Autoradiographic Study of the Turnover of Chromatin-Associated Phospholipids in Vicia Faba L.

SUMMARYAutoradiographic data from labelling studies on the incorporation of [14C] ethanolamine into nuclear phospholipids of meristem cells from roots of Vicia faba indicated the presence of a continuous labelling pattern for the nuclear membrane throughout the cell cycle. In contrast, a periodic labelling pattern was observed in chromatin of interphase nuclei which closely paralleled the synthesis pattern for DNA. This corroborated biochemical data from previously published studies of hepatocyte nuclear membranes and chromatin

Ion energy distribution measurements in rf and pulsed dc plasma discharges

A commercial retarding field analyzer is used to measure the time-averaged ion energy distributions of impacting ions at the powered electrode in a 13.56 MHz driven, capacitively coupled, parallel plate discharge operated at low pressure. The study is carried out in argon discharges at 10 mTorr where the sheaths are assumed to be collisionless. The analyzer is mounted flush with the powered electrode surface where the impacting ion and electron energy distributions are measured for a range of discharge powers. A circuit model of the discharge, in combination with analytical solutions for the ion energy distribution in radio-frequency sheaths, is used to calculate other important plasma parameters from the measured energy distributions. Radio-frequency compensated Langmuir probe measurements provide a comparison with the retarding field analyzer data. The time-resolved capability of the retarding field analyzer is also demonstrated in a separate pulsed dc magnetron reactor. The analyzer is mounted on the floating substrate holder and ion energy distributions of the impinging ions on a growing film, with 100 ns time resolution, are measured through a pulse period of applied magnetron power, which are crucial for the control of the microstructure and properties of the deposited films

Bacillus species at the Canberra Airport: A comparison of real-time polymerase chain reaction and massively parallel sequencing for identification

© 2018 Elsevier B.V. Anthrax, caused by the Gram-positive, spore forming bacterium Bacillus anthracis, is a disease with naturally occurring outbreaks in many parts of the world, primarily in domestic and wild herbivores. Due to the movement of people and stock, B. anthracis could, however, be at transportation hubs including airports. The continuous threat to national and international security from a biological agent release, or hoax attack, is a very real concern. Sensitive, robust and rapid (hours-day) methods to identify biological agents, including B. anthracis, and distinguish pathogenic from non-pathogenic species, is an essential cornerstone to national security. The aim of this project was to determine the presence of Bacillus species at the Canberra Airport using two massively parallel sequencing (MPS) approaches and compare with previous results using real-time polymerase chain reaction (qPCR). Samples were collected daily for seven days each month from August 2011–July 2012 targeting movement of people, luggage and freight into and out of the Canberra Airport. Extracted DNA was analysed using qPCR specific for B. anthracis. A subset of samples was analysed using two MPS approaches. Approach one, using the Ion PGM™ (Thermo Fisher Scientific; TFS) and an in-house assay, targeted the two B. anthracis virulence plasmids (cya and capB genes) and a single conserved region of the 16S rRNA gene. Approach two, using the Ion S5™ (TFS) and the commercial Ion 16S™ Metagenomics Kit (TFS), targeted multiple regions within the bacterial 16S rRNA gene. Overall there was consistency between the two MPS approaches and between MPS and qPCR, however, MPS was more sensitive, particularly for plasmid detection. Whilst the broad-range 16S genomic target(s) used in both MPS approaches in this study was able to generate a metagenomic fingerprint of the bacterial community at the Canberra Airport, it could not resolve Bacillus species beyond the level of the Bacillus cereus group. The inclusion of B. anthracis virulence plasmid targets in the in-house assay did allow for the potential presumptive identifications of pathogenic species. No plasmid targets were in the Ion 16S™ Metagenomics Kit. This study shows the choice of target(s) is key in MPS assay development and should be carefully considered to ensure the assay is fit for purpose, whether as an initial screening (presumptive) or a more specific (but not entirely confirmatory) test. Identification approaches may also benefit from a combination of MPS and qPCR as each has benefits and limitations

Retarding field energy analyser ion current calibration and transmission

International audienceAccurate measurement of ion current density and ion energy distributions (IED) is often critical for plasma processes in both industrial and research settings. Retarding field energy analyzers (RFEA) have been used to measure IEDs because they are considered accurate, relatively simple and cost effective. However, their usage for critical measurement of ion current density is less common due to difficulties in estimating the proportion of incident ion current reaching the current collector through the RFEA retarding grids. In this paper an RFEA has been calibrated to measure ion current density from an ion beam at pressures ranging from 0.5 to 50.0 mTorr. A unique method is presented where the currents generated at each of the retarding grids and the RFEA upper face are measured separately, allowing the reduction in ion current to be monitored and accounted for at each stage of ion transit to the collector. From these I-V measurements a physical model is described. Subsequently, a mathematical description is extracted which includes parameters to account for grid transmissions, upper face secondary electron emission and collisionality. Pressure-dependant calibration factors can be calculated from least mean square best fits of the collector current to the model allowing quantitative measurement of ion current density

Identity and technology: Organizational control of knowledge-intensive work

Much has been written about the functioning of managerial ideologies in identity-based organizational control. However, less attention has been given to the role of information and communication technologies (ICTs) and identity defined by a technological discourse in regulating knowledge-intensive work. The purpose of this research is to examine the roles of identity and ICTs in the control of knowledge-intensive work. A case study of a technology service organization reveals that the construction and consumption of a technologist identity operate as organizational control, and that ICTs enable the functioning of a dialectic of technological control. This study also demonstrates the paradoxical nature of work knowledge that both empowers and controls knowledge-workers

Microbiota-related Changes in Bile Acid & Tryptophan Metabolism are Associated with Gastrointestinal Dysfunction in a Mouse Model of Autism

peer-reviewedAutism spectrum disorder (ASD) is one of the most prevalent neurodevelopmental conditions worldwide. There is growing awareness that ASD is highly comorbid with gastrointestinal distress and altered intestinal microbiome, and that host-microbiome interactions may contribute to the disease symptoms. However, the paucity of knowledge on gut-brain axis signaling in autism constitutes an obstacle to the development of precision microbiota-based therapeutics in ASD. To this end, we explored the interactions between intestinal microbiota, gut physiology and social behavior in a BTBR T+ Itpr3tf/J mouse model of ASD. Here we show that a reduction in the relative abundance of very particular bacterial taxa in the BTBR gut – namely, bile-metabolizing Bifidobacterium and Blautia species, - is associated with deficient bile acid and tryptophan metabolism in the intestine, marked gastrointestinal dysfunction, as well as impaired social interactions in BTBR mice. Together these data support the concept of targeted manipulation of the gut microbiota for reversing gastrointestinal and behavioral symptomatology in ASD, and offer specific plausible targets in this endeavor.The APC Microbiome Institute is a research institute funded by Science Foundation Ireland (SFI) through the Irish Government's National Development Plan. J.F·C, T.G.D, C.S., S.A.J. and C.G.M.G. are supported by SFI (Grant Nos. SFI/12/RC/2273). S.A.J is also funded by SFI-EU 16/ERA-HDHL/3358. J.F·C, C.S. and T.G.D have research support from Mead Johnson, Cremo, 4D Pharma, Suntory Wellness, and Nutricia. J.F.C, C.S., T.G.D and G.C. have spoken at meetings sponsored by food and pharmaceutical companies

Resistance of Trichoplusia ni to Bacillus thuringiensis Toxin Cry1Ac Is Independent of Alteration of the Cadherin-Like Receptor for Cry Toxins

Alteration of binding sites for Bacillus thuringiensis (Bt) toxins in insect midgut is the major mechanism of high-level resistance to Bt toxins in insects. The midgut cadherin is known to be a major binding protein for Bt Cry1A toxins and linkage of Bt-resistance to cadherin gene mutations has been identified in lepidopterans. The resistance to Bt toxin Cry1Ac evolved in greenhouse populations of Trichoplusia ni has been identified to be associated with the down-regulation of an aminopeptidase N (APN1) gene by a trans-regulatory mechanism and the resistance gene has been mapped to the locus of an ABC transporter (ABCC2) gene. However, whether cadherin is also involved with Cry1Ac-resistance in T. ni requires to be understood. Here we report that the Cry1Ac-resistance in T. ni is independent of alteration of the cadherin. The T. ni cadherin cDNA was cloned and the cadherin sequence showed characteristic features known to cadherins from Lepidoptera. Various T. ni cadherin gene alleles were identified and genetic linkage analysis of the cadherin alleles with Cry1Ac-resistance showed no association of the cadherin gene with the Cry1Ac-resistance in T. ni. Analysis of cadherin transcripts showed no quantitative difference between the susceptible and Cry1Ac-resistant T. ni larvae. Quantitative proteomic analysis of midgut BBMV proteins by iTRAQ-2D-LC-MS/MS determined that there was no quantitative difference in cadherin content between the susceptible and the resistant larvae and the cadherin only accounted for 0.0014% (mol%) of the midgut BBMV proteins, which is 1/300 of APN1 in molar ratio. The cadherin from both the susceptible and resistant larvae showed as a 200-kDa Cry1Ac-binding protein by toxin overlay binding analysis, and nano-LC-MS/MS analysis of the 200-kDa cadherin determined that there is no quantitative difference between the susceptible and resistant larvae. Results from this study indicate that the Cry1Ac-resistance in T. ni is independent of cadherin alteration

The professional knowledge that counts in Australian contemporary early childhood teacher education

Australia is typical of many western countries where the provision of quality early childhood services has become a government priority. The government initiatives in Australia include repeated demands for 'well-qualified' early childhood educators. As a result of these demands the preservice preparation of early childhood educators is under intense scrutiny. This scrutiny raises many questions regarding the knowledge base considered to be essential for early childhood educators and leads to further questions about who has the authority to produce this knowledge. This article explores these questions by firstly examining some of the ways Australian early childhood teacher education is situated within the current knowledge environment. This is followed by a discussion regarding the debates about what early childhood educators 'need to know'. The third section of the article traces some of the historical features of Australian early childhood teacher education, for the author argues that contemporary questions about 'which' knowledge is to be included in early childhood teacher education are best understood alongside their historical precedents. The article concludes by considering the implications of the debates for contemporary early childhood teacher education and suggests that a way forward involves reconsidering the traditional binary between theory and practical knowledge

Human neutrophil clearance of bacterial pathogens triggers anti-microbial gamma delta T cell responses in early infection

Human blood Vc9/Vd2 T cells, monocytes and neutrophils share a responsiveness toward inflammatory chemokines and are rapidly recruited to sites of infection. Studying their interaction in vitro and relating these findings to in vivo observations in patients may therefore provide crucial insight into inflammatory events. Our present data demonstrate that Vc9/Vd2 T cells provide potent survival signals resulting in neutrophil activation and the release of the neutrophil chemoattractant CXCL8 (IL-8). In turn, Vc9/Vd2 T cells readily respond to neutrophils harboring phagocytosed bacteria, as evidenced by expression of CD69, interferon (IFN)-c and tumor necrosis factor (TNF)-a. This response is dependent on the ability of these bacteria to produce the microbial metabolite (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMB-PP), requires cell-cell contact of Vc9/Vd2 T cells with accessory monocytes through lymphocyte function-associated antigen-1 (LFA-1), and results in a TNF-a dependent proliferation of Vc9/Vd2 T cells. The antibiotic fosmidomycin, which targets the HMB-PP biosynthesis pathway, not only has a direct antibacterial effect on most HMB-PP producing bacteria but also possesses rapid anti-inflammatory properties by inhibiting cd T cell responses in vitro. Patients with acute peritoneal-dialysis (PD)-associated bacterial peritonitis – characterized by an excessive influx of neutrophils and monocytes into the peritoneal cavity – show a selective activation of local Vc9/Vd2 T cells by HMB-PP producing but not by HMB-PP deficient bacterial pathogens. The cd T celldriven perpetuation of inflammatory responses during acute peritonitis is associated with elevated peritoneal levels of cd T cells and TNF-a and detrimental clinical outcomes in infections caused by HMB-PP positive microorganisms. Taken together, our findings indicate a direct link between invading pathogens, neutrophils, monocytes and microbe-responsive cd T cells in early infection and suggest novel diagnostic and therapeutic approaches.Martin S. Davey, Chan-Yu Lin, Gareth W. Roberts, Sinéad Heuston, Amanda C. Brown, James A. Chess, Mark A. Toleman, Cormac G.M. Gahan, Colin Hill, Tanya Parish, John D. Williams, Simon J. Davies, David W. Johnson, Nicholas Topley, Bernhard Moser and Matthias Eber