Hyperpolarised xenon production via Rb and Cs optical pumping applied to functional lung MRI

Hyperpolarisation encompasses a multitude of methods to increase the species' spin polarisation for nuclear magnetic resonance (NMR) and magnetic resonance imaging (MRI) applications. Hyperpolarised 129Xe is produced via spin-exchange optical-pumping (SEOP). Firstly, electronic spins of alkali metal vapour are polarised via absorption of circularly polarised light. Alkali metal polarisation is subsequently transferred to noble gas nuclei via collisions. Within this thesis, the SEOP process is examined by probing the kinetics of the 129Xe polarisation build up. A combination of diagnostic techniques are used including low field NMR to measure 129Xe polarisation (PXe) at different spatial positions, near-IR optical absorption to give a global estimate of the alkali metal polarisation, and in situ Raman spectroscopy to spatially monitor the energy transport processes by detecting the internal gas temperatures (TN2). TN2 values were found to be dramatically elevated above oven thermocouple readings, with observations of up to 1000 K for an oven heated to only 400 K. Internal gas temperatures are presented for the first time along the length of the optical cell, showing spatial temperature and PXe variations during steady state and rubidium runaway conditions. Two contrasting methods of Raman spectroscopy are examined: a conventional orthogonal arrangement of detection and excitation optics, where intrinsic spatial filtering of the probe laser is utilised; and a newly designed inline module with all components in the same optical plane. Optical filtering is used to reduce the Rayleigh scattering and the probe laser line. This new inline device is presented herein and has a 23 fold improvement in signal to noise enabling increased accuracy and precision of `real-time' temperature monitoring. Rubidium, caesium and a rubidium/caesium hybrid are compared as the alkali metal of choice in the SEOP process. Caesium has a higher spin-exchange cross-section with 129Xe, thus a system is envisaged where current Rb D1 lasers in many polarisers can be utilised with a Rb/Cs hybrid to gain improvements in polarisation rates or levels. Xenon polarisations are shown up to 50% for a hybrid cell. Finally, preparatory experiments crucial to the imminent lung imaging study are presented, including measurements of PXe at low and high magnetic fields. In addition, polariser technology is examined including the current Nottingham device and an open-source consortium polariser

Hyperpolarised xenon production via Rb and Cs optical pumping applied to functional lung MRI

Hyperpolarisation encompasses a multitude of methods to increase the species' spin polarisation for nuclear magnetic resonance (NMR) and magnetic resonance imaging (MRI) applications. Hyperpolarised 129Xe is produced via spin-exchange optical-pumping (SEOP). Firstly, electronic spins of alkali metal vapour are polarised via absorption of circularly polarised light. Alkali metal polarisation is subsequently transferred to noble gas nuclei via collisions. Within this thesis, the SEOP process is examined by probing the kinetics of the 129Xe polarisation build up. A combination of diagnostic techniques are used including low field NMR to measure 129Xe polarisation (PXe) at different spatial positions, near-IR optical absorption to give a global estimate of the alkali metal polarisation, and in situ Raman spectroscopy to spatially monitor the energy transport processes by detecting the internal gas temperatures (TN2). TN2 values were found to be dramatically elevated above oven thermocouple readings, with observations of up to 1000 K for an oven heated to only 400 K. Internal gas temperatures are presented for the first time along the length of the optical cell, showing spatial temperature and PXe variations during steady state and rubidium runaway conditions. Two contrasting methods of Raman spectroscopy are examined: a conventional orthogonal arrangement of detection and excitation optics, where intrinsic spatial filtering of the probe laser is utilised; and a newly designed inline module with all components in the same optical plane. Optical filtering is used to reduce the Rayleigh scattering and the probe laser line. This new inline device is presented herein and has a 23 fold improvement in signal to noise enabling increased accuracy and precision of `real-time' temperature monitoring. Rubidium, caesium and a rubidium/caesium hybrid are compared as the alkali metal of choice in the SEOP process. Caesium has a higher spin-exchange cross-section with 129Xe, thus a system is envisaged where current Rb D1 lasers in many polarisers can be utilised with a Rb/Cs hybrid to gain improvements in polarisation rates or levels. Xenon polarisations are shown up to 50% for a hybrid cell. Finally, preparatory experiments crucial to the imminent lung imaging study are presented, including measurements of PXe at low and high magnetic fields. In addition, polariser technology is examined including the current Nottingham device and an open-source consortium polariser

Near-unity nuclear polarization with an open-source 129Xe hyperpolarizer for NMR and MRI

The exquisite NMR spectral sensitivity and negligible reactivity of hyperpolarized xenon-129 (HP129Xe) make it attractive for a number of magnetic resonance applications; moreover, HP129Xe embodies an alternative to rare and nonrenewable 3He. However, the ability to reliably and inexpensively produce large quantities of HP129Xe with sufficiently high 129Xe nuclear spin polarization (PXe) remains a significant challenge—particularly at high Xe densities. We present results from our “open-source” large-scale (∼1 L/h) 129Xe polarizer for clinical, preclinical, and materials NMR and MRI research. Automated and composed mostly of off-the-shelf components, this “hyperpolarizer” is designed to be readily implementable in other laboratories. The device runs with high resonant photon flux (up to 200 W at the Rb D1 line) in the xenon-rich regime (up to 1,800 torr Xe in 500 cc) in either single-batch or stopped-flow mode, negating in part the usual requirement of Xe cryocollection. Excellent agreement is observed among four independent methods used to measure spin polarization. In-cell PXe values of ∼90%, ∼57%, ∼50%, and ∼30% have been measured for Xe loadings of ∼300, ∼500, ∼760, and ∼1,570 torr, respectively. PXe values of ∼41% and ∼28% (with ∼760 and ∼1,545 torr Xe loadings) have been measured after transfer to Tedlar bags and transport to a clinical 3 T scanner for MR imaging, including demonstration of lung MRI with a healthy human subject. Long “in-bag” 129Xe polarization decay times have been measured (T1 ∼38 min and ∼5.9 h at ∼1.5 mT and 3 T, respectively)—more than sufficient for a variety of applications

Shiga Toxin–producing Escherichia coli Strains Negative for Locus of Enterocyte Effacement

The ehx plasmids of these strains are highly related, which suggests acquisition of the large plasmid was central to the strains’ emergence

Analysis of the Legionella longbeachae Genome and Transcriptome Uncovers Unique Strategies to Cause Legionnaires' Disease

Legionella pneumophila and L. longbeachae are two species of a large genus of bacteria that are ubiquitous in nature. L. pneumophila is mainly found in natural and artificial water circuits while L. longbeachae is mainly present in soil. Under the appropriate conditions both species are human pathogens, capable of causing a severe form of pneumonia termed Legionnaires' disease. Here we report the sequencing and analysis of four L. longbeachae genomes, one complete genome sequence of L. longbeachae strain NSW150 serogroup (Sg) 1, and three draft genome sequences another belonging to Sg1 and two to Sg2. The genome organization and gene content of the four L. longbeachae genomes are highly conserved, indicating strong pressure for niche adaptation. Analysis and comparison of L. longbeachae strain NSW150 with L. pneumophila revealed common but also unexpected features specific to this pathogen. The interaction with host cells shows distinct features from L. pneumophila, as L. longbeachae possesses a unique repertoire of putative Dot/Icm type IV secretion system substrates, eukaryotic-like and eukaryotic domain proteins, and encodes additional secretion systems. However, analysis of the ability of a dotA mutant of L. longbeachae NSW150 to replicate in the Acanthamoeba castellanii and in a mouse lung infection model showed that the Dot/Icm type IV secretion system is also essential for the virulence of L. longbeachae. In contrast to L. pneumophila, L. longbeachae does not encode flagella, thereby providing a possible explanation for differences in mouse susceptibility to infection between the two pathogens. Furthermore, transcriptome analysis revealed that L. longbeachae has a less pronounced biphasic life cycle as compared to L. pneumophila, and genome analysis and electron microscopy suggested that L. longbeachae is encapsulated. These species-specific differences may account for the different environmental niches and disease epidemiology of these two Legionella species

Legionella pneumophila strain 130b possesses a unique combination of type IV secretion systems and novel Dot/Icm secretion system effector proteins

Legionella pneumophila is a ubiquitous inhabitant of environmental water reservoirs. The bacteria infect a wide variety of protozoa and, after accidental inhalation, human alveolar macrophages, which can lead to severe pneumonia. The capability to thrive in phagocytic hosts is dependent on the Dot/Icm type IV secretion system (T4SS), which translocates multiple effector proteins into the host cell. In this study, we determined the draft genome sequence of L. pneumophila strain 130b (Wadsworth). We found that the 130b genome encodes a unique set of T4SSs, namely, the Dot/Icm T4SS, a Trb-1-like T4SS, and two Lvh T4SS gene clusters. Sequence analysis substantiated that a core set of 107 Dot/Icm T4SS effectors was conserved among the sequenced L. pneumophila strains Philadelphia-1, Lens, Paris, Corby, Alcoy, and 130b. We also identified new effector candidates and validated the translocation of 10 novel Dot/Icm T4SS effectors that are not present in L. pneumophila strain Philadelphia-1. We examined the prevalence of the new effector genes among 87 environmental and clinical L. pneumophila isolates. Five of the new effectors were identified in 34 to 62% of the isolates, while less than 15% of the strains tested positive for the other five genes. Collectively, our data show that the core set of conserved Dot/Icm T4SS effector proteins is supplemented by a variable repertoire of accessory effectors that may partly account for differences in the virulences and prevalences of particular L. pneumophila strains. Copyright © 2010, American Society for Microbiology. All Rights Reserved

The design, launch and assessment of a new volunteer-based plant monitoring scheme for the United Kingdom

Volunteer-based plant monitoring in the UK has focused mainly on distribution mapping; there has been less emphasis on the collection of data on plant communities and habitats. Abundance data provide different insights into ecological pattern and allow for more powerful inference when considering environmental change. Abundance monitoring for other groups of organisms is well-established in the UK, e.g. for birds and butterflies, and conservation agencies have long desired comparable schemes for plants. We describe a new citizen science scheme for the UK (the ‘National Plant Monitoring Scheme’; NPMS), with the primary aim of monitoring the abundance of plants at small scales. Scheme development emphasised volunteer flexibility through scheme co-creation and feedback, whilst retaining a rigorous approach to design. Sampling frameworks, target habitats and species, field methods and power are all described. We also evaluate several outcomes of the scheme design process, including: (i) landscape-context bias in the first two years of the scheme; (ii) the ability of different sets of indicator species to capture the main ecological gradients of UK vegetation; and, (iii) species richness bias in returns relative to a professional survey. Survey rates have been promising (over 60% of squares released have been surveyed), although upland squares are under-represented. Ecological gradients present in an ordination of an independent, unbiased, national survey were well-represented by NPMS indicator species, although further filtering to an entry-level set of easily identifiable species degraded signal in an ordination axis representing succession and disturbance. Comparison with another professional survey indicated that different biases might be present at different levels of participation within the scheme. Understanding the strengths and limitations of the NPMS will guide development, increase trust in outputs, and direct efforts for maintaining volunteer interest, as well as providing a set of ideas for other countries to experiment with

Comparative study of in situ N2 rotational Raman spectroscopy methods for probing energy thermalisation processes during spin-exchange optical pumping

Spin-exchange optical pumping (SEOP) has been widely used to produce enhancements in nuclear spin polarisation for hyperpolarised noble gases. However, some key fundamental physical processes underlying SEOP remain poorly understood, particularly in regards to how pump laser energy absorbed during SEOP is thermalised, distributed and dissipated. This study uses in situ ultra-low frequency Raman spectroscopy to probe rotational temperatures of nitrogen buffer gas during optical pumping under conditions of high resonant laser flux and binary Xe/N2 gas mixtures. We compare two methods of collecting the Raman scattering signal from the SEOP cell: a conventional orthogonal arrangement combining intrinsic spatial filtering with the utilisation of the internal baffles of the Raman spectrometer, eliminating probe laser light and Rayleigh scattering, versus a new in-line modular design that uses ultra-narrowband notch filters to remove such unwanted contributions. We report a ~23-fold improvement in detection sensitivity using the in-line module, which leads to faster data acquisition and more accurate real-time monitoring of energy transport processes during optical pumping. The utility of this approach is demonstrated via measurements of the local internal gas temperature (which can greatly exceed the externally measured temperature) as a function of incident laser power and position within the cell

XeNA: an automated ‘open-source’ 129Xe hyperpolarizer for clinical use

Here we provide a full report on the construction, components, and capabilities of our consortium’s “open-source” large-scale (~ 1 L/h) 129Xe hyperpolarizer for clinical, pre-clinical, and materials NMR/MRI (Nikolaou et al., Proc. Natl. Acad. Sci. USA, 110, 14150 (2013)). The ‘hyperpolarizer’ is automated and built mostly of off-the-shelf components; moreover, it is designed to be cost-effective and installed in both research laboratories and clinical settings with materials costing less than $125,000. The device runs in the xenon-rich regime (up to 1800 Torr Xe in 0.5 L) in either stopped-flow or single-batch mode—making cryo-collection of the hyperpolarized gas unnecessary for many applications. In-cell 129Xe nuclear spin polarization values of ~ 30%–90% have been measured for Xe loadings of ~ 300–1600 Torr. Typical 129Xe polarization build-up and T1 relaxation time constants were ~ 8.5 min and ~ 1.9 h respectively under our spin-exchange optical pumping conditions; such ratios, combined with near-unity Rb electron spin polarizations enabled by the high resonant laser power (up to ~ 200 W), permit such high PXe values to be achieved despite the high in-cell Xe densities. Importantly, most of the polarization is maintained during efficient HP gas transfer to other containers, and ultra-long 129Xe relaxation times (up to nearly 6 h) were observed in Tedlar bags following transport to a clinical 3 T scanner for MR spectroscopy and imaging as a prelude to in vivo experiments. The device has received FDA IND approval for a clinical study of chronic obstructive pulmonary disease subjects. The primary focus of this paper is on the technical/engineering development of the polarizer, with the explicit goals of facilitating the adaptation of design features and operative modes into other laboratories, and of spurring the further advancement of HP-gas MR applications in biomedicine