The growth of cognition: Free energy minimization and the embryogenesis of cortical computation.

The assumption that during cortical embryogenesis neurons and synaptic connections are selected to form an ensemble maximising synchronous oscillation explains mesoscopic cortical development, and a mechanism for cortical information processing is implied by consistency with the Free Energy Principle and Dynamic Logic. A heteroclinic network emerges, with stable and unstable fixed points of oscillation corresponding to activity in symmetrically connected, versus asymmetrically connected, sets of neurons. Simulations of growth explain a wide range of anatomical observations for columnar and non-columnar cortex, superficial patch connections, and the organization and dynamic interactions of neurone response properties. An antenatal scaffold is created, upon which postnatal learning can establish continuously ordered neuronal representations, permitting matching of co-synchronous fields in multiple cortical areas to solve optimization problems as in Dynamic Logic. Fast synaptic competition partitions equilibria, minimizing "the curse of dimensionality", while perturbations between imperfectly partitioned synchronous fields, under internal reinforcement, enable the cortex to become adaptively self-directed. As learning progresses variational free energy is minimized and entropy bounded.

Effect of cold plasma on polyphenol oxidase inactivation in cloudy apple juice and on the quality parameters of the juice during storage.

Direct cold plasma treatment has been investigated as an alternative non-thermal technology as a means of maintaining and improving quality of fresh cloudy apple juice. Process variables studied included type of plasma discharge, input voltage and treatment time on polyphenol oxidase (PPO) inactivation. Spark discharge plasma at 10.5 kV for 5 min was the best treatment, with near total inactivation of PPO achieved, although good PPO inactivation was also recorded using shorter treatment times. Residual activity (RA) of PPO was 16 and 27.6% after 5 and 4 min of treatment respectively. This PPO inactivation was maintained throughout the storage trials, but decreased with samples treated for a shorter time. Plasma treatment improved key quality parameters of Golden delicious cloudy apple juice, with retention of critical quality parameters during extended storage trials. Color was the most noticeable change, which was enhanced with retention of a greener color. An increase of 69 and 64% was obtained in the total phenolic content after 4 and 5 min of treatment, respectively. Therefore, cold plasma was demonstrated to be a good alternative to traditional heat treatments for enhanced quality retention of fresh cloudy apple juice and over its storage.

A high-quality genome sequence of Rosa chinensis to elucidate ornamental traits.

Rose is the world's most important ornamental plant, with economic, cultural and symbolic value. Roses are cultivated worldwide and sold as garden roses, cut flowers and potted plants. Roses are outbred and can have various ploidy levels. Our objectives were to develop a high-quality reference genome sequence for the genus Rosa by sequencing a doubled haploid, combining long and short reads, and anchoring to a high-density genetic map, and to study the genome structure and genetic basis of major ornamental traits. We produced a doubled haploid rose line ('HapOB') from Rosa chinensis 'Old Blush' and generated a rose genome assembly anchored to seven pseudo-chromosomes (512 Mb with N50 of 3.4 Mb and 564 contigs). The length of 512 Mb represents 90.1-96.1% of the estimated haploid genome size of rose. Of the assembly, 95% is contained in only 196 contigs. The anchoring was validated using high-density diploid and tetraploid genetic maps. We delineated hallmark chromosomal features, including the pericentromeric regions, through annotation of transposable element families and positioned centromeric repeats using fluorescent in situ hybridization. The rose genome displays extensive synteny with the Fragaria vesca genome, and we delineated only two major rearrangements. Genetic diversity was analysed using resequencing data of seven diploid and one tetraploid Rosa species selected from various sections of the genus. Combining genetic and genomic approaches, we identified potential genetic regulators of key ornamental traits, including prickle density and the number of flower petals. A rose APETALA2/TOE homologue is proposed to be the major regulator of petal number in rose. This reference sequence is an important resource for studying polyploidization, meiosis and developmental processes, as we demonstrated for flower and prickle development. It will also accelerate breeding through the development of molecular markers linked to traits, the identification of the genes underlying them and the exploitation of synteny across Rosaceae.

Microbiological interactions with cold plasma.

There is a diverse range of microbiological challenges facing the food, healthcare and clinical sectors. The increasing and pervasive resistance to broad-spectrum antibiotics and health-related concerns with many biocidal agents drives research for novel and complementary antimicrobial approaches. Biofilms display increased mechanical and antimicrobial stability and are the subject of extensive research. Cold plasmas (CP) have rapidly evolved as a technology for microbial decontamination, wound healing and cancer treatment, owing to the chemical and bio-active radicals generated known collectively as reactive oxygen and nitrogen species. This review outlines the basics of CP technology and discusses the interactions with a range of microbiological targets. Advances in mechanistic insights are presented and applications to food and clinical issues are discussed. The possibility of tailoring CP to control specific microbiological challenges is apparent. This review focuses on microbiological issues in relation to food- and healthcare-associated human infections, the role of CP in their elimination and the current status of plasma mechanisms of action.

Different visual stimuli affect body reorientation strategies during sidestepping.

Sidestepping in response to unplanned stimuli is a high-risk maneuver for anterior cruciate ligament (ACL) injuries. Yet, differences in body reorientation strategies between high- and low-level soccer players prior to sidestepping in response to quasi-game-realistic vs non-game-realistic stimuli, remain unknown. Fifteen high-level (semi-professional) and 15 low-level (amateur) soccer players responded to a quasi-game-realistic one-defender scenario (1DS) and two-defender scenario (2DS), and non-game-realistic arrow-planned condition (AP) and arrow-unplanned condition (AUNP). The AP, 1DS, 2DS to AUNP represented increasing time constraints to sidestep. Selected biomechanics from the penultimate step to foot-off were assessed using a mixed-model (stimuli × skill) ANOVA (P < 0.05). Step length decreased in the defender scenarios compared with the arrow conditions. Support foot placement increased laterally, away from mid-pelvis, with increasing temporal constraints. Greater trunk lateral flexion in the 1DS, 2DS, and AUNP has been associated with ACL injury onsets. Higher level players pushed off closer to their pelvic midline at initial foot contact in the 2DS especially. Higher level perception of game-realistic visual information could have contributed to this safer neuromuscular strategy that, when understood better, could potentially be trained in lower level players to reduce ACL injury risk associated with dangerous sidestepping postures.

Assessing stress responses to atmospheric cold plasma exposure using Escherichia coli knock-out mutants.

AIMS: This study investigated the effect of atmospheric cold plasma (ACP) exposure-induced stress on microbial inactivation patterns and the regulation of genes involved in the microbial stress response in conjunction with key processing parameters of exposure time and post-treatment storage time. METHODS AND RESULTS: Cell suspensions of Escherichia coli BW 25113 and its isogenic knock-out mutants in rpoS, soxR, soxS, oxyR and dnaK genes were treated with high-voltage ACP in a sealed package for 1, 3 and 5 min followed by 0-, 1- and 24-h post-treatment storage. Reactive oxygen species (ROS) densities and colony formation were determined. ΔrpoS strain showed higher microbial reduction and greater cell permeability than other mutants, while ΔoxyR only showed this effect after 5 min of treatment. With increased post-treatment storage time, ΔsoxS and ΔsoxR had increased sensitivity and resistance respectively. ΔdnaK cell suspensions had much higher ROS than other strains and showed increased sensitivity with 24 h post-treatment storage. CONCLUSIONS: RpoS and oxyR genes have both short-term and long-term regulatory effects under plasma stress. However, knocking out dnaK gene had an immediate response on ROS scavenging and long-term repairing mechanisms. ΔsoxR and ΔsoxS had different responses to ACP treatment with the increase in post-treatment time in relation to clearance of reactive species implying the different characteristics and functions as subunits. SIGNIFICANCE AND IMPACT OF THE STUDY: By comparing the response of mutants under ACP exposure to key processing parameters, the mechanism of microbial inactivation was partly revealed with respect to cellular regulation and repairing genes.

Mechanisms of Inactivation by High-Voltage Atmospheric Cold Plasma Differ for Escherichia coli and Staphylococcus aureus.

Atmospheric cold plasma (ACP) is a promising nonthermal technology effective against a wide range of pathogenic microorganisms. Reactive oxygen species (ROS) play a crucial inactivation role when air or other oxygen-containing gases are used. With strong oxidative stress, cells can be damaged by lipid peroxidation, enzyme inactivation, and DNA cleavage. Identification of ROS and an understanding of their role are important for advancing ACP applications for a range of complex microbiological issues. In this study, the inactivation efficacy of in-package high-voltage (80 kV [root mean square]) ACP (HVACP) and the role of intracellular ROS were investigated. Two mechanisms of inactivation were observed in which reactive species were found to either react primarily with the cell envelope or damage intracellular components. Escherichia coli was inactivated mainly by cell leakage and low-level DNA damage. Conversely, Staphylococcus aureus was mainly inactivated by intracellular damage, with significantly higher levels of intracellular ROS observed and little envelope damage. However, for both bacteria studied, increasing treatment time had a positive effect on the intracellular ROS levels generated.

Influence of high voltage atmospheric cold plasma process parameters and role of relative humidity on inactivation of Bacillus atrophaeus spores inside a sealed package.

BACKGROUND: Non-thermal plasma has received much attention for elimination of microbial contamination from a range of surfaces. AIM: This study aimed to determine the effect of a range of dielectric barrier discharge high voltage atmospheric cold plasma (HVACP) parameters for inactivation of Bacillus atrophaeus spores inside a sealed package. METHODS: A sterile polystyrene Petri dish containing B. atrophaeus spore strip (spore population 2.3 × 10(6)/strip i.e. 6.36 log10/strip) was placed in a sealed polypropylene container and was subjected to HVACP treatment. The HVACP discharge was generated between two aluminium plate electrodes using a high voltage of 70kVRMS. The effects of process parameters, including treatment time, mode of exposure (direct/indirect), and working gas types, were evaluated. The influence of relative humidity on HVACP inactivation efficacy was also assessed. The inactivation efficacy was evaluated using colony counts. Optical absorption spectroscopy (OAS) was used to assess gas composition following HVACP exposure. FINDINGS: A strong effect of process parameters on inactivation was observed. Direct plasma exposure for 60s resulted in ≥6 log10 cycle reduction of spores in all gas types tested. However, indirect exposure for 60s resulted in either 2.1 or 6.3 log10 cycle reduction of spores depending on gas types used for HVACP generation. The relative humidity (RH) was a critical factor in bacterial spore inactivation by HVACP, where a major role of plasma-generated species other than ozone was noted. Direct and indirect HVACP exposure for 60s at 70% RH recorded 6.3 and 5.7 log10 cycle reduction of spores, respectively. CONCLUSION: In summary, a strong influence of process parameters on spore inactivation was noted. Rapid in-package HVACP inactivation of bacterial spores within 30-60s demonstrates the promising potential application for reduction of spores on medical devices and heat-sensitive materials.

Atmospheric cold plasma inactivation of Escherichia coli, Salmonella enterica serovar Typhimurium and Listeria monocytogenes inoculated on fresh produce.

Atmospheric cold plasma (ACP) represents a potential alternative to traditional methods for non-thermal decontamination of foods. In this study, the antimicrobial efficacy of a novel dielectric barrier discharge ACP device against Escherichia coli, Salmonella enterica Typhimurium and Listeria monocytogenes inoculated on cherry tomatoes and strawberries, was examined. Bacteria were spot inoculated on the produce surface, air dried and sealed inside a rigid polypropylene container. Samples were indirectly exposed (i.e. placed outside plasma discharge) to a high voltage (70 kVRMS) air ACP and subsequently stored at room temperature for 24 h. ACP treatment for 10, 60 and 120 s resulted in reduction of Salmonella, E. coli and L. monocytogenes populations on tomato to undetectable levels from initial populations of 3.1, 6.3, and 6.7 log10 CFU/sample, respectively. However, an extended ACP treatment time was necessary to reduce bacterial populations attached on the more complex surface of strawberries. Treatment time for 300 s resulted in reduction of E. coli, Salmonella and L. monocytogenes populations by 3.5, 3.8 and 4.2 log10 CFU/sample, respectively, and also effectively reduced the background microflora of tomatoes.

Role of lateral and feedback connections in primary visual cortex in the processing of spatiotemporal regularity - a TMS study.

Our human visual system exploits spatiotemporal regularity to interpret incoming visual signals. With a dynamic stimulus sequence of four collinear bars (predictors) appearing consecutively toward the fovea, followed by a target bar with varying contrasts, we have previously found that this predictable spatiotemporal stimulus structure enhances target detection performance and its underlying neural process starts in the primary visual cortex (area V1). However, the relative contribution of V1 lateral and feedback connections in the processing of spatiotemporal regularity remains unclear. In this study we measured human contrast detection of a briefly presented foveal target that was embedded in a dynamic collinear predictor-target sequence. Transcranial magnetic stimulation (TMS) was used to selectively disrupt V1 horizontal and feedback connections in the processing of predictors. The coil was positioned over a cortical location corresponding to the location of the last predictor prior to target onset. Single-pulse TMS at an intensity of 10% below phosphene thresholdwas delivered at 20 or 90ms after the predictor onset. Our analysis revealed that the delivery of TMS at both time windows equally reduced, but did not abolish, the facilitation effect of the predictors on target detection. Furthermore, if the predictors' ordination was randomized to suppress V1 lateral connections, the TMS disruption was significantly more evident at 20ms than at 90-ms time window. We suggest that both lateral and feedback connections contribute to the encoding of spatiotemporal regularity in V1. These findings develop understanding of how our visual system exploits spatiotemporal regularity to facilitate the efficiency of visual perception.

In situ production of human ß defensin-3 in lager yeasts provides bactericidal activity against beer-spoiling bacteria under fermentation conditions.

AIMS: To examine the use of a natural antimicrobial peptide, human ß-defensin-3 (HBD3), as a means of preventing spoilage from bacterial contamination in brewery fermentations and in bottled beer. METHODS AND RESULTS: A chemically synthesised HBD3 peptide was tested for bactericidal activity against common Gram-positive and Gram-negative beer-spoiling bacteria, including species of Lactobacillus, Pediococcus and Pectinatus. The peptide was effective at the µmol l(-1) range in vitro, reducing bacterial counts by 95%. A gene construct encoding a secretable form of HBD3 was integrated into the genome of the lager yeast Saccharomyces pastorianus strain CMBS-33. The integrated gene was expressed under fermentation conditions and was secreted from the cell into the medium, but a significant amount remains associated with yeast cell surface. We demonstrate that under pilot-scale fermentation conditions, secreted HBD3 possesses bactericidal activity against beer-spoiling bacteria. Furthermore, when added to bottled beer, a synthetic form of HBD3 reduces the growth of beer-spoiling bacteria. CONCLUSIONS: Defensins provide prophylactic protection against beer-spoiling bacteria under brewing conditions and also in bottled beer. SIGNIFICANCE AND IMPACT OF THE STUDY: The results have direct application to the brewing industry where beer spoilage due to bacterial contamination continues to be a major problem in breweries around the world.

Bacterial inactivation by high-voltage atmospheric cold plasma: influence of process parameters and effects on cell leakage and DNA.

AIMS: This study investigated a range of atmospheric cold plasma (ACP) process parameters for bacterial inactivation with further investigation of selected parameters on cell membrane integrity and DNA damage. The effects of high voltage levels, mode of exposure, gas mixture and treatment time against Escherichia coli and Listeria monocytogenes were examined. METHODS AND RESULTS: 10(8) CFU ml(-1) E. coli ATCC 25922, E. coli NCTC 12900 and L. monocytogenes NCTC11994 were ACP-treated in 10 ml phosphate-buffered saline (PBS). Working gas mixtures used were air (gas mix 1), 90% N2 + 10% O2 (gas mix 2) and 65% O2 + 30% CO2 + 5% N2 (gas mix 3). Greater reduction of viability was observed for all strains using higher voltage of 70 kVRMS and with working gas mixtures with higher oxygen content in combination with direct exposure. Indirect ACP exposure for 30 s inactivated below detection level both E. coli strains. L. monocytogenes inactivation within 30 s was irrespective of the mode of exposure. Leakage was assessed using A260 absorbance, and DNA damage was monitored using PCR and gel electrophoresis. Membrane integrity was compromised after 5 s, with noticeable DNA damage also dependent on the target cell after 30 s. CONCLUSIONS: Plasma treatment was effective for inactivation of challenge micro-organisms, with a greater sensitivity of L. monocytogenes noted. Different damage patterns were observed for the different bacterial strains attributed to the membrane structure and potential resistance mechanisms. SIGNIFICANCE AND IMPACT OF THE STUDY: Using atmospheric air as working gas resulted in useful inactivation by comparison with high nitrogen or high oxygen mix. The mechanism of inactivation was a function of treatment duration and cell membrane characteristics, thus offering potential for optimized process parameters specific to the microbial challenge.

Atmospheric cold plasma inactivation of Escherichia coli in liquid media inside a sealed package.

AIMS: The main objective of this study was to determine the inactivation efficacy of dielectric barrier discharge atmospheric cold plasma (DBD-ACP) generated inside a sealed package for Escherichia coli ATCC 25922. METHODS AND RESULTS: A plasma discharge was generated between two circular aluminium electrodes at 40 kV. E. coli suspensions (10(7) CFU ml(-1)) in either maximum recovery diluent (MRD) or phosphate buffered saline (PBS) were treated in a 96-well microtitre plate inside a sealed package. The effects of treatment time, post-treatment storage time, either direct or indirect samples exposure to the plasma discharge and suspension media were studied. Regardless of the media tested, 20 s of direct and 45 s of indirect plasma treatment resulted in complete bacterial inactivation (7 log CFU ml(-1)). At the lower plasma treatment times (10-30 s) investigated, the effects of suspension media and mode of exposure on the inactivation efficacy were evident. The inactivation efficacy was also influenced by the post-treatment storage time. CONCLUSIONS: It was demonstrated that the novel DBD-ACP can inactivate high concentrations of E. coli suspended in liquids within sealed packages in seconds. SIGNIFICANCE AND IMPACT OF THE STUDY: A key advantage of this in-package nonthermal novel disinfection approach is the elimination of post-processing contamination.

Focus on systemic lupus erythematosus in indigenous Australians: towards a better understanding of autoimmune diseases.

The incidence and prevalence of autoimmune diseases such as rheumatoid arthritis, primary Sjögren syndrome, scleroderma and systemic lupus erythematosus (SLE) varies with geography and ethnicity. For example, SLE is reported to be more common in populations such as African-Caribbeans and Indigenous Australians (IA). As well as socio-economic status, variation in severity of disease may also show ethnic variability. The initial presentation of SLE in IA, in the context of a unique genetic background and distinctive environmental influences, is often florid with a recurring spectrum of clinical phenotypes. These clinical observations suggest a unique pathway for autoimmunity pathogenesis in this population. For instance, the high prevalence of bacterial infections in IA, particularly group A streptococcus, may be a potential explanation not only for increased incidence and prevalence of SLE but also the commonly florid acute disease presentation and propensity for rapidly progressive end organ threatening disease. This article will review the state of research in autoimmune disease of IA, consider key findings related to autoimmune disease in this population and propose a model potentially to explain the involvement of innate immunity and chronic infection in autoimmune disease pathogenesis. Ultimately, understanding of SLE at this level could affect management and result in personalised and targeted therapies to improve the health status of IA as well as better understanding of SLE pathogenesis per se.

Zygomycosis over-infection during voriconazole therapy for aspergillosis in a heart transplant patient, successfully treated with liposomal amphotericin and posaconazole.

Aspergillosis and zygomycosis are life-threatening fungal infections in immunocompromised patients. We report a heart transplant recipient with an early pulmonary invasive aspergillosis successfully treated with association of voriconazole and caspofungin. Zygomycosis sinusitis, which was diagnosed while he still was on voriconazole therapy, was successfully treated with the use of combination antifungal therapy including liposomal amphotericin plus posaconazole and conservative surgical debridement.

Assessing the microbial oxidative stress mechanism of ozone treatment through the responses of Escherichia coli mutants.

AIMS: To investigate the effect of the oxidative stress of ozone on the microbial inactivation, cell membrane integrity and permeability and morphology changes of Escherichia coli. METHODS AND RESULTS: Escherichia coli BW 25113 and its isogenic mutants in soxR, soxS, oxyR, rpoS and dnaK genes were treated with ozone at a concentration of 6 µg ml⁻¹ for a period up to 240 s. A significant effect of ozone exposure on microbial inactivation was observed. After ozonation, minor effects on the cell membrane integrity and permeability were observed, while scanning electron microscopy analysis showed slightly altered cell surface structure. CONCLUSIONS: The results of this study suggest that cell lysis was not the major mechanism of microbial inactivation. The deletion of oxidative stress-related genes resulted in increased susceptibility of E. coli cells to ozone treatment, implying that they play an important role for protection against the radicals produced by ozone. However, DnaK that has previously been shown to protect against oxidative stress did not protect against ozone treatment in this study. Furthermore, RpoS was important for the survival against ozone. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides important information about the role of oxidative stress in the responses of E. coli during ozonation.

The incidence of spinal cord ischaemia following thoracic and thoracoabdominal aortic endovascular intervention.

OBJECTIVES: To determine the incidence and risk factors for spinal cord ischaemia (SCI) following thoracic and thoracoabdominal aortic intervention. METHODS: A prospective database of all thoracic and thoracoabdominal aortic interventions between 2001 and 2009 was used to investigate the incidence of SCI. All elective and emergency cases for all indications were included. Logistic regression was used to investigate which factors were associated with SCI. RESULTS: 235 patients underwent thoracic aortic stent grafting; 111(47%) thoracic aortic stent-grafts alone, with an additional 14(6%) branched or fenestrated thoracic grafts, 30(13%) arch hybrid procedures and 80(34%) visceral hybrid surgical and endovascular procedures. The global incidence of SCI for all procedures was 23/235 (9.8%) and this included emergency indications (ruptured TAAA and acute complex dissections) but the incidence varied considerably between types of procedures. Of the 23 cases, death occurred in 4 patients but recovery of function was seen in 6. Thus, permanent paraplegia occurred in 13/235 (5.5%) patients. Of the nine pre-specified factors investigated for association with SCI, only percentage of aortic coverage was significantly associated with the incidence of SCI; adjusted odds ratio per 10% increase in aorta covered=1.78[95% CI 1.18-2.71], p=0.007. The procedures in patients who developed SCI took longer (463.5 versus 307.2 minutes) and utilised more stents (4 versus 2). CONCLUSION: SCI following thoracic and thoracoabdominal aortic endovascular intervention is associated with the proportion of aorta covered. The degree of risk varies between different types of procedure and this should be carefully considered in both selection and consenting of patients.

Inactivation of Escherichia coli by ozone treatment of apple juice at different pH levels.

This research investigated the efficacy of gaseous ozone on the inactivation of Escherichia coli ATCC 25922 and NCTC 12900 strains in apple juice of a range of pH levels, using an ozone bubble column. The pH levels investigated were 3.0, 3.5, 4.0, 4.5 and 5.0. Apple juice inoculated with E. coli strains (10(6)CFU/mL) was treated with ozone gas at a flow rate of 0.12L/min and ozone concentration of 0.048 mg/min/mL for up to 18 min. Results show that inactivation kinetics of E. coli by ozone were affected by pH of the juice. The ozone treatment duration required for achieving a 5-log reduction was faster (4 min) at the lowest pH than at the highest pH (18 min) studied. The relationship between time required to achieve 5log reduction (t(5d)) and pH for both strains was described mathematically by two exponential equations. Ozone treatment appears to be an effective process for reducing bacteria in apple juice and the required applied treatment for producing a safe apple juice is dependant on its acidity level.