Modelling the Valuesphere and the Ecosphere:

Methods for Life Cycle Impact Assessment have to cope with two critical aspects, the uncertainty in values and the (unknown) system behaviour. LCA methodology should cope explicitly with these subjective elements. A structured aggregation procedure is proposed that differentiates between the technosphere and the ecosphere and embeds them in the valuesphere. LCA thus becomes a decision support system that models and combines these three spheres. We introduce three structurally identical types of LCA, each based on one coherent but different set of values. These sets of values can be derived from the Cultural Theory and are labeled as ‘egalitarian’, ‘individualistic’, and ‘hierarchic’. Within Life Cycle Impact Assessment, a damage oriented assessment model is complemented with both a newly developed precautionary indicator designed to address unknown damage and an indicator for the manageability of environmental damages. The indicators for unknown damage and for manageability complete the set of indicators judged to be relevant by decision makers. The weights given to these indicators are also value-dependent. The framework proposed here answers the criticisms that present LCA methodology does not strictly enough separate subjective from objective elements and that it fails to accurately model environmental impacts.     

An Overview on the Enhancement of Enantioselectivity and Stability of Microbial Epoxide Hydrolases

Epoxide hydrolases (EHs; 3.3.2.x) catalyze the enantioselective ring opening of racemic epoxides to the corresponding enantiopure vicinal diols and remaining equivalent unreacted epoxides. These epoxides and diols are used for the synthesis of chiral drug intermediates. With an upsurge in the methods for identification of novel microbial EHs, a lot of EHs have been discovered and utilized for kinetic resolution of racemic epoxides. However, there is still a constraint on the account of limited EHs being successfully applied on the preparative scale for industrial biotransformations. This limitation has to be overcome before application of identified functional EHs on large scale. Many strategies such as optimizing reaction media, immobilizing EHs and laboratory-scale directed evolution of EHs have been adopted for enhancing the industrial potential of EHs. In this review, these approaches have been highlighted which can serve as a pathway for the enrichment of already identified EHs for their application on an industrial scale in future studies.     

Enhancement of the Stability of Insoluble Calcium Particles Using a Phospholipid Coating

Calcium is one of the most important elements in the human body. Insoluble calcium particles are often used in calcium-fortified food products, such as calcium-fortified milk, dairy beverages or protein powders. However, their suspension may be unstable often leading to precipitation in such products. In this study, three different kinds of insoluble calcium particles, i.e. hydroxyapatite (HA), tricalcium phosphate (TCP) and calcium carbonate (CaCO₃) were coated with amphiphilic phospholipids using a solvent-exchange method. Suspension stability of these insoluble calcium particles was effectively improved with phospholipid coating, especially for HA and TCP, as more phospholipids were coated on the surface of these two calcium particles than CaCO₃. Phospholipid coating increased the electrostatic repulsions between particles, preventing the particles from aggregating and precipitating. In addition, the digestibility of phospholipid-coated insoluble calcium particles was tested in simulated gastric juice, and the dissolution time of these insoluble calcium particles was prolonged through phospholipid coating.     

Hydrophobicity Density Profiles to Predict Thermal Stability Enhancement in Proteins

A hydrophobicity density is defined for a protein through its hydrophobicity tensor (similar to the inertia tensor), by using the Eisenberg hydrophobicity scale of the hydrophobic amino acids of a protein. This allows calculation of the radii of the corresponding hydrophobic ellipsoid of a protein and thus subsequently of its hydrophobic density. A hydrophobicity density profile is then obtained by simulating point mutations of each amino acid of a protein either to a high hydrophobicity value or to zero hydrophobicity. It is found that an increase in the hydrophobic density of the protein correlates with an increase of its mid-point transition temperature. From this profile it is possible to determine the amino acids or domain stretches in a protein that are most amenable to mutation in order to increase the thermal stability. The model is tested to predict the thermostabilisation effects of two mutations in a β-glucanase: M29G and M29F. This model is compared with other hydrophobicity-related profiles described by other authors.Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

Stability Enhancement for Peptide Analysis by Electrospray Using the Triple Quadrupole Mass Spectrometer

Electrospray ionization sources, used with triple quadrupole mass spectrometers from PE/Sciex (API III+), Micromass (Quattro II), and Finnigan (TSQ 7000), were modified with a 35-gauge stainless steel needle. The dimensions of the needle were 63 μm i.d. by 145 μm o.d. with variable length, depending on the specific instrument. This modification led to enhanced signal stability, improved signal/noise ratios, and lowered sample consumption for a wide range of peptides. Stable baselines were observed with flow rates in the range of 50 nL/min to 5 μL/min. An alternative design, based on a metal wire housed within a fused silica capillary, led to the most stable signals of all during infusion, but caused excessive peak broadening with capillary chromatography. The Finnigan interface was further modified with an external postcolumn addition tee, used in conjunction with capillary liquid chromatography columns of 30 and 50 μm internal diameter. The best results with the modified Finnigan interface were acquired using the 50-μm column at a flow rate of 150 to 200 nL/min.     

Biogenic amines in the regulation of wakefulness and sleep

Neurophysiological, neurochemical and neuropharmacological evidence indicates that cerebral monoamines are important regulators of wakefulness and sleep besides cerebral amino acid-ergic and peptidergic systems. The cerebral monoamines noradrenaline, dopamine and acetylcholine are positively involved in electroencephalographic aspects of waking and paradoxical or REM sleep. A high level of noradrenergic transmission facilitates waking, and a lower, moderate level facilitates REM sleep. Serotonin is involved in the regulation of synthesis, storage and release of sleep inducing factors, and in the gating mechanisms of REM sleep. Histamine neurons play a role in the regulation of vigilance during waking state. These neurotransmitter systems are important targets for drug actions.     

Dietary Folate and Biogenic Amines in the CNS

Abnormal biogenic amine biosynthesis has been observed in humans and animals with endogenous and exogenous disturbances in folate metabolism. In an attempt to study this interaction biochemically, rats were depleted or repleted with folate for 10 weeks. Folate levels in depleted animals in serum and CSF correlated with stores in liver and brain, respectively. In depleted or repleted animals, there was no significant effect on biogenic amine metabolism in the CNS, as determined by quantitation of biogenic amines in brain and their respective metabolites in brain and CSF. These results are contrary to studies by other investigators. We suspect, however, that specific genetic defects in folate metabolism do result in impaired biogenic amine metabolism and probably at the level of disturbed biopterin cofactor functions.     

Biogenic Amines in the Vomeronasal Organ

The vomeronasal organ of frog and mouse was investigated forthe presence and content of serotonin and catecholamines bymeans of high-performance liquid chromatography. Measurableamounts of serotonin, adrenaline and noradrenaline were foundin the vomeronasal organ of adult individuals of both species.The amine content varied with sex of adult frogs and mice andsexual maturity of mice. In preliminary experiments, acute exposureto male urine containing pheromone affected the amine contentin the vomeronasal organ of adult female mice. These data suggestthat functional sex dimorphism is present in the vomeronasalorgan, and biochemical changes therein take place accordingto stage of sexual maturity. The role of biogenic amines inthe vomeronasal organ deserves further study. Chem. Senses 22:439–445, 1997.     

Biogenic Amines in the Taste Organ

The presence and content of biogenic amines in taste disk-bearingfungiform papillae of the frog, Rana esculenta, the only availablemodel of an isolated taste organ, were verified by means ofHPLC. Fungiform papillae were found to contain measurable amountsof serotonin, epinephrine and norepinephrine. The amounts ofserotonin and epinephrine were significantly higher in fungiformpapillae than in the general mucosa of the tongue. Moreover,the epinephrine content of fungiform papillae was found to differacross the tongue, in accordance with previous physiologicalstudies showing an inhomogeneous response of different tongueregions to taste stimuli. Ultrastructural and histochemicalinvestigations confirmed the presence of catecholamine and serotonin.The latter was found to be contained mainly in the basal cellsof the frog taste disk. These results extend previous qualitativedata on the presence of biogenic amines in taste chemoreceptors.Chem. Senses 20: 329–335, 1995.     

Ecosphere,biosphere, or Gaia? What to call the global ecosystem

The terms biosphere, ecosphere, and Gaia are used as names for the global ecosystem. However, each has more than one meaning. Biosphere can mean the totality of living things residing on the Earth, the space occupied by living things, or life and life-support systems (atmosphere, hydrosphere, lithosphere, and pedosphere). Ecosphere is used as a synonym of biosphere and as a term for zones in the universe where life as we know it should be sustainable. Gaia is similar to biosphere (in the sense of life and life-support systems) and ecosphere (in the sense of biosphere as life and life-support systems), but, in its most extreme form, refers to the entire planet as a living entity. A case is made for avoiding the term Gaia (at least as a name for the planetary ecosystem), restricting biosphere to the totality of living things, and adopting the ecosphere as the most apt name for the global ecosystem.     

Interface Modification of Tin Oxide Electron-Transport Layer for the Efficiency and Stability Enhancement of Organic Solar Cells

A small molecular phenanthroline derivative Phen-NaDPO (3-[6-(diphenylphosphinyl)-2-naphthalenyl]-1,10-Phenanthroline) to modify tin oxide (SnO₂) electron-transport layer (ETL) in organic solar cells is employed. Quantum chemistry calculations and experimental results show that Phen-NaDPO can interact with SnO₂, thereby effectively passivating the surface defects, reducing the work function and improving the electrical conductivity of SnO₂, leading to more efficient electron extraction and transport in the organic solar cells (OSCs). Moreover, upon the Phen-NaDPO modification, the decreased surface energy of SnO₂ ETL accounts for enhanced exciton dissociation and charge transport, due to the more ordered molecular organizations of the active layers. Consequently, the inverted OSCs involving Phen-NaDPO/SnO₂ ETLs exhibit an enhanced power conversion efficiency of 17.06% (PM6:Y6) and 18.31% (PM6:L8-BO), which is the highest efficiency for SnO₂ ETL-based binary solar cells to date. Furthermore, the devices based on Phen-NaDPO/SnO₂ ETL show better device stability (storage stability, photostability and humid stability), with T₈₀ exceeding 200 h encapsulated under light irradiation and 400 h without encapsulation in high-humidity ambient condition. These results demonstrate that the modification of SnO₂ using wide-band highly stable conjugated small molecules is very promising for simultaneously improve the efficiencies and device stability of OSCs.     

Enhancement of the Stability of a Prolipase from Rhizopus oryzae toward Aldehydes by Saturation Mutagenesis

A prolipase from Rhizopus oryzae (proROL) was engineered in order to increase its stability toward lipid oxidation products such as aldehydes with the aim of improving its performance in oleochemical industries. Out of 22 amino acid residues (15 Lys and 7 His) prone to react with aldehydes, 6 Lys and all His residues (except for the catalytic histidine) were chosen and subjected to saturation mutagenesis. In order to quickly and reliably identify stability mutants within the resulting libraries, active variants were prescreened by an activity staining method on agar plates. Active mutants were expressed in Escherichia coli Origami in a 96-well microtiterplate format, and a stability test using octanal as a model deactivating agent was performed. The most stable histidine mutant (H201S) conferred a stability increase of 60%, which was further enhanced to 100% by combination with a lysine mutant (H201S/K168I). This increase in stability was also confirmed for other aldehydes. Interestingly, the mutations did not affect specific activity, as this was still similar to the wild-type enzyme.     

Stability Enhancement of Drug Layered Pellets in a Fixed Dose Combination Tablet

The purpose of this research was to develop a stable fixed dose combination tablet for a model DPP-IV inhibitor and metformin hydrochloride. The dipeptidyl peptidase IV (DPP-IV) inhibitor was particularly challenging to formulate due to its significant chemical instability and moisture sensitivity. Various formulation strategies were investigated and placed on accelerated stability to determine the lead approach and critical quality attributes. The lead formulation investigated was a drug layered pellet containing the DPP-IV inhibitor, which was further coated with various seal coats and moisture barriers, then compressed into a tablet with compression aids and granulated metformin hydrochloride. The investigations revealed that the drug layered pellets compressed into a fixed dose combination tablet yielded a unique stability enhancement. The stability was highly dependent on the final tablet water content and could be further improved by the addition of moisture barrier coatings. A fundamental understanding of the key critical quality attributes for the fixed dose combination product containing a DPP-IV inhibitor and metformin hydrochloride as an oral solid dosage form were established. This research identified a formulation approach to enable a successful commercial product to be developed.     

Biogenic capsules made of proteins and lipids

Multilayer biogenic capsules with a micrometer scale were fabricated by self-assembly of proteins and lipids at the interface of emulsion droplets. The optical microscopy images demonstrate that spherical capsules at a fluid interface have uniform walls and the dried capsules possess a high mechanical strength. The hollow shells obtained provide a novel class of assembly with encapsulating drug molecules based on the layer-by-layer technique.     

Biogenic amines in the two-spotted spider mite

Whole body extracts of the two-spotted spider mite (Tetranychus urticae Koch) were analyzed using a 16-channel electrochemical array high performance liquid chromatography (HPLC)-based detection system that allows the simultaneous isolation and identification of a variety of biogenic amines. The spider mite extracts were found to contain the biogenic amines octopamine, dopamine, and 5-hydroxytryptamine (5-HT), as well as several precursors and metabolites including tyrosine, tyramine, tryptophan, and N-acetyl octopamine. Differences in the levels of biogenic amines were observed between eggs and the adult stages and between males and females. This is the first direct determination of biogenic amines in the Tetranychidae and the first demonstration of 5-HT in any mite species. © 1994 Wiley-Liss, Inc.