Role of the yndE genes in the germination of Bacillus licheniformis spores

The risk of food poisoning and illness is an important area of research. How to prevent the presence of microorganisms in food products and the production of toxins causing illness is of large interest. Spore forming bacteria of the genera Bacillus cause problems in food production industry due to their role in pathogenesis in both humans and domestic animals. They also represent a significant food spoilage problem in the food industry where heat treatment is insufficient to kill the spores, while competition from other vegetative bacteria is eliminated. Later years, the demand for low processed refrigerated food with long durability has become a big issue. To maintain the quality of the food, the sterilization treatments need to be effective but have minimal impact on the foods sensory qualities. Knowledge of the germination receptors, present in the spores inner membrane, and their role in initiating the germination process has been shown of major importance for understanding the germination behavior of spores. Bacillus licheniformis, has been associated with food borne outbreaks in cooked meats and vegetables, raw milk, and industrially produced baby food. One of the germination receptors of B.licheniformis is encoded by the yndDEF operon. Our unpublished results demonstrate that the Ynd germination receptor is involved in germination in response to L-alanine, L-cysteine and L-valine. In contrast to other Bacillus species, like Bacillus subtilis, B.licheniformis seems to have three homologous yndE genes, yndE1, yndE2 and yndE3. The function of the yndE genes is unknown. This project aims to characterize the three homologous yndE genes in order to understand their role in germination of B.licheniformis spores. Attempts were made to construct separate null mutants in all three yndE homologs but only the yndE3 gene was successfully deleted. Analysis of the germination properties of the yndE3 mutant in response to L-alanine, L-cysteine and L-valine showed that the mutant strain exhibited a strongly reduced germination rate in response to L-cysteine. The germination responses to L-alanine and L-valine were also slightly reduced. This result indicate that the yndE3 gene is essential for efficient L-cysteine induced germination and suggest that the other yndE genes may be involved in Ynd dependent germination in response to L-alanine and L-valine. To our knowledge this is the first functional characterization of the role of paralogous germination receptor subunits encoded by a single germination receptor operon.Sykdomsutbrudd relatert til matforgiftning er et viktig forskningsfelt. Hvordan man kan hindre vekst av mikroorganismer i matvarer, samt toksin-produksjon, som fører til sykdom, er av stor interesse. Bakterier i Bacillusslekten, som danner sporer, er et stort problem i matproduksjon grunnet den sykdomsfremkallende rollen de har hos mennesker og husdyr. Sporedannere representer også et stort problem for matvareindustrien, siden varmebehandling ikke bare er utilstrekkelig i forhold til å drepe disse, men i tillegg eliminerer konkurransen ved å drepe andre ikke sporedannende bakterier. I de senere årene har etterspørselen etter lite prosessert mat med lang holdbarhet økt. For å kunne imøtekomme disse kravene er det nødvendig med effektive metoder for sterilisering som ikke påvirker kvaliteten på maten. Kunnskap om germineringsreseptorer, som er lokalisert i den indre membranen hos sporer, og deres rolle i indusering av germinerings prosessen har vist seg å ha stor betydning for vår forståelse av germineringsprosessen. Bacillus licheniformis, har blitt knyttet til bakterieutbrudd i kjøtt og grønnsaker som er kokt, melk og industrielt fremstilt barnemat. En av germineringsreseptorene til B.licheniformis er kodet av yndDEF operonet. Våre upubliserte resultater viser at Ynd germinerings reseptoren er involvert i germinering indusert av L-alanin, L-cystein og L-valin. I motsetning til andre Bacillus arter, som Bacillus subtilis, ser det ut til at B.licheniformis har tre homologe yndE gener, yndE1, yndE2og yndE3. Funksjonen til yndE genene er ukjent. Dette prosjektet har som mål å karakterisere de tre homologe yndE genene, for å forstå rollen de har germineringen av B.licheniformis sporer.Det ble forsøkt å konstruere egne null-mutanter i alle tre yndE homologene, men bare delesjon av yndE3 genet ble vellykket. Analysering av germinering i yndE3 mutanten, indusert av L-alanin, L-cystein og L-valin, viste at mutant stamen fikk en sterkt redusert germinerings respons til L-cystein. Germineringsresponsen til L-alanin og L-valin ble også svakt redusert. Dette resultatet indikerer at yndE3 genet er avgjørende for effektiv germinering indusert med L-cystein og kan muligens tyde på at de andre yndE genene kan være involvert i yndavhengig germinering indusert av L-alanin og L-valin. Dette er den første funksjonelle kartleggingen av rollen til paraloge germinerings reseptor underenheter,kodet av ett enkelt germinerings respetor operon vi er kjent medM-M

Antibiotics with Interleukin-15 inhibition reduces joint inflammation and bone erosions but not cartilage destruction in Staphylococcus aureus-induced arthritis

Background: Staphylococcus aureus-induced arthritis causes rapid joint destruction, often leading to disabling joint damage despite antibiotics. We have previously shown that IL-15 inhibition without antibiotics is beneficial in S. aureus-induced arthritis. We therefore hypothesized that inhibition of IL-15, in combination with antibiotics, might represent a useful therapy that would both reduce inflammation and joint destruction, but preserve the host's ability to clear the infection. Methods: Female wildtype C57BL/6 mice were intravenously inoculated with the TSST-1-producing LS-1 strain of S. aureus with 0.8x108 S. aureus LS-1/mouse. Three days later the treatment was started consisting of cloxacillin followed by flucloxacillin, together with either anti-IL-15 antibodies (aIL-15ab) or control antibodies. Outcomes included survival, weight change, bacterial clearance, and joint damage. Results: The addition of aIL-15ab to antibiotics in S. aureus-induced arthritis reduced synovitis and bone erosions compared to controls. The number of bone-resorbing osteoclasts in the joints was reduced, whereas cartilage destruction was not significantly altered. Importantly, the combination therapy did not adversely affect the clinical outcome of S. aureus-induced arthritis, such as survival, weight change or compromise the host's ability to clear the infection. Conclusions: As the clinical outcome of S. aureus-induced arthritis was not affected, the addition of aIL-15ab to antibiotics ought to be safe. Taken together, the combination of aIL-15ab and antibiotics is a beneficial, but not optimal, treatment of S. aureus-induced arthritis as it reduces synovitis and bone erosions but has a limited effect on cartilage destruction

Sweet flowers are slow and weeds make haste: anthropogenic dispersal of plants via garden and construction soil

This is the final version of the article. Available from Oxford University Press via the DOI in this record.Anthropogenic activities are increasingly responsible for the dispersal of plants. Of particular concern is anthropogenic dispersal of problematic invasive non-native plants. A common dispersal vector is the movement of soil containing seeds or rhizomes. Housing development and domestic gardening activities cause large quantities of soil to be moved, and understanding the role of these activities is critical for informing policy and management to reduce the spread of problematic plants. Here, by collecting soil samples being moved for housing development and domestic gardening, and observing the species that germinated from these samples, we determined the quantities and invasive status of plants moved. From our samples nearly 2000 individuals representing 90 species germinated. Our results suggest that given the quantity of topsoil needed to cover an average-sized UK garden (190 m2 ), there could be 2.2 million and c.2 million viable seeds in soil sourced from housing developments and gardens, respectively. In both housing development and garden samples, native species were more abundant and species-rich than non-native naturalised and invasive species. Buddleia (an invasive) was the most common species overall and in garden samples; this is likely due to multiple traits that adapt it to dispersal, such as prolific seed production. The abundance of invasive and naturalised species was significantly higher in garden than in housing development samples, suggesting that informal movement of soil between gardens poses a greater risk of spreading invasive plants than commercial sources. Consequences for models predicting future distributions of plants, and strategies to mitigate anthropogenic dispersal of problematic plants are considered.This project was funded by the University of Exeter and the Animal and Plant Health Agency. We are grateful to all who gave samples for this study

The Fundamental Plane in RX J0142.0+2131: a galaxy cluster merger at z=0.28

We present the Fundamental Plane (FP) in the z = 0.28 cluster of galaxies RX J0142.0+2131. There is no evidence for a difference in the slope of the FP when compared with the Coma cluster, although the internal scatter is larger. On average, stellar populations in RX J0142.0+2131 have rest-frame V-band mass-to-light ratios (M/L_V) 0.29+-0.03 dex lower than in Coma. This is significantly lower than expected for a passively-evolving cluster formed at z_f=2. Lenticular galaxies have lower average M/L_V and a distribution of M/L_V with larger scatter than ellipticals. Lower mass-to-light ratios are not due to recent star formation: our previous spectroscopic observations of RX J0142.0+2131 E/S0 galaxies showed no evidence for significant star-formation within the past ~4 Gyr. However, cluster members have enhanced alpha-element abundance ratios, which may act to decrease M/L_V. The increased scatter in the RX J0142.0+2131 FP reflects a large scatter in M/L_V implying that galaxies have undergone bursts of star formation over a range of epochs. The seven easternmost cluster galaxies, including the second brightest member, have M/L_V consistent with passive evolution and z_f = 2. We speculate that RX J0142.0+2131 is a cluster-cluster merger where the galaxies to the east are yet to fall into the main cluster body or have not experienced star formation as a result of the merger.Comment: 4 pages, 2 figures, accepted for publication in ApJ Letter

Exploring needs, barriers to, and facilitators of rehabilitation exercise following revision hip replacement - a grounded theory study

Purpose: Evidence on rehabilitation after revision total hip replacement (THR) is inadequate and development of rehabilitation interventions is warranted. Even so, little is known about patients’ experiences with revision THR rehabilitation. This study aimed to explore patients' rehabilitation exercise experiences after revision THR.Materials and methods: Using constructivist grounded theory, we conducted semi-structured qualitative interviews with twelve patients with completed or almost completed rehabilitation exercise after revision THR. Data collection and analysis were a constant comparative process conducted in three phases; initial, focused, and theoretical.Findings: From the data, we generated a substantial theory of the participant’s circumstances and ability to integrate rehabilitation exercise into their everyday life after revision THR. Four categories were constructed based on patients’ experiences in different contexts: hesitance, fear avoidance, self-commitment, and fidelity.Conclusions: This study highlighted that patients’ expectations, past experiences, attitudes, trusts, engagement, and circumstances interact to influence engagement and adherence to rehabilitation exercise and described four categories relating to the integration of THR rehabilitation exercise into their everyday life. Clinicians should be aware of and account for these categories during rehabilitation exercise. Tailored individual rehabilitation exercise interventions and clinician approaches to optimize commitment and adherence are needed among patients with revision THR

Comparison of deep inspiration breath hold and free breathing intensity modulated proton therapy of locally advanced lung cancer

Background and purpose: For locally advanced non-small cell lung cancer (LA-NSCLC), intensity-modulated proton therapy (IMPT) can reduce organ at risk (OAR) doses compared to intensity-modulated radiotherapy (IMRT). Deep inspiration breath hold (DIBH) reduces OAR doses compared to free breathing (FB) in IMRT. In IMPT, differences in dose distributions and robustness between DIBH and FB are unclear. In this study, we compare DIBH to FB in IMPT, and IMPT to IMRT. Materials and methods: Fortyone LA-NSCLC patients were prospectively included. 4D computed tomography images (4DCTs) and DIBH CTs were acquired for treatment planning and during weeks 1 and 3 of treatment. A new system for automated robust planning was developed and used to generate a FB and a DIBH IMPT plan for each patient. Plans were compared in terms of dose-volume parameters and normal tissue complication probabilities (NTCPs). Dose recalculations on repeat CTs were used to compare inter-fraction plan robustness. Results: In IMPT, DIBH reduced median lungs Dmean from 9.3 Gy(RBE) to 8.0 Gy(RBE) compared to FB, and radiation pneumonitis NTCP from 10.9 % to 9.4 % (p &lt; 0.001). Inter-fraction plan robustness for DIBH and FB was similar. Median NTCPs for radiation pneumonitis and mortality were around 9 percentage points lower with IMPT than IMRT (p &lt; 0.001). These differences were much larger than between FB and DIBH within each modality. Conclusion: DIBH IMPT resulted in reduced lung dose and radiation pneumonitis NTCP compared to FB IMPT. Inter-fraction robustness was comparable. OAR doses were far lower in IMPT than IMRT.</p

Comparison of deep inspiration breath hold and free breathing intensity modulated proton therapy of locally advanced lung cancer

Background and purpose: For locally advanced non-small cell lung cancer (LA-NSCLC), intensity-modulated proton therapy (IMPT) can reduce organ at risk (OAR) doses compared to intensity-modulated radiotherapy (IMRT). Deep inspiration breath hold (DIBH) reduces OAR doses compared to free breathing (FB) in IMRT. In IMPT, differences in dose distributions and robustness between DIBH and FB are unclear. In this study, we compare DIBH to FB in IMPT, and IMPT to IMRT. Materials and methods: Fortyone LA-NSCLC patients were prospectively included. 4D computed tomography images (4DCTs) and DIBH CTs were acquired for treatment planning and during weeks 1 and 3 of treatment. A new system for automated robust planning was developed and used to generate a FB and a DIBH IMPT plan for each patient. Plans were compared in terms of dose-volume parameters and normal tissue complication probabilities (NTCPs). Dose recalculations on repeat CTs were used to compare inter-fraction plan robustness. Results: In IMPT, DIBH reduced median lungs Dmean from 9.3 Gy(RBE) to 8.0 Gy(RBE) compared to FB, and radiation pneumonitis NTCP from 10.9 % to 9.4 % (p &lt; 0.001). Inter-fraction plan robustness for DIBH and FB was similar. Median NTCPs for radiation pneumonitis and mortality were around 9 percentage points lower with IMPT than IMRT (p &lt; 0.001). These differences were much larger than between FB and DIBH within each modality. Conclusion: DIBH IMPT resulted in reduced lung dose and radiation pneumonitis NTCP compared to FB IMPT. Inter-fraction robustness was comparable. OAR doses were far lower in IMPT than IMRT.</p

Trauma Mechanisms and Injuries Associated with Go-Karting

Annually, approximately 600 patients seek medical attention after go-kart accidents in the Netherlands. A large variability in injury patterns can be encountered. Knowledge of the trauma mechanisms of go-kart accidents and insight into the associated injuries is limited and requires improvement. Such additional knowledge may lead to customized trauma protocols for patients with a high index of suspicion on go-kart injuries. Research into trauma mechanisms may also lead to implementation of improved or additional safety measures for go-karting, involving both the go-karts itself as well as prerequisites to the go-kart tracks and qualifications for the drivers. The main trauma mechanisms involved in go-kart accidents, and three cases to illustrate the variety of injuries are described in the current manuscript