Insect cell conditioned medium contains an endoglycosidase able to liberate chondroitin sulfate chains from aggrecan.

Culture medium obtained from baculovirus-infected High Five insect cells contains an endoglycosidase activity capable of releasing chondroitin sulfate chains from aggrecan, decorin and biglycan. Release appears to occur by cleavage within the linkage region of the chondroitin sulfate chain, but not all chains are amenable to release. Culture medium from Sf9 insect cells does not contain this activity. The endoglycosidase may become a useful reagent for biochemical research for releasing intact chondroitin sulfate chains from proteoglycans. It may also represent an impediment to such research when baculovirus systems are used to generate recombinant proteoglycans.     

A global life cycle assessment for primary zinc production

Purpose

The purpose of this study was to update the average environmental impacts of global primary zinc production using a life cycle assessment (LCA) approach. This study represents the latest contribution from zinc producers, which historically established the first life cycle inventory for primary zinc production in 1998 (Western Europe) and the first global LCA-based cradle-to-gate study for zinc concentrate and special high-grade zinc (SHG; 99.99 %) in 2009. Improvements from the previous studies were realized through expanded geographical scope and range of production technologies.

Methods

The product system under study (SHG zinc) was characterized by collecting primary data for the relevant production processes, including zinc ore mining and concentration, transportation of the zinc concentrate, and zinc concentrate smelting. This data was modeled in GaBi 6 and complemented with background data from the GaBi 2013 databases to create the cradle-to-gate LCA model. Allocation was used to distribute the inputs and outputs among the various co-products produced during the production process, with mass of metal content being the preferred allocation approach, when applicable.

Results and discussion

In total, this global study includes primary data from 24 mines and 18 smelters, which cover 4.7?×?10⁶ MT of zinc concentrate and 3.4?×?10⁶ MT of SHG zinc, representing 36 and 27 % of global production, respectively. While the LCA model generated a full life cycle inventory, selected impact categories and indicators are reported in this article (global warming potential, acidification potential, eutrophication potential, photochemical ozone creation potential, ozone creation potential, and primary energy demand). The results show that SHG zinc has a primary energy demand of 37,500 MJ/t and a climate change impact of 2600 kg CO₂-eq./t. Across all impact categories and indicators reported here, around 65 % of the burden are associated with smelting, 30 % with mining and concentration, and 5 % with transportation of the concentrate. Sensitivity analyses were carried out for the allocation method (total mass versus mass of metal content) and transportation of zinc concentrate.

Conclusions

This study generated updated LCA information for the global production of SHG zinc, in line with the metal industry’s current harmonization efforts. Through the provision of unit process information for zinc concentrate and SHG zinc production, greater transparency is achieved. Technological and temporal representativeness was deemed to be high. Geographical representativeness, however, was found to be moderate to low. Future studies should focus on increasing company participation from underrepresented regions.

     

Interaction of a self-assembling peptide with oligonucleotides: complexation and aggregation

Molecular interaction of a self-assembling peptide, EAK16-II, to single- and double-stranded oligodeoxynucleotides (ODNs) was investigated under various solution conditions. The molecular events leading to EAK-ODN complexation and further aggregation were elucidated using a series of spectroscopic and microscopic methods. Despite the ability to self-assemble, EAK molecules bind to ODN molecules first upon mixing, resulting in EAK-ODN complexes. The complexes further associate to form EAK-ODN aggregates. A method based on UV-Vis absorption and centrifugation was developed to quantify the fraction of ODNs in the aggregates. The results were used to construct binding isotherms via a binding density function analysis. To compare the effects of different pH values and nucleotide types, the modified noncooperative McGhee and von Hippel model was used to extract binding parameters from the binding isotherms. The binding constant of EAK to ODNs was higher at pH 4 than at pH 7, and no binding was observed at pH 11, indicating that the interaction involved is primarily electrostatic in nature. EAK bound more strongly to single-stranded ODNs. The EAK-ODN aggregates were further visualized using atomic force microscopy; their size distribution as a function of EAK concentration was monitored by dynamic light scattering. The timescale for the EAK-ODN aggregation was on the order of minutes by fluorescence anisotropy and steady-state light scattering experiments. Fluorescence quenching experiments demonstrated that the ODNs in the aggregates were less accessible to the solvent, demonstrating a potential of oligonucleotide encapsulation by the self-assembling peptide.     

DNA bending by EcoRI DNA methyltransferase accelerates base flipping but compromises specificity.

EcoRI DNA methyltransferase was previously shown to bend its cognate DNA sequence by 52 degrees and stabilize the target adenine in an extrahelical orientation. We describe the characterization of an EcoRI DNA methyltransferase mutant in which histidine 235 was selectively replaced with asparagine. Steady-state kinetic and thermodynamic parameters for the H235N mutant revealed only minor functional consequences: DNA binding affinity (KDDNA) was reduced 10-fold, and kcat was decreased 30%. However, in direct contrast to the wild type enzyme, DNA bending within the mutant enzyme-DNA complexes was not observed by scanning force microscopy. The bending-deficient mutant showed enhanced discrimination against the methylation at nontarget sequence DNA. This enhancement of enzyme discrimination was accompanied by a change in the rate-limiting catalytic step. No presteady-state burst of product formation was observed, indicating that the chemistry step (or prior event) had become rate-limiting for methylation. Direct observation of the base flipping transition showed that the lack of burst kinetics was entirely due to slower base flipping. The combined data show that DNA bending contributes to the correct assembly of the enzyme-DNA complex to accelerate base flipping and that slowing the rate of this precatalytic isomerization can enhance specificity.     

Age-related changes in the structure of proteoglycan link proteins present in normal human articular cartilage.

Reduced-minus-oxidized difference spectra were recorded on particle preparations of the cyanobacterium Anacystis nidulans. Physiological oxidation of anaerobic membranes was effected either by O2 or by light. In both cases the spectral changes observed in the 550-570nm region were essentially the same. The results were confirmed by dual-wavelength spectrophotometry. It is concluded that a membrane-bound cytochrome f-b complex participates in both respiratory and photosynthetic elevtron transport.     

Animal disease and human trauma: the psychosocial implications of the 2001 UK foot and mouth disease disaster

The 2001 UK foot and mouth disease (FMD) crisis is commonly understood to have been a nonhuman animal problem, an economic industrial crisis that was resolved after eradication. By using a different lens, a longitudinal ethnographic study of the health and social consequences of the epidemic, the research reported here indicates that 2001 was a human tragedy as well as an animal one. In a diary-based study, it can be seen that life after the FMD crisis was accompanied by distress, feelings of bereavement, fear of a new disaster, loss of trust in authority and systems of control, and the undermining of the value of local knowledge. Diverse groups experienced distress well beyond the farming community. Such distress remained largely invisible to the range of “official” inquiries into the disaster. That an FMD epidemic of the scale of 2001 could happen again in a developed country is a deeply worrying prospect, but it is to be hoped that contingency plans are evolving along with enhanced understanding of the human, animal, and financial cost.     

Caenorhabditis elegans mutant allele identification by whole-genome sequencing

Identification of the molecular lesion in Caenorhabditis elegans mutants isolated through forward genetic screens usually involves time-consuming genetic mapping. We used Illumina deep sequencing technology to sequence a complete, mutant C. elegans genome and thus pinpointed a single-nucleotide mutation in the genome that affects a neuronal cell fate decision. This constitutes a proof-of-principle for using whole-genome sequencing to analyze C. elegans mutants.