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1.

Purpose

The crude palm oil (CPO) extraction is normally done by a wet extraction process, and wastewater treatment of the wet process emits high levels of greenhouse gases (GHGs). A dry process extracts mixed palm oil (MPO) from palm fruit without using water and has no GHG emissions from wastewater treatment. This work is aimed at determining the GHG emissions of a dry process and at evaluating GHG savings on changing from wet to dry process, including land use change (LUC) effects.

Methods

Life cycle assessment from cradle to gate was used. The raw material is palm fruits. The dry process includes primary production, oil room, and utilities. MPO is the main product, while palm cake and fine palm residue are co-products sold for animal feed. Case studies were undertaken without and with carbon stocks of firewood and of nitrogen recycling at plantations from fronds. Allocations by mass, economic, and heating values were conducted. The trading of GHG emissions from co-products to GHG emissions from animal feed was assessed. The GHG emissions or savings from direct LUC (dLUC) and from indirect LUC (iLUC) effects and for the change from wet to dry process were determined.

Results and discussion

Palm fruit and firewood were the major GHG emission sources. Nitrogen recycling on plantations from fronds significantly affects the GHG emissions. With the carbon stocks, the GHG emissions allocated by energy value were 550 kg CO2 eq/t MPO. The GHG emissions were affected by ?3 to 37% for the change from wet to dry process. When the plantation area was increased by 1 ha and the palm oil extraction was changed from wet to dry process, and the change included dLUC and iLUC, the GHG savings ranged from ?0.94 to 5.08 t CO2 eq/ha year. The iLUC was the main GHG emission source. The GHG saving mostly originated from the change of extraction process and from the dLUC effect. Based on the potential use of biodiesel production from oil palm, during 2015–2036 in Thailand, when the extraction process was changed and dLUC and iLUC effects were included, the saving in GHG emissions was estimated to range from ?35,454 to 274,774 t CO2 eq/year.

Conclusions

The change of palm oil extraction process and the LUC effects could minimize the GHG emissions from the palm oil industry. This advantage encourages developing policies that support the dry extraction process and contribute to sustainable developments in palm oil production.
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2.

Introduction

Untargeted metabolomics is a powerful tool for biological discoveries. To analyze the complex raw data, significant advances in computational approaches have been made, yet it is not clear how exhaustive and reliable the data analysis results are.

Objectives

Assessment of the quality of raw data processing in untargeted metabolomics.

Methods

Five published untargeted metabolomics studies, were reanalyzed.

Results

Omissions of at least 50 relevant compounds from the original results as well as examples of representative mistakes were reported for each study.

Conclusion

Incomplete raw data processing shows unexplored potential of current and legacy data.
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3.

Background

In this study, we optimized the process for enhancing amylase production from Pseudomonas balearica VITPS19 isolated from agricultural lands in Kolathur, India.

Methods

Process optimization for enhancing amylase production from the isolate was carried out by Response Surface Methodology (RSM) with optimized chemical and physical sources using Design expert v.7.0. A central composite design was used to evaluate the interaction between parameters. Interaction between four factors–maltose (C-source), malt extract (Nsource), pH, and CaCl2 was studied.

Results

The factors pH and CaCl2 concentration were found to affect amylase production. Validation of the experiment showed a nearly twofold increase in alpha amylase production.

Conclusion

Amylase production was thus optimized and increased yield was achieved.
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4.

Introduction

Mammalian cells like Chinese hamster ovary (CHO) cells are routinely used for production of recombinant therapeutic proteins. Cells require a continuous supply of energy and nutrients to sustain high cell densities whilst expressing high titres of recombinant proteins. Cultured mammalian cells are primarily dependent on glucose and glutamine metabolism for energy production.

Objectives

The TCA cycle is the main source of energy production and its continuous flow is essential for cell survival. Modulated regulation of TCA cycle can affect ATP production and influence CHO cell productivity.

Methods

To determine the key metabolic reactions of the cycle associated with cell growth in CHO cells, we transiently silenced each gene of the TCA cycle using RNAi.

Results

Silencing of at least four TCA cycle genes was detrimental to CHO cell growth. With an exception of mitochondrial aconitase (or Aco2), all other genes were associated with ATP production reactions of the TCA cycle and their resulting substrates can be supplied by other anaplerotic and cataplerotic reactions. This study is the first of its kind to have established key role of aconitase gene in CHO cells. We further investigated the temporal effects of aconitase silencing on energy production, CHO cell metabolism, oxidative stress and recombinant protein production.

Conclusion

Transient silencing of mitochondrial aconitase inhibited cell growth, reduced ATP production, increased production of reactive oxygen species and reduced cell specific productivity of a recombinant CHO cell line by at least twofold.
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5.

Purpose

The aim of this research is to reveal the overall money flows and physical flows of the livestock and feed production supply chain in Thailand in order to analyze the resource use and cost and assess material use efficiency of the whole supply chain. Another aim is to evaluate the options to improve and evaluate trade-off between economic and environmental performance.

Methods

This research conducted material and monetary flow modeling using material flow analysis (MFA) and input output analysis (IOA). Data collected from the Thai Input-Output Tables 2005 were employed to develop the monetary flow model. Direct and indirect resource consumption (energy, water, and land use) and turnover along the supply chain were assessed using environmentally extended input-output analysis model (EEIOA). Scenario analysis with improvement options was applied to the model to evaluate the effectiveness of the improvement options.

Results and discussion

One third of energy and water consumption were from the animal farm itself. The rest were from feed production and upstream raw material production. Land use in the system was mainly from maize and paddy field. Feed conversion ratio improvement should play an important role in the strategy for resource efficiency and reduce environmental impact in the whole supply chain. Energy intensity reduction, the best option in overall energy reduction, is the policy that the government is pushing to be implemented in all sectors, and it can also easily be applied along with the other options. Therefore, it should be applied with the other options for improvement.

Conclusions

The results from monetary flow and physical material flow can visually show the holistic view of the Thai livestock production supply chain quantitatively and allow the stakeholders to understand the economic structure of the supply chain system. This can enable the decision makers to analyze the interrelation effect and impact of changing one sector demand or changing resource efficiency to impact other sectors in the system.
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6.

Purpose

Bivalve production is an important aquaculture activity worldwide, but few environmental assessments have focused on it. In particular, bivalves’ ability to extract nutrients from the environment by intensely filtering water and producing a shell must be considered in the environmental assessment.

Methods

LCA of blue mussel bouchot culture (grown out on wood pilings) in Mont Saint-Michel Bay (France) was performed to identify its impact hotspots. The chemical composition of mussel flesh and shell was analyzed to accurately identify potential positive effects on eutrophication and climate change. The fate of mussel shells after consumption was also considered.

Results and discussion

Its potential as a carbon-sink is influenced by assumptions made about the carbon sequestration in wooden bouchots and in the mussel shell. The fate of the shells which depends on management of discarded mussels and household waste plays also an important role. Its carbon-sink potential barely compensates the climate change impact induced by the use of fuel used for on-site transportation. The export of N and P in mussel flesh slightly decreases potential eutrophication. Environmental impacts of blue mussel culture are determined by the location of production and mussel yields, which are influenced by marine currents and the distance to on-shore technical base.

Conclusions

Bouchot mussel culture has low environmental impacts compared to livestock systems, but the overall environmental performances depend on farming practices and the amount of fuel used. Changes to the surrounding ecosystem induced by high mussel density must be considered in future LCA studies.
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7.

Aims

The objective of this study was to investigate how plants maintain productivity under a limited supply of water and N along the topographical soil water and N gradients in semi-arid forests.

Methods

We investigated forest structure and productivity, N cycling, and water and N use by plants at three different slope positions in a forested area near an arid boundary on a loess plateau in China.

Results

Net primary production (NPP) and aboveground N uptake decreased as soil water and/or N availability decreased on upper slopes; however, NPP and aboveground N uptake were only slightly lower than those of more humid forest ecosystems. Water use efficiency (WUE), N use efficiency (NUE), and fine root biomass increased as soil water and/or N supply decreased with altitude. High NUE was linked to higher N mean residence time, caused by higher N resorption efficiency rather than increasing N productivity.

Conclusions

Our results suggest that NPP and N uptake can be maintained by increasing WUE and NUE and increasing fine root biomass in water and N co-limited semi-arid forest ecosystems near arid boundaries. Such changes in resource use and acquisition strategy can affect production and N cycling via plant-soil feedback systems.
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8.

Objectives

To enhance succinic acid production in Corynebacterium glutamicum by increasing the supply of NADH and the rate of glucose consumption by decreasing H+-ATPase activity.

Results

A mutant of C. glutamicum NC-3-1 with decreased H+-ATPase activity was constructed. This increased the rate of glycolysis and the supply of NADH. Fermentation of C. glutamicum NC-3-1 gave 39 % higher succinic acid production (113 and 81 g/l), a 29 % higher succinic acid yield (0.94 and 0.73 g succinic acid/g glucose) and decreased by-products formation compared to that of C. glutamicum NC-3 in 5 l bioreactor.

Conclusion

The point mutation in C. glutamicum NC-3-1 increased the rate of glycolysis and resulted in higher succinic acid production, higher succinic acid yield and significantly decreased formation of by-products.
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9.

Objective

To examine the feasibility of chitosan as an alternative transfection reagent candidate for protein expression in Bm5 cells and silkworm larvae using recombinant BmNPV bacmid DNA.

Result

Chitosan 100 and recombinant Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid DNA, in amino group/phosphate group (N/P) ratios of 0.1–10, were used for formation of chitosan/DNA nanocomplexes. The chitosan/BmNPV bacmid DNA nanocomplexes showed higher specific activity of GFPuv-β1,3-N-acetylglucosaminyltransferase 2 (β3GnT2) fusion protein (GGT2) expressed in silkworm larvae than DMRIE-C, a conventional silkworm transfection reagent. In particular, the composition of chitosan and BmNPV bacmid DNA nanocomplexes formed by an N/P ratio of 8 or 10, respectively, showed the highest specific activity of β3GnT2 in the silkworm larvae hemolymph. In addition, three different proteins were expressed in silkworm larvae to the same extent using chitosan as that using DMRIE-C.

Conclusion

This is the first finding that chitosan/BmNPV bacmid DNA nanocomplexes can rival the performance of commercially available transfection reagents for the expression of recombinant proteins in Bm5 cells and silkworm larvae.
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10.

Introduction

Swine dysentery caused by Brachyspira hyodysenteriae is a production limiting disease in pig farming. Currently antimicrobial therapy is the only treatment and control method available.

Objective

The aim of this study was to characterize the metabolic response of porcine colon explants to infection by B. hyodysenteriae.

Methods

Porcine colon explants exposed to B. hyodysenteriae were analyzed for histopathological, metabolic and pro-inflammatory gene expression changes.

Results

Significant epithelial necrosis, increased levels of l-citrulline and IL-1α were observed on explants infected with B. hyodysenteriae.

Conclusions

The spirochete induces necrosis in vitro likely through an inflammatory process mediated by IL-1α and NO.
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11.

Introduction

Collecting feces is easy. It offers direct outcome to endogenous and microbial metabolites.

Objectives

In a context of lack of consensus about fecal sample preparation, especially in animal species, we developed a robust protocol allowing untargeted LC-HRMS fingerprinting.

Methods

The conditions of extraction (quantity, preparation, solvents, dilutions) were investigated in bovine feces.

Results

A rapid and simple protocol involving feces extraction with methanol (1/3, M/V) followed by centrifugation and a step filtration (10 kDa) was developed.

Conclusion

The workflow generated repeatable and informative fingerprints for robust metabolome characterization.
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12.

Objectives

To engineer Yarrowia lipolytica for improving the heterologous production of campesterol (a key precursor to manufacture pharmaceutical steroids).

Results

By screening 7-dehydrocholesterol reductase (DHCR7) from diverse species, DHCR7 from Danio rerio was the best candidate for campesterol synthesis. Overexpression of ACL (ATP: citrate lyase) or POX2 (peroxisome acyl-CoA oxidase 2) were key to improving campesterol production. The highest yield of campesterol was 942 mg/l was with the strain overexpressing POX2 in a 5 l bioreactor via high cell density fermentation process with a restricted supply of carbon sourc, sunflower seed oil.

Conclusions

A promising platform to synthesize downstream steroid drugs was established. Efficient approaches were provided to improve the production of desired molecules in Y. lipolytica with high oil utilization efficiency.
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13.

Introduction

Data sharing is being increasingly required by journals and has been heralded as a solution to the ‘replication crisis’.

Objectives

(i) Review data sharing policies of journals publishing the most metabolomics papers associated with open data and (ii) compare these journals’ policies to those that publish the most metabolomics papers.

Methods

A PubMed search was used to identify metabolomics papers. Metabolomics data repositories were manually searched for linked publications.

Results

Journals that support data sharing are not necessarily those with the most papers associated to open metabolomics data.

Conclusion

Further efforts are required to improve data sharing in metabolomics.
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14.

Aims

Progress in improving the internal phosphorus utilisation efficiency of crops has been limited, which may be due to poor screening methods that allow differences in P uptake among genotypes grown in soil to mask genotypic differences in shoot biomass produced per unit of shoot P (PUE). We investigated alternative soil and hydroponic screening methods for their capacity to produce a consensus ranking of genotypes with regard to PUE.

Methods

Six rice genotypes previously identified in hydroponic screening studies as being high, intermediate or low in PUE were screened using multi P rate hydroponic and soil-based experiments.

Results

Comparisons made at each rate of soil-P supply produced estimates of PUE strongly biased by P uptake differences among genotypes. Using multiple-rate data to derive response functions per genotype showed that similar P content was achieved at different rates of P supply but that high-PUE genotypes clearly separated from intermediate- and low-PUE genotypes if equal P content was used. Ranking analysis suggested that results obtained from soil agreed well with those from the hydroponic study.

Conclusions

PUE was significantly influenced by genotype and P supply, but there was no significant genotype x P supply interaction. Hence, we conclude that screening genotypes using hydroponics at one or two P supply levels is the most cost- and time effective means to screen large numbers of rice genotypes for PUE.
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15.

Introduction

Raspberries are becoming increasingly popular due to their reported health beneficial properties. Despite the presence of only trace amounts of anthocyanins, yellow varieties seems to show similar or better effects in comparison to conventional raspberries.

Objectives

The aim of this work is to characterize the metabolic differences between red and yellow berries, focussing on the compounds showing a higher concentration in yellow varieties.

Methods

The metabolomic profile of 13 red and 12 yellow raspberries (of different varieties, locations and collection dates) was determined by UPLC–TOF-MS. A novel approach based on Pearson correlation on the extracted ion chromatograms was implemented to extract the pseudospectra of the most relevant biomarkers from high energy LC–MS runs. The raw data will be made publicly available on MetaboLights (MTBLS333).

Results

Among the metabolites showing higher concentration in yellow raspberries it was possible to identify a series of compounds showing a pseudospectrum similar to that of A-type procyanidin polymers. The annotation of this group of compounds was confirmed by specific MS/MS experiments and performing standard injections.

Conclusions

In berries lacking anthocyanins the polyphenol metabolism might be shifted to the formation of a novel class of A-type procyanidin polymers.
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16.

Objectives

To achieve biosynthesis of caffeoylmalic acid from glucose in engineered Escherichia coli.

Results

We constructed the biosynthetic pathway of caffeoylmalic acid in E. coli by co-expression of heterologous genes RgTAL, HpaBC, At4CL2 and HCT2. To enhance the production of caffeoylmalic acid, we optimized the tyrosine metabolic pathway of E. coli to increase the supply of the substrate caffeic acid. Consequently, an E. coliE. coli co-culture system was used for the efficient production of caffeoylmalic acid. The final titer of caffeoylmalic acid reached 570.1 mg/L.

Conclusions

Microbial production of caffeoylmalic acid using glucose has application potential. In addition, microbial co-culture is an efficient tool for producing caffeic acid esters.
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17.

Background, Goal, and Scope

Currently, only 40%, or 44.5 million metric tons, of coal combustion products (CCPs) generated in the United States each year by electric utilities are diverted from disposal in landfills or surface impoundments and recycled. Despite promising economic and environmental savings, there has been scant attention devoted to assessing life cycle impacts of CCP disposal and beneficial use. The objective of this paper is to present a life cycle inventory considering two cases of CCP management, including the stages of coal mining and preparation, coal combustion, CCP disposal, and CCP beneficial use. Six beneficial uses were considered: concrete production, structural fills, soil amendments, road construction, blasting grit and roofing granules, and wallboard.

Methods

Primary data for raw material inputs and emissions of all stages considered were obtained from surveys and site visits of coal-burning utilities in Florida conducted in 2002, and secondary data were obtained from various published sources and from databases available in SimaPro 5.1 (PRé Consultants, Amersfoort, The Netherlands).

Results

Results revealed that 50 percent of all CCPs produced, or 108 kg per 1,000 kg of coal combusted, are diverted for application in a beneficial use; however, the relative amounts sold by each utility is dependent on the process operating parameters, air emission control devices, and resulting quality of CCP. Diversion of 50% of all CCPs to beneficial use applications yields a decrease in the total raw materials requirements (with the exception of gravel and iron) and most emissions to air, water, and land, as compared to 100% disposal.

Discussion

The greatest reduction of raw materials was attributed to replacing Portland cement with fly ash, using bottom ash as an aggregate in concrete production and road construction in place of natural materials, and substituting FGD gypsum for natural gypsum in wallboard. The use of fly ash as cementitious material in concrete also promised significant reductions in emissions, particularly the carbon dioxide that would be generated from Portland cement production. Beneficial uses of fly ash and gypsum showed reductions of emissions to water (particularly total dissolved solids) and emissions of metals to land, although these reductions were small compared to simply diverting 50% of all CCPs from landfills or surface impoundments.

Conclusions

This life cycle inventory (LCI) provides the foundation for assessing the impacts of CCP disposal and beneficial use. Beneficial use of CCPs is shown here to yield reductions in raw material requirements and various emissions to all environmental compartments, with potential tangible savings to human health and the environment.

Recommendations and Perspectives

Extension of this life cycle inventory to include impact assessment and sensitivity analysis will enable a determination of whether the savings in emissions reported here actually result in significant improvements in environmental and human health impacts.
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18.

Purpose

This paper explores the issue of fairness in global supply chains. Taking the Western European clothing supply chain as a case study, we demonstrate how applying a normative indicator in Social Life Cycle Assessment (SLCA) can contribute academic and practical insights into debates on fairness. To do so, we develop a new indicator that addresses some of the limitations of the living wage for SLCA.

Methods

We extend the standard form of living wage available for developing countries to include income tax and social security contributions. We call this extension ‘living labour compensation’. Using publically available data, we estimate net living wages, gross living wages, and living labour compensation rates for Brazil, Russia, India, and China (BRIC) in 2005. We then integrate living labour compensation rates into an input-output framework, which we use to compare living labour compensation and actual labour compensation in the BRIC countries in the Western European clothing supply chain in 2005.

Results and discussion

We find that in 2005, actual labour compensation in the Western European clothing supply chain was around half of the living labour compensation level, with the greatest difference being in the Agricultural sector. Therefore, we argue that BRIC pay in the Western European clothing supply chain was unfair. Furthermore, our living labour compensation estimates for BRIC in 2005 are ~ 35% higher than standard living wage estimates. Indeed, adding income taxes and employee social security contributions alone increases the living wage by ~ 10%. Consequently, we argue there is a risk that investigations based on living wages are not using a representative measure of fairness from the employee’s perspective and are substantially underestimating the cost of living wages from an employer’s perspective. Finally, we discuss implications for retailers and living wage advocacy groups.

Conclusions

Living labour compensation extends the living wage, maintaining its strengths and addressing key weaknesses. It can be estimated for multiple countries from publically available data and can be applied in an input-output framework. Therefore, it is able to provide a normative assessment of fairness in complex global supply chains. Applying it to the Western European clothing supply chain, we were able to show that pay for workers in Brazil, Russia, India, and China is unfair, and draw substantive conclusions for practice.
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19.

Background

In recent years the visualization of biomagnetic measurement data by so-called pseudo current density maps or Hosaka-Cohen (HC) transformations became popular.

Methods

The physical basis of these intuitive maps is clarified by means of analytically solvable problems.

Results

Examples in magnetocardiography, magnetoencephalography and magnetoneurography demonstrate the usefulness of this method.

Conclusion

Hardware realizations of the HC-transformation and some similar transformations are discussed which could advantageously support cross-platform comparability of biomagnetic measurements.
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20.

Background

One of the most common side effects of the immunosuppressive drug tacrolimus (FK506) is the increased risk of new-onset diabetes mellitus. However, the molecular mechanisms underlying this association have not been fully clarified.

Methods

We studied the effects of the therapeutic dose of tacrolimus on mitochondrial fitness in beta-cells.

Results

We demonstrate that tacrolimus impairs glucose-stimulated insulin secretion (GSIS) in beta-cells through a previously unidentified mechanism. Indeed, tacrolimus causes a decrease in mitochondrial Ca2+ uptake, accompanied by altered mitochondrial respiration and reduced ATP production, eventually leading to impaired GSIS.

Conclusion

Our observations individuate a new fundamental mechanism responsible for the augmented incidence of diabetes following tacrolimus treatment. Indeed, this drug alters Ca2+ fluxes in mitochondria, thereby compromising metabolism-secretion coupling in beta-cells.
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