In vitro and in vivo detection of microbial gene expression in bioactivated scaffolds seeded with cyanobacteria.

Dermal replacement materials bioactivated with cyanobacteria have shown promising potential for wound regeneration. To date, extraction of cyanobacteria RNA from seeded scaffolds has not been described. The aim of this study was to develop a method to isolate total RNA from bioactivated scaffolds and to propose a new approach in determining living bacteria based on real-time PCR. Transgenic Synechococcus sp. PCC 7002 (tSyn7002) were seeded in liquid cultures or scaffolds for dermal regeneration in vitro and in vivo for 7 days. RNA was extracted with a 260/280 ratio of ≥2. The small subunit of the 30S ribosome in prokaryotes (16S) and RNAse P protein (rnpA) were validated as reference transcripts for PCR analysis. Gene expression patterns differed in vitro and in vivo. Expression of 16S was significantly upregulated in scaffolds in vitro, as compared to liquid cultures, whilst rnpA expression was comparable. In vivo, both 16S and rnpA showed reduced expression compared to in vitro (16S: in vivo Ct value 13.21 ± 0.32, in vitro 12.44 ± 0.42; rnpA in vivo Ct value 19.87 ± 0.41, in vitro 17.75 ± 1.41). Overall, the results demonstrate rnpA and 16S expression after 7 days of implantation in vitro and in vivo, proving the presence of living bacteria embedded in scaffolds using qPCR.

Centromedial amygdala is more relevant for phobic confrontation relative to the bed nucleus of stria terminalis in patients with spider phobia.

Recent studies indicate differential involvement of the centromedial amygdala (CM) and the bed nucleus of the stria terminalis (BNST) during processing (anticipation and confrontation) of threat stimuli. Here, temporal predictability was shown to be a relevant factor. In this study, we want to investigate the relevance of these effects, which were found in healthy subjects, for anxiety disorders. Therefore, we investigated the differential involvement of CM and BNST in the anticipation and confrontation of phobic stimuli under variation of temporal predictability in spider phobia. 21 patients with spider phobia and 21 healthy controls underwent a temporally predictable/unpredictable phobic and neutral anticipation and confrontation paradigm using functional magnetic resonance imaging (fMRI) and ROI analyses. During the anticipation phase, healthy controls showed higher CM and BNST activity during the predictable compared with the unpredictable condition compared with the anxiety patients. During a confrontation phase that followed the anticipation phase, CM was more activated than BNST during the phobic compared with the neutral confrontation. While this effect was independent of threat predictability in patients, healthy controls showed higher activation in the CM compared with the BNST only during the predictable spider confrontation compared with the predictable bird confrontation. The results contribute to a better understanding of the separate roles of the CM and BNST during phobic processes. The CM was found to be more relevant to phobic confrontation in patients with spider phobia compared with the BNST.

BNST and amygdala activation to threat: Effects of temporal predictability and threat mode.

Recent animal and human studies highlight the uncertainty about the onset of an aversive event as a crucial factor for the involvement of the centromedial amygdala (CM) and bed nucleus of the stria terminalis (BNST) activity. However, studies investigating temporally predictable or unpredictable threat anticipation and confrontation processes are rare. Furthermore, the few existing fMRI studies analyzing temporally predictable and unpredictable threat processes used small sample sizes or limited fMRI paradigms. Therefore, we measured functional brain activity in 109 predominantly female healthy participants during a temporally predictable-unpredictable threat paradigm, which aimed to solve limited aspects of recent studies. Results showed higher BNST activity compared to the CM during the cue indicating that the upcoming confrontation is aversive relative to the cue indicating an upcoming neutral confrontation. Both the CM and BNST showed higher activity during the confrontation with unpredictable and aversive stimuli, but the reaction to aversive confrontation relative to neutral confrontation was stronger in the CM compared to the BNST. Additional modulation analyses by NPSR1 rs324981 genotype revealed higher BNST activity relative to the CM in unpredictable anticipation relative to predictable anticipation in T-carriers compared to AA carriers. Our results indicate that during the confrontation with aversive or neutral stimuli, temporal unpredictability modulates CM and BNST activity. Further, there is a differential activity concerning threat processing, as BNST is more involved when focussing on fear-related anticipation processes and CM is more involved when focussing on threat confrontation.

Thermophysical properties of liquid Zr52.5Cu17.9Ni14.6Al10Ti5-prospects for bulk metallic glass manufacturing in space.

Bulk metallic glasses are of critical interest for a wide range of applications, including their use in spacecraft gearboxes and mechanisms due to their excellent low-temperature, unlubricated wear resistance. Also of interest, is the potential for in-space manufacturing of metal alloys and the use of microgravity to determine fundamental thermophysical properties to inform ground-based modeling and experimentation. In this work, a Zr-based bulk metallic glass was processed in the electromagnetic levitator ISS-EML to determine undercooling, electrical resistivity, specific heat capacity, surface tension, and viscosity. A 6.5 mm sphere was vitrified during the processing, resulting in the first bulk metallic glass manufactured on board the international space station (ISS).

Neural correlates of trauma-related single word processing in posttraumatic stress disorder.

Interpersonal violence (IPV) is one of the most frequent causes for the development of posttraumatic stress disorder (PTSD) in women. One key component in PTSD is altered processing of trauma-related cues, leading to emotional symptoms. In the everyday environment, words with trauma-associated semantic content represent typical, albeit abstract, trauma-related stimuli for patients suffering from PTSD. However, the functional neuroanatomy associated with processing single trauma-related words in IPV-PTSD is not understood. The present event-related functional magnetic resonance imaging study investigated the neural basis of trauma-related word processing in women with IPV-PTSD relative to healthy controls (HC) during a non-emotional vigilance task in which the emotional content of the words was task-irrelevant. On the behavioral level, trauma-related relative to neutral word stimuli evoked more unpleasant feelings, higher arousal as well as anxiety in IPV-PTSD patients as compared to HC. Functional imaging data showed hyperactivation to trauma-related versus neutral words in the basolateral amygdala (BLA) and cortical language-processing regions (inferior frontal gyrus, posterior cingulate cortex, angular/supramarginal gyrus) in IPV-PTSD compared to HC. These results propose a role of the BLA in hypervigilant responding to verbal trauma associated cues in IPV-PTSD. Furthermore, the particular involvement of cortical language-processing regions indicates enhanced processing of trauma-related words in brain regions associated with analysis and memory of verbal material. Taken together, our findings suggest that both subcortical and cortical mechanisms contribute to automatic responsivity to verbal trauma cues in PTSD.

Three-dimensional imaging of shear bands in bulk metallic glass composites.

The mechanism of the increase in ductility in bulk metallic glass matrix composites over monolithic bulk metallic glasses is to date little understood, primarily because the interplay between dislocations in the crystalline phase and shear bands in the glass could neither be imaged nor modelled in a validated way. To overcome this roadblock, we show that shear bands can be imaged in three dimensions by atom probe tomography from density variations in the reconstructed atomic density, which density-functional theory suggests being a local-work function effect. Imaging of near-interface shear bands in Ti48 Zr20 V12 Cu5 Be15 bulk metallic glass matrix composite permits measurement of their composition, thickness, branching and interactions with the dendrite interface. These results confirm that shear bands here nucleate from stress concentrations in the glass due to intense, localized plastic deformation in the dendrites rather than intrinsic structural inhomogeneities.

Radical formation at the gallium nitride nanowire-electrolyte interface by photoactivated charge transfer.

We investigated the transfer of photogenerated charge carriers from GaN nanowires into a surrounding electrolyte by electron paramagnetic resonance (EPR) and fluorescence spectroscopy. Using 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) as a spin trap we find that the formation of hydroxyl radicals dominates in acidic, neutral and moderately basic environments, while in an electrolyte with a pH of 13.5 the superoxide formation becomes detectable. We explain the two processes considering the redox potentials for radical formation in the electrolyte as well as the positions of the conduction and valence bands. The role of surface band bending and surface states in the semiconductor is discussed.

Self-interaction correction in a real-time Kohn-Sham scheme: access to difficult excitations in time-dependent density functional theory.

We present a real-time Kohn-Sham propagation scheme for the self-interaction correction (SIC). The multiplicative Kohn-Sham potential is constructed in real-time and real-space based on the generalized optimized effective potential equation. We demonstrate that this approach yields promising results for a wide range of test systems, including hydrogen terminated silicon clusters, conjugated molecular chains, and molecular charge-transfer systems. We analyze the nature of excitations by calculating transition densities from the time evolution and by evaluating the time-dependent exchange-correlation potential. A properly constructed Kohn-Sham SIC potential shows a time-dependent field-counteracting behavior. These favorable characteristics of the exchange-correlation potential may be lost in approximations such as the SIC-Slater potential.

Kohn-Sham self-interaction correction in real time.

We present a solution scheme for the time-dependent Kohn-Sham self-interaction correction. Based on the generalized optimized effective potential approach, the multiplicative Kohn-Sham potential is constructed in real time and real space for the self-interaction corrected local density approximation. Excitations of different character, including charge-transfer excitations that had been regarded as prime examples for the failure of standard time-dependent density functionals, are described correctly by this approach. We analyze the time-dependent exchange-correlation potential and density, revealing features that are decisive for the correct description of the response.

[Experience in contrast vision testing]. / Erfahrungen mit der Untersuchung des Kontrastsehens.

INTRODUCTION: Since July 2011 driving licensing in Germany requires contrast vision to be tested. Recommendations are available for testing under mesopic conditions only. This study is aimed to provide recommendations for photopic testing. METHOD: 169 persons were tested both under mesopic (Mesotest) and photopic (Optovist) conditions. RESULTS: With the Mesotest the setting without glare and the requirement to recognize contrast 1:5 or with glare contrast 1:23 proved to be suitable. With the Optovist visual acuity size 0.4 without and 0.25 with glare proved to be adequate, if the requirement is to recognize the limit of 5 %. DISCUSSION: Not unexpectedly, there is no perfect congruence between the tests. We therefore recommend that the screening methods used in industrial medicine need to be sufficiently sensitive and specific, unfortunately this is met only by some of the conditions supplied with the instruments.

A powerful computational crystallography method to study ice polymorphism.

Classical molecular dynamics (MD) simulations are employed as a tool to investigate structural properties of ice crystals under several temperature and pressure conditions. All ice crystal phases are analyzed by means of a computational protocol based on a clustering approach following standard MD simulations. The MD simulations are performed by using a recently published classical interaction potential for oxygen and hydrogen in bulk water, derived from neutron scattering data, able to successfully describe complex phenomena such as proton hopping and bond formation/breaking. The present study demonstrates the ability of the interaction potential model to well describe most ice structures found in the phase diagram of water and to estimate the relative stability of 16 known phases through a cluster analysis of simulated powder diagrams of polymorphs obtained from MD simulations. The proposed computational protocol is suited for automated crystal structure identification.

[Traumatic rupture of the brachial artery due to a closed elbow joint dislocation]. / Traumatische Ruptur der A. brachialis durch eine geschlossene Ellenbogengelenkluxation.

A rare complication of a brachial artery rupture as a result of a closed elbow dislocation is presented. If vascular damage is suspected angiography must be carried out and if signs of ischemia are present on the affected arm an immediate operational intervention with vascular reconstruction is necessary. The problems associated with this type of injury are discussed on the basis of the current literature.

Electrochemistry-mass spectrometry for mechanistic studies and simulation of oxidation processes in the environment.

Electrochemistry (EC) coupled to mass spectrometry (MS) has already been successfully applied to metabolism research for pharmaceutical applications, especially for the oxidation behaviour of drug substances. Xenobiotics (chemicals in the environment) also undergo various conversions; some of which are oxidative reactions. Therefore, EC-MS might be a suitable tool for the investigation of oxidative behaviour of xenobiotics. A further evaluation of this approach to environmental research is presented in the present paper using sulfonamide antibiotics. The results with sulfadiazine showed that EC-MS is a powerful tool for the elucidation of the oxidative degradation mechanism within a short time period. In addition, it was demonstrated that EC-MS can be used as a fast and easy method to model the chemical binding of xenobiotics to soil. The reaction of sulfadiazine with catechol, as a model substance for organic matter in soil, led to the expected chemical structure. Finally, by using EC-MS a first indication was obtained of the persistence of a component under chemical oxidation conditions for the comparison of the oxidative stability of different classes of xenobiotics. Overall, using just a few examples, the study demonstrates that EC-MS can be applied as a versatile tool for mechanistic studies of oxidative degradation pathways of xenobiotics and their possible interaction with soil organic matter as well as their oxidative stability in the environment. Further studies are needed to evaluate the full range of possibilities of the application of EC-MS in environmental research.

Effectiveness of budesonide nebulising suspension compared to disodium cromoglycate in early childhood asthma.

OBJECTIVE: The optimal treatment for early childhood asthma remains controversial. Budesonide (BUD) has shown superiority over placebo in infants, and over disodium cromoglycate (DSCG) in children aged > 2 years. The aim of this double-blind, randomised, parallel-group study was to compare the effectiveness of nebulised BUD and DSCG in asthmatic children aged < 36 months. RESEARCH DESIGN AND METHODS: 82 infants (mean age 18.0 months [range, 11.6-31.2 months]) with suspected asthma (three exacerbations of dyspnoea and wheezing during the past 12 months, with one or more exacerbations in the past 3 months) were treated for 3 months with nebulised BUD (Pulmicort Respules) 0.5 mg/2 mL bid or DSCG 20 mg/2 mL tid. Follow-up was at 6 months. MAIN OUTCOME MEASURES AND RESULTS: Patients treated with BUD had a lower exacerbation rate than DSCG-treated patients after 3 months of treatment (5.4% vs. 31.7%; p = 0.003) and towards the end of follow-up (30% vs. 49%; p = 0.062). During treatment, days without cough were 80% and 65% for BUD and DSCG, respectively (p = 0.014), and nights without cough were 89% and 78%, respectively (p = 0.016). Side-effects were mild and of similar frequency in both groups. CONCLUSIONS: Inhaled nebulised BUD was well tolerated and more effective than nebulised DSCG in reducing the incidence of asthma exacerbations and days with symptoms. These beneficial effects of BUD were maintained throughout the 6-month follow-up.

Increasing cardiac output by fluid loading: effects on indocyanine green plasma disappearance rate and splanchnic microcirculation.

BACKGROUND: Sufficient cardiac pre-load for maintaining adequate cardiac output is a major goal in the treatment of critically ill patients. We studied the effects of increasing cardiac output by fluid loading on the indocyanine green plasma disappearance rate (ICG-PDR) and gastric mucosal regional CO2 tension (PRco2) as an indicator of splanchnic microcirculation. METHODS: With approval by our ethics committee and written consent, we studied post-operatively 12 patients (1 female, 11 males; 66 +/- 13 years) with elective coronary artery bypass grafting (n = 10) or aortic valve replacement (n = 2). All patients had received pulmonary artery and left atrial catheterization previously for clinical indications. Cardiac output and filling pressures were measured immediately after intensive care unit (ICU) admission and 1 h after the beginning of fluid loading. RESULTS: Overall, 630 +/- 130 ml of 6% hydroxyethylstarch (130 kDa) was infused with the splanchnic perfusion pressure remaining constant. Norepinephrine and epinephrine dosages were unchanged. The cardiac index increased significantly from 2.8 +/- 0.7 to 3.5 +/- 0.6 l/min/m2 and the stroke volume index from 30 +/- 7 to 38 +/- 8 ml/m2. ICG-PDR showed no significant change, i.e. from 21.2 +/- 6.5 to 21.6 +/- 6.5%/min. Gastric mucosal PRco2 and the Pco2 gap (difference between regional and end-tidal CO2 tension) were constant, i.e. changed from 5.1 +/- 0.8 to 5.5 +/- 1.1 kPa and from 0.9 +/- 0.5 to 1.0 +/- 0.7 kPa, respectively. CONCLUSION: Increasing cardiac output to supranormal values by fluid loading is not associated with a significant change in ICG-PDR or gastric mucosal PRco2.

A third blind test of crystal structure prediction.

Following the interest generated by two previous blind tests of crystal structure prediction (CSP1999 and CSP2001), a third such collaborative project (CSP2004) was hosted by the Cambridge Crystallographic Data Centre. A range of methodologies used in searching for and ranking the likelihood of predicted crystal structures is represented amongst the 18 participating research groups, although most are based on the global minimization of the lattice energy. Initially the participants were given molecular diagrams of three molecules and asked to submit three predictions for the most likely crystal structure of each. Unlike earlier blind tests, no restriction was placed on the possible space group of the target crystal structures. Furthermore, Z' = 2 structures were allowed. Part-way through the test, a partial structure report was discovered for one of the molecules, which could no longer be considered a blind test. Hence, a second molecule from the same category (small, rigid with common atom types) was offered to the participants as a replacement. Success rates within the three submitted predictions were lower than in the previous tests - there was only one successful prediction for any of the three ;blind' molecules. For the ;simplest' rigid molecule, this lack of success is partly due to the observed structure crystallizing with two molecules in the asymmetric unit. As in the 2001 blind test, there was no success in predicting the structure of the flexible molecule. The results highlight the necessity for better energy models, capable of simultaneously describing conformational and packing energies with high accuracy. There is also a need for improvements in search procedures for crystals with more than one independent molecule, as well as for molecules with conformational flexibility. These are necessary requirements for the prediction of possible thermodynamically favoured polymorphs. Which of these are actually realised is also influenced by as yet insufficiently understood processes of nucleation and crystal growth.

[Extended hemodynamic monitoring using transpulmonary thermodilution Influence of various factors on the accuracy of the estimation of intrathoracic blood volume and extravascular lung water in critically ill patients]. / Erweitertes hämodynamisches Monitoring mithilfe der transpulmonalen Thermodilution. Einfluss verschiedener Faktoren auf die Genauigkeit der Abschätzung von intrathorakalem Blutvolumen und extravaskulärem Lungenwasser bei kritisch kranken Patienten.

BACKGROUND: The transpulmonary thermal-dye dilution (TDD) is the clinical gold standard for measurement of intrathoracic blood volume (ITBV) and extravascular lung water (EVLW). Recently, experimental and clinical studies found that ITBV and EVLW can be derived reliably by single transpulmonary thermodilution (TD), however, dependency from various factors of lung function has been discussed. PATIENTS AND METHODS: We retrospectively analyzed data from 174 critically ill patients who had undergone hemodynamic monitoring by TDD. We calculated the differences of simultaneous TD and TDD determined by ITBV or EVLW measurements. Bias and standard deviations (SD) were calculated and extended Bland-Altman regression analyses were performed. Association of the differences and the potential confounders gender, age, PEEP, p(a)O(2)/F(I)O(2) ratio and the corresponding TDD measurement were analyzed by multiple linear regression and LOWESS regression. RESULTS: Overall results (bias+/-SD) were 1+/-58 ml/m(2) for ITBV and 0.0+/-1.4 ml/kg bw for EVLW, but TD overestimated TDD results in patients with higher ITBV and underestimated those for lower ITBV values. The deviations were small, and the dependence of ITBV could be explained completely by dependence on PEEP and age. However, low values of EVLW are overestimated by TD. CONCLUSION: Single transpulmonary thermodilution for estimation of ITBV or EVLW is reliable in patients with severe lung injury for normal and higher values.

The distribution of the unoccupied volume in glassy polymers.

The aim of this paper is to present an improved method to describe the unoccupied volume in glassy polymers. The method is able to treat atoms with non-spherical symmetry. The fineness of the raster points scanning the unit cell can be as small as 0.01 nm. This method was used to determine the unoccupied volume of molecular dynamic simulations of poly(amide imide) unit cells by probing it with a tracer atom. Since the reachable unoccupied volume strongly depends on the tracer radius, we used tracer radii between 0.03 and 0.17 nm. The poly(amide imide)s were used in the present work because they are well characterized, especially there already exist free volume data determined by positron lifetime experiments.

Sample preparation techniques for the determination of natural 15N/14N variations in amino acids by gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS).

In order to identify natural nitrogen isotope variations of biologically important amino acids four derivatization reactions (t-butylmethylsilylation, esterification with subsequent trifluoroacetylation, acetylation and pivaloylation) were tested with standard mixtures of 17 proteinogenic amino acids and plant (moss) samples using GC-C-IRMS. The possible fractionation of the nitrogen isotopes, caused for instance by the formation of multiple reaction products, was investigated. For biological samples, the esterification of the amino acids with subsequent trifluoroacetylation is recommended for nitrogen isotope ratio analysis. A sample preparation technique is described for the isotope ratio mass spectrometric analysis of amino acids from the non-protein (NPN) fraction of terrestrial moss. 14N/15N ratios from moss (Scleropodium spec.) samples from different anthropogenically polluted areas were studied with respect to ecotoxicologal bioindication.

Efficacy of dry extract of ivy leaves in children with bronchial asthma--a review of randomized controlled trials.

OBJECTIVES: To investigate if extracts from dried ivy leaves (Hedera helix L.) are effective in the treatment of chronic airway obstruction in children suffering from bronchial asthma. DESIGN: Systematic review of trials documented in the literature with re-analysis of original data. TRIALS: 5 randomized controlled trials investigating the efficacy of ivy leaf extract preparations in chronic bronchitis, 3 of which were conducted in children and met our selection criteria. One compared ivy leaf extract cough drops to placebo, one compared suppositories to drops and one tested syrup against drops. MAIN OUTCOME MEASURES: Body-plethysmographic and spirometric measures. RESULTS: Drops were significantly superior to placebo in reducing airway resistance (primary outcome measure; p = 0.04 two-sided) and descriptively superior in all other 'objective' measures. For syrup and suppositories, at least 54%, resp. 35% of the effect against placebo were preserved. CONCLUSIONS: The trials included in this review indicate that ivy leaf extract preparations have effects with respect to an improvement of respiratory functions of children with chronic bronchial asthma, but more far-reaching conclusions can hardly be drawn because of a meagre database, including the fact that only one primary trial included a placebo control. Further research, particularly into the long-term efficacy of the herbal extract, is needed.