First Observation of Self-Amplified Spontaneous Emission in a Free-Electron Laser at 109 nm Wavelength

We present the first observation of Self-Amplified Spontaneous Emission (SASE) in a free-electron laser (FEL) in the Vacuum Ultraviolet regime at 109 nm wavelength (11 eV). The observed free-electron laser gain (approx. 3000) and the radiation characteristics, such as dependency on bunch charge, angular distribution, spectral width and intensity fluctuations all corroborate the existing models for SASE FELs.Comment: 6 pages including 6 figures; e-mail: [email protected]

An AFM study of solid-phase bilayers of unsaturated PC lipids and the lateral distribution of the transmembrane model peptide WALP23 in these bilayers

An altered lipid packing can have a large influence on the properties of the membrane and the lateral distribution of proteins and/or peptides that are associated with the bilayer. Here, it is shown by contact-mode atomic force microscopy that the surface topography of solid-phase bilayers of PC lipids with an unsaturated cis bond in their acyl chains shows surfaces with a large number of line-type packing defects, in contrast to the much smoother surfaces observed for saturated PC lipids. Di-n:1-PC (n = 20, 22, 24) and (16:0,18:1)-PC (POPC) were used. Next, the influence of an altered lipid environment on the lateral distribution of the single α-helical model peptide WALP23 was studied by incorporating the peptide in the bilayers of di-n:1-PC (n = 20, 22, 24) and (16:0,18:1)-PC unsaturated lipids. The presence of WALP23 leads to an increase in the number of packing defects but does not lead to the formation of the striated domains that were previously observed in bilayers of saturated PC lipids and WALP. This is ascribed to the less efficient lateral lipid packing of the unsaturated lipids, while the increase in packing defects is probably an indirect effect of the peptide. Finally, the fact that an altered lipid packing affects the distribution of WALP23 is also confirmed in an additional experiment where the solvent TFE (2,2,2-trifluorethanol) is added to bilayers of di-16:0-PC/WALP23. At 3.5 vol% TFE, the previous striated ordering of the peptide is abolished and replaced by loose lines

Point Mutations in Aβ Result in the Formation of Distinct Polymorphic Aggregates in the Presence of Lipid Bilayers

A hallmark of Alzheimer's disease (AD) is the rearrangement of the β-amyloid (Aβ) peptide to a non-native conformation that promotes the formation of toxic, nanoscale aggregates. Recent studies have pointed to the role of sample preparation in creating polymorphic fibrillar species. One of many potential pathways for Aβ toxicity may be modulation of lipid membrane function on cellular surfaces. There are several mutations clustered around the central hydrophobic core of Aβ near the α-secretase cleavage site (E22G Arctic mutation, E22K Italian mutation, D23N Iowa mutation, and A21G Flemish mutation). These point mutations are associated with hereditary diseases ranging from almost pure cerebral amyloid angiopathy (CAA) to typical Alzheimer's disease pathology with plaques and tangles. We investigated how these point mutations alter Aβ aggregation in the presence of supported lipid membranes comprised of total brain lipid extract. Brain lipid extract bilayers were used as a physiologically relevant model of a neuronal cell surface. Intact lipid bilayers were exposed to predominantly monomeric preparations of Wild Type or different mutant forms of Aβ, and atomic force microscopy was used to monitor aggregate formation and morphology as well as bilayer integrity over a 12 hour period. The goal of this study was to determine how point mutations in Aβ, which alter peptide charge and hydrophobic character, influence interactions between Aβ and the lipid surface. While fibril morphology did not appear to be significantly altered when mutants were prepped similarly and incubated under free solution conditions, aggregation in the lipid membranes resulted in a variety of polymorphic aggregates in a mutation dependent manner. The mutant peptides also had a variable ability to disrupt bilayer integrity

Self-consistent field theory for the interactions between keratin intermediate filaments

Background: Keratins are important structural proteins found in skin, hair and nails. Keratin Intermediate Filaments are major components of corneocytes, nonviable horny cells of the Stratum Corneum, the outermost layer of skin. It is considered that interactions between unstructured domains of Keratin Intermediate Filaments are the key factor in maintaining the elasticity of the skin. Results: We have developed a model for the interactions between keratin intermediate filaments based on self-consistent field theory. The intermediate filaments are represented by charged surfaces, and the disordered terminal domains of the keratins are represented by charged heteropolymers grafted to these surfaces. We estimate the system is close to a charge compensation point where the heteropolymer grafting density is matched to the surface charge density. Using a protein model with amino acid resolution for the terminal domains, we find that the terminal chains can mediate a weak attraction between the keratin surfaces. The origin of the attraction is a combination of bridging and electrostatics. The attraction disappears when the system moves away from the charge compensation point, or when excess small ions and/or NMF-representing free amino acids are added. Conclusions: These results are in concordance with experimental observations, and support the idea that the interaction between keratin filaments, and ultimately in part the elastic properties of the keratin-containing tissue, is controlled by a combination of the physico-chemical properties of the disordered terminal domains and the composition of the medium in the inter-filament region. Keywords: Stratum corneum, Skin keratins, Intermediate filaments, Unstructured terminal domains, Bridging attractio

Molecular dynamics simulations reveal that AEDANS is an inert fluorescent probe for the study of membrane proteins

Computer simulations were carried out of a number of AEDANS-labeled single cysteine mutants of a small reference membrane protein, M13 major coat protein, covering 60% of its primary sequence. M13 major coat protein is a single membrane-spanning, α-helical membrane protein with a relatively large water-exposed region in the N-terminus. In 10-ns molecular dynamics simulations, we analyze the behavior of the AEDANS label and the native tryptophan, which were used as acceptor and donor in previous FRET experiments. The results indicate that AEDANS is a relatively inert environmental probe that can move unhindered through the lipid membrane when attached to a membrane protein

Patient outcomes with positive pressure versus spontaneous ventilation in non-paralysed adults with the laryngeal mask

Purpose: To compare patient outcomes for positive pressure ventilation (PPV) and spontaneous ventilation (SV) in non-paralysed patients with the LMA using either isoflurane or sevoflurane anaesthesia. Methods: One hundred and sixty-four adult patients were studied. Anaesthesia was with fentanyl-propofol and NO 66% in O with 0.75 MAC isoflurane or sevoflurane and either PPV or SV. Positive pressure ventilation was with tidal volumes of 6-8 ml·kg. Peak airway pressures were < 15 cm HO. Patients were evaluated for airway problems, cardiorespiratory effects, and anaesthesia emergence times. Results: There were no failed episodes of PPV or SV. Gastric insufflation was not detected by epigastric auscultation. Airway problems and cardiovascular effects were similar among groups. During maintenance: SpO2 was greater in the PPV group than in the SV group (98.4 vs 97%, P < 0.001); also, (P(ET)CO) (34 vs 43 mmHg) and the respiratory rate (RR) (15 vs 19 min) were higher and the minute ventilation(MV) (5.7 vs 7.2 L) were lower in the SV groups (P < 0.0001). Shorter times to LMA removal and orientation were observed in the sevoflurane groups (P < 0.0001). Conclusions: Patients outcome is similar for SV and PPV in non-paralysed adult patients with the LMA. Isoflurane and sevoflurane at 0.75 MAC provide suitable conditions for maintenance and emergence, but emergence is more rapid with sevoflurane

Direct measurement of mucosal pressures exerted by cuff and non-cuff portions of tracheal tubes with different cuff volumes and head and neck positions

We measured directly mucosal pressures against the cuff and non-cuff portions of the tracheal tube in different head-neck positions and tested the reliability of calculated mucosal pressures, in vivo intracuff pressures and cuff volume as determinants of directly measured mucosal pressures. We studied 10 anaesthetized, paralysed adult patients. An 8.5-mm, high volume, low pressure PVC tracheal tube was used. Microchip sensors were attached to three cuff locations (anterior, lateral and posterior) and two non-cuff locations (anterior tip and anterior aspect of the tube, 5 cm proximal to the cuff). Directly measured mucosal pressures, in vivo intracuff pressures and calculated mucosal pressures (in vivo minus in vitro intracuff pressures) were determined after brief inflation (< 15 s) to 0, 5, 10 and 15 ml. In vivo intracuff pressures were then set at 30 mm Hg and the measurements repeated, first in the neutral position and then with the head-neck extended, flexed and rotated. Cuff mucosal pressures were highest anteriorly and lowest posteriorly. Non-cuff mucosal pressures did not vary with cuff volume and were approximately 15 mm Hg. Compared with the neutral position, in vivo intracuff pressures were higher in the rotated, extended and flexed positions. Compared with the neutral position, mucosal pressure increased on the anterior aspect of the tube in the flexed position by 22 mm Hg (P = 0.003), at the anterior tip in the extended position by 11 mm Hg (P = 0.002) and at the anterior tip (5 mm Hg, P = 0.05) and lateral aspect of the cuff (5 mm Hg, P = 0.03) in the rotated position. In vivo intracuff pressures and calculated mucosal pressures were moderate predictors of measured mucosal pressures; cuff volume was a poor predictor. We conclude that tracheal mucosal pressures were highest anteriorly, that non-cuff portions of the tube exerted substantial mucosal pressures and that the rotated position caused a greater increase in tracheal mucosal pressure than the extended or flexed position. Indirect methods of measuring mucosal pressure were of moderate predictive value