The NILE Project — Advances in the Conversion of Lignocellulosic Materials into Ethanol

NILE ("New Improvements for Lignocellulosic Ethanol") was an integrated European project (2005-2010) devoted to the conversion of lignocellulosic raw materials to ethanol. The main objectives were to design novel enzymes suitable for the hydrolysis of cellulose to glucose and new yeast strains able to efficiently converting all the sugars present in lignocellulose into ethanol. The project also included testing these new developments in an integrated pilot plant and evaluating the environmental and socio-economic impacts of implementing lignocellulosic ethanol on a large scale. Two model raw materials – spruce and wheat straw – both preconditioned with similar pretreatments, were used. Several approaches were explored to improve the saccharification of these pretreated raw materials such as searching for new efficient enzymes and enzyme engineering. Various genetic engineering methods were applied to obtain stable xylose- and arabinose-fermenting Saccharomyces cerevisiae strains that tolerate the toxic compounds present in lignocellulosic hydrolysates. The pilot plant was able to treat 2 tons of dry matter per day, and hydrolysis and fermentation could be run successively or simultaneously. A global model integrating the supply chain was used to assess the performance of lignocellulosic ethanol from an economical and environmental perspective. It was found that directed evolution of a specific enzyme of the cellulolytic cocktail produced by the industrial fungus, Trichoderma reesei, and modification of the composition of this cocktail led to improvements of the enzymatic hydrolysis of pretreated raw material. These results, however, were difficult to reproduce at a large scale. A substantial increase in the ethanol conversion yield and in specific ethanol productivity was obtained through a combination of metabolic engineering of yeast strains and fermentation process development. Pilot trials confirmed the good behaviour of the yeast strains in industrial conditions as well as the suitability of lignin residues as fuels. The ethanol cost and the greenhouse gas emissions were highly dependent on the supply chain but the best performing supply chains showed environmental and economic benefits. From a global standpoint, the results showed the necessity for an optimal integration of the process to co-develop all the steps of the process and to test the improvements in a flexible pilot plant, thus allowing the comparison of various configurations and their economic and environmental impacts to be determined. <br> Le projet NILE, acronyme de "New Improvements for Lignocellulosic Ethanol", était un projet européen (2005-2010) consacré à la conversion des matières premières lignocellulosiques en éthanol. Ses principaux objectifs étaient de concevoir de nouvelles enzymes adaptées à l’hydrolyse de la cellulose en glucose et de nouvelles souches de levure capables de convertir efficacement tous les sucres présents dans la lignocellulose en éthanol. Une autre partie du projet consistait à tester ces nouveaux systèmes dans une installation pilote et à évaluer les impacts environnementaux et socio-économiques de la production et utilisation à grande échelle d’éthanol lignocellulosique. Deux matières premières modèles (l’épicéa et la paille de blé) prétraitées de façon semblable, ont été étudiées. Différentes approches ont été tentées pour améliorer la saccharification de ces matières premières, par exemple, la recherche de nouvelles enzymes efficaces ou l’ingénierie d’enzymes. Plusieurs stratégies d’ingénierie génétique ont été utilisées pour obtenir des souches stables de Saccharomyces cerevisiae capables de fermenter le xylose et l’arabinose, et de tolérer les composés toxiques présents dans les hydrolysats lignocellulosiques. L’installation pilote pouvait traiter 2 tonnes de matières sèches par jour, et l’hydrolyse et la fermentation pouvaient être menées successivement ou simultanément. Un modèle global intégrant la chaîne d’approvisionnement en matière première a servi à évaluer les performances économiques et environnementales de la production d’éthanol lignocellulosique. L’évolution dirigée d’une enzyme du cocktail cellulolytique produit par le champignon Trichoderma reesei, et la modification de la composition de ce cocktail améliorent l’hydrolyse enzymatique des matières premières prétraitées. Cependant, ces résultats n’ont pu être reproduits à grande échelle. Le rendement de conversion et la productivité spécifique en éthanol ont été sensiblement augmentés grâce à l’ingénierie métabolique des souches de levure et au développement d’un procédé optimal de fermentation. Les essais en pilote ont confirmé le bon comportement de ces souches de levure en conditions industrielles ainsi que la possibilité d’utiliser les résidus riches en lignine comme combustible. Le coût de production de l’éthanol et le bilan des émissions de gaz à effet de serre étaient très dépendants des sources d’énergie utilisées. D’un point de vue plus global, les résultats ont montré que l’optimisation du procédé nécessite de codévelopper toutes les étapes de façon intégrée et de valider les améliorations dans une installation pilote, afin notamment de pouvoir comparer différentes configurations et d’en déterminer les effets sur l’économie du procédé et ses impacts environnementaux

Epicycles and Poincar\'{e} Resonances in General Relativity

The method of geodesic deviations provides analytic approximations to geodesics in arbitrary background space-times. As such the method is a useful tool in many practical situations. In this note we point out some subtleties in the application of the method related to secular motions, in first as well as in higher order. In particular we work out the general second-order contribution to bound orbits in Schwarzschild space-time and show that it provides very good analytical results all the way up to the innermost stable circular orbit.Comment: 24 pages, 4 figure

Key principle of the efficient running, swimming, and flying

Empirical observations indicate striking similarities among locomotion in terrestrial animals, birds, and fish, but unifying physical grounds are lacking. When applied to efficient locomotion, the analytical mechanics principle of minimum action yields two patterns of mechanical similarity via two explicit spatiotemporal coherent states. In steady locomotory modes, the slow muscles determining maximal optimum speeds maintain universal intrinsic muscular pressure. Otherwise, maximal speeds are due to constant mass-dependent stiffness of fast muscles generating a uniform force field, exceeding gravitation. Being coherent in displacements, velocities and forces, the body appendages of animals are tuned to natural propagation frequency through the state-dependent elastic muscle moduli. Key words: variational principle of minimum action (04.20.Fy), locomotion (87.19.ru), biomechanics (87.85.G-).Comment: Submitted to the Europhysical Letter

Oral rehydration therapies in Senegal, Mali, and Sierra Leone: A spatial analysis of changes over time and implications for policy

Background: Oral rehydration solution (ORS) is a simple intervention that can prevent childhood deaths from severe diarrhea and dehydration. In a previous study, we mapped the use of ORS treatment subnationally and found that ORS coverage increased over time, while the use of home-made alternatives or recommended home fluids (RHF) decreased, in many countries. These patterns were particularly striking within Senegal, Mali, and Sierra Leone. It was unclear, however, whether ORS replaced RHF in these locations or if children were left untreated, and if these patterns were associated with health policy changes.Methods: We used a Bayesian geostatistical model and data from household surveys to map the percentage of children with diarrhea that received (1) any ORS, (2) only RHF, or (3) no oral rehydration treatment between 2000 and 2018. This approach allowed examination of whether RHF was replaced with ORS before and after interventions, policies, and external events that may have impacted healthcare access.Results: We found that RHF was replaced with ORS in most Sierra Leone districts, except those most impacted by the Ebola outbreak. In addition, RHF was replaced in northern but not in southern Mali, and RHF was not replaced anywhere in Senegal. In Senegal, there was no statistical evidence that a national policy promoting ORS use was associated with increases in coverage. In Sierra Leone, ORS coverage increased following a national policy change that abolished health costs for children.Conclusions: Children in parts of Mali and Senegal have been left behind during ORS scale-up. Improved messaging on effective diarrhea treatment and/or increased ORS access such as through reducing treatment costs may be needed to prevent child deaths in these areas

Improving resolution of public health surveillance for human Salmonella enterica serovar Typhimurium infection: 3 years of prospective multiple-locus variable-number tandem-repeat analysis (MLVA)

<p>Abstract</p> <p>Background</p> <p>Prospective typing of <it>Salmonella enterica </it>serovar Typhimurium (STM) by multiple-locus variable-number tandem-repeat analysis (MLVA) can assist in identifying clusters of STM cases that might otherwise have gone unrecognised, as well as sources of sporadic and outbreak cases. This paper describes the dynamics of human STM infection in a prospective study of STM MLVA typing for public health surveillance.</p> <p>Methods</p> <p>During a three-year period between August 2007 and September 2010 all confirmed STM isolates were fingerprinted using MLVA as part of the New South Wales (NSW) state public health surveillance program.</p> <p>Results</p> <p>A total of 4,920 STM isolates were typed and a subset of 4,377 human isolates was included in the analysis. The STM spectrum was dominated by a small number of phage types, including DT170 (44.6% of all isolates), DT135 (13.9%), DT9 (10.8%), DT44 (4.5%) and DT126 (4.5%). There was a difference in the discriminatory power of MLVA types within endemic phage types: Simpson's index of diversity ranged from 0.109 and 0.113 for DTs 9 and 135 to 0.172 and 0.269 for DTs 170 and 44, respectively. 66 distinct STM clusters were observed ranging in size from 5 to 180 cases and in duration from 4 weeks to 25 weeks. 43 clusters had novel MLVA types and 23 represented recurrences of previously recorded MLVA types. The diversity of the STM population remained relatively constant over time. The gradual increase in the number of STM cases during the study was not related to significant changes in the number of clusters or their size. 667 different MLVA types or patterns were observed.</p> <p>Conclusions</p> <p>Prospective MLVA typing of STM allows the detection of community outbreaks and demonstrates the sustained level of STM diversity that accompanies the increasing incidence of human STM infections. The monitoring of novel and persistent MLVA types offers a new benchmark for STM surveillance.</p> <p>A part of this study was presented at the MEEGID × (Molecular Epidemiology and Evolutionary Genetics of Infectious Diseases) Conference, 3-5 November 2010, Amsterdam, The Netherlands</p

Loss of Biological Diversity: A Global Crisis Requiring International Solutions: A Report to the National Science Board

Executive Summary Biological diversity refers to the variety and variability among living organisms and the ecological complexes in which they occur. Diversity can be defined as the number of different items and their relative frequency. For biological diversity, these items are organized at many levels, ranging from complete ecosystems to the chemical structures that are the molecular basis of heredity. Thus, the term encompasses different ecosystems, species, genes, and their relative abundance (OTA, 1987). There is an ongoing, unprecedented loss of the variety as well as absolute numbers of organisms-from the smallest microorganism to the largest and most spectacular of mammals. Loss of tropical moist forests, which contain over half the total species of organisms, has been well documented by scientists and is now widely reported in the media. Many other ecosystems are also threatened; as human populations and their support systems expand, natural ecosystems at all latitudes are altered or converted. At its meeting on October 15, 1987, the National Science Board concluded that the world\u27s decreasing biological diversity is a critical scientific issue requiring immediate attention. The National Science Board\u27s Committee on International Science was asked to study the scientific and international aspects of the decline of biological diversity and to recommend a course of action. This report describes what the National Science Foundation (NSF) can do to influence the U.S. science and education base, articulates where international scientific cooperation is needed, and suggests roles for other agencies and organizations (both national and international) which have scientific, educational, and management responsibilities. The current disappearance of biota has several causes: the destruction or degradation of entire ecosystems; the accelerating loss of individual species from communities or ecosystems as a result of human disturb;mce; and the loss of genetically distinct parts of populations due to human-induced selective pressures. Although not all parts of the planet are equally affected, the problem is global, and human activities are the primary cause. The loss of biological diversity is important because human existence depends on the biological resources of 1 the earth. Human prosperity is based very largely on the ability to utilize biological diversity: to take advantage of the properties of plants, animals, fungi, and microorganisms for food, clothing, medicine, and shelter. Scientific knowledge about the earth\u27s biological diversity has huge gaps. This lack of information hampers society\u27s ability either to estimate the magnitude of the problem or to prevent further losses. It is impossible to identify all the biological resources at risk, since there is no complete inventory of all the life forms on earth. Approximately 1.4 million species have been given scientific names, but estimates of actual numbers range from 5 million to 80 million species. Although knowledge of some taxa is extensive, the vast majority of groups are largely unknown. The current wave of extinction is destroying both known biotic resources and those still undiscovered. As is proving to be the case with most environmental problems, neither the loss of biological diversity nor its solution is the exclusive province of any one nation. International cooperation is necessary to develop both scientific knowledge and successful mitigation and management strategies. The root causes of the problem include sociological and economic processes which operate on an global scale; a thorough understanding will require investigation and elucidation of both biological and non-biological components. There are several reasons for increasing National Science Foundation (NSF) involvement in biodiversity studies: the economic and social importance of biodiversity (and the risk of opportunity lost due to accelerating extinction); the contributions such leadership can make toward to conservation of biological diversity; the important role of such studies in the international growth of science, especially in tropical countries; the potential impact of such studies on the future course of biology as a whole; and enhancing public awareness of the issues. NSF should assume a scientific leadership position with respect to agencies in the U.S. and throughout the world. By insisting on the central importance of biodiversity, the NSF could encourage collaborative support for the actions recommended below. 1. The Committee believes that the role of the NSF is clear-NSF should, as a matter of National Science Board Policy, provide leadership to undertake the inventory of the world\u27s biodiversity. 2. The scientific basis for conservation biology, restoration ecology, and environmental management must be strengthened. 3. Educational and public awareness programs related to biodiversity need increased support. 4. The economic and social aspects of the biodiversity crisis need additional study. 5. Enhance support for developing country scientists and institutions for biodiversity research and conservation

Investigation of an Escherichia coli O145 outbreak in a child day-care centre - extensive sampling and characterization of eae- and stx1-positive E. coli yields epidemiological and socioeconomic insight

<p>Abstract</p> <p>Background</p> <p>On October 29^th2009 the health authorities in the city of Trondheim, Norway were alerted about a case of Shiga toxin-positive <it>E. coli </it>(STEC) O145 in a child with bloody diarrhoea attending a day-care centre. Symptomatic children in this day-care centre were sampled, thereby identifying three more cases. This initiated an outbreak investigation.</p> <p>Methods</p> <p>A case was defined as a child attending the day-care centre, in whom <it>eae- </it>and <it>stx</it>₁- but not <it>stx</it>₂-positive <it>E. coli </it>O145:H28 was diagnosed from a faecal sample, with multilocus variable number of tandem repeat analysis (MLVA) profile identical to the index isolate. All 61 children, a staff of 14 in the day-care centre, and 74 close contacts submitted faecal samples. Staff and parents were interviewed about cases' exposure to foods and animals. Faecal samples from 31 ewes from a sheep herd to which the children were exposed were analyzed for <it>E. coli </it>O145.</p> <p>Results</p> <p>Sixteen cases were identified, from which nine presented diarrhoea but not haemolytic uremic syndrome (HUS). The attack rate was 0.26, and varied between age groups (0.13-0.40) and between the three day-care centre departments (0.20-0.50), and was significantly higher amongst the youngest children. Median duration of shedding was 20 days (0-71 days). Children were excluded from the day-care centre during shedding, requiring parents to take compassionate leave, estimated to be a minimum total of 406 days for all cases. Atypical enteropathogenic <it>E. coli </it>(aEPEC) were detected among 14 children other than cases. These isolates were genotypically different from the outbreak strain. Children in the day-care centre were exposed to faecal pollution from a sheep herd, but <it>E. coli </it>O145 was not detected in the sheep.</p> <p>Conclusions</p> <p>We report an outbreak of <it>stx</it>₁- and <it>eae-</it>positive STEC O145:H28 infection with mild symptoms among children in a day-care centre. Extensive sampling showed occurrence of the outbreak strain as well as other STEC and aEPEC strains in the outbreak population. MLVA-typing of the STEC-isolates strongly indicates a common source of infection. The study describes epidemiological aspects and socioeconomic consequences of a non-O157 STEC outbreak, which are less commonly reported than O157 outbreaks.</p

Quantum Averaging I: Poincar\'e--von Zeipel is Rayleigh--Schr\"odinger

An exact analogue of the method of averaging in classical mechanics is constructed for self--adjoint operators. It is shown to be completely equivalent to the usual Rayleigh--Schr\"odinger perturbation theory but gives the sums over intermediate states in closed form expressions. The anharmonic oscillator and the Henon--Heiles system are treated as examples to illustrate the quantum averaging method.Comment: 12 pages, LaTeX, to appear in Journ. Phys.

A genomic biomarker signature can predict skin sensitizers using a cell-based in vitro alternative to animal tests

<p>Abstract</p> <p>Background</p> <p>Allergic contact dermatitis is an inflammatory skin disease that affects a significant proportion of the population. This disease is caused by an adverse immune response towards chemical haptens, and leads to a substantial economic burden for society. Current test of sensitizing chemicals rely on animal experimentation. New legislations on the registration and use of chemicals within pharmaceutical and cosmetic industries have stimulated significant research efforts to develop alternative, human cell-based assays for the prediction of sensitization. The aim is to replace animal experiments with in vitro tests displaying a higher predictive power.</p> <p>Results</p> <p>We have developed a novel cell-based assay for the prediction of sensitizing chemicals. By analyzing the transcriptome of the human cell line MUTZ-3 after 24 h stimulation, using 20 different sensitizing chemicals, 20 non-sensitizing chemicals and vehicle controls, we have identified a biomarker signature of 200 genes with potent discriminatory ability. Using a Support Vector Machine for supervised classification, the prediction performance of the assay revealed an area under the ROC curve of 0.98. In addition, categorizing the chemicals according to the LLNA assay, this gene signature could also predict sensitizing potency. The identified markers are involved in biological pathways with immunological relevant functions, which can shed light on the process of human sensitization.</p> <p>Conclusions</p> <p>A gene signature predicting sensitization, using a human cell line in vitro, has been identified. This simple and robust cell-based assay has the potential to completely replace or drastically reduce the utilization of test systems based on experimental animals. Being based on human biology, the assay is proposed to be more accurate for predicting sensitization in humans, than the traditional animal-based tests.</p