Towards an ecological understanding of the killer trait – A reproducible protocol for testing its impact on freshwater ciliates

Paramecium strains with the ability to kill other paramecia often harbour intracellular bacteria belonging to the genera Caedibacter or Caedimonas. Central structures of this killer trait are refractile bodies (R-bodies) produced by the endosymbionts. Once ingested by a sensitive Paramecium, R-bodies presumably act as delivery system for an unidentified toxin which causes the death of endosymbiont-free paramecia while those infected gain resistance from their symbionts. The killer trait is therefore considered as competitive advantage for the hosts of R-body producers. While its effectiveness against paramecia is well documented, the effects on other aquatic ciliates are much less studied.In order to address the broadness of the killer trait, a reproducible killer test assay considering the effects on predatory ciliates (Climacostomum virens and Dileptus jonesi) as well as potential bacterivorous Paramecium competitors (Dexiostoma campyla, Euplotes aediculatus, Euplotes woodruffi, and Spirostomum teres) as possibly susceptible species was established. All used organisms were molecularly characterized to increase traceability and reproducibility. The absence of any lethal effects in both predators and competitors after exposure to killer paramecia strongly suggests a narrow action range for the killer trait. Thus, R-body producing bacteria provide their host with a complex, costly strategy to outcompete symbiont-free congeners only.     

Biofilm research using calorimetry – a marriage made in heaven?

Aggregated bacteria growing on a surface, so-called biofilms, play an important role in technical processes like wastewater treatment, bioremediation, or bioprocessing. In contrast, problems arise when biofilm growth results in undesired processes that may cause huge financial losses, e.g., clogged pipes, microbially influenced corrosion or pathogenic contamination. For observation purposes and to develop efficient control strategies, real-time monitoring tools for biofilms are required. Among the large variety of tools used in biofilm research, calorimetry is rarely applied, even though many characteristics qualify it for biofilm investigation and monitoring. Calorimetric measurements are non-invasive and non-destructive, and can be applied to nearly any kind of samples (including heterogeneous or turbid solutions) without the need of special sample preparation. Online and real-time data acquisition reduces the labor and facilitates high-throughput measurements. The following article is meant to introduce and promote calorimetry as a future tool in biofilm research. It attempts an assessment of common, existent monitoring tools and specifically addresses the potential of calorimetry in this field.     

Towards National Surgical Surveillance in the UK – A Pilot Study

Objective

The Bristol heart inquiry in the United Kingdom (UK) highlighted the lack of standards for evaluating surgical performance and quality. In 2009, the World Health Organisation (WHO) proposed six standardised metrics for surgical surveillance. This is the first study to collect and analyse such metrics from a cohort of National Health Service (NHS) Trusts in England, helping to determine their feasibility and utility in measuring surgical performance, its impact on public health and mortality, and for tracking surgical trends over time.

Methods

Freedom of Information Act 2000 (FOI) requests for WHO standardised surgical metrics were made to 36 NHS Trusts in England during July to November 2010. Additional data on Hospital Standardised Mortality Ratio (HSMR), Patient Safety Score and Abdominal Aortic Aneurysm (AAA) volume and mortality was obtained from Dr Foster Health and The Guardian Newspaper. Analysis was performed using mixed-effect logistic regression.

Results

30/36 trusts responded (83%). During 2005–9, 5.4 million operations were performed with a 24.2% increase in annual number of operations. This rising volume within hospitals was associated with lower mortality ratios. A 10% increase in operative volume was associated with a lower day of surgery death rate (DDR OR = 0.94, p = 0.056) and post-operative inpatient 30-day mortality (PDR30 OR = 0.93, p = 0.001). For every 10,000 more operations that an NHS Trust does, a 4% drop in PDR30 mortality was achieved. A 10% increase in the volume of elective AAAs was associated with lower elective AAA (OR = 0.96, p = 0.032) and emergency AAA (OR = 0.95, p = 0.009) PDR30 mortality. Lower DDR mortality was noted for emergency AAA mortality (OR = 0.95, p = 0.025) but not elective AAAs (OR = 0.97, p = 0.116).

Conclusion

Standarised surgical metrics can provide policy makers and commissioners with valuable summary data on surgical performance allowing for statistical process control of a complex intervention. This study has shown their collection is feasible albeit using FOI and the first to show a statistically significant volume-outcome relationship for surgery as a whole within hospitals. It adds weight to the argument that patients are safer in larger hospitals or those that become larger by growing their patient base. Together with other measures, such metrics can help build a picture of surgical surveillance in the UK and potentially lead us to safer surgery.     

Hyperspectral modeling of ecological indicators – A new approach for monitoring former military training areas

Military areas are valuable habitats and refuges for rare and endangered plants and animals. We developed a new approach applying innovative methods of hyperspectral remote sensing to bridge the existing gap between remote sensing technology and the demands of the nature conservation community. Remote sensing has already proven to be a valuable monitoring instrument. However, the approaches lack the consideration of the demands of applied nature conservation which includes the legal demands of the EU Habitat Directive. Following the idea of the Vital Signs Monitoring in the USA, we identified a subset of the highest priority monitoring indicators for our study area. We analyzed continuous spectral response curves and tested the measurability of N = 19 indicators on the basis of complexity levels aggregated from extensive vegetation assemblages. The spectral differentiability for the floristic as well as faunistic indicators revealed values up to 100% accuracy. We point out difficulties when it comes to distinguishing faunistic habitat requirements of several species adapted to dry open landscapes, which in this case results in Overall accuracy of 67, 87–95, and 35% in the error matrix. In summary, we provide an applicable and feasible method to facilitating monitoring military areas by hyperspectral remote sensing in the following.     

Radiofrequency Ablation for Benign Thyroid Nodules – A Look Towards the Future of Interventional Thyroidology

Abbreviations: FNA = fine-needle aspiration LAT = laser ablation therapy PEI = percutaneous ethanol injection RFA = radiofrequency ablation US = ultrasound     

Sperm cryopreservation in guppies and black mollies—A generalized freezing protocol for livebearers in Poeciliidae

In this study, we evaluated various parameters of sperm cryopreservation in two livebearers, guppies (Poecilia reticulata) and black mollies (P. latipinna). Our results suggested a common freezing protocol for the guppies and mollies: suspend sperm in Hanks’ balanced salt solution (HBSS) at 300 mOsm/kg, use 14% glycerol as cryoprotectant, cool at 25 °C/min, and thaw at 40 °C in a water bath for 7 s. Live young were produced from females inseminated with frozen-thawed sperm in both species. In guppies, percent fertilization (F) and the number of embryos (N) produced with cryopreserved sperm (F = 50%, N = 74, from 26 females) were similar to those of fresh controls (F = 54%, N = 61, from 22 females). Interestingly, this same freezing protocol has been used successfully for sperm cryopreservation in green swordtails Xiphophorus helleri, and platyfish of X. couchianus with post-thaw motility as high as 80%. All these species belong to the family of Poeciliidae, and their sperm are similar in morphology exhibiting the absence of acrosome, elongate sperm head, and the long mitochondrial sheaths. Besides their internal fertilization reproduction mode, these fish are also small in size (2–4 cm) and live in a freshwater environment. Sperm cryopreservation in fish has been generally recognized as species specific, and new protocols are required for new species. However, results presented in this study suggested otherwise. Thus, sperm cryopreservation methods optimized for one species may be applicable to others if they are taxonomical closely related species with similar sperm morphology and reproduction mode. Considering the enormous number of fish species on the planet, development of generalized sperm freezing protocols for species in groups could have additional advantages for genetic conservation.     

A protocol for absolute dose verification of SBRT/SRS treatment plans using Gafchromic™ EBT-XD films

PurposeTo provide a practical protocol for absolute dose verification of stereotactic body radiotherapy (SBRT) and stereotactic radiosurgery (SRS) treatment plans, based on our clinical experience. It aims to be a concise summary of the main aspects to be considered when establishing an accurate film dosimetry system.MethodsProcedures for film calibration and conversion to dose are described for a dosimetry system composed of Gafchromic™ EBT-XD films and a flatbed document scanner. Factors that affect the film-scanner response are also reviewed and accounted for. The accuracy of the proposed methodology was assessed by taking a set of strips irradiated to known doses and its applicability is illustrated for ten SBRT/SRS treatment plans. The film response was converted to dose using red and triple channel dosimetry. The agreement between the planned and measured dose distributions was evaluated using global gamma analysis with criteria of 3%/2mm 10% threshold (TH), 2%/2mm 10% TH, and 2%/2mm 20% TH.ResultsThe differences between the expected and determined doses from the strips analysis were 0.9 ± 0.6% for the red channel and 1.1 ± 0.7% for the triple channel method. Regarding the SBRT/SRS plans verification, the mean gamma passing rates were 99.5 ± 1.0% vs 99.6 ± 1.0% (3%/2mm 10% TH), 96.9 ± 3.5% vs 99.1 ± 1.3% (2%/2mm 10% TH) and 98.4 ± 1.8% vs 98.8 ± 1.5% (2%/2mm 20% TH) for red and triple channel dosimetry, respectively.ConclusionsThe proposed protocol allows for accurate absolute dose verification of SBRT/SRS treatment plans, applying both single and triple channel methods. It may work as a guide for users that intend to implement a film dosimetry system.     

Key evaluation framework for the impacts of urbanization on air environment – A case study

Pressure-State-Response (PSR) model is one of the most commonly used models for environmental impacts assessment. As an effective tool for strategic management and performance evaluation, Balanced Scorecard (BSC) has been widely applied in the management field. PSR and BSC share the similar principles such as systematic, causality and sustainability oriented. Furthermore, BSC features equilibrium and dynamics. It is a feasible and innovative approach to establish a comprehensive environmental assessment framework by combining BSC and PSR. By placing the customer indicator on the top layer and following the PSR principle on the second layer, one model was designed to examine the impact of urbanization on the air environment in Shandong Province from 2005 to 2009 with factor component analysis. The results showed that urbanization and air environmental sustainability has been an upward trend in Shandong. The sustainable development level is low in Shandong presently, which provides a large potential for further improvement. This study is a useful reference for policy makers in terms of economic and environmental management.     

A new method for examining the complexity and relationships of “timers” in developing systems

Simple methods are developed for analyzing the rate-limiting pathways, or “developmental timers,” for consecutive stages in a developing system. Two conditions are first defined for short and long timing to a developmental stage. Shifts are then performed at time intervals from short to long and long to short conditions. The total time to the stage (time under first condition plus time under second condition) is scored and plotted as a function of the time of shift, resulting in two plots, one for shifts from the short to long condition, and the other for shifts from the long to short condition. Each plot is then analyzed for the number of components, slopes of components, absolute times of origins and termini of components, and discontinuities between components. This information is then used (1) to distinguish between single- and multiple-component timers, (2) to assess the sensitivity of each timer component to the change in the environmental condition employed in the method, including reversibility, (3) to test for the addition of a new timer component under long conditions, and (4) to test for an identity change of a timer component between short and long conditions. These interpretations in turn provide a minimum estimate of the complexity of the rate-limiting pathway to a developmental stage, temporally define major transition points between timer components, and provide some insight into the nature of timer components. By characterizing the rate-limiting pathway from the origin of a developmental program for each consecutive stage in that program, distinctions can also be made between single, parallel, sequential, and branching timer relationships. From these interpretations, a detailed temporal “map” of the rate-limiting program can be generated for any developmental system in which consecutive stages can be reproducibly monitored with time.     

Deer antler – A novel model for studying organ regeneration in mammals

Deer antler is the only mammalian organ that can fully grow back once lost from its pedicle – the base from which it grows. Therefore, antlers probably offer the most pertinent model for studying organ regeneration in mammals. This paper reviews our current understanding of the mechanisms underlying regeneration of antlers, and provides insights into the possible use for human regenerative medicine. Based on the definition, antler renewal belongs to a special type of regeneration termed epimorphic. However, histological examination failed to detect dedifferentiation of any cell type on the pedicle stump and the formation of a blastema, which are hallmark features of classic epimorphic regeneration. Instead, antler regeneration is achieved through the recruitment, proliferation and differentiation of the single cell type in the pedicle periosteum (PP). The PP cells are the direct derivatives of cells resident in the antlerogenic periosteum (AP), a tissue that exists in prepubertal deer calves and can induce ectopic antler formation when transplanted elsewhere on the deer body. Both the AP and PP cells express key embryonic stem cell markers and can be induced to differentiate into multiple cell lineages in vitro and, therefore, they are termed antler stem cells, and antler regeneration is a stem cell-based epimorphic regeneration. Comparisons between the healing process on the stumps from an amputated mouse limb and early regeneration of antlers suggest that the stump of a mouse limb cannot regenerate because of the limited potential of periosteal cells in long bones to proliferate. If we can impart a greater potential of these periosteal cells to proliferate, we might at least be able to partially regenerate limbs lost from humans. Taken together, a greater understanding of the mechanisms that regulate the regeneration of antlers may provide a valuable insight to aid the field of regenerative medicine.This article is part of a Directed Issue entitled: Regenerative Medicine: the challenge of translation.     

Towards an updated ESTRO-EFOMP core curriculum for education and training of medical physics experts in radiotherapy – A survey of current education and training practice in Europe

PurposeESTRO-EFOMP intend to update the core curriculum (CC) for education and training of medical physicists in radiotherapy in line with the European Commission (EC) guidelines on Medical Physics Experts (MPE), the CanMEDS methodology and recent developments in radiotherapy. As input, a survey of the current structure of radiotherapy MPE national training schemes (NTS) in Europe was carried out.MethodsA 35-question survey was sent to all European medical physics national societies (NS) with a focus on existence of an NTS, its format and duration, required entry-level education, and financial support for trainees.ResultsTwenty-six of 36 NS responded. Twenty had an NTS. Minimum required pre-training education varied from BSc in physics or related sciences (5/2) to MSc in medical physics, physics or related sciences (6/5/2) with 50–210 ECTS in fundamental physics and mathematics. The training period varied from 1 to 5 years (median 3 years with 50% dedicated to radiotherapy). The ratio of time spent on university lectures versus hospital training was most commonly 25%/75%. In 14 of 20 countries with an NTS, a research project was mandatory. Residents were paid in 17 of 20 countries. The recognition was mostly obtained by examination. Medical physics is recognised as a healthcare profession in 19 of 26 countries.ConclusionsThe NTS entrance level, duration and curriculum showed significant variations. This survey serves to inform the design of the updated CC to define a realistic minimum training level for safe and effective practice aiming at further harmonization in line with EC guidelines.     

Prefabrication of 3D Cartilage Contructs: Towards a Tissue Engineered Auricle – A Model Tested in Rabbits

The reconstruction of an auricle for congenital deformity or following trauma remains one of the greatest challenges in reconstructive surgery. Tissue-engineered (TE) three-dimensional (3D) cartilage constructs have proven to be a promising option, but problems remain with regard to cell vitality in large cell constructs. The supply of nutrients and oxygen is limited because cultured cartilage is not vascular integrated due to missing perichondrium. The consequence is necrosis and thus a loss of form stability. The micro-surgical implantation of an arteriovenous loop represents a reliable technology for neovascularization, and thus vascular integration, of three-dimensional (3D) cultivated cell constructs. Auricular cartilage biopsies were obtained from 15 rabbits and seeded in 3D scaffolds made from polycaprolactone-based polyurethane in the shape and size of a human auricle. These cartilage cell constructs were implanted subcutaneously into a skin flap (15×8 cm) and neovascularized by means of vascular loops implanted micro-surgically. They were then totally enhanced as 3D tissue and freely re-implanted in-situ through microsurgery. Neovascularization in the prefabricated flap and cultured cartilage construct was analyzed by microangiography. After explantation, the specimens were examined by histological and immunohistochemical methods. Cultivated 3D cartilage cell constructs with implanted vascular pedicle promoted the formation of engineered cartilaginous tissue within the scaffold in vivo. The auricles contained cartilage-specific extracellular matrix (ECM) components, such as GAGs and collagen even in the center oft the constructs. In contrast, in cultivated 3D cartilage cell constructs without vascular pedicle, ECM distribution was only detectable on the surface compared to constructs with vascular pedicle. We demonstrated, that the 3D flaps could be freely transplanted. On a microangiographic level it was evident that all the skin flaps and the implanted cultivated constructs were well neovascularized. The presented method is suggested as a promising alternative towards clinical application of engineered cartilaginous tissue for plastic and reconstructive surgery.     

Conservation strategies for threatened medicinal plant – Barleria prionitis L. – using in vitro and ex vitro propagation techniques

Over-utilisation and continuous depletion of medicinal plants have affected their supply and loss of genetic diversity. Hence the current study is based on conservation strategies for threatened medicinal plants with special reference to Barleria prionitis L. using in vitro and ex vitro propagation techniques. We have developed here a protocol for plant regeneration of Barleria prionitis L. We have also developed an efficient system of vegetative propagation of Barleria prionitis L. through stem cuttings using revive rooting hormones. These studies can be useful for conservation strategies of this important medicinal plant.     

Is wildlife research useful for wildlife conservation in the tropics? A review for Borneo with global implications

The urgency of the tropical biodiversity crisis continues to be a major justification for wildlife research and its funding. To examine the benefits of this research for on-the-ground conservation, we focused on Borneo, where conservation has a long history and we have direct experience. We compiled, categorized and evaluated 284 publications from a broad variety of sources, 153 from peer-reviewed journals. We found that few studies address threats to species and fewer still provide input for or guidance to effective management. We consider various reasons for these shortcomings. Research is seldom judged on its relevance to pragmatic problem solving. Furthermore, many research programs lack the necessary long-term vision and organizational structure for useful applied research. We consulted conservation leaders about our conclusions and all responses suggest that our concerns are not unique to Borneo but reflect wider problems. We conclude that conservation research across most of the tropics is failing to address conservation needs. We make a number of recommendations based on our findings. Conservation biologists should place a higher priority on addressing practical conservation needs and goals.     

Can the problem of hybridization in threatened species be evaluated using a fieldwork research? A case study in snapdragons

Hybridization, natural or artificial, is considered disadvantageous for species biodiversity when it threatens the population integrity of endangered species. Frequently, studies investigating whether hybridization poses a legitimate risk to rare species are based on genetic data obtained in molecular biology laboratories. In this study, we used field research to approach the problem that hybridization could cause for the viability of a population of a rare species and to be able to propose the most appropriate initial conservation strategy. Specifically, using the model genus Antirrhinum, the reproductive barriers between the rare A. pulverulentum and its common congener A. litigiosum have been analysed under the reproductive isolation index (RI). A. pulverulentum had a high value of total RI, indicating that there are barriers to gene flow from A. litigiosum towards this species, and also had a high value for the intrinsic RI, reflecting a low inherent capacity for production of hybrid plants; in addition, the possibility of successful backcrosses between this species with hybrids produced from A. litigiosum ovules were low, given the high intrinsic RI of A. litigiosum. These data indicate the current existence of strong and permanent barriers to hybridization between the two species, suggesting that hybridization does not seem to be a serious problem for the conservation of A. pulverulentum in the studied population, nor for the near future. This study shows how the RI index can provide useful information for conservation purposes and proposes different management recommendations.     

A climate goal–based,multicriteria method for system evaluation in life cycle assessment

The International Journal of Life Cycle Assessment - Our ability to restrict global warming to the established objectives in the Paris Agreement depends on the metrics used to evaluate the climate...