Effectiveness, safety and quality-of-life effects of guselkumab and ustekinumab in patients with psoriasis: Week 104 results from the non-interventional, prospective, German multicentre PERSIST study.

BACKGROUND: PERSIST was a prospective, non-interventional, real-world study of guselkumab and ustekinumab in adult patients with moderate-to-severe plaque psoriasis in Germany. OBJECTIVES: To examine effectiveness, safety and quality-of-life (QoL) outcomes to Week (W) 104 of treatment with guselkumab and ustekinumab in patients with moderate-to-severe plaque psoriasis. METHODS: Patients (≥18 years of age) received guselkumab or ustekinumab as per routine clinical practice. Outcomes to W104 were examined separately in guselkumab and ustekinumab recipients. An ad hoc exploratory analysis of outcomes with guselkumab versus ustekinumab was also performed following propensity score matching. RESULTS: Overall, 302 and 313 patients received guselkumab and ustekinumab, respectively. Patients in both cohorts experienced improvements in disease activity and QoL that were maintained to W104, with 64.7% and 63.6% of guselkumab- and 54.6% and 64.4% of ustekinumab-treated patients achieving a Psoriasis Area and Severity Index (PASI) 90 response and a Dermatology Life Quality Index (DLQI) 0/1 score, respectively. Propensity score matching yielded well-balanced baseline characteristics except for prior biologic use, which was higher in guselkumab versus ustekinumab recipients (51.7% vs. 32.0%). Achievement of PASI ≤1 at W104 was more common in guselkumab versus ustekinumab recipients (58.7% vs. 49.7%). The W104 PASI90 response rate was 65.6% with guselkumab and 56.0% with ustekinumab; corresponding rates for PASI100 were 44.3% and 28.5%. In guselkumab recipients, response rates were higher in biologic-naïve versus biologic-experienced patients (PASI90, 77.1% vs. 53.4%; PASI100, 55.0% vs. 33.0%). A high level of response for QoL outcomes was observed for both treatments. CONCLUSIONS: Ustekinumab and guselkumab led to improvements in physician-assessed and patient-reported outcomes that were sustained for up to 2 years, with no new safety signals identified. Following propensity score matching, greater improvements in PASI outcomes were observed with guselkumab versus ustekinumab. Improvements with guselkumab were highest in biologic-naïve patients, highlighting the value of early treatment.

GABA (γ-Aminobutyric Acid) Uptake Via the GABA Permease GabP Represses Virulence Gene Expression in Pseudomonas syringae pv. tomato DC3000.

The nonprotein amino acid γ-aminobutyric acid (GABA) is the most abundant amino acid in the tomato (Solanum lycopersicum) leaf apoplast and is synthesized by Arabidopsis thaliana in response to infection by the bacterial pathogen Pseudomonas syringae pv. tomato DC3000 (hereafter called DC3000). High levels of exogenous GABA have previously been shown to repress the expression of the type III secretion system (T3SS) in DC3000, resulting in reduced elicitation of the hypersensitive response (HR) in the nonhost plant tobacco (Nicotiana tabacum). This study demonstrates that the GABA permease GabP provides the primary mechanism for GABA uptake by DC3000 and that the gabP deletion mutant ΔgabP is insensitive to GABA-mediated repression of T3SS expression. ΔgabP displayed an enhanced ability to elicit the HR in young tobacco leaves and in tobacco plants engineered to produce increased levels of GABA, which supports the hypothesis that GABA uptake via GabP acts to regulate T3SS expression in planta. The observation that P. syringae can be rendered insensitive to GABA through loss of gabP but that gabP is retained by this bacterium suggests that GabP is important for DC3000 in a natural setting, either for nutrition or as a mechanism for regulating gene expression. [Formula: see text] Copyright © 2016 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

[Body contouring]. / Körperkonturierung.

In our contemporary postmodern society, a modified perception of the human body is accompanied by an increasing demand for body shaping procedures. The treatment needs to be effective but it is just as important that they are safe and can be easily integrated into the daily working and routine schedule. While the options for minimally invasive volume addition are largely limited to injectable implants based on hyaluronic acid or autologous fat, a multitude of options are available for volume reduction. Before deciding on the method of choice, the following needs to be considered: which indications need to be treated, the extent of the reduction in volume and how much pain and possible undesired reactions the patient is prepared to accept.

Tracheostomy, ventilation and anterior cervical surgery: Timing and Complications.

BACKGROUND: Cervical spine injured patients often require prolonged ventilatory support due to intercostal paralysis and recurrent chest infections. This may necessitate tracheotomy. Concern exists around increased complications when anterior cervical spine surgery and tracheotomies are performed. OBJECTIVE: The primary aim of this study was to evaluate the effect of tracheostomy in anterior cervical surgery patients in term of complications. In addition, the aetiology of trauma and incidence of anterior surgery and ventilation in this patient group was assessed. METHOD: Patients undergoing anterior cervical surgery and requiring ventilation were identified from the unit's prospectively maintained database. These patients were further sub-divided into whether they had a tracheotomy or not. The aetiology of injury and incidence of complications were noted both from the database and a case note review. RESULTS: Of the 1829 admissions over an 8.5 year period, 444 underwent anterior cervical surgery. Of the 112 that required ventilation, 72 underwent tracheotomy. Motor vehicle accidents, followed by falls, were the most frequent cause of injury. There was a bimodal incidence of tracheostomy insertion, the day of spine surgery and 6-8 days later. There was no difference in the general complication rate between the two groups. With regards to specific complications attributable to the surgical approach/tracheotomy, there was no statistically significant difference. The timing of the tracheotomy also had no effect on complication rate. Although the complications occurred mostly in the formal insertion group as opposed to the percutaneous insertion group, this was most likely due to selection bias. CONCLUSION: Anterior cervical surgery and subsequent tracheostomy are safe despite the intuitive concerns. Timing does not affect the incidence of complications and there is no reason to delay the insertion of the tracheostomy. Ventilation in general is associated with increased complications rather than the tracheostomy tube per se.

Strategies for investigating the plant metabolic network with steady-state metabolic flux analysis: lessons from an Arabidopsis cell culture and other systems.

Steady-state (13)C metabolic flux analysis (MFA) is currently the experimental method of choice for generating flux maps of the compartmented network of primary metabolism in heterotrophic and mixotrophic plant tissues. While statistically robust protocols for the application of steady-state MFA to plant tissues have been developed by several research groups, the implementation of the method is still far from routine. The effort required to produce a flux map is more than justified by the information that it contains about the metabolic phenotype of the system, but it remains the case that steady-state MFA is both analytically and computationally demanding. This article provides an overview of principles that underpin the implementation of steady-state MFA, focusing on the definition of the metabolic network responsible for redistribution of the label, experimental considerations relating to data collection, the modelling process that allows a set of metabolic fluxes to be deduced from the labelling data, and the interpretation of flux maps. The article draws on published studies of Arabidopsis cell cultures and other systems, including developing oilseeds, with the aim of providing practical guidance and strategies for handling the issues that arise when applying steady-state MFA to the complex metabolic networks encountered in plants.

Pathways and fluxes: exploring the plant metabolic network.

The transition from a pathway-centred view of plant metabolism to a network-wide perspective is still incomplete. Further progress in this direction requires tools to facilitate the structural description of the network on the basis of fully annotated genomes, techniques for modelling the properties of the network, and experimental methods for constraining the models and verifying their outputs. It also requires a focus on metabolic flux as the key to understanding the regulation of metabolic activity and the relationship between the inputs and outputs of the network. Progress is being made on several fronts and this Special Issue on 'Pathways and fluxes: exploring the plant metabolic network' describes current developments in the genomic reconstruction of metabolic networks, the application of flux-balance analysis to such networks, kinetic modelling, and both steady-state-and non-steady state isotope-based measurements of multiple fluxes in the network of central carbon metabolism. The papers also highlight insights that can be obtained from pathway analysis, particularly in relation to the thermodynamic and kinetic efficiency of the predicted and observed flux distributions.

Response of cytoplasmic pH to anoxia in plant tissues with altered activities of fermentation enzymes: application of methyl phosphonate as an NMR pH probe.

BACKGROUND AND AIMS: Acidification of the cytoplasm is a commonly observed response to oxygen deprivation in plant tissues that are intolerant of anoxia. The response was monitored in plant tissues with altered levels of lactate dehydrogenase (LDH) and pyruvate decarboxylase (PDC) with the aim of assessing the contribution of the targeted enzymes to cytoplasmic pH (pH(cyt)) regulation. METHODS: The pH(cyt) was measured by in vivo (31)P nuclear magnetic resonance (NMR) spectroscopy using methyl phosphonate (MeP) as a pH probe. The potential toxicity of MeP was investigated by analysing its effect on the metabolism of radiolabelled glucose. KEY RESULTS: MeP accumulated to detectable levels in the cytoplasm and vacuole of plant tissues exposed to millimolar concentrations of MeP, and the pH-dependent (31)P NMR signals provided a convenient method for measuring pH(cyt) values in tissues with poorly defined signals from the cytoplasmic inorganic phosphate pool. Pretreatment of potato (Solanum tuberosum) tuber slices with 5 mm MeP for 24 h did not affect the metabolism of [U-(14)C]glucose or the pattern of (14)CO(2) release from specifically labelled [(14)C]-substrates. Time-courses of pH(cyt) measured before, during and after an anoxic episode in potato tuber tissues with reduced activities of LDH, or in tobacco (Nicotiana tabacum) leaves with increased activities of PDC, were indistinguishable from their respective controls. CONCLUSIONS: MeP can be used as a low toxicity (31)P NMR probe for measuring intracellular pH values in plant tissues with altered levels of fermentation enzymes. The measurements on transgenic tobacco leaves suggest that the changes in pH(cyt) during an anoxic episode are not dominated by fermentation processes; while the pH changes in the potato tuber tissue with reduced LDH activity show that the affected isozymes do not influence the anoxic pH response.

Influence of Elevated Fructose-2,6-Bisphosphate Levels on Starch Mobilization in Transgenic Tobacco Leaves in the Dark.

The aim of this work was to study the effect of elevated fructose-2,6-bisphosphate (Fru-2,6-bisP) levels on carbohydrate metabolism in leaves in the dark. In transgenic tobacco (Nicotiana tabacum L.) lines containing mammalian 6-phosphofructo-2-kinase activity there is an inverse relationship between the level of Fru-2,6-bisP in leaves and the rate of starch breakdown in the dark. Estimates of the flux response coefficient for the rate of net starch degradation with respect to changes in Fru-2,6-bisP level are -0.57 for whole leaves and -0.69 to -0.89 for excised leaf discs. We suggest that this decrease in the net rate of starch breakdown is caused, at least in part, by stimulation of unidirectional starch synthesis. Measurements of the levels of metabolic intermediates and the metabolism of [U-14C]glucose indicate that the stimulation of starch synthesis in the dark is a result of high Fru-2,6-bisP levels, increasing the 3-phosphoglycerate:inorganic phosphate ratio in leaves. We argue that the observed response to changes in the level of Fru-2,6-bisP are effected through activation of pyrophosphate:fructose-6-phosphate 1-phosphotransferase. However, the extent to which changes in Fru-2,6-bisP influence starch metabolism in wild-type plants is not known.

Essential Arginyl Residue at the Active Site of Pyrophosphate:Fructose 6-Phosphate 1-Phosphotransferase from Potato (Solanum tuberosum) Tuber.

The aim of this work was to test the proposal that the active site of pyrophosphate:fructose 6-phosphate 1-phosphotransferase (PFP) contains an essential arginyl residue. Enzyme activity was inhibited equally in the glycolytic and gluconeogenic directions by arginine-modifying reagents. The second-order rate constants for 2,3-butanedione and phenylglyoxal were 13.1 [plus or minus] 0.45 and 55.3 [plus or minus] 1.3 M-1 min-1, respectively. The corresponding values for the kinetic order of inactivation by these modifying reagents were 0.84 [plus or minus] 0.049 for 2,3-butanedione and 0.89 [plus or minus] 0.052 for phenylglyoxal. The substrates, fructose 6-phosphate and pyrophosphate, and a range of substrate analogs protected the enzyme from inactivation by 2,3-butanedione. These data suggest that modification of no more than one arginyl residue at, or close to, the active site is required to inhibit the enzyme. This result supports the proposal that the active site of PFP in plants is equivalent to that of the bacterial ATP-phosphofructokinase (S.M. Carlisle, S.D. Blakeley, S.M. Hemmingsen, S.J. Trevanion, T. Hiyoshi, N.J. Kruger, and D.T. Dennis [1990] J Biol Chem 265: 18366-18371).

Strategies for metabolic flux analysis in plants using isotope labelling.

Flux measurements through metabolic pathways generate insights into the integration of metabolism, and there is increasing interest in using such measurements to quantify the metabolic effects of mutation and genetic manipulation. Isotope labelling provides a powerful approach for measuring metabolic fluxes, and it gives rise to several distinct methods based on either dynamic or steady-state experiments. We discuss the application of these methods to photosynthetic and non-photosynthetic plant tissues, and we illustrate the different approaches with an analysis of the pathways interconverting hexose phosphates and triose phosphates. The complicating effects of the pentose phosphate pathway and the problems arising from the extensive compartmentation of plant cell metabolism are considered. The non-trivial nature of the analysis is emphasised by reference to invalid deductions in earlier work. It is concluded that steady-state isotopic labelling experiments can provide important information on the fluxes through primary metabolism in plants, and that the combination of stable isotope labelling with detection by nuclear magnetic resonance is particularly informative.

Errors and artifacts in coupled spectrophotometric assays of enzyme activity.

This paper considers problems associated with the accurate determination of enzyme activity using coupled spectrophotometric assays. Criteria for establishing optimum assay conditions and ensuring that the coupled assay accurately reflects enzyme activity are presented. The susceptibility of such assays to interference is illustrated by five instances in which contamination of specific assay components has produced misleading estimates of phosphofructokinase and pyrophosphate:fructose 6-phosphate phosphotransferase activity. Such artifacts have resulted in publication of spurious biochemical and physiological conclusions. These examples suggest that problems associated with contaminants are likely to be widespread in coupled spectrophotometric assays, and are likely to confound interpretation of the measurements. Strategies for identifying artifacts resulting from contaminants in coupled assays are proposed.

The bradford method for protein quantitation.

A rapid and accurate method for the estimation of protein concentration is essential in many fields of protein study. The Lowry method ( Chapter 1 in vol. 1 of this series) has been widely used, but is susceptible to interference from a wide range of compounds commonly present in biological extracts. Although interference can be avoided by trichloracetic acid precipitation of the protein prior to assay, this lengthens the procedure.

Purification of immunoglobulins using protein a-sepharose.

Most immunochemical techniques used in molecular biology rely on a specific class of antibodies, immunoglobulin G (IgG). In mammals, IgG antibodies are produced during the secondary humoral response and contribute about 80% of the serum immunoglobulin. Factors affecting the experimental production of antibodies have been discussed in detail (1). In this chapter we describe a method for the rapid and efficient purification of IgG to near homogeneity from rabbit serum. This is achieved by affinity chromatography using protein A.

Immunodetection of proteins on "western" blots using (125)i-labeled protein a.

Immunoblotting provides a simple and effective method for identifying specific antigens in a complex mixture of proteins. Initially, the constituent polypeptides are separated using SDS-polyacrylamide gel electrophoresis, or a similar technique, and then are transferred either electrophoretically or by diffusion onto a nitrocellulose filter. The transfer of proteins from gels onto membranes is discussed in vol. 1 of this series and Chapters 28 and 29 , this volume. Once immobilized on a sheet of nitrocellulose, specific polypeptides can be identified using antibodies that bind to antigens retained on the filter. In this chapter we describe conditions appropriate for binding antibodies to immobilized proteins and a method for locating these antibody-antigen complexes using (125)I-labeled protein A. These methods are based on those of Burnett (1) and Towbin et al. (2).

Improving the quality of health services organization structure by reengineering: circular design and clinical case impact in an academic medical center.

Innovation to improve the quality of structure and process in health care organization is reported in this case example of change in an academic medical center. Interactive planning and the circular organization design concept were the driving principles and methods. This report presents the needs for and initial obstructions to change, planning and project design work, a description of the change process, and illustrative accomplishments to date--two cases, one of conscious sedation policy and one of nuisance pages. Evaluative criteria for judging the progress and lessons of the project regarding key design characteristics also are included.

[Physicians, internet and new knowledge. Utilization and efficiency of new continuing education media in the hospital]. / Arzte, Internet und neues Wissen. Nutzung und Effizienz von neuen Weiterbildungsmedien im Krankenhaus.

BACKGROUND AND OBJECTIVES: The application of new information technologies will have a lasting impact on the physicians' working environment. Not only will classic learning methods change, but also ways of communication and strategies to find relevant information. This survey is designed to demonstrate current methods of and demand for continuing medical education and communication. METHODS: 110 participating physicians (26% female, 74% male) in nine different regional hospitals were asked regarding "continuing medical education", "communication" and "use of computerized technology". RESULTS: Most physicians regard their colleagues, scientific journals, textbooks and scientific meetings as most useful. The internet, scientific staff at research institutions and pharmaceutical representatives were valued as less useful. Physicians regarded scientific basis, speed and user-friendliness as the most important features of potential data and knowledge sources. Case specificity, German language and interactivity were viewed as less important. Most physicians expressed a growing need for education regarding use of e-mail as a communication tool and internet research techniques. CONCLUSION: Continuing medical education is in strong demand. The successful introduction of modern information technologies into physicians' daily work may and should be enhanced by specifically tailored training courses.

Evaluating a critical care internship program.

The authors describe the evaluation of their Critical Care Internship Program. The program was evaluated by job knowledge and clinical performance of graduate nurses compared to nurses with past critical care or medical-surgical experience.

Quo vadis? - Monitoring Campylobacter in Germany.

Campylobacter is a poorly recognized foodborne pathogen, leading the statistics of bacterially caused human diarrhoea in Europe during the last years. In this review, we present qualitative and quantitative German data obtained in the framework of specific monitoring programs and from routine surveillance. These also comprise recent data on antimicrobial resistances of food isolates. Due to the considerable reduction of in vitro growth capabilities of stressed bacteria, there is a clear discrepancy between the detection limit of Campylobacter by cultivation and its infection potential. Moreover, antimicrobial resistances of Campylobacter isolates established during fattening of livestock are alarming, since they constitute an additional threat to human health. The European Food Safety Authority (EFSA) discusses the establishment of a quantitative limit for Campylobacter contamination of broiler carcasses in order to achieve an appropriate level of protection for consumers. Currently, a considerable amount of German broiler carcasses would not comply with this future criterion. We recommend Campylobacter reduction strategies to be focussed on the prevention of fecal contamination during slaughter. Decontamination is only a sparse option, since the reduction efficiency is low and its success depends on the initial contamination concentration.