Effect of natural products on the production and activity of Clostridium difficile toxins in vitro

Clostridium difficile infection is a toxin-mediated disease of the colon. C. difficile virulence is primarily attributed to the production of toxin A and toxin B; thus this study was aimed to investigate the effect of a range of natural products on the production and activity of C. difficile toxins in vitro. Twenty-two natural products were investigated against four C. difficile strains. The activity of products against toxins was determined using Vero and HT-29 cells cytotoxicity and neutral red uptake assays. The indirect effect of products on toxin-mediated cytotoxicity was determined using the same cell lines. The effect of seven products on toxin production by C. difficile was determined using ELISA. Zingerone (0.3 mg/ml) protected both cell lines from C. difficile cytopathic effects, confirmed by the neutral red uptake assay (P \u3c 0.05). Three Leptospermum honeys (4% w/v), fresh onion bulb extract (12.5% v/v) and trans-cinnamaldehyde (0.005% v/v) all reduced toxin production and activity significantly (P ≤ 0.023). Garlic clove powder (4.7 mg/ml) only reduced toxin activity (P ≤ 0.047). Overall, several natural products had activity against C. difficile toxins in vitro encouraging further investigation against C. difficile toxins in vivo

Genome analysis of Clostridium difficile PCR ribotype 014 lineage in Australian pigs and humans reveals a diverse genetic repertoire and signatures of long-range interspecies transmission

Clostridium difficile PCR ribotype (RT) 014 is well-established in both human and porcine populations in Australia, raising the possibility that C. difficile infection (CDI) may have a zoonotic or foodborne etiology. Here, whole genome sequencing and high-resolution core genome phylogenetics were performed on a contemporaneous collection of 40 Australian RT014 isolates of human and porcine origin. Phylogenies based on MLST (7 loci, STs 2, 13, and 49) and core orthologous genes (1260 loci) showed clustering of human and porcine strains indicative of very recent shared ancestry. Core genome single nucleotide variant (SNV) analysis found 42 % of human strains showed a clonal relationship (separated by ≤ 2 SNVs in their core genome) with one or more porcine strains, consistent with recent inter-host transmission. Clones were spread over a vast geographic area with 50 % of the human cases occurring without recent healthcare exposure. These findings suggest a persistent community reservoir with long-range dissemination, potentially due to agricultural recycling of piggery effluent. We also provide the first pan-genome analysis for this lineage, characterizing its resistome, prophage content, and in silico virulence potential. The RT014 is defined by a large “open” pan-genome (7587 genes) comprising a core genome of 2296 genes (30.3 % of the total gene repertoire) and an accessory genome of 5291 genes. Antimicrobial resistance genotypes and phenotypes varied across host populations and ST lineages and were characterized by resistance to tetracycline [tetM, tetA(P), tetB(P) and tetW], clindamycin/erythromycin (ermB), and aminoglycosides (aph3-III-Sat4A-ant6-Ia). Resistance was mediated by clinically important mobile genetic elements, most notably Tn6194 (harboring ermB) and a novel variant of Tn5397 (harboring tetM). Numerous clinically important prophages (Siphoviridae and Myoviridae) were identified as well as an uncommon accessory gene regulator locus (agr3). Conservation in the pathogenicity locus and S-layer correlated with ST affiliation, further extending the concept of clonal C. difficile lineages. This study provides novel insights on the genetic variability and strain relatedness of C. difficile RT014, a lineage of emerging One Health importance. Ongoing molecular and genomic surveillance of strains in humans, animals, food, and the environment is imperative to identify opportunities to reduce the overall CDI burden

Early social environment influences the behaviour of a family-living lizard

Financial support for this research was provided by the Australian Research Council (DP130102998; grant to M.J.W. and R.B.W.), Natural Sciences and Engineering Research Council of Canada (scholarship to J.L.R.), the Australasian Society for the Study of Animal Behaviour, and Macquarie University. D.W.A.N. was supported by an ARC Discovery Early Career Research Award (DE150101774) and UNSW Vice Chancellors Fellowship.Early social environment can play a significant role in shaping behavioural development. For instance, in many social mammals and birds, isolation rearing results in individuals that are less exploratory, shyer, less social and more aggressive than individuals raised in groups. Moreover, dynamic aspects of social environments, such as the nature of relationships between individuals, can also impact the trajectory of development. We tested if being raised alone or socially affects behavioural development in the family-living tree skink, Egernia striolata. Juveniles were raised in two treatments: alone or in a pair. We assayed exploration, boldness, sociability and aggression repeatedly throughout each juvenile's first year of life, and also assessed social interactions between pairs to determine if juveniles formed dominant–subordinate relationships. We found that male and/or the larger skinks within social pairs were dominant. Developing within this social environment reduced skink growth, and subordinate skinks were more prone to tail loss. Thus, living with a conspecific was costly for E. striolata. The predicted negative effects of isolation failed to materialize. Nevertheless, there were significant differences in behavioural traits depending on the social environment (isolated, dominant or subordinate member of a pair). Isolated skinks were more social than subordinate skinks. Subordinate skinks also became more aggressive over time, whereas isolated and dominant skinks showed invariable aggression. Dominant skinks became bolder over time, whereas isolated and subordinate skinks were relatively stable in their boldness. In summary, our study is evidence that isolation rearing does not consistently affect behaviour across all social taxa. Our study also demonstrates that the social environment plays an important role in behavioural development of a family-living lizard.Publisher PDFPeer reviewe

Volume 13. Article 3. Hydrographic and biological studies of Block Island Sound.

https://elischolar.library.yale.edu/bulletin_yale_bingham_oceanographic_collection/1150/thumbnail.jp

Low and High-Power Inductive Pulsed Plasma Thruster Development Testing at NASA-MSFC

The inductive pulsed plasma thruster (IPPT) is an electromagnetic plasma accelerator that has been identified in NASA roadmaps as an enabling propulsion technology for some niche low-power missions and for high-power in-space propulsion needs. The IPPT is an electrodeless space propulsion device where a capacitor is charged to an initial voltage and then discharged producing a high current pulse through a coil. The field produced by this pulse ionizes propellant, inductively driving current in a plasma located near the face of the coil. Once the plasma is formed it can be accelerated and expelled at a high exhaust velocity by the electromagnetic Lorentz body force arising from the interaction of the induced plasma current and the magnetic field produced by the current in the coil. Thrusters of this type possess many demonstrated and potential benefits that make them worthy of continued investigation. The electrodeless nature of these thrusters eliminates the lifetime and contamination issues associated with electrode erosion in conventional electric thrusters. Also, a wider variety of propellants are accessible when compatibility with metallic electrodes in no longer an issue. IPPTs have been successfully operated using propellants like ammonia, hydrazine, and CO2, and there is no fundamental reason why they would not operate on other in situ propellants like H2O. It is well-known that pulsed accelerators can maintain constant specific impulse (I(sub sp)) and thrust efficiency (eta(sub t)) over a wide range of input power levels by adjusting the pulse rate to hold the discharge energy per pulse constant. It has also been demonstrated that an inductive pulsed plasma thruster can operate in a regime where eta(sub t) is relatively constant over a wide range of I(sub sp) values (3000-8000 s). Finally, thrusters in this class have operated in single-pulse mode at high energy per pulse, and by increasing the pulse rate they offer the potential to process very high levels of power using a single thruster. There has been significant previous research on IPPTs designed around a planar-coil (flat-plate) geometry. The most notable of these was the Pulsed Inductive Thruster (PIT), with the PIT MkV presently representing the state-of- the-art in pulsed high-power IPPT technological development. In this paper, we focus on two planar-geometry devices that operate at significantly different power levels. Most work performed at NASA-Marshall Space Flight Center (MSFC) has, to date, focused on lower power thruster operation (approx. = 10s to 100s of J/pulse, up to 2-2.5 kW average power throughput) and previously described. The most recent work aimed to assemble a device that could be tested in cyclic mode on a thrust-stand, and which could augment the existing data set for IPPTs. In addition, the thruster was designed to serve as a test-bed for solid state switching circuitry and pulsed gas valves, with the modular design of the device allowing for variation in or upgrades to test configuration. Recently, MSFC obtained on loan from the Georgia Institute of Technology (Atlanta, GA) the PIT MkVI, successor to the PIT MkV. The MkV and MkVI are similar in design with much of the hardware from the former, specifically the capacitors and spark-gap switches, being reused in the latter. The coil is similar in geometry but has bent copper rods used in the latest iteration in place of the Litz wire windings found in the MkV. The MkVI master switch for the spark gaps is located in the vacuum chamber contained within a sealed, pressurized vessel fastened to the back of the thruster. This is different from the MkV where many capacitor charging lines and spark gap-triggering delay lines ran to the thruster from a master trigger located outside the vacuum chamber. The MkVI was damaged during testing soon after its fabrication was completed. The thruster arrived at MSFC still-damaged and mostly disassembled into many individual pieces. The device has been repaired, with a few additional design changes implemented after discussions with the late Prof. Lovberg regarding the initial testing results and issues encountered. In the present work, we present results from testing of both the small IPPT and the larger MkVI thruster. The smaller device (Fig. 1) is tested on a thrust stand on multiple gases to demonstrate its capability to operate in a repetition-rate mode and serve as a IPPT technology-development testbed. The larger MkVI (Fig. 2) is operated for the first time in its newly reconstituted state, demonstrating full-power pulsed operation and, for the first time, repetition-rate operation of a high-power IPPT. The additional upgrades required for synchronous operation of all the pulsed systems in single-pulse and repetition-rate mode are described in detail

Inductive Pulsed Plasma Thruster Development Testing at NASA-MSFC

The inductive pulsed plasma thruster (IPPT) is an electromagnetic plasma accelerator that has been identified in NASA roadmaps as an enabling propulsion technology for some niche low-power missions and for high-power in-space propulsion needs. The IPPT is an electrodeless space propulsion device where a capacitor is charged to an initial voltage and then discharged producing a high current pulse through a coil. The field produced by this pulse ionizes propellant, inductively driving current in a plasma located near the face of the coil. Once the plasma is formed it can be accelerated and expelled at a high exhaust velocity by the electromagnetic Lorentz body force arising from the interaction of the induced plasma current and the magnetic field produced by the current in the coil. Thrusters of this type possess many demonstrated and potential benefits that make them worthy of continued investigation. The electrodeless nature of these thrusters eliminates the lifetime and contamination issues associated with electrode erosion in conventional electric thrusters. Also, a wider variety of propellants are accessible when compatibility with metallic electrodes in no longer an issue. IPPTs have been successfully operated using propellants like ammonia, hydrazine, and CO2, and there is no fundamental reason why they would not operate on other in situ propellants like H2O. It is well-known that pulsed accelerators can maintain constant specific impulse (I(sub sp)) and thrust efficiency ((sub t)) over a wide range of input power levels by adjusting the pulse rate to hold the discharge energy per pulse constant. It has also been demonstrated that an inductive pulsed plasma thruster can operate in a regime where (sub t) is relatively constant over a wide range of I(sub sp) values (3000-8000 s). Finally, thrusters in this class have operated in single-pulse mode at high energy per pulse, and by increasing the pulse rate they offer the potential to process very high levels of power using a single thruster. There has been significant previous research on IPPTs designed around a planar-coil (flat-plate) geometry. The most notable of these was the Pulsed Inductive Thruster (PIT), with the PIT MkV presently representing the state of- the-art in pulsed high-power IPPT technological development. In this paper, we focus on two planar-geometry devices that operate at significantly different power levels. Most work performed at NASA-Marshall Space Flight Center (MSFC) has, to date, focused on lower power thruster operation ( 10s to 100s of J/pulse, up to 2-2.5 kW average power throughput) and previously described in Refs. [5,6]. The most recent work aimed to assemble a device that could be tested in cyclic mode on a thrust-stand, and which could augment the existing data set for IPPTs. In addition, the thruster was designed to serve as a test-bed for solid state switching circuitry and pulsed gas valves, with the modular design of the device allowing for variation in or upgrades to test configuration. Recently, MSFC obtained on loan from the Georgia Institute of Technology (Atlanta, GA) the PIT MkVI, successor to the PIT MkV. The MkV and MkVI are similar in design with much of the hardware from the former, specifically the capacitors and spark-gap switches, being reused in the latter. The coil is simliar in geometry but has bent copper rods used in the latest iteration in place of the Litz wire windings found in the MkV. The MkVI master switch for the spark gaps is located in the vacuum chamber contained within a sealed, pressurized vessel fastened to the back of the thruster. This is different from the MkV where many capacitor charging lines and spark gap-triggering delay lines ran to the thruster from a master trigger located outside the vacuum chamber. The MkVI was damaged during testing soon after its fabrication was completed. The thruster arrived at MSFC still-damaged and mostly disassembled into many individual pieces. The device has been repaired, with a few additional design changes implemented after discussions with the late Prof. Lovberg regarding the initial testing results and issues encountered

Novice Lifters Exhibit A More Kyphotic Lifting Posture Than Experienced Lifters In Straight-Leg Lifting

As torso flexion and repetitive lifting are known risk factors for low back pain and injury, it is important to investigate lifting techniques that might reduce injury during repetitive lifting. By normalizing lumbar posture to a subject’s range of motion (ROM), as a function of torso flexion, this research examined when subjects approached their range of motion limits during dynamic lifting tasks. For this study, it was hypothesized that experienced lifters would maintain a more neutral lumbar angle relative to their range of motion, while novice lifters would approach the limits of their lumbar ROM during the extension phase of a straight-leg lift. The results show a statistically significant difference in lifting patterns for these two groups supporting this hypothesis. The novice group maintained a much more kyphotic lumbar angle for both the flexion (74% of the lumbar angle ROM) and extension phases (86% of the lumbar angle ROM) of the lifting cycle, while the experienced group retained a more neutral curvature throughout the entire lifting cycle (37% of lumbar angle ROM in flexion and 48% of lumbar angle ROM in extension). By approaching the limits of their range of motion, the novice lifters could be at greater risk of injury by placing greater loads on the supporting soft tissues of the spine. Future research should examine whether training subjects to assume more neutral postures during lifting could indeed lower injury risks

IFN-γ-producing CD4+ T cells promote experimental cerebral malaria by modulating CD8+ T cell accumulation within the brain.

It is well established that IFN-γ is required for the development of experimental cerebral malaria (ECM) during Plasmodium berghei ANKA infection of C57BL/6 mice. However, the temporal and tissue-specific cellular sources of IFN-γ during P. berghei ANKA infection have not been investigated, and it is not known whether IFN-γ production by a single cell type in isolation can induce cerebral pathology. In this study, using IFN-γ reporter mice, we show that NK cells dominate the IFN-γ response during the early stages of infection in the brain, but not in the spleen, before being replaced by CD4(+) and CD8(+) T cells. Importantly, we demonstrate that IFN-γ-producing CD4(+) T cells, but not innate or CD8(+) T cells, can promote the development of ECM in normally resistant IFN-γ(-/-) mice infected with P. berghei ANKA. Adoptively transferred wild-type CD4(+) T cells accumulate within the spleen, lung, and brain of IFN-γ(-/-) mice and induce ECM through active IFN-γ secretion, which increases the accumulation of endogenous IFN-γ(-/-) CD8(+) T cells within the brain. Depletion of endogenous IFN-γ(-/-) CD8(+) T cells abrogates the ability of wild-type CD4(+) T cells to promote ECM. Finally, we show that IFN-γ production, specifically by CD4(+) T cells, is sufficient to induce expression of CXCL9 and CXCL10 within the brain, providing a mechanistic basis for the enhanced CD8(+) T cell accumulation. To our knowledge, these observations demonstrate, for the first time, the importance of and pathways by which IFN-γ-producing CD4(+) T cells promote the development of ECM during P. berghei ANKA infection

Clostridium difficile ribotype 017–characterization, evolution and epidemiology of the dominant strain in Asia

Clostridium difficile ribotype (RT) 017 is an important toxigenic C. difficile RT which, due to a deletion in the repetitive region of the tcdA gene, only produces functional toxin B. Strains belonging to this RT were initially dismissed as nonpathogenic and circulated largely undetected for almost two decades until they rose to prominence following a series of outbreaks in the early 2000s. Despite lacking a functional toxin A, C. difficile RT 017 strains have been shown subsequently to be capable of causing disease as severe as that caused by strains producing both toxins A and B. While C. difficile RT 017 strains can be found in almost every continent today, epidemiological studies suggest that the RT is endemic in Asia and that the global spread of this MLST clade 4 lineage member is a relatively recent event. C. difficile RT 017 transmission appears to be mostly from human to human with only a handful of reports of isolations from animals. An important feature of C. difficile RT 017 strains is their resistance to several antimicrobials and this has been documented as a possible factor driving multiple outbreaks in different parts of the world. This review summarizes what is currently known regarding the emergence and evolution of strains belonging to C. difficile RT 017 as well as features that have allowed it to become an RT of global importance

The Soft-Excess in Mrk 509: Warm Corona or Relativistic Reflection?

We present the analysis of the first NuSTAR observations ($\sim 220$ ks), simultaneous with the last SUZAKU observations ($\sim 50$ ks), of the active galactic nucleus of the bright Seyfert 1 galaxy Mrk 509. The time-averaged spectrum in the $1-79$ keV X-ray band is dominated by a power-law continuum ($\Gamma\sim 1.8-1.9$), a strong soft excess around 1 keV, and signatures of X-ray reflection in the form of Fe K emission ($\sim 6.4$ keV), an Fe K absorption edge ($\sim 7.1$ keV), and a Compton hump due to electron scattering ($\sim 20-30$ keV). We show that these data can be described by two very different prescriptions for the soft excess: a warm ($kT\sim 0.5-1$ keV) and optically thick ($\tau\sim10-20$) Comptonizing corona, or a relativistically blurred ionized reflection spectrum from the inner regions of the accretion disk. While these two scenarios cannot be distinguished based on their fit statistics, we argue that the parameters required by the warm corona model are physically incompatible with the conditions of standard coronae. Detailed photoionization calculations show that even in the most favorable conditions, the warm corona should produce strong absorption in the observed spectrum. On the other hand, while the relativistic reflection model provides a satisfactory description of the data, it also requires extreme parameters, such as maximum black hole spin, a very low and compact hot corona, and a very high density for the inner accretion disk. Deeper observations of this source are thus necessary to confirm the presence of relativistic reflection, and to further understand the nature of its soft excess.Comment: Accepted for publication in ApJ, 18 pages, 7 figure