A fluorescence in situ staining method for investigating spores and vegetative cells of Clostridia by confocal laser scanning microscopy and structured illuminated microscopy

Non-pathogenic spore-forming Clostridia are of increasing interest due to their application in biogas production and their capability to spoil different food products. The life cycle for Clostridium includes a spore stage that can assist in survival under environmentally stressful conditions, such as extremes of temperature or pH. Due to their size, spores can be investigated by a range of microscopic techniques, many of which involve sample pretreatment. We have developed a quick, simple and non-destructive fluorescent staining procedure that allows a clear differentiation between spores and vegetative cells and effectively stains spores, allowing recovery and tracking in subsequent experiments. Hoechst 34580, Propidium iodide and wheat germ agglutinin WGA 488 were used in combination to stain four strains of Clostridia at different life cycle stages. Staining was conducted without drying the sample, preventing changes induced by dehydration and cells observed by confocal laser scanner microscopy or using a super-resolution microscope equipped with a 3D-structured illumination module. Dual staining with Hoechst/Propidium iodide differentiated spores from vegetative cells, provided information on the viability of cells and was successfully applied to follow spore production induced by heating. Superresolution microscopy of spores probed by Hoechst 34580 also allowed chromatin to be visualised. Direct staining of a cheese specimen using Nile Red and Fast Green allowed in situ observation of spores within the cheese and their position within the cheese matrix. The proposed staining method has broad applicability and can potentially be applied to follow Clostridium spore behaviour in a range of different environments

Effect of temperature on the microstructure of fat globules and the immunoglobulin-mediated interactions between fat and bacteria in natural raw milk creaming

Natural creaming of raw milk is the first step in production of Grana Padano and Parmigiano Reggiano Protected Denomination of Origin cheeses. This process decreases the fat content and plays an important role in the removal of clostridia species that may cause late-blowing defects in ripened cheeses. Partial coalescence of fat globules-that may influence fat behavior in cheese making and affect the microstructure of fat in the final cheese product-was observed at creaming temperatures higher than 22\uc2\ub0C by confocal laser scanning microscopy. The widespread practice of heating of milk at 37\uc2\ub0C before creaming at 8\uc2\ub0C resulted in important changes in the size distribution of fat globules in raw milk, potentially altering the ability of fat to entrap clostridia spores. We investigated the role of immunoglobulin classes in both the clustering of fat globules and the agglutination of Clostridium tyrobutyricum to fat globules during creaming. Immunogold labeling and transmission electron microscopy showed that IgA and IgM but not IgG were involved in both clustering and agglutination. Both vegetative cells and spores were clearly shown to agglutinate to fat droplets, a process that was suppressed by thermal denaturation of the immunoglobulins. The debacterization of raw milk through natural creaming was improved by the addition of purified immunoglobulins. Overall, these findings provide not only a better understanding of the phenomena occurring during the natural creaming but also practical insights into how the process of creaming may be optimized in cheese production plants

Distribution of Salmonella enterica isolates from human cases in Italy, 1980 to 2011

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Parametric instabilities and their control in advanced interferometer GW detectors

A detailed simulation of Advanced LIGO test mass optical cavities shows that parametric instabilities will excite acoustic modes in the test masses in the frequency range 28-35 kHz and 64-72 kHz. Using nominal Advanced LIGO optical cavity parameters with fused silica test masses, parametric instability excites 7 acoustic modes in each test mass, with parametric gain R up to 7. For the alternative sapphire test masses only 1 acoustic mode is excited in each test mass with R ~ 2. Fine tuning of the test mass radii of curvature cause the instabilities to sweep through various modes with R as high as ~2000. Sapphire test mass cavities can be tuned to completely eliminate instabilities using thermal g-factor tuning with negligible degradation of the noise performance. In the case of fused silica test mass, instabilities can be minimized but not eliminated.Comment: 5 pages, 4 figure

Three-mode opto-acoustic parametric interactions with coupled cavity

We theoretically analyze three-mode opto-acoustic parametric interactions in a coupled Fabry-Perot cavity, where one acoustic mode interacts with two optical modes. We show explicitly that extra degrees of freedom in a coupled cavity allow explorations of both parametric instability and cooling regimes with high parametric gain in a single table top experiment. This work can motivate experimental realizations of the three-mode parametric instability which might be an issue in next-generation gravitational-wave detectors with high optical-power cavities, helping in the development of better models, and in developing techniques for controls. In addition, we show that the same scheme can be implemented in the resolved-sideband acoustic-mode cooling.Comment: 15 pages, 7 figure

Observations of sulfur dioxide uptake and new particle formation in a midlatitude cumulus cloud

International audienceAirborne measurements, obtained during the Asian Aerosol Characterisation Experiment (ACE-Asia), of SO2 and condensation nuclei (CN) concentrations were made in the local environment of a cumulus cloud band. Conserved quantities, wet equivalent potential temperature ?q, and total water content Q, were used to identify the sources of air detrained on the downwind side of the cumulus band. It was found that ~65% of the detrained air originated from below cloud base and the remainder was air that had been entrained from the free troposphere upwind of the cloud and subsequently been detrained. Calculation of the sources of the detrained air parcels enabled a prediction of the concentration of SO2 and CN, assuming that SO2 and CN experienced no processing within cloud. A comparison of the predicted concentration of SO2 and CN was made with those observed. The concentration of SO2 observed was less than predicted and the amount of SO2 scavenged within cloud was calculated. The CN concentration observed was also less than predicted and, moreover, inclusion of the loss of CN to cloud droplets due to Brownian scavenging resulted in an enhanced decrease of the number concentration of CN predicted. Clear air regions around the cloud exhibited no indication of being a major source of new particles. It was concluded that new particles were formed within cloud

Pyridazinediones and amino acid receptors: theoretical studies, design, synthesis, and evaluation of novel analogues

http://www.pharmacol.usyd.edu.au/thesis This thesis is primarily concerned with a class of chemical compounds known as pyridazinediones, being 6-membered aromatic rings containing two adjacent nitrogen atoms (pyridazine), doubly substituted with oxygen. In particular, the work focuses on pyridazine-3,6-diones, derivatives of maleic hydrazide (1). Understanding of the chemistry of these compounds is extended, using theoretical and synthetic techniques. This thesis is also concerned with two very important classes of receptors which bind amino acids in the brain: firstly, the inhibitory GABA receptor, which binds g-aminobutyric acid (GABA) (2) in vivo, and for which muscimol (3) is an agonist of the GABAA subclass; secondly, Excitatory Amino Acid (EAA) receptors, which bind glutamate (4) in vivo, and in particular the AMPA subclass, for which (S)-2-amino-3-(3-hydroxy-5-methylisoxazol-4-yl)propionic acid (AMPA) (5) is an agonist. The connection between pyridazinediones and amino acid receptors is the design, synthesis, and evaluation of structures based on pyridazinediones as potential GABA and EAA receptor ligands. Techniques of theoretical chemistry, molecular modelling, synthetic chemistry, and in vitro pharmacology are used to explore pyridazine-3,6-dione derivatives as ligands

Errors in chromosome segregation during oogenesis and early embryogenesis

Errors in chromosome segregation occurring during human oogenesis and early embryogenesis are very common. Meiotic chromosome development during oogenesis is subdivided into three distinct phases. The crucial events, including meiotic chromosome pairing and recombination, take place from around 11 weeks until birth. Oogenesis is then arrested until ovulation, when the first meiotic division takes place, with the second meiotic division not completed until after fertilization. It is generally accepted that most aneuploid fetal conditions, such as trisomy 21 Down syndrome, are due to maternal chromosome segregation errors. The underlying reasons are not yet fully understood. It is also clear that superimposed on the maternal meiotic chromosome segregation errors, there are a large number of mitotic errors taking place post-zygotically during the first few cell divisions in the embryo. In this chapter, we summarise current knowledge of errors in chromosome segregation during oogenesis and early embryogenesis, with special reference to the clinical implications for successful assisted reproduction

Rheological properties of honey from Burkina Faso: loss modulus and complex viscosity modelling

This is an Author's Accepted Manuscript of an article published in International Journal of Food Properties, 19, 11, 2575-2586 © Taylor & Francis, available online at: http://www.tandfonline.com/10.1080/10942912.2015.1136938[EN] This study evaluated the rheological behaviour of Burkina Faso honey and the use of exponential and polynomial models to predict the influence of chemical composition and temperature on the viscoelastic parameters: complex viscosity (ç*) and loss modulus (G ). Samples were first characterized by evaluating: water activity, 5-hydroxy methyl furfural (HMF), sugars (fructose, glucose and sucrose), electrical conductivity, moisture and colour. Dynamic rheological properties were obtained at different temperatures (5, 10, 15, 20, 25, 30 and 40°C). All the honeys displayed Newtonian behaviour. Complex viscosity and loss modulus can be predicted based on the chemical composition and temperature using polynomial models (R2> 98.00%).The authors thank the Universitat Politecnica de Valencia for funding the project "Seguridad alimentaria en la Region Suroeste de Burkina Faso. Capacitacion en manipulacion, transformacion y conservacion de alimentos locales, y contra la malnutricion infantil (BONALIMENT2013)", in the framework of the Program ADSIDEO of the Area de Cooperacio al Desenvolupament.Escriche Roberto, MI.; Oroian, M.; Visquert Fas, M.; Gras Romero, ML.; Vidal Brotons, DJ. (2016). Rheological properties of honey from Burkina Faso: loss modulus and complex viscosity modelling. International Journal of Food Properties. 19(11):2575-2586. doi:10.1080/10942912.2015.1136938S25752586191

Real time- and control software for the new orbit measurement system for the CERN SPS

The 240 channel SPS Orbit acquisition system is implemented on a PowerPC under the LynxOS operating system, making use of multi threaded real-time capabilities. The acquired data is transferred efficiently by DMA via the PCI bus into the main memory. System configuration aspects were implemented in a Broker architecture, where individual threads communicate with an Oracle database and the acquisition systems. This Broker hides the implementation details of the front-end systems. A versatile configuration client is provided in Java, to provide both local graphical user interfaces and remote WWW access using a dedicated gateway to the SL equipment layer. The timing diagnostics of the acquisition system are provided in a LabView application integrating oscilloscope control and channel multiplex control. This paper describes in detail the technical solutions implemented and reports on the arguments, which have led to particular choices