Investigation of `glitches' in the energy spectrum induced by single-crystal diamond compound X-ray refractive lenses.

Single-crystal diamond stands out among all the candidate materials that could be exploited to fabricate compound refractive lenses (CRLs) owing to its extremely stable properties. Among all related experimental features, beam divergence, χ-angles relative to the incoming beam in Eulerian geometry and different positions of the X-ray beam relative to the lens geometry may influence the transmission energy spectrum of CRLs. In addition, the orientation of the single-crystal diamond sample may also affect the glitches significantly. To verify these initial assumptions, two experiments, an energy scan and an ω-scan, were set up by employing a polished diamond plate consisting of five biconcave lenses. The results show that beam divergence does not affect the spectrum, nor do χ-angles when ω is set to zero. Nevertheless, different incident positions have an appreciable effect on the transmission spectrum, in particular the `strengths' of the glitches. This is attributed to absorption. The ω-scan setup is capable of determining the so-called orientation matrix, which may be used to predict both `energy positions' and `strengths' of the glitches.

Cationic lipids bearing succinic-based, acyclic and macrocyclic hydrophobic domains: Synthetic studies and in vitro gene transfer.

In this communication we describe the construction of four succinic-based cationic lipids, their formulation with plasmid DNA (pDNA), and an evaluation of their in vitro gene delivery into Chinese hamster ovarian (CHO-K1) cells. The cationic lipids employed in this work possess either a dimethylamine or trimethylamine headgroup, and a macrocyclic or an acyclic hydrophobic domain composed of, or derived from two 16-atom, succinic-based acyl chains. The synthesized lipids and a co-lipid of neutral charge, either cholesterol or 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), were formulated in an overall 3:2 cationic-to-neutral lipid molar ratio, then complexed with plasmid DNA (pDNA). The relative transfection performance was evaluated via a comparison between matched versus mismatched formulations defined by the rigidity relationship between the lipids employed. Gel electrophoresis was used to characterize the binding of the lipid formulations with plasmid DNA and the relative degree of plasmid degradation using a DNase I degradation assay. Small angle X-ray diffraction (SAXD) was employed to characterize the packing morphology of the lipid-DNA complexes. In general, the succinic unit embedded within the hydrophobic domain of the cationic lipids was found to improve lipid hydration. The transfection assays revealed a general trend in which mismatched formulations that employed a rigid lipid combined with a non-rigid (or flexible) lipid, outperformed the matched formulations. The results from this work suggest that the design of the cationic lipid structure and the composition of the lipoplex formulation play key roles in governing the transfection performance of nonviral gene delivery agents.

Next generation macrocyclic and acyclic cationic lipids for gene transfer: Synthesis and in vitro evaluation.

Previously we reported the synthesis and in vitro evaluation of four novel, short-chain cationic lipid gene delivery vectors, characterized by acyclic or macrocyclic hydrophobic regions composed of, or derived from, two 7-carbon chains. Herein we describe a revised synthesis of an expanded library of related cationic lipids to include extended chain analogues, their formulation with plasmid DNA (pDNA) and in vitro delivery into Chinese hamster ovarian (CHO-K1) cells. The formulations were evaluated against each other based on structural differences in the hydrophobic domain and headgroup. Structurally the library is divided into four sets based on lipids derived from two 7- or two 11-carbon hydrophobic chains, C7 and C11 respectively, which possess either a dimethylamine or a trimethylamine derived headgroup. Each set includes four cationic lipids based on an acyclic or macrocyclic, saturated or unsaturated hydrophobic domain. All lipids were co-formulated with the commercial cationic lipid 1,2-dimyristoyl-sn-glycero-3-ethylphosphocholine (EPC) in a 1:1 molar ratio, along with one of two distinct neutral co-lipids, cholesterol or 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE) in an overall cationic-to-neutral lipid molar ratio of 3:2. Binding of lipid formulations with DNA, and packing morphology associated with the individual lipid-DNA complexes were characterized by gel electrophoresis and small angle X-ray diffraction (SAXD), respectively. As a general trend, lipoplex formulations based on mismatched binary cationic lipids, composed of a shorter C7 lipid and the longer lipid EPC (C14), were generally associated with higher transfection efficiency and lower cytotoxicity than their more closely matched C11/EPC binary lipid formulation counterparts. Furthermore, the cyclic lipids gave transfection levels as high as or greater than their acyclic counterparts, and formulations with cholesterol exhibited higher transfection and lower cytotoxicity than those formulated with DOPE. A number of the lipid formulations with cholesterol as co-lipid performed as well as, or better than Lipofectamine 2000™ and EPC, the two positive controls employed in these studies. These results suggest that our novel cyclic and acyclic cationic lipid vectors are effective nonviral gene transfer agents that warrant further investigation.

The atomic anisotropic displacement tensor ­ completing the picture.

A simplified approach for calculating the equivalent isotropic displacement parameter is presented and the transformation property of the tensor representation U to point-group operations is analysed. Complete tables have been compiled for the restrictions imposed upon the tensor owing to the site symmetry associated with all special positions as listed in Hahn [(2011), International Tables for Crystallography, Vol. A, Space-group Symmetry, 5th revised ed. Chichester: John Wiley and Sons, Ltd].

Novel cationic polyene glycol phospholipids as DNA transfer reagents--lack of a structure-activity relationship due to uncontrolled self-assembling processes.

Cationic glycol phospholipids were synthesized introducing chromophoric, rigid polyenoic C20:5 and C30:9 chains next to saturated flexible alkyl chains of variable lengths C6-20:0. Surface properties and liposome formation of the amphiphilic compounds were determined, the properties of liposome/DNA complexes (lipoplexes) were established using three formulations (no co-lipid, DOPE as a co-lipid, or cholesterol as a co-lipid), and the microstructure of the best transfecting compounds inspected using small angle X-ray diffraction to explore details of the partially ordered structures of the systems that constitute the series. Transfection and cytotoxicity of the lipoplexes were evaluated by DNA delivery to Chinese hamster ovary (CHO-K1) cells using the cationic glycerol phospholipid 1,2-dioleoyl-sn-glycero-3-ethylphosphocholine (EPC) as a reference compound. The uncontrollable self-association of the molecules in water resulted in aggregates and liposomes of quite different sizes without a structure-property relationship. Likewise, adding DNA to the liposomes gave rise to unpredictable sized lipoplexes, which, again, transfected without a structure-activity relationship. Nevertheless, one compound among the novel lipids (C30:9 chain paired with a C20:0 chain) exhibited comparable transfection efficiency and toxicity to the control cationic lipid EPC. Thus, the presence of a rigid polyene chain in this best performing achiral glycol lipid did not have an influence on transfection compared with the chiral glycerolipid reference ethyl phosphocholine EPC with two flexible saturated C14 chains.

The benefit of the European User Community from transnational access to national radiation facilities.

Transnational access (TNA) to national radiation sources is presently provided via programmes of the European Commission by BIOSTRUCT-X and CALIPSO with a major benefit for scientists from European countries. Entirely based on scientific merit, TNA allows all European scientists to realise synchrotron radiation experiments for addressing the Societal Challenges promoted in HORIZON2020. In addition, by TNA all European users directly take part in the development of the research infrastructure of facilities. The mutual interconnection of users and facilities is a strong prerequisite for future development of the research infrastructure of photon science. Taking into account the present programme structure of HORIZON2020, the European Synchrotron User Organization (ESUO) sees considerable dangers for the continuation of this successful collaboration in the future.

Aspects of nonviral gene therapy: correlation of molecular parameters with lipoplex structure and transfection efficacy in pyridinium-based cationic lipids.

This study seeks correlations between the molecular structures of cationic and neutral lipids, the lipid phase behavior of the mixed-lipid lipoplexes they form with plasmid DNA, and the transfection efficacy of the lipoplexes. Synthetic cationic pyridinium lipids were co-formulated (1:1) with the cationic lipid 1,2-dimyristoyl-sn-glycero-3-ethylphosphocholine (EPC), and these lipids were co-formulated (3:2) with the neutral lipids 1,2-dioleoyl-sn-glycero-3-phosphatidylethanolamine (DOPE) or cholesterol. All lipoplex formulations exhibited plasmid DNA binding and a level of protection from DNase I degradation. Composition-dependent transfection (beta-galactosidase and GFP) and cytotoxicity was observed in Chinese hamster ovarian-K1 cells. The most active formulations containing the pyridinium lipids were less cytotoxic but of comparable activity to a Lipofectamine 2000™ control. Molecular structure parameters and partition coefficients were calculated for all lipids using fragment additive methods. The derived shape parameter values correctly correlated with observed hexagonal lipid phase behavior of lipoplexes as derived from small-angle X-ray scattering experiments. A transfection index applicable to hexagonal phase lipoplexes derived from calculated parameters of the lipid mixture (partition coefficient, shape parameter, lipoplex packing) produced a direct correlation with transfection efficiency.

Three-beam resonant X-ray diffraction in germanium - Laue transmission cases.

Perturbation of the two-beam diffracted power owing to the influence of a third lattice node has been examined for various three-beam cases in a small finite germanium crystal in the vicinity of the K-absorption edge. Although the crystal was slightly imperfect, the main parts of the experimental results are very well described within the framework of the fundamental theory of X-ray diffraction in conjunction with Cromer-Liberman calculations for the resonant scattering terms. Beam divergence and dynamical block size are treated as adjustable parameters in the analysis. Observed changes in the three-beam profile asymmetry are mainly attributed to size and not to resonance effects associated with the triplet phase sum of the involved reflections. Close to the absorption edge there is however some evidence indicating that f' values should be reduced in magnitude compared to the tabulated ones.

Three-beam diffraction in an elastically strained crystal plate.

Based on the Takagi-Taupin equations, an analytical expression for the three-beam diffraction profile function in an elastically strained crystal plate is derived. The strain parameters appear in the shape functions governing both triplet-phase dependent and independent terms in the solution. Simulations show that three-beam profiles are significantly influenced. With increasing strain any phase information in the profiles is obscured and subsequently lost.

[Postoperative urinary retention]. / Postoperativ urinretensjon.

BACKGROUND: Retention of urine is a common postoperative problem. MATERIALS AND METHODS: We present two patients with postoperative urinary retention. Current opinions on monitoring and treatment are reviewed. RESULTS: One male postoperative patient was transferred from the postanaesthesia care unit after orthopaedic surgery under spinal anaesthesia. Shortly thereafter he collapsed. Hypotension and bradycardia were treated with intravenous ephedrine and atropine. Urinary retention was suspected and the bladder catheterised for 1,000 ml urine. A follow-up examination revealed no sequelae. A female patient was operated for haemorrhoids under epidural anaesthesia. The second postoperative day she was catheterised for 1,500 ml of urine. Two years later she had an ovarian cyst removed under general anaesthesia. Postoperative urinary retention developed with bladder overdistension (volume > 1,000 ml). A follow-up examination showed rest urine of 25 to 75 ml, and a chronic bladder dysfunction. The incidence of postoperative urinary retention varies depending upon type of surgery, gender, age, and preoperative history of urinary tract dysfunction. Method of anaesthesia or postoperative pain treatment is not correlated to incidence of urinary retention, but excessive intravenous volume load during the operation is. INTERPRETATION: Postoperative urinary retention should be monitored in all patients after surgery. All patients with history of difficulties with micturition need close supervision to avoid urinary bladder overdistention.

Three-beam X-ray diffraction in a semi-infinite perfect crystal.

The approach to multibeam X-ray diffraction by Shen [Acta Cryst. (1986). A42, 525-533] has been carefully re-examined for three-beam Laue-Laue diffraction in a perfect non-absorbing semi-infinite crystal. A modified version of Shen's theory is shown to be equivalent to the Takagi-Taupin formalism. The perturbation of the two-beam integrated power when a third beam is being excited is thus expressed by non-diverging terms. Both Aufhellung and Umweganregung terms are recovered.