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Over the past decades, there has been extensive debate about whether or not ‘irrealis’, in the sense of a general notion of unrealized state of affairs, plays a role in the grammatical organization of the world’s languages. This paper discusses various facts pertaining to the encoding of unrealized states of affairs cross-linguistically, including facts about person marking, switch-reference, negation, and the encoding of states of affairs that failed to take place in the past or may possibly take place in the future. These facts show that there are three issues that have generally failed to be kept distinct in the debate on ‘irrealis’: the fact that the notion of unrealized state of affairs can be used to describe particular grammatical patterns, the fact that this notion plays a role in the mechanisms that bring about these patterns, and the fact that this notion corresponds to a grammatical category of particular languages. This problem is representative of a widespread approach in linguistic analysis, one where the description of observed grammatical patterns and the formulation of explanatory hypotheses about these patterns are combined together. These two levels are independent of each other, and mixing the two may lead linguists to address the wrong theoretical issues.

▼ This review aims to cover the state of the art of renewable feedstock use in materials production using photopolymerization processes. This area of investigation is an emerging field of research, and it combines biosourced molecules with a cheap and rapid radiative processing method that avoids any emission of volatile organic compounds. The main classes of naturally occurring molecules and macromolecules such as lipids, amino acids, carbohydrates, polyenes, etc. are detailed. The way they are used or integrated in photopolymerizable systems are described in relation to their applications: coatings, biomaterials, biodegradable drug delivery systems, microelectronics or optoelectronics. This critical review takes into account the reactivity of the various compounds as well as their cytotoxicity, biodegradability and finally their end uses.

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This review aims at highlighting on recent developments in preparation, characterization, properties, crystallization behaviors, melt rheology, processing, and future applications possibilities of biodegradable polymers and their layered silicate nanocomposites. These materials are attracting considerable interest in materials science research. Montmorillonite and hectorite are among the most commonly used smectite-type layered silicates for the preparation of nanocomposites. In their pristine form they are hydrophilic in nature, and this property makes them very difficult to disperse into biodegradable polymer matrices. The most common strategy to overcome this difficulty is to replace the interlayer clay cations with quarternized ammonium or phosphonium cations, preferably with long alkyl chains.

A wide range of biodegradable polymer matrices is described in this review with a special emphasis on polylactide because of more eco-friendliness from its origin as contrast to the fully petroleum-based biodegradable polymers and control of carbon dioxide balance after their composting.

Preparative techniques include (i) intercalation of polymers or prepolymers from solution, (ii) in situ intercalative polymerization method, and (iii) melt intercalation method.

This new family of composite materials frequently exhibits remarkable improvements of mechanical and material properties when compared with virgin polymers or conventional micro- and macro-composites. Improvements can include a high storage modulus both in solid and molten states, increased tensile and flexural properties, a decrease in gas permeability and flammability, increased heat distortion temperature and thermal stability, increase in the biodegradation rate, and so forth.

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The evolution of vapour phase sampling of the volatile fraction of vegetable matrices, or of products directly related to them, over the period 1996–2007 is reviewed. High concentration capacity headspace (HCC-HS) and dynamic headspace (D-HS) techniques, that is headspace sampling approaches where the analytes in the vapour phase are concentrated into a sorbent, an adsorbent or a solvent, are considered. Advantages, disadvantages and applications to the vegetable field of several successful techniques based on these approaches are critically presented, including in-tube sorptive extraction (INCAT, HS-SPDE), headspace sorptive extraction (HSSE), solid-phase aroma concentrate extraction (SPACE), large surface area HCC-HS sampling (MESI, MME, HS-STE), headspace liquid-phase microextraction (HS-LPME) and dynamic headspace samplings (D-HS). The developments necessary to overcome some of the limits of the above approaches and techniques are also discussed in view of their application to new fields.

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Vitamin C (ascorbate, AsA) is the most abundant water-soluble antioxidant in plants. Ascorbate provides the first line of defense against damaging reactive oxygen species (ROS), and helps protect plant cells from many factors that induce oxidative stress, including wounding, ozone, high salinity, and pathogen attack. Plant defenses against these stresses are also dependent upon jasmonates (JAs), a class of plant hormones that promote ROS accumulation. Here, we review evidence showing that wounding and JAs influence AsA accumulation in various plant species, and we report new data from Arabidopsis and tomato testing the influence of JAs on AsA levels in wounded and unwounded plants. In both species, certain mutations that impair JA metabolism and signaling influence foliar AsA levels, suggesting that endogenous JAs may regulate steady-state AsA. However, the impact of wounding on AsA accumulation was similar in JA mutants and wild type controls, indicating that this wound response does not require JAs. Our findings also indicate that the effects of wounding and JAs on AsA accumulation differ between species; these factors both enhanced AsA accumulation in Arabidopsis, but depressed AsA levels in tomato. These results underscore the importance of obtaining data from more than one model species, and demonstrate the complexity of AsA regulation.

▼ The microbiota of Westerners is significantly reduced in comparison to rural individuals living a similar lifestyle to our Paleolithic forefathers but also to that of other free-living primates such as the chimpanzee. The great majority of ingredients in the industrially produced foods consumed in the West are absorbed in the upper part of small intestine and thus of limited benefit to the microbiota. Lack of proper nutrition for microbiota is a major factor under-pinning dysfunctional microbiota, dysbiosis, chronically elevated inflammation, and the production and leakage of endotoxins through the various tissue barriers. Furthermore, the over-comsumption of insulinogenic foods and proteotoxins, such as advanced glycation and lipoxidation molecules, gluten and zein, and a reduced intake of fruit and vegetables, are key factors behind the commonly observed elevated inflammation and the endemic of obesity and chronic diseases, factors which are also likely to be detrimental to microbiota. As a consequence of this lifestyle and the associated eating habits, most barriers, including the gut, the airways, the skin, the oral cavity, the vagina, the placenta, the blood-brain barrier, etc., are increasingly permeable. Attempts to recondition these barriers through the use of so called 'probiotics', normally applied to the gut, are rarely successful, and sometimes fail, as they are usually applied as adjunctive treatments, e.g. in parallel with heavy pharmaceutical treatment, not rarely consisting in antibiotics and chemotherapy. It is increasingly observed that the majority of pharmaceutical drugs, even those believed to have minimal adverse effects, such as proton pump inhibitors and anti-hypertensives, in fact adversely affect immune development and functions and are most likely also deleterious to microbiota. Equally, it appears that probiotic treatment is not compatible with pharmacological treatments. Eco-biological treatments, with plant-derived substances, or phytochemicals, e.g. curcumin and resveratrol, and pre-, pro- and syn-biotics offers similar effects as use of biologicals, although milder but also free from adverse effects. Such treatments should be tried as alternative therapies; mainly, to begin with, for disease prevention but also in early cases of chronic diseases. Pharmaceutical treatment has, thus far, failed to inhibit the tsunami of endemic diseases spreading around the world, and no new tools are in sight. Dramatic alterations, in direction of a paleolithic-like lifestyle and food habits, seem to be the only alternatives with the potential to control the present escalating crisis. The present review focuses on human studies, especially those of clinical relevance.

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Farm mechanization has been progressively increasing in Thailand for the past decades. Consumption and abuse of energy intensive inputs, machinery and agro-chemicals is increasingly propagated into agricultural production systems. Effects of energy intensive input utilization and farm technologies are directly associated especially with farm economic and atmospheric issues. This warrants the need of energy input–output analyses coupled with its environmental dimension. This paper presents the energy input–output analyses of different agricultural activities and fresh pond-culture (polyculture), for which data were collected from 46 rainfed integrated agricultural production systems (IAPSs) of 281 farm plots surveyed. Total energy consumption including non-renewable energy input (NREI), direct and indirect energy input, and system efficiency are calculated and compared for different crops. Resource-wise energy input utilization and energy consumed by farm operations are also discussed for different crops. Further, this study simultaneously relates energy consumption in agricultural production systems associated with their corresponding greenhouse gases (GHGs) emission – presented in terms of total carbon dioxide equivalent (CO₂e). Results reveal noticeable variations in energy consumption and CO₂e emissions from various agricultural production activities. The study reveals that the maximum energy consumer is cassava (32.4GJha⁻¹). Major energy input consumption for all productions are indicated by fossil fuel (diesel oil) as fresh pond-culture depended on fish feed. Transplanted rice provides the highest CO₂e emission (1112kgCO₂eha⁻¹) among crops, in which more than 50% is contributed by methane (CH₄).

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Farm mechanization has been progressively increasing in Thailand for the past decades. Consumption and abuse of energy intensive inputs, machinery and agro-chemicals is increasingly propagated into agricultural production systems. Effects of energy intensive input utilization and farm technologies are directly associated especially with farm economic and atmospheric issues. This warrants the need of energy input–output analyses coupled with its environmental dimension. This paper presents the energy input–output analyses of different agricultural activities and fresh pond-culture (polyculture), for which data were collected from 46 rainfed integrated agricultural production systems (IAPSs) of 281 farm plots surveyed. Total energy consumption including non-renewable energy input (NREI), direct and indirect energy input, and system efficiency are calculated and compared for different crops. Resource-wise energy input utilization and energy consumed by farm operations are also discussed for different crops. Further, this study simultaneously relates energy consumption in agricultural production systems associated with their corresponding greenhouse gases (GHGs) emission – presented in terms of total carbon dioxide equivalent (CO₂e). Results reveal noticeable variations in energy consumption and CO₂e emissions from various agricultural production activities. The study reveals that the maximum energy consumer is cassava (32.4GJha⁻¹). Major energy input consumption for all productions are indicated by fossil fuel (diesel oil) as fresh pond-culture depended on fish feed. Transplanted rice provides the highest CO₂e emission (1112kgCO₂eha⁻¹) among crops, in which more than 50% is contributed by methane (CH₄).

▼ We have calculated the economic value of forest hydrological services for Hoa Binh Hydroelectric Plant in Vietnam, which is a major power supplier for the capital Hanoi. Our valuation is based on measurements over a six-year period from 2001 to 2006 in 240 permanent sample plots in different vegetation types distributed throughout the watershed. We have synthesized the information with GIS, and carried out simulations with derived empirical models for different land use, electricity price and payment proportion scenarios. Our findings indicate that the economic value of forest hydrological services for electricity production ranges from 26.3 million USD to 85.5 million USD per year; and that the longevity of the hydroelectric plant can be prolonged by about 35-80 years, depending on the state of forest cover in the watershed.

▼ We have calculated the economic value of forest hydrological services for Hoa Binh Hydroelectric Plant in Vietnam, which is a major power supplier for the capital Hanoi. Our valuation is based on measurements over a six-year period from 2001 to 2006 in 240 permanent sample plots in different vegetation types distributed throughout the watershed. We have synthesized the information with GIS, and carried out simulations with derived empirical models for different land use, electricity price and payment proportion scenarios. Our findings indicate that the economic value of forest hydrological services for electricity production ranges from 26.3 million USD to 85.5 million USD per year; and that the longevity of the hydroelectric plant can be prolonged by about 35-80 years, depending on the state of forest cover in the watershed.

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Precision agriculture (PA) is an innovative, integrated and internationally standardized approach aiming to increase the efficiency of resource use and to reduce the uncertainty of decisions required to control variation on farms. Worldwide, investments in research and technology on PA have considerably increased during the past decades. Advances in agricultural research, combined with application of PA on grasslands, could provide major economic and environmental benefits. However, most of the available technology is currently being applied on arable land but not on grassland although the latter contributes substantially to agricultural land use as well as to capital and income. The authors hypothesise that heterogeneity of grassland among and within fields is a major constraint to PA application and that understanding such heterogeneity would enable to implement PA where competitive and cost efficient.

In this paper, current developments and future perspectives of digital image processing, remote sensing, yield measurement, and site-specific management on grassland are discussed. At fine scales, on-board sensors can provide information on sward properties that application techniques could respond to. At larger scales, remotely sensed information can provide digital maps of type and status of vegetation that allow a precise management, although the mix of spatial coverage and spatial resolution is not yet ideal. The site-specific management on arable land, however, has not yet proven its applicability in different environments. On grassland, some of these technologies are either already implemented as prototypes into research projects or under development with a short-term perspective to be introduced into practice.

Grassland scientists can significantly contribute to the current development and, in return, benefit from latest sensor technology and so enable rapid data acquisition and improvement of experimentation and knowledge. Also existing computer simulation models can contribute to make PA on grassland a success. Animal husbandry is strongly linked to grassland utilization and can also benefit from present PA technology through animal production and health control and animal guidance with appropriate electronic stimuli. Considerable interdisciplinary research is required to link existing knowledge on PA technology with grassland science and with animal science. This could mainly be accomplished by the interpretation of signal–object interaction that is needed to set up suitable configurations on grassland.

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Understanding fire is essential to improving forest management strategies. More specifically, an accurate knowledge of the spatial distribution of fuels is critical when analyzing, modelling and predicting fire behaviour. First, we review the main concepts and terminology associated with forest fuels and a number of fuel type classifications. Second, we summarize the main techniques employed to map fuel types starting with the most traditional approaches, such as field work, aerial photo interpretation or ecological modelling. We pay special attention to more contemporary techniques, which involve the use of remote sensing systems. In general, remote sensing systems are low-priced, can be regularly updated and are less time-consuming than traditional methods, but they are still facing important limitations. Recent work has shown that the integration of different sources of information and methods in a complementary way helps to overcome most of these limitations. Further research is encouraged to develop novel and enhanced remote sensing techniques.

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Report of the American Congress of Rehabilitation Medicine, Brain Injury-Interdisciplinary Special Interest Group, Disorders of Consciousness Task Force: Seel RT, Task Force Chair, Sherer M, Whyte J, Katz DI, Giacino JT, Rosenbaum AM, Hammond FM, Kalmar K, Pape TL, Zafonte R, Biester RC, Kaelin D, Kean J, Zasler N. Assessment scales for disorders of consciousness: evidence-based recommendations for clinical practice and research.

Objectives

To conduct a systematic review of behavioral assessment scales for disorders of consciousness (DOC); provide evidence-based recommendations for clinical use based on their content validity, reliability, diagnostic validity, and ability to predict functional outcomes; and provide research recommendations on DOC scale development and validation.

Data Sources

Articles published through March 31, 2009, using MEDLINE, CINAHL, Psychology and Behavioral Sciences Collection, Cochrane Database of Systematic Reviews, Database of Abstracts of Reviews of Effects, Cochrane Central Register of Controlled Trials, Biomedical Reference Collection, and PsycINFO. Thirteen primary terms that defined DOC were paired with 30 secondary terms that defined aspects of measurement. Scale names, abbreviations, and authors were also used as search terms. Task force members identified additional articles by using personal knowledge and examination of references in reviewed articles.

Study Selection

Primary criteria included the following: (1) provided reliability, diagnostic validity, and/or prognostic validity data; (2) examined a cohort, case control, or case series sample of persons with DOC who were age older than or equal to 18 years; and (3) assessed in an acute care or rehabilitation setting. Articles were excluded if peer review was not conducted, original data were not reported, or an English language article was not available. The initial search yielded 580 articles. After paired rater review of study abstracts, guideline development was based on 37 articles representing 13 DOC scales.

Data Extraction

Rater pairs classified studies addressing diagnostic and prognostic validity by using the American Academy of Neurology 4-tier level of evidence scheme, and reliability by using a task force–developed 3-tier evidence scheme. An independent quality review of ratings was conducted, and corrections were made.

Data Synthesis

The Coma Recovery Scale-Revised (CRS-R), Sensory Stimulation Assessment Measure (SSAM), Wessex Head Injury Matrix (WHIM), Western Neuro Sensory Stimulation Profile (WNSSP), Sensory Modality Assessment Technique (SMART), Disorders of Consciousness Scale (DOCS), and Coma/Near-Coma Scale (CNC) have acceptable standardized administration and scoring procedures. The CRS-R has excellent content validity and is the only scale to address all Aspen Workgroup criteria. The SMART, SSAM, WHIM, and WNSSP demonstrate good content validity, containing items that could distinguish persons who are in a vegetative state, are in a minimally conscious state (MCS), or have emerged from MCS. The Full Outline of UnResponsiveness Score (FOUR), WNSSP, CRS-R, Comprehensive Levels of Consciousness Scale (CLOCS), and Innsbruck Coma Scale (INNS) showed substantial evidence of internal consistency. The FOUR and the CRS-R showed substantial evidence of good interrater reliability. Evidence of diagnostic validity and prognostic validity in brain injury survivor samples had very high levels of potential bias because of methodologic issues such as lack of rater masking.

Conclusions

The CRS-R may be used to assess DOC with minor reservations, and the SMART, WNSSP, SSAM, WHIM, and DOCS may be used to assess DOC with moderate reservations. The CNC may be used to assess DOC with major reservations. The FOUR, INNS, Glasgow-Liege Coma Scale, Swedish Reaction Level Scale-1985, Loewenstein Communication Scale, and CLOCS are not recommended at this time for bedside behavioral assessment of DOC because of a lack of content validity, lack of standardization, and/or unproven reliability.

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Report of the American Congress of Rehabilitation Medicine, Brain Injury-Interdisciplinary Special Interest Group, Disorders of Consciousness Task Force: Seel RT, Task Force Chair, Sherer M, Whyte J, Katz DI, Giacino JT, Rosenbaum AM, Hammond FM, Kalmar K, Pape TL, Zafonte R, Biester RC, Kaelin D, Kean J, Zasler N. Assessment scales for disorders of consciousness: evidence-based recommendations for clinical practice and research.

Objectives

To conduct a systematic review of behavioral assessment scales for disorders of consciousness (DOC); provide evidence-based recommendations for clinical use based on their content validity, reliability, diagnostic validity, and ability to predict functional outcomes; and provide research recommendations on DOC scale development and validation.

Data Sources

Articles published through March 31, 2009, using MEDLINE, CINAHL, Psychology and Behavioral Sciences Collection, Cochrane Database of Systematic Reviews, Database of Abstracts of Reviews of Effects, Cochrane Central Register of Controlled Trials, Biomedical Reference Collection, and PsycINFO. Thirteen primary terms that defined DOC were paired with 30 secondary terms that defined aspects of measurement. Scale names, abbreviations, and authors were also used as search terms. Task force members identified additional articles by using personal knowledge and examination of references in reviewed articles.

Study Selection

Primary criteria included the following: (1) provided reliability, diagnostic validity, and/or prognostic validity data; (2) examined a cohort, case control, or case series sample of persons with DOC who were age older than or equal to 18 years; and (3) assessed in an acute care or rehabilitation setting. Articles were excluded if peer review was not conducted, original data were not reported, or an English language article was not available. The initial search yielded 580 articles. After paired rater review of study abstracts, guideline development was based on 37 articles representing 13 DOC scales.

Data Extraction

Rater pairs classified studies addressing diagnostic and prognostic validity by using the American Academy of Neurology 4-tier level of evidence scheme, and reliability by using a task force–developed 3-tier evidence scheme. An independent quality review of ratings was conducted, and corrections were made.

Data Synthesis

The Coma Recovery Scale-Revised (CRS-R), Sensory Stimulation Assessment Measure (SSAM), Wessex Head Injury Matrix (WHIM), Western Neuro Sensory Stimulation Profile (WNSSP), Sensory Modality Assessment Technique (SMART), Disorders of Consciousness Scale (DOCS), and Coma/Near-Coma Scale (CNC) have acceptable standardized administration and scoring procedures. The CRS-R has excellent content validity and is the only scale to address all Aspen Workgroup criteria. The SMART, SSAM, WHIM, and WNSSP demonstrate good content validity, containing items that could distinguish persons who are in a vegetative state, are in a minimally conscious state (MCS), or have emerged from MCS. The Full Outline of UnResponsiveness Score (FOUR), WNSSP, CRS-R, Comprehensive Levels of Consciousness Scale (CLOCS), and Innsbruck Coma Scale (INNS) showed substantial evidence of internal consistency. The FOUR and the CRS-R showed substantial evidence of good interrater reliability. Evidence of diagnostic validity and prognostic validity in brain injury survivor samples had very high levels of potential bias because of methodologic issues such as lack of rater masking.

Conclusions

The CRS-R may be used to assess DOC with minor reservations, and the SMART, WNSSP, SSAM, WHIM, and DOCS may be used to assess DOC with moderate reservations. The CNC may be used to assess DOC with major reservations. The FOUR, INNS, Glasgow-Liege Coma Scale, Swedish Reaction Level Scale-1985, Loewenstein Communication Scale, and CLOCS are not recommended at this time for bedside behavioral assessment of DOC because of a lack of content validity, lack of standardization, and/or unproven reliability.

▼ Here we describe small mobilizable vectors based on the Escherichia coli plasmids pK18 and pK19. We combined the useful properties of the pK plasmids (e.g., multiple cloning site, lacZα. fragment, sequencing with M13 primers) with the broad-host-range transfer machinery of plasmid RP4 and a modified sacB gene from Bacillus subtilis. The new pK derivatives can be transferred by RP4-mediated conjugation into a wide range of Gram^- and Gram^+ bacteria, and should facilitate gene disruption and allelic exchange by homologous recombination. As an application example, the generation of a denned deletion of the hom-thrB genes in the chromosome of the Gram^+ bacterium Corynebacterium glutamicum is presented.

▼ A series of broad-host-range expression and lac fusion vectors, based on RSF1010 derivatives, was constructed. The expression vectors contain various promoters (pNm, plac, ptac and pS1) for expression of foreign genes. The efficiency of the promoters was determined in Escherichia coli, Rhizobium meliloti, Rhizobium leguminosarum and Pseudomonas putida by β-galactosidase activity measurements. Of the promoters assayed in E. coli, the most effective is the tac promoter, whereas in soil bacteria the appropriate promoter for overexpression of foreign genes is the Nm^R promoter. The Gm^R gene, serving as a selectable marker for the plasmids, was efficiently expressed in R. meliloti as revealed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and thus, pGm was also used to construct an expression vector. The translational fusion vectors allow the identification and characterization of promoter-carrying cloned fragments on the translational level, whereas the transcriptional fusion vectors can be used to identify and to study promoters on cloned fragments. All lac fusion vectors contain the E. coli lacZ gene or the complete lac operon facilitating quantification of expression.

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The interaction between methylmercury (MeHg) and marine fatty acids is of great interest in risk benefit assessments, and is increasingly important also in fish nutrition when fish oil is replaced with vegetable oils. To assess the effects of replacing fish oil (high dietary n−3/n−6 ratio) by soybean oil (low dietary n−3/n−6 ratio) in combination with MeHg, Atlantic salmon (Salmo salar L.) were fed experimental diets (fish oil (FO) or vegetable oil (VO) based, with or without the addition of MeHg) for 3 months.

As expected replacing dietary fish oil by soybean oil had greater effect on the fatty acid composition in white muscle compared to brain. In brain, the relative levels of 18:2 n−6 and 20:4 n−6 increased in all lipid classes in fish fed the VO based diets whereas the levels of marine omega-3 PUFA (polyunsaturated fatty acid) were unaffected. Different dietary lipid sources did not affect the accumulation of mercury in brain nor muscle. However, exposure to dietary MeHg resulted in decreased levels of 20:4 n−6 (arachidonic acid; ARA) in phosphatidylinositol in brain of VO fed fish. The decreased amounts of ARA in VO fish is suggested to be caused by interaction between high n−6 level and MeHg.

▼ The algorithms in the literature focusing to predict tissue:blood PC (P_tb) for environmental chemicals and tissue:plasma PC based on total (K_p) or unbound concentration (K_pu) for drugs differ in their consideration of binding to hemoglobin, plasma proteins and charged phospholipids. The objective of the present study was to develop a unified algorithm such that P_tb, K_p and K_pu for both drugs and environmental chemicals could be predicted. The development of the unified algorithm was accomplished by integrating all mechanistic algorithms previously published to compute the PCs. Furthermore, the algorithm was structured in such a way as to facilitate predictions of the distribution of organic compounds at the macro (i.e. whole tissue) and micro (i.e. cells and fluids) levels. The resulting unified algorithm was applied to compute the rat P_tb, K_p or K_pu of muscle (n=174), liver (n=139) and adipose tissue (n=141) for acidic, neutral, zwitterionic and basic drugs as well as ketones, acetate esters, alcohols, aliphatic hydrocarbons, aromatic hydrocarbons and ethers. The unified algorithm reproduced adequately the values predicted previously by the published algorithms for a total of 142 drugs and chemicals. The sensitivity analysis demonstrated the relative importance of the various compound properties reflective of specific mechanistic determinants relevant to prediction of PC values of drugs and environmental chemicals. Overall, the present unified algorithm uniquely facilitates the computation of macro and micro level PCs for developing organ and cellular-level PBPK models for both chemicals and drugs.

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Mutant analysis includes approaches that range from traditional screening of mutant populations (forward genetics), to identifying mutations in known genes (reverse genetics), to examining the effects of site-specific mutations that encode modified proteins. All these methodologies have been applied to study jasmonate synthesis and signaling, and their use has led to important discoveries. The fad3 fad7 fad8 mutant of Arabidopsis, and other mutants defective in jasmonate synthesis, revealed the roles of jasmonate in flower development and plant defense against necrotrophic fungal pathogens. The coi1 mutant identified the F-box protein that is now known to be the receptor for jasmonoyl-isoleucine, the active form of jasmonate hormone. Investigations of how JASMONATE-ZIM DOMAIN (JAZ) proteins bind to COI1 and facilitate jasmonate perception have relied on the jai3 mutant, on JAZΔJas constructs, and on site-specific mutations in the Jas and ZIM domains of these proteins.

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The fuel economy and mass emission of an automotive spark ignition vehicle fuelled with the methane enriched biogas (93% CH₄) and base CNG (89.14%) were evaluated using chassis dynamometer under modified Indian driving cycle (MIDC). The transient emissions were also measured with respect to time. The emissions such as CO, HC and NO_x are marginally higher with the enriched biogas than base CNG. But, the experimental results indicate that the vehicle’s emission with the enriched biogas fuel meets to the BS IV Emission Norms. There is no significant change in fuel economy of the vehicle fuelled with the enriched biogas (24.11km/kg) as compared to base CNG (24.38km/kg). The experimental data indicates transient emission characteristics (CO, HC and NO_x) of the vehicle with respect to time for both fuels are very higher with urban cycle (low speed) than extra urban cycle (high speed) and the findings could be useful to find out scopes of further improvement in transient emission and performance of the engine. As the methane enriched biogas gives similar performance like fossil CNG, the enriched biogas could be used as an auto fuel for spark ignition vehicles.

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The fuel economy and mass emission of an automotive spark ignition vehicle fuelled with the methane enriched biogas (93% CH₄) and base CNG (89.14%) were evaluated using chassis dynamometer under modified Indian driving cycle (MIDC). The transient emissions were also measured with respect to time. The emissions such as CO, HC and NO_x are marginally higher with the enriched biogas than base CNG. But, the experimental results indicate that the vehicle’s emission with the enriched biogas fuel meets to the BS IV Emission Norms. There is no significant change in fuel economy of the vehicle fuelled with the enriched biogas (24.11km/kg) as compared to base CNG (24.38km/kg). The experimental data indicates transient emission characteristics (CO, HC and NO_x) of the vehicle with respect to time for both fuels are very higher with urban cycle (low speed) than extra urban cycle (high speed) and the findings could be useful to find out scopes of further improvement in transient emission and performance of the engine. As the methane enriched biogas gives similar performance like fossil CNG, the enriched biogas could be used as an auto fuel for spark ignition vehicles.

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The effect of N availability on photosynthetic capacity, growth parameters and yield was studied in field-grown durum-wheat plants at both the leaf and canopy levels. Two contrasting nitrogen levels (120 and 0kgha⁻¹⁾ were assayed in a randomised block design with nine replicates each. Total biomass was measured at anthesis and yield and its agronomical components at maturity. Photosynthetic measurements were performed 2 weeks after anthesis in two plots of each N treatment. Flag leaves were measured, using a LI-COR 6400 combined with the chlorophyll fluorescence meter, and the whole canopy by measuring CO₂ and H₂O fluxes in an innovative canopy-chamber system. We showed a clear increase in photosynthetic gas exchange and chlorophyll contents with N fertilisation at both canopy and leaf levels. As a consequence the increase in yield as response to N fertilisation seems the result of a larger green leaf area combined with a higher photosynthetic capacity of the leaves attributable to an increase in the maximum carboxylation velocity of Rubisco. Moreover gas-exchange measurements of the flag leaf during grain filling seem to provide a realistic characterisation, not just of the photosynthetic performance of the crop, but also about the impact of N availability on yield. Thus, measurements performed on the flag leaf matched those at the canopy level, with proportional increases in terms of gas exchange and chlorophyll content, providing a fast, cheap and reliable estimation of canopy photosynthesis and the grain yield attained by the crop.

▼ The present study was performed to identify possible genotoxicity induced by organic extracts from particulate matter in the exhaust of two typical diesel engines run on diesel fuel and neat heated fuel-grade rapeseed oil: a Cummins ISBe4 engine tested using the World Harmonized Steady State Test Cycle (WHSC) and modified Engine Steady Cycle (ESC) and a Zetor 1505 engine tested using the Non-Road Steady State Cycle (NRSC). In addition, biodiesel B-100 (neat methylester of rapeseed oil) was tested in the Cummins engine run on the modified ESC. Diluted exhaust was sampled with high-volume samplers on Teflon coated filters. Filters were extracted with dichlormethane (DCM) and DNA adduct levels induced by extractable organic matter (EOM) in an acellular assay of calf thymus DNA coupled with ^3^2P-postlabeling in the presence and absence of rat liver microsomal S9 fraction were employed. Simultaneously, the chemical analysis of 12 priority PAHs in EOM, including 7 carcinogenic PAHs (c-PAHs) was performed. The results suggest that diesel emissions contain substantially more total PAHs than rapeseed oil emissions (for the ESC) or that these concentrations were comparable (for the WHSC and NRSC), while c-PAHs levels were comparable (for the ESC) or significantly higher (for the WHSC and NRSC) for rapeseed oil emissions. DNA adduct levels induced by diesel and rapeseed oil derived EOM were comparable, but consistently slightly higher for diesel than for rapeseed oil. Highly significant correlations were found between 12 priority PAHs concentrations and DNA adduct levels (0.980; p<0.001) and these correlations were even stronger for c-PAHs (0.990; p<0.001). Metabolic activation by the microsomal S9 fraction resulted in several fold higher genotoxicity, suggesting a major contribution of PAHs to genotoxicity. Directly acting compounds, other than c-PAHs, and not requiring S9 to exhibit DNA reactivity were also significant. Generally, DNA adduct levels were more dependent on the type of engine and the test cycle than on the fuel. Our findings suggest that the genotoxicity of particulate emissions from the combustion of rapeseed oil is significant and is comparable to that from the combustion of diesel fuel. A more detailed study is ongoing to verify and extent these preliminary findings.

▼ The repeat in toxin (Rtx) of an environmental isolate ATCC 7966 of Aeromonas hydrophila consists of six genes (rtxACHBDE) organized in an operon similar to the gene organization found for the Rtx of the Vibrio species. The first gene in this operon (rtxA) encodes an exotoxin in vibrios, while other genes code for proteins needed for proper activation of RtxA and in secretion of this toxin from Vibrio cholerae. However, the RtxA of ATCC 7966, as well as from the clinical isolate SSU of A. hydrophila, was exclusively expressed and produced during co-infection of this pathogen with the host, e.g., HeLa cells, indicating that rtxA gene expression required host cell contact. Within the RtxA, an actin cross-linking domain (ACD) exists and to investigate the functionality of this domain, several truncated versions of ACD were generated to discern its minimal biological active region. Such genetically modified genes encoding ACD, which were truncated on either the NH₂? or the COOH? terminal, as well as on both ends, were expressed from a bidirectional promoter of the pBI-enhanced green fluorescent protein (EGFP) vector in a HeLa-Tet-Off cell system. We demonstrated that only the full-length ACD of RtxA from A. hydrophila catalyzed the covalent cross-linking of the host cellular actin, whereas the ACD truncated on the NH₂?, COOH? or both ends did not exhibit such actin cross-linking characteristics. Further, we showed that the full-length ACD of A. hydrophila RtxA disrupted the actin cytoskeleton of HeLa cells, resulting in their rounding phenotype. Finally, our data provided evidence that the full-length ACD of RtxA induced host cell apoptosis. Our study is the first to report that A. hydrophila possesses a functional RtxA having an ACD that contributes to the host cell apoptosis, and hence could represent a potential virulence factor of this emerging human pathogen.

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In the following works, Mathieu Lehanneur readdresses our understanding of domestic spaces through the environmental systems that control and condition them. Such spaces are generally conditioned by centralised heating, cooling and ventilation systems that lump together many sensory qualities, including primary thermal controls as well as by-products of scent and sound, into a singular, mechanical system that fails to engage any of them fully. Lehanneur’s projects look to separate these senses into systems distributed throughout the home, engaging each one specifically and locally, in an attempt to redefine the domestic spaces they are located within. Copyright © 2009 John Wiley & Sons, Ltd.

▼ Evidence is presented to indicate that a PvuII fragment of approx. 2 kb isolated from the 39-kb IncN-group plasmid pCU1 contains all plasmid-borne determinants for stable maintenance as an extrachromosomal element in Escherichia coli K-12. The fragment was sequenced. The features of this sequence include a group of 13 direct tandem repeats of 37 bp and a second group of two other direct repeats of 30 bp flanking a third partial member of this group. In addition, for a 19-bp sequence that overlaps a member of this second group, there are inverted repeats that straddle the members of the first group. There are three open reading frames within the fragment. We compare features of this sequence with that of other plasmid replicons and draw attention to similar and to dissimilar features.

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The aim of this study was to evaluate the toxicology safety of a genetically modified (GM) Bacillus thuringiensis with vegetative insecticidal protein (VIP) gene. Acute and subacute toxicity studies by using its powder preparation were conducted in Wistar rats. The result of the acute study showed the no-observable-adverse-effect level (NOAEL) of this GM B. thuringiensis powder preparation was greater than 5000mg/kg body weight (BW). In the subacute study, the data analysis of body weight gain, food and water consumptions, clinical observations, haematology, serum biochemistry, organ weight ratios and histopathological findings did not show significant differences between control and treated groups. These results proved the NOAEL of this GM B. thuringiensis powder preparation in subacute test was greater than 5000mg/kg BW. Since both the acute and subacute oral toxicity were not detected at the highest dose recommended by OECD guidelines, this GM B. thuringiensis could be generally regarded as safe for use in bio-pesticide industry.

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Manual and low-tech well drilling techniques have potential to assist in reaching the United Nations’ millennium development goal for water in sub-Saharan Africa. This study used publicly available geospatial data in a regression tree analysis to predict groundwater depth in the Zinder region of Niger to identify suitable areas for manual well drilling. Regression trees were developed and tested on a database for 3681 wells in the Zinder region. A tree with 17 terminal leaves provided a range of ground water depth estimates that were appropriate for manual drilling, though much of the tree’s complexity was associated with depths that were beyond manual methods. A natural log transformation of groundwater depth was tested to see if rescaling dataset variance would result in finer distinctions for regions of shallow groundwater. The RMSE for a log-transformed tree with only 10 terminal leaves was almost half that of the untransformed 17 leaf tree for groundwater depths less than 10m. This analysis indicated important groundwater relationships for commonly available maps of geology, soils, elevation, and enhanced vegetation index from the MODIS satellite imaging system.

▼ As initial steps toward understanding the regulation and function of the stringent starvation protein (SSP) of Escherichia coli, we have isolated the ssp gene (encoding SSP), defined the operon in which ssp is found, and created insertion-deletion mutations of the ssp gene in recBC, sbc and recD strains by linear DNA transformation. During attempts to move the insertion-deletion structure to other strains by P1 transduction, we found that P1 was unable to form plaques on hosts lacking an intact ssp gene. The Δssp mutation, however, did not affect transduction of the Δssp strains and mutant strains were able to support lysogenic P1. When P1 lytic growth was induced, an increase in P1 DNA was detected without lysis or plaque formation. Examination of proteins synthesized in the Δssp host during induction revealed the absence of P1 late gene products. Also, the apparent continued synthesis of early gene products during late time points was observed in the Δssp host. The results reported here suggest that the defect in P1 lytic growth brought about by the absence of SSP occurs at the point at which bacteriophage P1 shifts from early to late gene expression. We also report the results of experiments on stable RNA synthesis following amino acid (aa) starvation induced by serine hydroxamate, and experiments on stable RNA synthesis following resupplementation of a limiting aa. These experiments show that SSP is not involved in stable RNA synthesis. Additionally, complementation studies have shown that ssp is identical to the previously described pog gene of E. coli.

▼ The WRKY transcription factors are involved in plant resistance against both biotrophic and necrotrophic pathogens. Arabidopsis WRKY46 is specifically induced by salicylic acid (SA) and biotrophic pathogen Pseudomonas syringae infection. To determine its possible roles in plant defense and elucidate potential functional redundancy with structurally related WRKY70 and WRKY53, we examined loss-of-function T-DNA insertion single, double and triple mutants, as well as gain-of-function transgenic WRKY46 over-expressing plants in response to P. syringae. WRKY46 over-expressing plants were more resistant to P. syringae. In contrast, pathogen-infected wrky46wrky70, wrky46wrky53 double mutants and wrky46wrky70wrky53 triple mutants showed increased susceptibility to this pathogen, with increased bacterial growth and more severe disease symptoms. The contrasting responses of gain-of-function plants and loss-of-function mutants were correlated with increased or reduced expression of defense-related PR1 gene. Expression studies of WRKY46, WRKY70, and WRKY53 in various defense-signaling mutants suggested that they are partially involved in SA-signaling pathway. In addition, our findings demonstrated negative cross-regulation among these three genes. These results indicate that WRKY46, WRKY70, and WRKY53 positively regulate basal resistance to P. syringae; and that they play overlapping and synergetic roles in plant basal defense.