In vivo analysis of staphylococcus aureus-infected mice reveals differential temporal and spatial expression patterns of fhuD2

Staphylococcus aureus is an opportunistic human pathogen and a major cause of invasive infections such as bacteremia, endocarditis, pneumonia and wound infections. FhuD2 is a staphylococcal lipoprotein involved in the uptake of iron-hydroxymate and is under the control of the iron uptake regulator Fur. The protein is part of an investigational multi-component vaccine formulation that has shown protective efficacy in several murine models of infection. Even though fhuD2 expression was shown to be upregulated in murine kidneys infected with S. aureus, it is unknown whether the bacterium undergoes increased iron deprivation during prolonged infection. Furthermore, different infection niches of S. aureus might provide different environments and iron availability resulting in different fhuD2 expression pattern within different host organs. To address these questions, we characterized the in vitro expression of the fhuD2 gene and confirmed Fur-dependent iron-regulation of its expression. We further investigated its expression in mice infected with a bioluminescent reporter strain of S. aureus expressing the luciferase operon under the control of the fhuD2 promoter. The emission of bioluminescence in different organs was followed over a seven-day time course, as well as quantitative real-time PCR analysis of the RNA transcribed from the endogenous fhuD2 gene. Using this approach, we could show that fhuD2 expression was induced during infection in all organs analyzed and that differences in expression were observed in the temporal expression profiles, and between infected organs. Our data suggest that S. aureus undergoes increased iron deprivation during progression of infection in diverse host organs and accordingly induces dedicated iron acquisition mechanisms. Since FhuD2 plays a central role in providing the pathogen with the required iron, further knowledge of the patterns of fhuD2 expression in vivo during infection is instrumental in better defining the role of this antigen in S. aureus pathogenesis and as a vaccine antigen

Acquisition of the Sda1-encoding bacteriophage does not enhance virulence of the serotype M1 Streptococcus pyogenes strain SF370

The resurgence of invasive disease caused by Streptococcus pyogenes (group A Streptococcus [GAS]) in the past 30 years has paralleled the emergence and global dissemination of the highly virulent M1T1 clone. The GAS M1T1 clone has diverged from the ancestral M1 serotype by horizontal acquisition of two unique bacteriophages, encoding the potent DNase Sda1/SdaD2 and the superantigen SpeA, respectively. The phage-encoded DNase promotes escape from neutrophil extracellular traps and is linked to enhanced virulence of the M1T1 clone. In this study, we successfully used in vitro lysogenic conversion to transfer the Sda1-encoding phage from the M1T1 clonal strain 5448 to the nonclonal M1 isolate SF370 and determined the impact of this horizontal gene transfer event on virulence. Although Sda1 was expressed in SF370 lysogens, no capacity of the phage-converted strain to survive human neutrophil killing, switch to a hyperinvasive covRS mutant form, or cause invasive lethal infection in a humanized plasminogen mouse model was observed. This work suggests that the hypervirulence of the M1T1 clone is due to the unique synergic effect of the M1T1 clone bacteriophage-specific virulence factor Sda1 acting in concert with the M1T1 clone-specific genetic scaffold

Acceptability to patients, carers and clinicians of an mHealth platform for the management of Parkinson's disease (PD_Manager): study protocol for a pilot randomised controlled trial.

BACKGROUND: Parkinson's disease is a degenerative neurological condition causing multiple motor and non-motor symptoms that have a serious adverse effect on quality of life. Management is problematic due to the variable and fluctuating nature of symptoms, often hourly and daily. The PD_Manager mHealth platform aims to provide a continuous feed of data on symptoms to improve clinical understanding of the status of any individual patient and inform care planning. The objectives of this trial are to (1) assess patient (and family carer) perspectives of PD_Manager regarding comfort, acceptability and ease of use; (2) assess clinician views about the utility of the data generated by PD_Manager for clinical decision making and the acceptability of the system in clinical practice. METHODS/DESIGN: This trial is an unblinded, parallel, two-group, randomised controlled pilot study. A total of 200 persons with Parkinson's disease (Hoehn and Yahr stage 3, experiencing motor fluctuations at least 2 h per day), with primary family carers, in three countries (110 Rome, 50 Venice, Italy; 20 each in Ioannina, Greece and Surrey, England) will be recruited. Following informed consent, baseline information will be gathered, including the following: age, gender, education, attitudes to technology (patient and carer); time since Parkinson's diagnosis, symptom status and comorbidities (patient only). Randomisation will assign participants (1:1 in each country), to PD_Manager vs control, stratifying by age (1 ≤ 70 : 1 > 70) and gender (60% M: 40% F). The PD_Manager system captures continuous data on motor symptoms, sleep, activity, speech quality and emotional state using wearable devices (wristband, insoles) and a smartphone (with apps) for storing and transmitting the information. Control group participants will be asked to keep a symptom diary covering the same elements as PD_Manager records. After a minimum of two weeks, each participant will attend a consultation with a specialist doctor for review of the data gathered (by either means), and changes to management will be initiated as indicated. Patients, carers and clinicians will be asked for feedback on the acceptability and utility of the data collection methods. The PD_Manager intervention, compared to a symptom diary, will be evaluated in a cost-consequences framework. DISCUSSION: Information gathered will inform further development of the PD_Manager system and a larger effectiveness trial. TRIAL REGISTRATION: ISRCTN Registry, ISRCTN17396879 . Registered on 15 March 2017

Plane model of fluid interface rupture in an electric field

Modeling of an air-fluid interface in an electric field is presented. Specifically, equilibrium of the interface under the dominant forces—electric stress, surface tension, and pressure—is investigated. Since interface shape and equilibrium are related, the shape of an electrified interface is also addressed. To determine the electric stress, an analytical expression for the electric field in the vicinity of the interface is determined. The operating point of the interface is shown to exist in a three-dimensional parameter space that is divided by a critical surface into equilibrium, quasiequilibrium, and nonequilibrium subdomains. The three parameters are applied voltage, electrode separation, and pressure difference. Interface size, counterelectrode size, and fluid properties are also considered. The subdomain in which the operating point resides defines the important characteristics of the interface. The operating point moves within, and transfers between, equilibrium subdomains, and points on the critical surface represent “rupture points” of the interface. The final shape of the interface is solved iteratively using this equilibrium model. Interfaces emitting an electrospray can have a range of apex angles, and it is shown that the magnitude of this angle impacts equilibrium. It is revealed that the excess pressure difference term is critical in determining the interface shape specifically the cone generatrix and that minimization of the potential energy of all forces can be used to predict the magnitude of the apex angle and pressure immediately after interface rupture. The equilibrium model is important from an operational and optimization perspective, as it is useful to predict the conditions required to break equilibrium and transfer to a quasiequilibrium state i.e., an electrospray, and the conditions necessary to maintain quasiequilibrium once it is formed.NSER

On the Pulsed and Transitional Behavior of an Electrified Fluid Interface

Journal of Fluids Engineering Copyright © 2009 by ASME SEPTEMBER 2009, Vol. 131 / 091202-1Transient modes of an electrified fluid interface are investigated, specifically, (a) intermittent or pulsed cone-jet mode and (b) smooth and abrupt transitions of the interface in response to a step voltage. These modes were studied experimentally by capturing the motion of the interface and measuring the emitted ion current (via electrospray) as they occur. The observed phenomena are described using an analytical model for the equilibrium of an electrified fluid interface, and the effect of operational parameters on the transient modes is discussed. Pressure, which is related to the supplied flow rate, significantly influences the behavior of the transient modes. It is useful to understand transient modes so they can be avoided in applications that require a stable electrospray. However, with improved knowledge, the modes studied here can assist in the development of specialized applications. DOI: 10.1115/1.3203203NSER

Application of an Equilibrium Model for an Electrified Fluid Interface—Electrospray Using a PDMS Microfluidic Device

An experimental investigation of an electrified fluid interface is presented. The experimental findings are related to a previously developed analytical model of Gubarenko , which is used to determine when a fluidic interface under electrical stress is in equilibrium, and to observations reported in the literature. The effect of key parameters on causing the interface to rupture, form, and maintain an electrospray is investigated. The experimental results reveal the dependence of interface shape on operational parameters, the impact of the interface apex angle on equilibrium, the conditions that cause either dripping mode or cone-jet mode, and the structure of operational domains. This paper confirms predictions made using the analytical model, including the range of parameters that cause the onset and steadiness of a quasi-equilibrium (electrospray) state of the interface. Testing is performed using an electrospray emitter chip fabricated from two layers of Polydimethylsiloxane and one layer of glass. The model and experimental results assist in design decisions for electrospray emitters. Applications of electrified interfaces (electrosprays) are found in mass spectrometry, microfluidics, material deposition, and colloidal thrusters for propulsion