Moving the research forward: The best of british biology using the tractable model system dictyostelium discoideum

The social amoeba Dictyostelium discoideum provides an excellent model for research across a broad range of disciplines within biology. The organism diverged from the plant, yeast, fungi and animal kingdoms around 1 billion years ago but retains common aspects found in these kingdoms. Dictyostelium has a low level of genetic complexity and provides a range of molecular, cellular, biochemical and developmental biology experimental techniques, enabling multidisciplinary studies to be carried out in a wide range of areas, leading to research breakthroughs. Numerous laboratories within the United Kingdom employ Dictyostelium as their core research model. This review introduces Dictyostelium and then highlights research from several leading British research laboratories, covering their distinct areas of research, the benefits of using the model, and the breakthroughs that have arisen due to the use of Dictyostelium as a tractable model system

Do Staphylococcus epidermidis genetic clusters predict isolation sources?

Staphylococcus epidermidis is a ubiquitous colonizer of human skin and a common cause of medical device-associated infections. The extent to which the population genetic structure of S. epidermidis distinguishes commensal from pathogenic isolates is unclear. Previously, Bayesian clustering of 437 multilocus sequence types (STs) in the international database revealed a population structure of six genetic clusters (GCs) that may reflect the species' ecology. Here, we first verified the presence of six GCs, including two (GC3 and GC5) with significant admixture, in an updated database of 578 STs. Next, a single nucleotide polymorphism (SNP) assay was developed that accurately assigned 545 (94%) of 578 STs to GCs. Finally, the hypothesis that GCs could distinguish isolation sources was tested by SNP typing and GC assignment of 154 isolates from hospital patients with bacteremia and those with blood culture contaminants and from nonhospital carriage. GC5 was isolated almost exclusively from hospital sources. GC1 and GC6 were isolated from all sources but were overrepresented in isolates from nonhospital and infection sources, respectively. GC2, GC3, and GC4 were relatively rare in this collection. No association was detected between fdh-positive isolates (GC2 and GC4) and nonhospital sources. Using a machine learning algorithm, GCs predicted hospital and nonhospital sources with 80% accuracy and predicted infection and contaminant sources with 45% accuracy, which was comparable to the results seen with a combination of five genetic markers (icaA, IS256, sesD [bhp], mecA, and arginine catabolic mobile element [ACME]). Thus, analysis of population structure with subgenomic data shows the distinction of hospital and nonhospital sources and the near-inseparability of sources within a hospital

Anti-emetic effects of thalidomide: Evidence, mechanism of action, and future directions

The rationale for using thalidomide (THD) as a treatment for nausea and vomiting during pregnancy in the late 1950s appears to have been based on its sedative or hypnotic properties. In contrast to contemporaneous studies on the anti-emetic activity of phenothiazines, we were unable to identify publications reporting preclinical or clinical evaluation of THD as an anti-emetic. Our survey of the literature revealed a clinical study in 1965 showing THD reduced vomiting in cancer chemotherapy which was substantiated by similar studies from 2000, particularly showing efficacy in the delayed phase of chemotherapy-induced nausea and vomiting. To identify the mechanism(s) potentially involved in thalidomide's anti-emetic activity we reviewed its pharmacology in the light of nausea and vomiting mechanisms and their pharmacology with a particular emphasis on chemotherapy and pregnancy. The process identified the following potential mechanisms: reduced secretion of Growth Differentiation Factor 15, suppression of inflammation/prostaglandin production, downregulation of cytotoxic drug induced upregulation of iNOS, and modulation of BK (KCa1.1) channels and GABAA/glutamate transmission at critical points in the emetic pathways (nucleus tractus solitarius, area postrema). We propose ways to investigate these hypothesized mechanisms and discuss the associated challenges (e.g., objective quantification of nausea) in addition to some of the more general aspects of developing novel drugs to treat nausea and vomiting

All You Need Is Fats-for Seizure Control: Using Amoeba to Advance Epilepsy Research

Since the original report of seizure control through starvation in the 1920s, the ketogenic diet has been considered an energy-related therapy. The diet was assumed to be functioning through the effect of reduced carbohydrate intake regulating cellular energy state, thus giving rise to seizure control. From this assumption, the generation of ketones during starvation provided an attractive mechanism for this altered energy state; however, many years of research has sought and largely failed to correlate seizure control and ketone levels. Due to this focus on ketones, few studies have examined a role for free fatty acids, as metabolic intermediates between the triglycerides provided in the diet and ketones, in seizure control. Recent discoveries have now suggested that the medium-chain fats, delivered through the medium-chain triglyceride (MCT) ketogenic diet, may provide a key therapeutic mechanism of the diet in seizure control. Here we describe an unusual pathway leading to this discovery, beginning with the use of a tractable non-animal model—Dictyostelium, through to the demonstration that medium-chain fats play a direct role in seizure control, and finally the identification of a mechanism of action of these fats and related congeners leading to reduced neural excitability and seizure control

Epitope-specific humoral responses to human cytomegalovirus glycoprotein-B vaccine with MF59 adjuvant in seropositive solid organ transplant recipients: anti-AD2 levels correlate with protection from viraemia

The human cytomegalovirus (HCMV) virion envelope protein glycoprotein B (gB) is essential for viral entry and represents a major target for humoral responses following infection. Previously, a phase-2 placebo-controlled clinical trial conducted in solid organ transplant candidates demonstrated that vaccination with gB plus MF59 adjuvant significantly increased gB ELISA antibody levels whose titer correlated directly with protection against post-transplant viremia. The aim of the current study was to investigate in more detail this protective humoral response in vaccinated seropositive transplant recipients. We focussed on four key antigenic domains (AD) of gB; AD1, AD2, AD4 and AD5 measuring antibody levels in patient sera and correlating these with post-transplant HCMV viremia. Vaccination of seropositive patients significantly boosted pre-existing antibody levels against the immunodominant region AD1 as well as against AD2, AD4 and AD5. A decreased incidence of viremia correlated with higher antibody titers against AD2 but not with antibody titers against the other three ADs. Overall, these data support the hypothesis that antibodies against AD2 are a major component of the immune protection of seropositives seen following vaccination with gB/MF59 vaccine and identify a correlate of protective immunity in allograft patients

Assessment of aeolian dust properties in the Port Hedland Area and implications for future air quality management strategies

Substantial amounts of dust are generated in Port Hedland, an iron ore handling port 1300 km north of Perth in Western Australia, with particulates in the air exceeding acceptable levels on 55 days during 2010. In addition, the incidence of respiratory hospitalisation is 30% higher in the Port Hedland region than in the rest of Western Australia. The iron ore handling operations are currently suspected to be the major cause. However the contribution from other sources is poorly documented and as the industry grows in the area so too does the number of possible sources. This pilot study reports the chemical composition of dust samples from 8 collection sites located up to 20 km from industrial facilities and compares the composition to that from 6 potential source locations. The samples were collected between the months of July and September 2010. Ion Beam Analysis was used to determine the chemical composition of the aeolian samples and Inductively Coupled Plasma – Atomic Emission Spectroscopy was used to determine the composition of the source samples. Elevated levels of Fe have been observed at all aeolian sample sites indicating widespread dispersion of iron ore dust; however when considered relative to Al, there appears to be a discrepancy between the composition of aeolian samples and iron ore products. This suggests a significant contribution from sources such as dredge spoil areas and areas disturbed by other infrastructure projects. Further study to determine the elemental make up of dust from the Port Hedland area is being undertaken to determine the contribution made by the various emission sources in the area in the event that acceptable levels of airborne particulate matter are exceeded. This will provide an accurate means of designing air quality management and dust abatement strategies for the town and the industry groups as industrial expansion occurs.© 2011-Clean Air Society of Australia & New Zealan

Phytocannabinoid-dependent mTORC1 regulation is dependent upon inositol polyphosphate multikinase activity

BACKGROUND AND PURPOSE: Cannabidiol (CBD) has been shown to differentially regulate the mechanistic target of rapamycin complex 1 (mTORC1) in preclinical models of disease, where it reduces activity in models of epilepsies and cancer and increases it in models of multiple sclerosis (MS) and psychosis. Here, we investigate the effects of phytocannabinoids on mTORC1 and define a molecular mechanism. EXPERIMENTAL APPROACH: A novel mechanism for phytocannabinoids was identified using the tractable model system, Dictyostelium discoideum. Using mouse embryonic fibroblasts, we further validate this new mechanism of action. We demonstrate clinical relevance using cells derived from healthy individuals and from people with MS (pwMS). KEY RESULTS: Both CBD and the more abundant cannabigerol (CBG) enhance mTORC1 activity in D. discoideum. We identify a mechanism for this effect involving inositol polyphosphate multikinase (IPMK), where elevated IPMK expression reverses the response to phytocannabinoids, decreasing mTORC1 activity upon treatment, providing new insight on phytocannabinoids' actions. We further validated this mechanism using mouse embryonic fibroblasts. Clinical relevance of this effect was shown in primary human peripheral blood mononuclear cells, where CBD and CBG treatment increased mTORC1 activity in cells derived from healthy individuals and decreased mTORC1 activity in cells derived from pwMS. CONCLUSION AND IMPLICATIONS: Our findings suggest that both CBD and the abundant CBG differentially regulate mTORC1 signalling through a mechanism dependent on the activity of the upstream IPMK signalling pathway, with potential relevance to the treatment of mTOR-related disorders, including MS

An Alternative Method for Thermal Cycling Test: Effect on the Marginal Microleakage and Bond Strength of Dental Polymer Bonded to Dentin

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)This study evaluated an alternative method for thermal cycling test on the microleakage and bond strength of the polymer-dentin bond. For the microleakage test the cavities were restored with a TEGDMA+UDMA+bis-EMA composite polymer light cured for 20 s. Samples were immersed in 2% methylene blue solution for 2 h and sectioned. Microleakage scores were submitted to Kruskal-Wallis test. For the shear bond strength test the adhesive was applied to dentin, photoactivated for 10 s and the composite polymer incrementally photoactivated. Samples were submitted to shear bond strength test in a machine with a cross-head speed of 0.5 mm/min and data were submitted to ANOVA and Tukey's test. Studied groups were: 1 - without thermocycling; 2 - thermocycled at 5 degrees C and 55 degrees C with intermediate bath at 37 degrees C; 3 - thermocycled at 5 degrees C and 37 degrees C; 4 - thermocycled at 37 degrees C and 55 degrees C; 5 - thermocycled at 5 degrees C and 55 degrees C (traditional test). Cold baths promoted greater microleakage when compared to control and hot bath, whereas control and hot bath were similar. Cold baths presented significant lower shear bond strength than those submitted to hot bath and control. It was concluded that the alternative method for thermal cycling test showed that cold temperatures increased the microleakage and decreased the bond strength of the polymeric adhesive.15610451049Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq