What is the best support surface in prevention and treatment, as of 2012, for a patient at risk and/or suffering from pressure ulcer sore? Developing French guidelines for clinical practice.

INTRODUCTION: The use of support surfaces in the prevention and treatment of pressure ulcers prevention is an important part of care for a patient at risk and/or suffering from sore(s). OBJECTIVES: Define which support surfaces to use in prevention and treatment of at-risk and/or pressure sore patients. METHODOLOGY: A systematic review of the literature querying the several Pascal Biomed, PubMed and Cochrane Library databases from 2000 through 2010. RESULTS (GRADE A): In prevention, a structured foam mattress is more efficient than a standard hospital mattress. An alternating pressure mattress is more effective than a visco-elastic mattress limiting the occurrence heel pressure ulcers, but those that do occur are more serious. A low-air-loss bed is more efficient than a mixed pulsating air mattress in prevention of heel pressure ulcers. Some types of sheepskin can reduce sacral pressure ulcer incidence in orthopedic patients. Use of an overlay on an operating table limits the occurrence of peroperative and postoperative pressure ulcers. An air-fluidized bed improves pressure ulcer healing. DISCUSSION: The data in the literature are not always relevant and do not suffice to dictate a clinician's choices. We are compelled to recognize the methodological limitations of many studies, the lack of corporate interest in conducting such studies and the relatively small number of available trials. However, the effectiveness of some support surfaces reaches a sufficient level of evidence, especially when they are associated with postural, hydration and nutritional measures. CONCLUSION: Support surfaces are recommended in prevention and treatment of patients at risk and/or already suffering from pressure ulcer, and their use should constitute part of an overall preventive or curative strategy.

Membrane proteins in four acts: function precedes structure determination.

Studies on four membrane protein systems, which combine information derived from crystal structures and biophysical studies have emphasized, as a precursor to crystallization, demonstration of functional activity. These assays have relied on sensitive spectrophotometric, electrophysiological, and microbiological assays of activity to select purification procedures that lead to functional complexes and with greater likelihood to successful crystallization: (I), Hetero-oligomeric proteins involved in electron transport/proton translocation. (1) Crystal structures of the eight subunit hetero-oligomeric trans-membrane dimeric cytochrome b(6)f complex were obtained from cyanobacteria using a protocol that allowed an analysis of the structure and function of internal lipids at specific intra-membrane, intra-protein sites. Proteolysis and monomerization that inactivated the complex and prevented crystallization was minimized through the use of filamentous cyanobacterial strains that seem to have a different set of membrane-active proteases. (2) An NADPH-quinone oxido-reductase isolated from cyanobacteria contains an expanded set of 17 monotopic and polytopic hetero-subunits. (II) ß-Barrel outer membrane proteins (OMPs). High resolution structures of the vitamin B(12) binding protein, BtuB, solved in meso and in surfo, provide the best example of the differences in such structures that were anticipated in the first application of the lipid cubic phase to membrane proteins [1]. A structure of the complex of BtuB with the colicin E3 and E2 receptor binding domain established a "fishing pole" model for outer membrane receptor function in cellular import of nuclease colicins. (III) A modified faster purification procedure contributed to significantly improved resolution (1.83Å) of the universal porin, OmpF, the first membrane protein for which meaningful 3D crystals have been obtained [2]. A crystal structure of the N-terminal translocation domain of colicin E3 complexed to OmpF established the role of OmpF as an import channel for colicin nuclease cytotoxins. (IV) α-Synuclein, associated with the etiology of Parkinson's Disease, is an example of a protein, which is soluble and disordered in solution, but which can assume an ordered predominantly α-helical conformation upon binding to membranes. When subjected in its membrane-bound form to a trans-membrane electrical potential, α-synuclein can form voltage-gated ion channels. Summary of methods to assay functions/activities: (i) sensitive spectrophotometric assay to measure electron transfer activities; (ii) hydrophobic chromatography to deplete lipids, allowing reconstitution with specific lipids for studies on lipid-protein interactions; (iii) microbiological screen to assay high affinity binding of colicin receptor domains to Escherichia coli outer membrane receptors; (iv) electrophysiology/channel analysis (a) to select channel-occluding ligands for co-crystallization with ion channels of OmpF, and (b) to provide a unique description of voltage-gated ion channels of α-synuclein.

Fabrication of ZnS nanoparticle chains on a protein template.

In the present study, we have exploited the properties of a fibrillar protein for the template synthesis of zinc sulfide (ZnS) nanoparticle chains. The diameter of the ZnS nanoparticle chains was tuned in range of ~30 to ~165 nm by varying the process variables. The nanoparticle chains were characterized by field emission scanning electron microscopy, UV-Visible spectroscopy, transmission electron microscopy, electron energy loss spectroscopy, and high-resolution transmission electron microscopy. The effect of incubation temperature on the morphology of the nanoparticle chains was also studied.

Biotemplated synthesis of metallic nanoparticle chains on an alpha-synuclein fiber scaffold.

Biomolecular templates provide an excellent potential tool for bottom-up device fabrication. Self-assembling alpha-synuclein protein fibrils, the formation of which has been linked to Parkinson's disease, have yet to be explored for potential device fabrication. In this paper, alpha-synuclein fibrils were used as a template for palladium (Pd), gold (Au) and copper (Cu) nanoparticle chains synthesis. Deposition over a range of conditions resulted in metal-coated fibers with reproducible average diameters between 50 and 200 nm. Active elemental palladium deposited on the protein fibrils is used as a catalyst for the electroless deposition of Au and Cu. Nanoparticle chains were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray energy dispersive spectrometry (XEDS), and electron energy loss spectrometry (EELS).

Protein-templated semiconductor nanoparticle chains.

Cadmium sulfide and lead sulfide semiconducting nanoparticle chains have been fabricated for the first time by exploiting a general property of proteins, amyloidogenicity. The diameter of the CdS and PbS nanowires was tuned in the range of ∼50 to ∼350 nm by changing the process parameters. The nanoparticle chains were characterized by field emission scanning electron microscopy, UV-visible spectroscopy, transmission electron microscopy, electron energy loss spectroscopy and high-resolution transmission electron microscopy.

Heterotopic ossification of the elbow in patients with burns. Results after early excision.

Heterotopic ossification which may develop around the elbow in patients with burns may lead to severe functional impairment. We describe the outcome of early excision of such heterotopic ossification in 28 patients (35 elbows), undertaken as soon as the patient's general and local condition allowed. The mean age at operation was 42 years. The mean area of burnt body surface was 49%. The mean pre-operative range of movement was 22 degrees in flexion/extension and 94 degrees in pronation/supination. The mean time between the burn and operation was 12 months with the median being 9.5. The mean follow-up period was for 21 months. At the last review, the mean range of movement was 123 degrees in flexion/extension and 160 degrees in pronation/supination. Clinical evidence of recurrence was seen in four patients, occurring within the first two months after operation. Nevertheless, three of these elbows gained 60 degrees or more in flexion/extension and in pronation/supination. Based on this experience, we recommend early surgical treatment of heterotopic ossification of the elbow in patients with severe burns.

Production and detection of cold antihydrogen atoms.

A theoretical underpinning of the standard model of fundamental particles and interactions is CPT invariance, which requires that the laws of physics be invariant under the combined discrete operations of charge conjugation, parity and time reversal. Antimatter, the existence of which was predicted by Dirac, can be used to test the CPT theorem-experimental investigations involving comparisons of particles with antiparticles are numerous. Cold atoms and anti-atoms, such as hydrogen and antihydrogen, could form the basis of a new precise test, as CPT invariance implies that they must have the same spectrum. Observations of antihydrogen in small quantities and at high energies have been reported at the European Organization for Nuclear Research (CERN) and at Fermilab, but these experiments were not suited to precision comparison measurements. Here we demonstrate the production of antihydrogen atoms at very low energy by mixing trapped antiprotons and positrons in a cryogenic environment. The neutral anti-atoms have been detected directly when they escape the trap and annihilate, producing a characteristic signature in an imaging particle detector.

[Burns and rehabilitation]. / Brûlure et rééducation.

Necessary principles of rehabilitation for burn patient are based on empirical findings recently corroborated by discoveries about healing pathophysiology. Risks are assessable immediately from the extensive, depth and situation of the burns, problems appear only if the dermis is affected: retraction, hypertrophy and losses of substances. To cutaneous problems it is necessary to add those linked to the prolonged immobilization and to complications of the resuscitation. To be effective, re-education has to be precocious, continuously suited to cicatricial processing and to the different therapeutic steps: resuscitation, surgical treatment, processing in a re-education and rehabilitation center, steady at home and processing of the sequelae. The processing rests on the repressive cloth port 23/24 hours during more of a year, the port of orthesis of immobilization and segmental posture (to stretch the dermis permanently) and the mobilization of articulations to avoid their stiffening. The cooperation of the patient is essential, it needs the share of therapies as well as the totality of problems and difficulties met by the patient, that they are physical, psychological, social, family or occupational. The steady has to be insured by a pluridisciplinarity team during at least the two necessary years for the cicatricial maturation.

Kinetic stabilization of the alpha-synuclein protofibril by a dopamine-alpha-synuclein adduct.

The substantia nigra in Parkinson's disease (PD) is depleted of dopaminergic neurons and contains fibrillar Lewy bodies comprising primarily alpha-synuclein. We screened a library to identify drug-like molecules to probe the relation between neurodegeneration and alpha-synuclein fibrilization. All but one of 15 fibril inhibitors were catecholamines related to dopamine. The inhibitory activity of dopamine depended on its oxidative ligation to alpha-synuclein and was selective for the protofibril-to-fibril conversion, causing accumulation of the alpha-synuclein protofibril. Adduct formation provides an explanation for the dopaminergic selectivity of alpha-synuclein-associated neurotoxicity in PD and has implications for current and future PD therapeutic and diagnostic strategies.

Vesicle permeabilization by protofibrillar alpha-synuclein: implications for the pathogenesis and treatment of Parkinson's disease.

Fibrillar alpha-synuclein is a component of the Lewy body, the characteristic neuronal inclusion of the Parkinson's disease (PD) brain. Both alpha-synuclein mutations linked to autosomal dominant early-onset forms of PD promote the in vitro conversion of the natively unfolded protein into ordered prefibrillar oligomers, suggesting that these protofibrils, rather than the fibril itself, may induce cell death. We report here that protofibrils differ markedly from fibrils with respect to their interactions with synthetic membranes. Protofibrillar alpha-synuclein, in contrast to the monomeric and the fibrillar forms, binds synthetic vesicles very tightly via a beta-sheet-rich structure and transiently permeabilizes these vesicles. The destruction of vesicular membranes by protofibrillar alpha-synuclein was directly observed by atomic force microscopy. The possibility that the toxicity of alpha-synuclein fibrillization may derive from an oligomeric intermediate, rather than the fibril, has implications regarding the design of therapeutics for PD.

Pig heart CoA transferase exists as two oligomeric forms separated by a large kinetic barrier.

Pig heart CoA transferase (EC 2.8.3.5) has been shown previously to adopt a homodimeric structure, in which each subunit has a molecular weight of 52 197 and consists of N- and C-domains linked by a hydrophilic linker or "hinge". Here we identify and characterize a second oligomeric form constituent in purified enzyme preparations, albeit at low concentrations. Both species catalyze the transfer of CoA with similar values for k(cat) and K(M). This second form sediments more rapidly than the homodimer under the conditions of conventional sedimentation velocity and active enzyme centrifugation. Apparent molecular weight values determined by sedimentation equilibrium and gel filtration chromatography are 4-fold greater than the subunit molecular weight, confirming that this form is a homotetramer. The subunits of both oligomeric forms are indistinguishable with respect to molecular mass, far-UV CD, intrinsic tryptophan fluorescence, and equilibrium unfolding. Dissociation of the homotetramer to the homodimer occurs very slowly in benign solutions containing high salt concentrations (0.25-2.0 M KCl). The homotetramer is fully converted to homodimer during refolding from denaturant at low protein concentrations. Disruption of the hydrophilic linker between the N- and C-domains by mutagenesis or mild proteolysis causes a decrease in the relative amount of the larger conformer. The homotetramer is stabilized by interactions involving the helical hinge region, and a substantial kinetic barrier hinders interconversion of the two oligomeric species under nondenaturing conditions.

Inhibition of fibrillization and accumulation of prefibrillar oligomers in mixtures of human and mouse alpha-synuclein.

Parkinson's disease (PD) is a neurodegenerative disorder attributed to the loss of dopaminergic neurons from the substantia nigra. Some surviving neurons are characterized by cytoplasmic Lewy bodies, which contain fibrillar alpha-synuclein. Two mutants of human alpha-synuclein (A53T and A30P) have been linked to early-onset, familial PD. Oligomeric forms of these mutants accumulate more rapidly and/or persist for longer periods of time than oligomeric, human wild-type alpha-synuclein (WT), suggesting a link between oligomerization and cell death. The amino acid sequences of the mouse protein and WT differ at seven positions. Mouse alpha-synuclein, like A53T, contains a threonine residue at position 53. We have assessed the conformational properties and fibrillogenicity of the murine protein. Like WT and the two PD mutants, mouse alpha-synuclein adopts a "natively unfolded" or disordered structure. However, at elevated concentrations, the mouse protein forms amyloid fibrils more rapidly than WT, A53T, or A30P. The fibrillization of mouse alpha-synuclein is slowed by WT and A53T. Inhibition of fibrillization leads to the accumulation of nonfibrillar, potentially toxic oligomers. The results are relevant to the interpretation of the phenotypes of transgenic animal models of PD and suggest a novel approach for testing the cause and effect relationship between fibrillization and neurodegeneration.

The effect of progesterone alone or in combination with estradiol on follicular dynamics, gonadotropin profiles, and estrus in beef cows following calf isolation and restricted suckling.

The effects of calf isolation and restricted suckling on LH pulse characteristics and interval to first ovulation (postpartum interval) were studied in 52 multiparous beef cows, with or without exogenous progesterone. At 30 d postpartum, cows were randomly allocated to one of four treatments (n = 13/treatment): 1) Ad lib, ad libitum access of cows to calves; 2) CI/RS, calf isolation/restricted suckling, where suckling was restricted to once daily; 3) CI/RS+P4, same as CI/RS but cows received an intravaginal progesterone-releasing device at calf isolation for 6 d; or 4) CI/RS+P4+E2, as CI/RS+P4 but the intravaginal progesterone-releasing device had a 10-mg estradiol capsule attached. Daily ovarian scanning and twice-daily blood sampling were performed from d 25 postpartum until the day of second ovulation. A random sample of cows from each treatment (n = 31 in total) were blood-sampled at 15-min intervals for 10 h on d 29, 32, 35, and 38. Ovulatory response to treatment was regarded as ovulation of either the dominant follicle growing at d 30 or the subsequent DF. There was a treatment x day effect (P = .09) on LH pulse frequency, but neither progesterone (CI/ RS+P4) nor progesterone and estradiol (CI/RS+P4+E2) treatment suppressed the calf isolation/restricted suckling-induced increase in LH pulse frequency. The estradiol capsule (CI/RS+P4+E2) delivered sufficient estradiol to delay new follicle wave emergence (treatment x stage; P < .001) and the associated preemergence increase in concentrations of FSH (treatment, P < .05) in cows treated at the postselection stage of follicle wave development, prolonging dominance of the dominant follicle present at treatment initiation (P < .001). The number of cows that ovulated in response to treatment was greater (P < .001) in cows with calf isolation/restricted suckling than in cows suckled ad libitum. Hence, cows assigned to the Ad lib treatment had a longer postpartum interval (P < .001) than cows of the other treatments. Exogenous progesterone treatment increased the frequency of cows exhibiting clinical signs of estrus at first ovulation (P < .001) and reduced the frequency of short estrous cycles (P < .001). We conclude that, in beef cows with calves, a 6-d progesterone treatment does not suppress the calf isolation/restricted suckling-induced increase in LH pulse frequency. Hence, on progesterone withdrawal, the LH pulse frequency is sufficient to stimulate first ovulation, accompanied by overt estrous expression and elimination of a short estrous cycle in most cows.

Amyloid fibrillogenesis: themes and variations.

Recent progress has improved our knowledge of how proteins form amyloid fibrils. Both 'natively unfolded' and globular proteins have been shown to initiate fibrillization by adopting a partially structured conformation. Oligomeric prefibrillar intermediates have been extensively characterized with respect to their morphology and temporal evolution. Three-dimensional models obtained using biophysical and computational methods have provided information about fibril structure. All of these advances suggest common features of self-assembly pathways, with subtle variations accounting for differences among distinct amyloid fibrils.

Acceleration of oligomerization, not fibrillization, is a shared property of both alpha-synuclein mutations linked to early-onset Parkinson's disease: implications for pathogenesis and therapy.

The Parkinson's disease (PD) substantia nigra is characterized by the presence of Lewy bodies containing fibrillar alpha-synuclein. Early-onset PD has been linked to two point mutations in the gene that encodes alpha-synuclein, suggesting that disease may arise from accelerated fibrillization. However, the identity of the pathogenic species and its relationship to the alpha-synuclein fibril has not been elucidated. In this in vitro study, the rates of disappearance of monomeric alpha-synuclein and appearance of fibrillar alpha-synuclein were compared for the wild-type (WT) and two mutant proteins, as well as equimolar mixtures that may model the heterozygous PD patients. Whereas one of the mutant proteins (A53T) and an equimolar mixture of A53T and WT fibrillized more rapidly than WT alpha-synuclein, the other (A30P) and the corresponding equimolar mixture with WT fibrillized more slowly. However, under conditions that ultimately produced fibrils, the A30P monomer was consumed at a comparable rate or slightly more rapidly than the WT monomer, whereas A53T was consumed even more rapidly. The difference between these trends suggested the existence of nonfibrillar alpha-synuclein oligomers, some of which were separated from fibrillar and monomeric alpha-synuclein by sedimentation followed by gel-filtration chromatography. Spheres (range of heights: 2-6 nm), chains of spheres (protofibrils), and rings resembling circularized protofibrils (height: ca. 4 nm) were distinguished from fibrils (height: ca. 8 nm) by atomic force microscopy. Importantly, drug candidates that inhibit alpha-synuclein fibrillization but do not block its oligomerization could mimic the A30P mutation and thus may accelerate disease progression.

Productive interactions between the two domains of pig heart CoA transferase during folding and assembly.

The enzyme CoA transferase from porcine heart (EC 2.8.3.5) is a homodimer; each subunit consists of two domains linked by a hydrophilic "hinge" region. We have prepared separate DNA segments encoding each of these domains. Incorporation of these two DNA segments within an operon or within two separate transcription units does not preclude the synthesis and assembly of CoA transferase in Escherichia coli. When the two domain fragments are produced and purified individually from separate cultures and subsequently mixed, enzyme activity accumulates to near wild-type levels only after a lengthy incubation. Each domain is more susceptible to aggregation than wild-type CoA transferase. Circular dichroism shows that, prior to mixing, the domains possess a different secondary structural profile compared to their counterparts in the native enzyme. However, mixing and incubation of the domains produces a complex with far-UV CD, fluorescence, and ultracentrifugation properties similar to those of wild-type CoA transferase. Finally, we show that the intact hydrophilic peptide which links the two domains is essential for the recovery of activity observed upon refolding of the denatured enzyme in vitro. These results indicate that the folding and assembly of pig heart CoA transferase require a productive interaction between its two domains, involving a substantial conformational rearrangement.

Identification of glutamate 344 as the catalytic residue in the active site of pig heart CoA transferase.

The enzyme CoA transferase (succinyl-CoA:3-ketoacid coenzyme A transferase [3-oxoacid CoA transferase], EC 2.8.3.5) is essential for the metabolism of ketone bodies in the mammalian mitochondrion. It is known that its catalytic mechanism involves the transient thioesterification of an active-site glutamate residue by CoA. As a means of identifying this glutamate within the sequence, we have made use of a fortuitous autolytic fragmentation that occurs at the active site when the enzyme-CoA covalent intermediate is heated. The presence of protease inhibitors has no effect on the extent of cleavage detectable by SDS-PAGE, supporting the view that this fragmentation is indeed autolytic. This fragmentation can be carried out on intact CoA transferase, as well as on a proteolytically nicked but active form of the enzyme. Because the resulting C-terminal fragment is blocked at its N-terminus by a pyroglutamate moiety, it is not amenable to direct sequencing by the Edman degradation method. As an alternative, we have studied a peptide (peptide D) generated specifically by autolysis of the nicked enzyme and predicted to have an N-terminus corresponding to the site of proteolysis and a C-terminus determined by the site of autolysis. This peptide was purified by reversed-phase HPLC and subsequently characterized by electrospray mass spectrometry. We have obtained a mass value for peptide D, from which it can be deduced that glutamate 344, known to be conserved in all sequenced CoA transferases, is the catalytically active amino acid. This information should prove useful to future mutagenesis work aimed at better understanding the active-site structure and catalytic mechanism of CoA transferase.