Research priorities for managing the impacts and dependencies of business upon food, energy, water and the environment.

Delivering access to sufficient food, energy and water resources to ensure human wellbeing is a major concern for governments worldwide. However, it is crucial to account for the 'nexus' of interactions between these natural resources and the consequent implications for human wellbeing. The private sector has a critical role in driving positive change towards more sustainable nexus management and could reap considerable benefits from collaboration with researchers to devise solutions to some of the foremost sustainability challenges of today. Yet opportunities are missed because the private sector is rarely involved in the formulation of deliverable research priorities. We convened senior research scientists and influential business leaders to collaboratively identify the top forty questions that, if answered, would best help companies understand and manage their food-energy-water-environment nexus dependencies and impacts. Codification of the top order nexus themes highlighted research priorities around development of pragmatic yet credible tools that allow businesses to incorporate nexus interactions into their decision-making; demonstration of the business case for more sustainable nexus management; identification of the most effective levers for behaviour change; and understanding incentives or circumstances that allow individuals and businesses to take a leadership stance. Greater investment in the complex but productive relations between the private sector and research community will create deeper and more meaningful collaboration and cooperation.

A kinetic study of reactions of calcium-containing molecules with O and H atoms: implications for calcium chemistry in the upper atmosphere.

This paper describes the kinetic study of a number of gas-phase reactions involving neutral Ca-containing species, many of which are important for describing the chemistry of meteor-ablated calcium in the Earth's upper atmosphere. Ca atoms were produced thermally in the upstream section of a fast flow tube, and then converted to the molecular species CaO, CaO(2), CaO(3), CaCO(3) or Ca(OH)(2) by the addition of appropriate reagents. Atomic O or H was added further downstream, and both Ca and CaO were detected at the downstream end of the flow tube by laser-induced fluorescence. The following rate coefficients were determined: k(CaO + O --> Ca + O(2)) = (3.1) x 10(-10) at 300 K and (1.3) x 10(-10) at 203 K; k(CaO(2) + O --> CaO + O(2)) = (2.2) x 10(-11) at 300 K and (1.6) x 10(-11) at 203 K; k(CaO(2) + H --> products, 298 K) = (1.2 +/- 0.6) x 10(-11); k(CaCO(3) + O --> CaO(2) + CO(2), 300 K) < or = 1.0 x 10(-12); k(CaCO(3) + H--> CaOH + CO(2), 298 K) > or = 2.8 x 10(-12) and < or = 3.6 x 10(-11); k(CaO(3) + H--> CaOH + O(2), 298 K) > or = 1.7 x 10(-11); k(Ca(OH)(2) + H --> CaOH + H(2)O, 298 K) > or = 1.1 x 10(-11); k(CaOH + H --> Ca + H(2)O, 298 K) > or = 1.1x 10(-11) cm(3) molecule(-1) s(-1). The kinetics of the reactions of Ca and CaO with NO(2) and N(2)O were also studied, yielding k(Ca + NO(2) --> CaO + NO) = (2.6 +/- 0.3) x 10(-10) at 300 K and (2.0 +/- 0.3) x 10(-10) at 203 K; k(CaO + NO(2) --> CaO(2) + NO) = (8.1 +/- 2.0) x 10(-10) at 300 K and (2.9 +/- 1.0) x 10(-10) at 202 K; k(CaO + N(2)O --> CaO(2) + N(2)) = (4.2 +/- 1.7) x 10(-11) at 300 K and (2.2 +/- 1.2) x 10(-12) at 206 K; k(CaO + H(2) --> Ca + H(2)O, 300 K) = (3.4 +/- 1.3) x 10(-10) cm(3) molecule(-1) s(-1). Electronic structure calculations of the relevant potential energy surfaces were performed to interpret the experimental results, and the atmospheric implications of these measurements are then discussed.

The role of molecular chaperones in human misfolding diseases.

Human misfolding diseases arise when proteins adopt non-native conformations that endow them with a tendency to aggregate and form intra- and/or extra-cellular deposits. Molecular chaperones, such as Hsp70 and TCP-1 Ring Complex (TRiC)/chaperonin containing TCP-1 (CCT), have been implicated as potent modulators of misfolding disease. These chaperones suppress toxicity of disease proteins and modify early events in the aggregation process in a cooperative and sequential manner reminiscent of their functions in de novo protein folding. Further understanding of the role of Hsp70, TRiC, and other chaperones in misfolding disease is likely to provide important insight into basic pathomechanistic principles that could potentially be exploited for therapeutic purposes.

N-terminal polyglutamine-containing fragments inhibit androgen receptor transactivation function.

Abstract Several neurodegenerative diseases, including Kennedy's disease (KD), are associated with misfolding and aggregation of polyglutamine (polyQ)-expansion proteins. KD is caused by a polyQ-expansion in the androgen receptor (AR), a key player in male sexual differentiation. Interestingly, KD patients often show signs of mild-to-moderate androgen insensitivity syndrome (AIS) resulting from AR dysfunction. Here, we used the yeast Saccharomyces cerevisiae to investigate the molecular mechanism behind AIS in KD. Upon expression in yeast, polyQ-expanded N-terminal fragments of AR lacking the hormone binding domain caused a polyQ length-dependent growth defect. Interestingly, while AR fragments with 67 Q formed large, SDS-resistant inclusions, the most pronounced toxicity was observed upon expression of 102 Q fragments which accumulated exclusively as soluble oligomers in the 100-600 kDa range. Analysis using a hormone-dependent luciferase reporter revealed that full-length polyQ-expanded AR is fully functional in transactivation, but becomes inactivated in the presence of the corresponding polyQ-expanded N-terminal fragment. Furthermore, the greatest impairment of AR activity was observed upon interaction of full-length AR with soluble AR fragments. Taken together, our results suggest that soluble polyQ-containing fragments bind to full-length AR and inactivate it, thus providing insight into the mechanism behind AIS in KD and possibly other polyglutamine diseases, such as Huntington's disease.

A kinetic study of Ca-containing ions reacting with O, O2, CO2 and H2O: implications for calcium ion chemistry in the upper atmosphere.

A series of gas-phase reactions involving molecular Ca-containing ions was studied by the pulsed laser ablation of a calcite target to produce Ca+ in a fast flow of He, followed by the addition of reagents downstream and detection of ions by quadrupole mass spectrometry. Most of the reactions that were studied are important for describing the chemistry of meteor-ablated calcium in the earth's upper atmosphere. The following rate coefficients were measured: k(CaO+ + O --> Ca+ + O2) = (4.2 +/- 2.8) x 10(-11) at 197 K and (6.3 +/- 3.0) x 10(-11) at 294 K; k(CaO+ + CO --> Ca+ + CO2, 294 K) = (2.8 +/- 1.5) x 10(-10); k(Ca+.CO2 + O2 --> CaO2+ + CO2, 294 K) = (1.2 +/- 0.5) x10(-10); k(Ca+.CO2 + H2O --> Ca+.H2O + CO2) = (13.0 +/- 4.0) x 10(-10); and k(Ca+.H2O + O2 --> CaO2+ + H2O, 294 K) = (4.0 +/- 2.5) x 10(-10) cm3 molecule(-1) s(-1). The quoted uncertainties are a combination of the 1 sigma standard errors in the kinetic data and the systematic errors in the models used to extract the rate coefficients. Rate coefficients were also obtained for the following recombination (also termed association) reactions in He bath gas: k(Ca+.CO2 + CO2 --> Ca+.(CO2)2, 294 K) = (2.6 +/- 1.0) x 10(-29); k(Ca+.H2O + H2O --> Ca+.(H2O)2) = (1.6 +/- 1.1) x 10(-27); and k(CaO2+ + O2 --> CaO2+.O2) < 1 x 10(-31) cm6 molecule(-2) s(-1). These recombination rate coefficients, as well as those for the ligand-switching reactions listed above, were then interpreted using a combination of high level quantum chemistry calculations and RRKM theory using an inverse Laplace transform solution of the master equation. The surprisingly slow reaction between CaO+ and O was explained using quantum chemistry calculations on the lowest 2A', 2A'' and 4A'' potential energy surfaces. These calculations indicate that reaction mostly occurs on the 2A' surface, leading to production of Ca+ (2S) + O2(1 Delta g). The importance of this reaction for controlling the lifetime of Ca+ in the upper mesosphere and lower thermosphere is then discussed.

Mitochondrial stress signaling: a pathway unfolds.

Disruption of protein homeostasis in mitochondria elicits a cellular response, which upregulates mitochondrial chaperones and other factors that serve to remodel the mitochondrial-folding environment. In a recent study, Haynes and colleagues uncovered a novel signal transduction pathway underlying this process. The upstream mitochondrial component of this pathway is an orthologue of Escherichia coli ClpP, which functions in the bacterial heat-shock response. These findings suggest that molecular aspects of stress sensing might be conserved between bacteria and mitochondria.

A kinetic study of the reactions of Ca+ ions with O3, O2, N2, CO2 and H2O.

The reactions between Ca(+)(4(2)S(1/2)) and O(3), O(2), N(2), CO(2) and H(2)O were studied using two techniques: the pulsed laser photo-dissociation at 193 nm of an organo-calcium vapour, followed by time-resolved laser-induced fluorescence spectroscopy of Ca(+) at 393.37 nm (Ca(+)(4(2)P(3/2)-4(2)S(1/2))); and the pulsed laser ablation at 532 nm of a calcite target in a fast flow tube, followed by mass spectrometric detection of Ca(+). The rate coefficient for the reaction with O(3) is essentially independent of temperature, k(189-312 K) = (3.9 +/- 1.2) x 10(-10) cm(3) molecule(-1) s(-1), and is about 35% of the Langevin capture frequency. One reason for this is that there is a lack of correlation between the reactant and product potential energy surfaces for near coplanar collisions. The recombination reactions of Ca(+) with O(2), CO(2) and H(2)O were found to be in the fall-off region over the experimental pressure range (1-80 Torr). The data were fitted by RRKM theory combined with quantum calculations on CaO(2)(+), Ca(+).CO(2) and Ca(+).H(2)O, yielding the following results with He as third body when extrapolated from 10(-3)-10(3) Torr and a temperature range of 100-1500 K. For Ca(+) + O(2): log(10)(k(rec,0)/cm(6) molecule(-2) s(-1)) = -26.16 - 1.113log(10)T- 0.056log(10)(2)T, k(rec,infinity) = 1.4 x 10(-10) cm(3) molecule(-1) s(-1), F(c) = 0.56. For Ca(+) + CO(2): log(10)(k(rec,0)/ cm(6) molecule(-2) s(-1)) = -27.94 + 2.204log(10)T- 1.124log(10)(2)T, k(rec,infinity) = 3.5 x 10(-11) cm(3) molecule(-1) s(-1), F(c) = 0.60. For Ca(+) + H(2)O: log(10)(k(rec,0)/ cm(6) molecule(-2) s(-1)) = -23.88 - 1.823log(10)T- 0.063log(10)(2)T, k(rec,infinity) = 7.3 x 10(-11)exp(830 J mol(-1)/RT) cm(3) molecule(-1) s(-1), F(c) = 0.50 (F(c) is the broadening factor). A classical trajectory analysis of the Ca(+) + CO(2) reaction is then used to investigate the small high pressure limiting rate coefficient, which is significantly below the Langevin capture frequency. Finally, the implications of these results for calcium chemistry in the mesosphere are discussed.

Translocation of mitochondrially synthesized Cox2 domains from the matrix to the intermembrane space.

The N-terminal and C-terminal domains of mitochondrially synthesized cytochrome c oxidase subunit II, Cox2, are translocated through the inner membrane to the intermembrane space (IMS). We investigated the distinct mechanisms of N-tail and C-tail export by analysis of epitope-tagged Cox2 variants encoded in Saccharomyces cerevisiae mitochondrial DNA. Both the N and C termini of a truncated protein lacking the Cox2 C-terminal domain were translocated to the IMS via a pathway dependent upon the conserved translocase Oxa1. The topology of this Cox2 variant, accumulated at steady state, was largely but not completely unaffected in mutants lacking proteins required for export of the C-tail domain, Cox18 and Mss2. C-tail export was blocked by truncation of the last 40 residues from the C-tail domain, indicating that sequence and/or structural features of this domain are required for its translocation. Mss2, a peripheral protein bound to the inner surface of the inner membrane, coimmunoprecipitated with full-length newly synthesized Cox2, whose leader peptide had already been cleaved in the IMS. Our data suggest that the C-tail domain is recognized posttranslationally by a specialized translocation apparatus after the N-tail has been translocated by Oxa1.

Identification of anti-prion compounds as efficient inhibitors of polyglutamine protein aggregation in a zebrafish model.

Several neurodegenerative diseases, including Huntington disease (HD), are associated with aberrant folding and aggregation of polyglutamine (polyQ) expansion proteins. Here we established the zebrafish, Danio rerio, as a vertebrate HD model permitting the screening for chemical suppressors of polyQ aggregation and toxicity. Upon expression in zebrafish embryos, polyQ-expanded fragments of huntingtin (htt) accumulated in large SDS-insoluble inclusions, reproducing a key feature of HD pathology. Real time monitoring of inclusion formation in the living zebrafish indicated that inclusions grow by rapid incorporation of soluble htt species. Expression of mutant htt increased the frequency of embryos with abnormal morphology and the occurrence of apoptosis. Strikingly, apoptotic cells were largely devoid of visible aggregates, suggesting that soluble oligomeric precursors may instead be responsible for toxicity. As in nonvertebrate polyQ disease models, the molecular chaperones, Hsp40 and Hsp70, suppressed both polyQ aggregation and toxicity. Using the newly established zebrafish model, two compounds of the N'-benzylidene-benzohydrazide class directed against mammalian prion proved to be potent inhibitors of polyQ aggregation, consistent with a common structural mechanism of aggregation for prion and polyQ disease proteins.

Kinetic study of the reaction Ca+ + N2O from 188 to 1207 K.

Ion-molecule reactions involving metallic species play a central role in the chemistry of planetary ionospheres and in many combustion processes. The kinetics of the Ca(+) + N(2)O --> CaO(+) + N(2) reaction was studied by the pulsed multiphoton dissociation at 193 nm of organo-calcium vapor in the presence of N(2)O, followed by time-resolved laser-induced fluorescence spectroscopy of Ca(+) at 393.37 nm (4(2)P(3/2) <-- 4(2)S(1/2)). This yielded k(188-1207 K) = 5.45 x 10(-11) (T/300 K)(0.53) exp(282 K/T) cm(3) molecule(-1) s(-1), with an estimated accuracy of +/-13% (188-600 K) and +/-27% (600-1207 K). The temperature dependence of this barrierless reaction, with a minimum in the rate coefficient between 400 and 600 K, appears to be explained by the role of N(2)O vibrational excitation. This is examined using a classical trajectory treatment on a potential energy surface calculated at the B3LYP/6-311+g(2d,p) level of theory.

Molecular chaperones of the Hsp110 family act as nucleotide exchange factors of Hsp70s.

Hsp70 molecular chaperones function in protein folding in a manner dependent on regulation by co-chaperones. Hsp40s increase the low intrinsic ATPase activity of Hsp70, and nucleotide exchange factors (NEFs) remove ADP after ATP hydrolysis, enabling a new Hsp70 interaction cycle with non-native protein substrate. Here, we show that members of the Hsp70-related Hsp110 family cooperate with Hsp70 in protein folding in the eukaryotic cytosol. Mammalian Hsp110 and the yeast homologues Sse1p/2p catalyze efficient nucleotide exchange on Hsp70 and its orthologue in Saccharomyces cerevisiae, Ssa1p, respectively. Moreover, Sse1p has the same effect on Ssb1p, a ribosome-associated isoform of Hsp70 in yeast. Mutational analysis revealed that the N-terminal ATPase domain and the ultimate C-terminus of Sse1p are required for nucleotide exchange activity. The Hsp110 homologues significantly increase the rate and yield of Hsp70-mediated re-folding of thermally denatured firefly luciferase in vitro. Similarly, deletion of SSE1 causes a firefly luciferase folding defect in yeast cells under heat stress in vivo. Our data indicate that Hsp110 proteins are important components of the eukaryotic Hsp70 machinery of protein folding.

Proteolytic cleavage of polyglutamine-expanded ataxin-3 is critical for aggregation and sequestration of non-expanded ataxin-3.

Spinocerebellar ataxia type 3 (SCA3), like other polyglutamine (polyQ) diseases, is characterized by the formation of intraneuronal inclusions, but the mechanism underlying their formation is poorly understood. Here, we tested the "toxic fragment hypothesis", which predicts that proteolytic production of polyQ-containing fragments from the full-length disease protein initiates the aggregation process associated with inclusion formation and cellular dysfunction. We demonstrate that the removal of the N-terminus of polyQ-expanded ataxin-3 (AT3) is required for aggregation in vitro and in vivo. Consistently, proteolytic cleavage of full-length, pathogenic AT3 initiates the formation of sodium dodecylsulfate-resistant aggregates in neuroblastoma cells. Although full-length AT3 does not readily aggregate on its own, it is susceptible to co-aggregation with polyQ-expanded AT3 fragments. Interestingly, interaction with soluble polyQ-elongated fragments causes a structural distortion of wild-type AT3 prior to the formation of stable co-aggregates. These results establish the critical role of C-terminal, proteolytic fragments of AT3 in the molecular pathomechanism of SCA3, in strong support of the toxic fragment hypothesis.

Vertex-linked ZnO2S2 tetrahedra in the oxysulfide BaZnOS: a new coordination environment for zinc in a condensed solid.

The wide-band-gap semiconductor BaZnOS adopts a high-symmetry modification of the SrZnO2 structure type and contains layers of vertex-linked ZnO2S2 tetrahedra, which represent a novel coordination environment for zinc in the solid state. BaZnOS: orthorhombic, space group Cmcm; a = 3.9619(2) angstroms, b = 12.8541(7) angstroms, c = 6.1175(4) angstroms, Z = 4. Diffuse-reflectance spectroscopy measurements reveal a direct band gap of 3.9(3) eV, consistent with the white color and the results of band structure calculations. The band gap is larger than those observed in ZnO and ZnS, consistent with the more ionic nature of BaZnOS. Attempts to dope this compound electronically have so far not proved possible.

Roles of molecular chaperones in protein misfolding diseases.

Human misfolding diseases result from the failure of proteins to reach their active state or from the accumulation of aberrantly folded proteins. The mechanisms by which molecular chaperones influence the development of these diseases is beginning to be understood. Mutations that compromise the activity of chaperones lead to several rare syndromes. In contrast, the more frequent amyloid-related neurodegenerative diseases are caused by a gain of toxic function of misfolded proteins. Toxicity in these disorders may result from an imbalance between normal chaperone capacity and production of dangerous protein species. Increased chaperone expression can suppress the neurotoxicity of these molecules, suggesting possible therapeutic strategies.

Mss51p promotes mitochondrial Cox1p synthesis and interacts with newly synthesized Cox1p.

The post-transcriptional role of Mss51p in mitochondrial gene expression is of great interest since MSS51 mutations suppress the respiratory defect caused by shy1 mutations. SHY1 is a Saccharomyces cerevisiae homolog of human SURF1, which when mutated causes a cytochrome oxidase assembly defect. We found that MSS51 is required for expression of the mitochondrial reporter gene ARG8(m) when it is inserted at the COX1 locus, but not when it is at COX2 or COX3. Unlike the COX1 mRNA-specific translational activator PET309, MSS51 has at least two targets in COX1 mRNA. MSS51 acts in the untranslated regions of the COX1 mRNA, since it was required to synthesize Arg8p when ARG8(m) completely replaced the COX1 codons. MSS51 also acts on a target specified by the COX1 coding region, since it was required to translate either COX1 or COX1:: ARG8(m) coding sequences from an ectopic COX2 locus. Mss51p was found to interact physically with newly synthesized Cox1p, suggesting that it could coordinate Cox1p synthesis with insertion into the inner membrane or cytochrome oxidase assembly.