselectivity. The wild-type enzyme displays a major 1,6 and minor 1,3 regioselectivity. We used random mutagenesis and staggered extension process (StEP) to obtain mutant enzymes displaying modified regioselectivity. We developed a screening procedure allowing first the elimination of AgaB mutants bearing the 1,6 regioselectivity and secondly the selection of those retaining a 1,3 regioselectivity. Our results show that, among the evolved enzymes that have lost most of their activity towards the 1,6 linkage both in hydrolysis and in synthesis, one (E901) has retained its 1,3 activity. However the transglycosylation level reached by this mutant is quite low versus that of the native enzyme. This work constitutes the first example of modification of glycosylhydrolase regioselectivity by directed evolution.

Identification by saturation mutagenesis of a single residue involved in the alpha-galactosidase AgaB regioselectivity.

The alpha-galactosidase AgaB of Bacillus stearothermophilus displays a major 1,6 and a minor 1,3 regioselectivity. The wild-type enzyme was subjected to directed evolution (random mutagenesis and in vitro recombination) using a double screening strategy based on the elimination of the 1,6 regioselectivity and the analysis by TLC of the transglycosylation products. One of the AgaB mutants (E500) exhibited a new 1,2 regioselectivity and a rather high level of transglycosylation. The corresponding gene contains 10 mutations compared to the agaB gene and we demonstrated by saturation mutagenesis that the G442R substitution strongly contributes to the emergence of this new regioselectivity. Moreover, other single point mutations at this position led to new mutants displaying other kinds of regioselectivity demonstrating the importance of this position in the subtle kinetic control of transglycosylation.

Structural evidence for a programmed general base in the active site of a catalytic antibody.

The crystal structure of the complex of a catalytic antibody with its cationic hapten at 1.9-A resolution demonstrates that the hapten amidinium group is stabilized through an ionic pair interaction with the carboxylate of a combining-site residue. The location of this carboxylate allows it to act as a general base in an allylic rearrangement. When compared with structures of other antibody complexes in which the positive moiety of the hapten is stabilized mostly by cation-pi interactions, this structure shows that the amidinium moiety is a useful candidate to elicit a carboxylate in an antibody combining site at a predetermined location with respect to the hapten. More generally, this structure highlights the advantage of a bidentate hapten for the programmed positioning of a chemically reactive residue in an antibody through charge complementarity to the hapten.

Mechanism of an antibody-catalysed allylic isomerization.

The catalytic antibody 4B2, which was generated against a substituted amidine 1, catalyses the allylic isomerization of beta, gamma-unsaturated ketones with an acceleration factor (k(cat)/k(uncat)) of 1.5x10(3). On the basis of the 'bait and switch' strategy, it was reasoned that the positively charged hapten could elicit, by charge complementarity, an acidic residue (Asp or Glu) in the antibody-binding site in the right position to catalyse this proton transfer reaction. The pH dependence curve of k(cat)/K(m) shows a bell-shaped feature with an optimum at approx. pH 4.5. By cloning and sequencing the light and heavy chains of the 4B2 antibody, we confirmed the presence of several Asp and Glu residues in the complementarity-determining region loops. The antibody catalyses the alpha-proton exchange on the same substrates, demonstrating the involvement of a dienol intermediate in the reaction mechanism. Kinetic studies with (2)H-NMR provide evidence that alpha-proton abstraction is stereospecific. Whether the process involves one or two acid/base residues in this simple proton transfer or whether it is a concerted mechanism is discussed.

Micromachined resonant temperature sensors: theoretical and experimental results.

Describes the study of quartz temperature sensors based on new bulk acoustic wave microresonators operating in thickness modes. First, we compare the thermal sensitivity and the electromechanical coupling coefficients of singly or doubly rotated cuts. These investigations allow us to select some cuts with both a good thermal sensitivity and piezoelectric characteristics. In the second part, emphasis is placed on the micromachining of resonators suspended by four bridges. These two theoretical considerations lead to the choice of three cuts. Experimental measurements are then presented. The temperature-frequency characteristics of the resonators are measured over the range 20 to 100 degrees C. Motional resistances and Q factors are determined at room temperature.

Micromachining of quartz plates: determination of a database by combined stereographic analysis and 3-D simulation of etching shapes.

Simple microstructures such as mesa and membrane are micromachined on singly and doubly rotated quartz plates. A stereographic analysis of 3-D etching shapes is used to recognize rhombohedral faces limiting micromachined structures. An iterative procedure is used to adjust the database in which first we generate extrema in the dissolution slowness for these faces, and second, we compare step-by-step theoretical 3-D etching shapes with experimental structures. Theoretical shapes for microstructures are found to be in satisfactory agreement with experiments.

Profile of the musculoskeletal pain suffered by textile tufting workers handling thread cones according to work, age and employment duration.

This paper studies the relationship between some socio-professional characteristics of workers (e.g. age, actual work done, experience in the job, overall time on job market, height) carrying out thread-cone handling tasks and their musculoskeletal pain profiles. Interviews were carried out with foremen and workers, and task analysis was performed. Self-administered questionnaires on work-related pain were filled out by 114 machine operators and creelers in four carpet-manufacturing companies. Statistics on nine individual and 25 pain-symptom characteristics were compiled. For data treatment, two statistical methods complemented one another: the Factorial Analysis of Correspondence (FAC) and the Hierarchical Ascendant Classification (HAC). Four classes of workers showing large differences one to another regarding time of employment on the job market, age, seniority in the company, and job experience were portrayed. No remarkable differences were found between the classes in relation to the proportion of workers showing body pain symptoms; it is greater than 50% in all classes (12 months). Moreover, in all classes, for several workers, the first region of pain (out of three possible) was reported as one persisting over the weekend. The group of workers from 25 to 35 years of age appears to be the one most seriously affected by musculoskeletal pain.

NMR, molecular modeling, and crystallographic studies of lentil lectin-sucrose interaction.

The conformational features of sucrose in the combining site of lentil lectin have been characterized through elucidation of a crystalline complex at 1.9-A resolution, transferred nuclear Overhauser effect experiments performed at 600 Mhz, and molecular modeling. In the crystal, the lentil lectin dimer binds one sucrose molecule per monomer. The locations of 229 water molecules have been identified. NMR experiments have provided 11 transferred NOEs. In parallel, the docking study and conformational analysis of sucrose in the combining site of lentil lectin indicate that three different conformations can be accommodated. Of these, the orientation with lowest energy is identical with the one observed in the crystalline complex and provides good agreement with the observed transferred NOEs. These structural investigations indicate that the bound sucrose has a unique conformation for the glycosidic linkage, close to the one observed in crystalline sucrose, whereas the fructofuranose ring remains relatively flexible and does not exhibit any strong interaction with the protein. Major differences in the hydrogen bonding network of sucrose are found. None of the two inter-residue hydrogen bonds in crystalline sucrose are conserved in the complex with the lectin. Instead, a water molecule bridges hydroxyl groups O2-g and O3-f of sucrose.

Modulation by dimethyl sulphoxide of the toxicity induced by cis-diamminedichloroplatinum in cultured thyrocytes.

Cis-diamminedichloroplatinum II (cisplatin) is an effective and widely used antitumour drug which induces many side-effects. We investigated the potency of dimethyl sulphoxide (DMSO) as an antidote to cisplatin-induced damage to cultured human thyrocytes. The effects of DMSO were studied on human thyroid cells cultured in monolayers and in follicles. Addition of 2% DMSO suppressed the cisplatin-induced decrease in intracellular lactate dehydrogenase, alpha-hydroxybutyryldehydrogenase and creatine kinase activities and in neutral red uptake. DMSO also partially or totally abolished the inhibition of cyclic AMP (cAMP), thyroglobulin (Tg) and triiodothyronine (T(3)) production observed with 7.5, 15 or 30 mum-cisplatin. In the presence of 75 mum-cisplatin, cAMP, Tg and T(3) secretion was always totally inhibited. The increase in thyroxine levels observed with cisplatin administration did not occur in the presence of DMSO. These data indicate that 2% DMSO interacted with cisplatin and therefore prevented the cytotoxic and inhibitory effects of cisplatin on cultured thyroid cells.

Effects of cisplatin on human thyrocytes in monolayer or follicle culture.

Cis-diamminedichloroplatinum (II) (cisplatin) is a widely used anticancer drug which induces many side-effects, but its action on the thyroid gland is still unknown. We have investigated the effects of this drug on human thyrocytes cultured in monolayers or in follicles and stimulated with 200 microU TSH/ml. After 72 h in culture, different concentrations of cisplatin (15, 30 and 75 microM) caused partial or total inhibition of cyclic AMP (cAMP), thyroglobulin (Tg) and tri-iodothyronine (T3) production, whereas thyroxine levels increased in the medium of thyrocytes cultured as follicles. Small doses of the drug did not affect thyrocyte production. Decreases in neutral-red uptake by thyroid cells and in intracellular lactate dehydrogenase, alpha-hydroxybutyryldehydrogenase and creatine phosphokinase activities were induced by 30 and 75 microM cisplatin. These data show that high concentrations of cisplatin had a cytotoxic effect on thyrocytes. Cisplatin also induced inhibition of the production of cAMP, Tg and T3.

Diastereotopic covalent binding of the natural inhibitor leupeptin to trypsin: detection of two interconverting hemiacetals by solution and solid-state NMR spectroscopy.

The naturally occurring peptidyl protease inhibitor leupeptin (N-acetyl-L-leucyl-L-leucyl-L-argininal) has been prepared labeled with 13C at the argininal carbonyl. 13C chemical shift data for the trypsin-leupeptin inhibitor complex in the pH range 3.0-7.6 reveal the presence of two pH-dependent covalent complexes, suggestive of two interconverting diastereomers at the new asymmetric tetrahedral center created by covalent addition of Ser195 to either side of the 13C-enriched aldehyde of the inhibitor. At pH 7 two signals are observable at delta 98.8 and delta 97.2 (84:16 ratio), while at pH 3.0 the latter signal predominates. In the selective proton 13C-edited NOE spectrum of the major diastereomer at pH 7.4, a strong NOE is observed between the hemiacetal proton of the inhibitor and the C2 proton of His57 of the enzyme, thus defining the stereochemistry of the high pH complex to the S configuration in which the hemiacetal oxygen resides in the oxyanion hole. pH titration studies further indicate that the 13C chemical shift of the S diastereomer follows a titration curve with a pKa of 4.69, the magnitude of which is consistent with direct titration of the hemiacetal oxygen. Similar pH-dependent chemical shifts were obtained by using CPMAS 13C NMR, providing evidence for the existence of the same diastereomeric equilibrium in the solid state.

Modulation of the functional properties of human thyrocytes in monolayer or follicle culture: effects of some anaesthetic drugs.

It is well known that some volatile anaesthetic drugs, such as halothane and isoflurane, alter the functions of the human thyroid gland, but the action of other anaesthetic drugs, such as thiopental, midazolam and ketamine, on thyroid function is still unknown. We have investigated the effects of these three drugs on the functional properties of human thyrocytes cultured in monolayers or follicles and stimulated by TSH. Thiopental, midazolam and ketamine induced total suppression or a partial reduction, depending on the dose administered, of cyclic AMP (cAMP), follicular thyroglobulin (Tg) and free tri-iodothyronine (FT3) production. In contrast, free thyroxine levels increased in the medium of thyrocytes cultured as follicles. Small doses of the drugs did not affect thyrocyte production. The inhibiting effect of thiopental, midazolam and ketamine on Tg and FT3 production seems to result from the inhibition of cAMP production and 5'-deiodinase.

Training in handling: an evaluative study.

Training programmes in handling have been one of the most used means of preventing work-related backache. The effectiveness of these programmes has not yet been demonstrated however, since they have rarely been evaluated. The purpose of this study was precisely to assess one of these programmes in the hospital sector which specifically dealt with the handling of patients. A field study was conducted and handling methods used by 32 trained orderlies were characterized using an observational grid developed and validated specifically to describe patient handling operations. The extent to which these orderlies used the handling methods taught was determined, and the observed deviations from these methods were characterized and interpreted. The results show that the handling principles taught (working with a straight back using the legs) were not frequently used in the workplace. Furthermore, the use of training was closely related to the type of handling carried out. In handling operations in which the effort includes a horizontal component (mainly those carried out in bed) the training was hardly used, while in vertical handling operations the taught principles were more frequently used. These results suggest that actual training is not well adapted to the handling of patients, particularly to horizontal handling carried out in the bed. Two main deficiencies in the actual programmes are pointed out. First, it is shown that, for reasons of physical constraints, training could not always be applied; second, the rationale of the taught principles could also be questioned, particularly the emphasis given to the use of the legs.

Calcium and spermine interaction with phospholipid bilayers: a 15N NMR study.

The binding of Ca2+ to membrane models composed of diplamitoylphosphatidylcholine (DPPC) and dipalmitoylphosphatidylethanolamine (DPPE) 15N-labeled in the polar head group was investigated at pH 8.5 and pH 9.4 by 15N-NMR spectroscopy. Both phospholipids exhibit a decrease in the chemical shift anisotropy, indicating changes of the order parameter of the C-N bond and decrease in half-height width. Binding of Ca2+ induces a chemical shift change for the DPPE signal which indicates a decrease in the pKa of the amino group. The binding of spermine was also investigated for mixed phase (DPPC/DPPE) at pH 8 and pH 9.4; a decrease in the DPPE pKa was also noted. The signals of both phospholipids are broadened and the line shapes are more complex because of the lower mobility and the higher steric bulk of this molecule. The results show the value of 15N-NMR in the study of mixed liposomes and indicate that the deprotonation of membrane surface could constitute a necessary step for fusion processes.

Magnetic resonance imaging studies on nude mice grafted with colorectal adenocarcinoma using gadolinium-labeled monoclonal antibody.

A monoclonal antibody (Ab) 19.9 specific for colorectal carcinoma was labeled with a high number of gadolinium (Gd) atoms for its potential application as a contrast agent in magnetic resonance imaging (MRI). The DTPA was conjugated to 19.9 Ab via the bicyclic DTPA anhydride method (c. DTPA) using c. DTPA/Ab molar ratios between 5 and 150. The aggregates present in great amount at high c. DTPA/Ab ratios were systematically removed. Then the exact number of DTPA effectively conjugated, the immunoreactivity of the resulting 111In-DTPA-Ab were measured. The number of DTPA conjugated per antibody can be increased 20 to 25 with only a little loss of immunoreactivity. The 19.9 antibody conjugated with 16 and 25 DTPA was labeled with 153GdCl3 for pharmacokinetic studies on xenografted nude mice and with nonradioactive gadolinium to measure ex vivo the effect on the relaxation time T1 of the tumor. We found a 15 to 20% decrease of T1 on the tumor. In vivo experiments using a Bruker system and the same animal model showed a difference in the tumor contrast after the injection of 2 mg of Gd-labeled Ab.

Gd-25 DTPA-MAb, a potential NMR contrast agent for MRI in the xenografted nude mouse: preliminary studies.

Monoclonal antibodies (MAbs) 19-9 and 73-3 specific for human colon adenocarcinoma were labelled with a high number of gadolinium atoms. Twenty five DTPA were chelated per MAb, with only slight loss of immunoreactivity. The NMR contrast agent Gd-25 DTPA-MAb 19-9 or 73-3 ([Gd] 17 mumole/kg, [MAb] 60 microM) was injected into nude mice bearing human colon adenocarcinoma (SW948). Tumours were removed 24 hr after injection and T1 was measured in vitro. T1 relaxation time varied according to MAb specificity against tumour targets; T1 decreased 20% for MAb 19-9 and MAb 73-3 with SW948 tumour. Imaging was performed with this model. Very good contrast was obtained 24 hr after Gd-25 DTPA-MAb injection.

Interactions between biliary lipid micelles and intestinal brush border membranes investigated by 1H and 31P nuclear magnetic resonance.

The effect of taurocholate and lecithin-cholesterol-taurocholate mixed micelles on the structure of isolated intestinal brush border membranes was investigated by nuclear magnetic resonance (NMR). Rabbit brush border membranes isolated by a Mg2+ precipitation step were chosen for this study because of their stability and integrity as revealed by 31P NMR. Incubation of taurocholate with the brush border membranes does not induce significant solubilization of these membranes even when the taurocholate/phospholipid ratio reaches 3.0. 1H NMR studies indicate that taurocholate is included in the membrane bilayer at low concentration (3 mM). However this biliary salt produces a size diminution of the vesicles when its concentration increases. Incorporation of lecithin or lecithin-cholesterol in micelles of taurocholate and subsequent incubation with brush border membranes lead simultaneously to a decrease in the 31P NMR isotropic/bilayer line ratio, and to an increase in delta sigma. These results indicate a protective effect of these compounds against lytic damage of taurocholate. Furthermore the equilibrium distribution of lecithin between mixed micelles and the membrane bilayer is strongly in favour of complete integration of micellar components in the bilayer. These data suggest that uptake of lipids from the micellar phase by isolated brush border membranes involves an interaction of the micelles with membranes followed by a fusion process.

Molecular order, dynamics, and ionization state of phosphatidylethanolamine bilayers as studied by 15N NMR.

Dipalmitoylphosphatidylethanolamine (DPPE) and dipalmitoylphosphatidylcholine (DPPC), 15N-labeled in the polar head group, were synthesized. The proton-decoupled 15N spectra of DPPC and DPPE in aqueous dispersion have exactly the form anticipated for powder line shapes governed by an axially symmetric shielding tensor. The chemical shift anisotropy (delta sigma) of DPPC is lower than 0.4 ppm at 30 degrees C and vanished when the temperature or the half-height line width is increased; DPPE always exhibits an asymmetric line shape, and 15N NMR spectra of DPPE are obtained at various temperatures and simulated to measure exactly the chemical shift anisotropy. At each temperature, the order parameter of the C-N bond segment is derivated from delta sigma and reveals that the average orientation of the C-N bond around the axis of rotation is near the "magic angle" (54.7 degrees). Isotropic correlation times are derived from T1, which are higher than values obtained for phosphatidylcholine by other nuclei. Arrhenius plots of T1 and T2 allowed us to calculate the activation energy for the motion of the DPPE and the DPPC C-N bond. The value of this activation energy for the DPPE (53 kJ/mol) is higher than the one found for the DPPC C-N bond (32 kJ/mol). These differences agree with the capacity of the ethanolamine head groups to bind noncovalently to their neighbors in the plane of the membrane surface. A direct titration curve of the amino group is achieved by the variation of the chemical shift with the bulk pH, and the interfacial pKa is calculated to be 11.1.(ABSTRACT TRUNCATED AT 250 WORDS)