Interaction of Arabidopsis Thaliana with Plasmodiophora Brassicae

Plasmodiophora brassicae is a protistan pathogen that attacks roots of brassicaceous plant species causing devastating disease. Resistance is characterised by restriction of the pathogen and susceptibility by the development of severely malformed roots (&lsquo;clubroots&rsquo;) and stunting of the plant that is associated with alterations in the synthesis of cytokinin and auxin hormones. We are examining the susceptible response in Arabidopsis and whether suppression of key resistance factors by the pathogen contributes to susceptibility. The interaction is being studied using a number of approaches including microscopy of the infection process and development of the pathogen within roots and host gene expression analysis. Quantitative PCR was used to confirm the timing of infection of roots and showed that infection occurred at day four and colonisation increased thereafter to high levels by 23 days after inoculation by which time roots were showing systemic abnormalities. To investigate the basis of this compatible interaction we have conducted a time course experiment following infection of a susceptible ecotype of Arabidopsis (Col-0) to examine whole genome geneexpression changes in the host. Differential gene expression analysis of inoculated versus control roots showed that a higher number of genes had altered expression levels at day four compared to that at day seven and at day ten. At day four the expression levels of several genes known to be important for recognition and signal transduction in resistant interactions and genes involved in the biosynthesis of lignin, phenylpropanoids and ethylene were suppressed. Suppression by P. brassicae of specific plant defence responses appears to be a key component of susceptibility in this system.<br /

Studies on physiologic specialisation of albugo candida causing white blister on broccoli in Victoria

White blister caused by oomycete Albugo candida (Pers. Ex. Lev.) Kuntze, (AC), is an important disease affecting many cruciferous hosts, including vegetable brassicas. The outbreaks of white blister in broccoli and cauliflower crops (Brassica oleracea var. italica and var. botrytis) in Southern Australia in the last three years led to restrictions on movement of fresh produce and seedlings from the disease-affected areas. Current classification of AC races is based on physiologic specialisation of this pathogen. Race 9 has been identified to cause white blister on B. oleracea in the USA. We report on specialisation of AC causing disease in Victorian broccoli crops and the use of molecular tools for the separation of AC races. In a glasshouse, 12 Brassicaceae species/varieties replicated 6 times, were inoculated twice at the fully developed cotyledon stage with a distilled water suspension of zoosporangia (1x104 per ml) collected from a single broccoli leaf. Two weeks after inoculation the incidence of white blister on cotyledons and seedling leaves of cauliflower, broccoli, black mustard and Indian mustard was 79.7, 78.4, 73.7 and 6.9% respectively. Cabbage plants were symptomless indicating that further specialisation of the pathogen may have occurred in Australia. High disease incidence among black mustard plants shows that the Australian isolate differs from overseas AC race 9. The interaction of a number of B. oleracea varieties to a range of AC isolates from various hosts will be investigated. Degenerate primers are now being used to amplify actin and &beta;-tubulin genes to identify race specific polymorphisms in AC isolates from three different hosts (wild radish, Chinese cabbage, and broccoli). Differing PCR amplification efficiencies from broccoli and wild radish isolates using degenerate actin primers indicates sequence differences in the two isolates. The fragments are now being cloned and sequenced for race comparison.<br /

Investigation of plasmodiophora brassicae (clubroot disease) in vegetable brassica using arabidopsis thaliana as a model system

Clubroot, caused by Plasmodiophora brassicae, is the most devastating soil-borne disease of vegetable brassicas. It occurs all over the world and is responsible for crop losses of up to 10% every year. In Australia, the disease is being managed effectively with chemicals and cultural practices, but ideally control can be improved in the long term by the introduction of resistant cultivars. The life cycle ofP. brassicae and mode of action of plant resistance has not been fully elucidated because of the technical difficulties of working with an obligate, soil-borne plant pathogen. However, Arabidopsis thaliana, which is a host ofP. brassicae, has great potential as a model system for studying the life cycle, the infection process and development of resistance. We have developed a sand-liquid-culture system for growing Arabidopsis that allows easy observation of all life stages and, most importantly, the primary plasmodial stages within the root hair. The method was first optimised for observations of the lifecycle of the pathogen in a susceptible Arabidopsis ecotype (Col-3) where all stages of the lifecycle have now been observed and characterised. Further screening of Arabidopsis ecotypes for disease resistance has utilised one of the most virulent Australian pathotypes of brassica (ECD number 16/19/31). To date, Arabidopsis ecotype Ta-0 has shown a level of tolerance to the disease even though the roots get infected. It has been reported earlier that resistance toP. brassicae in Arabidopsis is due to one or a small number of genes. To examine changes in gene expression during the early, critical stages of infection, RNA was extracted from the susceptible and resistant ecotypes at two time points, 4 days and 17 days after inoculation. Microarray analysis will be used to investigate genome wide changes in gene expression during infection but also to identify candidate genes that may confer resistance to Australian isolates of the pathogen.<br /

Phytochemical investigations on Artemisia alba Turra growing in the North-East of Italy

Artemisia alba Turra (Asteraceae) is an Euro-Mediterranean plant used in Veneto (North-East of Italy) as traditional medicine for the treatment of various diseases. A. alba is a taxonomically problematic species, characterized by common polymorphism leading to a quite high variability in secondary metabolites content. Nonetheless, the phytochemical knowledge on its phytoconstituents, especially non-volatile components, is limited. In the present paper, the phytochemical composition of a tincture obtained from the aerial parts of A. alba growing in Veneto is presented. Extensive chromatographic separations led to the isolation of three new sesquiterpene derivatives, whose structures were elucidated by 1D and 2D NMR experiments and mass spectrometry. Furthermore, flavonoid composition and volatile constituents of the tincture of A. alba were preliminary studied by HPLC-MSn and GC-MS, respectivel

Efficacy of pulsatile flow perfusion in adult cardiac surgery: Hemodynamic energy and vascular reactivity

Background: The role of pulsatile (PP) versus non-pulsatile (NP) flow during a cardiopulmonary bypass (CPB) is still debated. This study’s aim was to analyze hemodynamic effects, endothelial reactivity and erythrocytes response during a CPB with PP or NP. Methods: Fifty-two patients undergoing an aortic valve replacement were prospectively randomized for surgery with either PP or NP flow. Pulsatility was evaluated in terms of energy equivalent pressure (EEP) and surplus hemodynamic energy (SHE). Systemic (SVRi) and pulmonary (PVRi) vascular resistances, endothelial markers levels and erythrocyte nitric-oxide synthase (eNOS) activity were collected at different perioperative time-points. Results: In the PP group, the resultant EEP was 7.3% higher than the mean arterial pressure (MAP), which corresponded to 5150 ± 2291 ergs/cm3 of SHE. In the NP group, the EEP and MAP were equal; no SHE was produced. The PP group showed lower SVRi during clamp-time (p = 0.06) and lower PVRi after protamine administration and during first postoperative hours (p = 0.02). Lower SVRi required a higher dosage of norepinephrine in the PP group (p = 0.02). Erythrocyte eNOS activity results were higher in the PP patients (p = 0.04). Renal function was better preserved in the PP group (p = 0.001), whereas other perioperative variables were comparable between the groups. Conclusions: A PP flow during a CPB results in significantly lower SVRi, PVRi and increased eNOS production. The clinical impact of increased perioperative vasopressor requirements in the PP group deserves further evaluation

Is it possible to assess the best mitral valve repair in the individual patient? Preliminary results of a finite element study from magnetic resonance imaging data

ObjectivesFinite element modeling was adopted to quantitatively compare, for the first time and on a patient-specific basis, the biomechanical effects of a broad spectrum of different neochordal implantation techniques for the repair of isolated posterior mitral leaflet prolapse.MethodsCardiac magnetic resonance images were acquired from 4 patients undergoing surgery. A patient-specific 3-dimensional model of the mitral apparatus and the motion of the annulus and papillary muscles were reconstructed. The location and extent of the prolapsing region were confirmed by intraoperative findings, and the mechanical properties of the mitral leaflets, chordae tendineae and expanded polytetrafluoroethylene neochordae were included. Mitral systolic biomechanics was simulated under preoperative conditions and after 5 different neochordal procedures: single neochorda, double neochorda, standard neochordal loop with 3 neochordae of the same length and 2 premeasured loops with 1 common neochordal loop and 3 different branched neochordae arising from it, alternatively one third and two thirds of the entire length.ResultsThe best repair in terms of biomechanics was achieved with a specific neochordal technique in the single patient, according to the location of the prolapsing region. However, all techniques achieved a slight reduction in papillary muscle forces and tension relief in intact native chordae proximal to the prolapsing region. Multiple neochordae implantation improved the repositioning of the prolapsing region below the annular plane and better redistributed mechanical stresses on the leaflet.ConclusionsAlthough applied on a small cohort of patients, systematic biomechanical differences were noticed between neochordal techniques, potentially affecting their short- to long-term clinical outcomes. This study opens the way to patient-specific optimization of neochordal techniques

Optimal fishery with coastal catch

In many spatial resource models, it is assumed that an agent is able to harvest the resource over the complete spatial domain. However, agents frequently only have access to a resource at particular locations at which a moving biomass, such as fish or game, may be caught or hunted. Here, we analyze an infinite time‐horizon optimal control problem with boundary harvesting and (systems of) parabolic partial differential equations as state dynamics. We formally derive the associated canonical system, consisting of a forward–backward diffusion system with boundary controls, and numerically compute the canonical steady states and the optimal time‐dependent paths, and their dependence on parameters. We start with some one‐species fishing models, and then extend the analysis to a predator–prey model of the Lotka–Volterra type. The models are rather generic, and our methods are quite general, and thus should be applicable to large classes of structurally similar bioeconomic problems with boundary controls. Recommedations for Resource Managers Just like ordinary differential equation‐constrained (optimal) control problems and distributed partial differential equation (PDE) constrained control problems, boundary control problems with PDE state dynamics may be formally treated by the Pontryagin's maximum principle or canonical system formalism (state and adjoint PDEs). These problems may have multiple (locally) optimal solutions; a first overview of suitable choices can be obtained by identifying canonical steady states. The computation of canonical paths toward some optimal steady state yields temporal information about the optimal harvesting, possibly including waiting time behavior for the stock to recover from a low‐stock initial state, and nonmonotonic (in time) harvesting efforts. Multispecies fishery models may lead to asymmetric effects; for instance, it may be optimal to capture a predator species to protect the prey, even for high costs and low market values of the predators

Surgical management and outcomes in patients with acute type A aortic dissection and cerebral malperfusion

Objective: The study objective was to evaluate the surgical results in patients with acute type A aortic dissection and cerebral malperfusion. Methods: From 2000 to 2019, 234 patients with type A aortic dissection and cerebral malperfusion were stratified into 3 groups: 50 (21%) with syncope (group 1), 152 (65%) with persistent loss of focal neurological function (group 2), and 32 (14%) with coma (group 3). Results were evaluated and compared by univariable and multivariable analyses. Results: Median age was higher in group 1, and incidence of cardiogenic shock was higher in group 3. The femoral artery was the most common cannulation site, whereas the axillary artery was used in 18% of group 1, 30% of group 2, and 25% of group 3 patients (P = .337). Antegrade cerebral perfusion was performed in more than 80% of patients, and ascending aorta/arch replacement was performed in 40% of group 1, 27% of group 2, and 31% of group 3 (P = .21). In-hospital mortality was 18% in group 1, 27% in group 2, and 56% in group 3 (P = .001). Survival at 5 years is 57.0% in group 1, 57.7% in group 2, and 38.7% in group 3 (P = .0005). On multivariable analysis, age, cardiopulmonary bypass time, and group 3 versus group 2 were independent risk factors for mortality, whereas axillary cannulation was a protective factor. Conclusions: Patients with aortic dissection and cerebral malperfusion without preoperative coma showed acceptable mortality, and those with coma had a high in-hospital mortality regardless of the type of brain protection. Overall axillary artery cannulation appeared to be a protective factor

Influence of Myocardial Fibrosis on Left Ventricular Hypertrophy in Patients with Symptomatic Severe Aortic Stenosis.

Aim: It was the aim of our study to determine whether myocardial fibrosis influences physiologic or non-physiologic left ventricular (LV) hypertrophy in severe aortic stenosis. Methods: Myocardial fibrosis was evaluated using specimens taken from the ventricular septum in 79 patients submitted to aortic valve replacement because of symptomatic aortic stenosis. Patients were considered to have physiologic LV hypertrophy if end-systolic wall stress, evaluated by echocardiography, was 90 kdyn/cm(2) were considered to have non-physiologic hypertrophy. Results: Fibrosis tissue mass index was significantly inversely related with LV fractional shortening and directly related with LV diastolic and systolic diameter and LV mass index (LVMI). Patients with non-physiologic hypertrophy (n = 24) had a higher LVMI due to larger LV diastolic and systolic diameters with thinner wall, resulting in lower relative wall thickness. These patients had a higher fibrosis tissue mass index and impaired LV systolic and diastolic functions, as suggested by lower LV fractional shortening and higher mean wedge pressure. At follow-up of 7.4 \ub1 2.1 months, the LVMI and New York Heart Association class remained higher in patients with non-physiologic hypertrophy. Conclusions: Our study suggests a different quality of hypertrophies in patients with aortic stenosis, where myocardial fibrosis seems to be the critical abnormality that differentiates adaptive from maladaptive response to increased afterload