Cold Temperature Limits to Biodiesel Use under Present and Future Climates in North America.

Cold weather operability is sometimes a limiting factor in the use of biodiesel blends for transportation. Regional temperature variability can therefore influence biodiesel adoption, with potential economic and environmental implications. This study assesses present and future biodiesel cold weather operability limits in North America according to temperature data from weather stations, atmospheric reanalysis, and global climate models with highest resolution over Ontario, Canada. Future temperature projections using the RCP8.5 climate change scenario show increases in the potential duration for certain seasonal fuel blends. For example, biodiesel blends whose cloud point temperature is -9 °C may expand their duration by 3-7% in North America for nonwinter seasons according to projections for 2040. Cloud point specifications among supply orbits in Ontario increase up to +6 °C during nonwinter seasons, with most increases observed in Fall and Spring. In winter, however, the modeling suggests no change in Ontario cloud point specifications because the coldest temperatures by mid-century are not significantly warmer than the past climate normal according to our climate simulations. This study provides a quantitative analysis on biodiesel usage scenarios under a changing climate, including Ontario region geographic temperature clusters that could prove useful for biodiesel blend-related decision-making.

Environmental Aspects of Biotechnology.

A key motivation behind the development and adoption of industrial biotechnology is the reduction of negative environmental impacts. However, accurately assessing these impacts remains a formidable task. Environmental impacts of industrial biotechnology may be significant across a number of categories that include, but may not be limited to, nonrenewable resource depletion, water withdrawals and consumption, climate change, and natural land transformation/occupation. In this chapter, we highlight some key environmental issues across two broad areas: (a) processes that use biobased feedstocks and (b) industrial activity that is supported by biological processes. We also address further issues in accounting for related environmental impacts such as geographic and temporal scope, co-product management, and uncertainty and variability in impacts. Case studies relating to (a) lignocellulosic ethanol, (b) biobased plastics, and (c) enzyme use in the detergent industry are then presented, which illustrate more specific applications. Finally, emerging trends in the area of environmental impacts of biotechnology are discussed.

Principles of Renal Function Measurement (Limitations of Gotch's Kt/V).

Kt/V is a nondimensional number and a scaling parameter that has, with arbitrary definitions, been recast as a measure of dialysis by Gotch and Lysaght. This editorial discusses the concept of nondimensional numbers within the context of dialysis measurement, modeling, and medical evidence. It concludes that Gotch's Kt/V, Lysaght's Kt/V, and standardized Kt/V are not well suited to measure dialysis. An ideal dialysis measure would be proportional to toxins cleared by the kidney, portable with regard to dialysis modality, practical (largely devoid of calculations) and reflective of the pathology/physiology.

Le Kt/V est un nombre non dimensionnel et un paramètre de dispersion qui, selon des définitions arbitraires, est devenu une mesure de l'efficacité de la dialyse avec Gotch et Lysaght. Le présent éditorial discute du concept de nombre non dimensionnel dans le contexte de la mesure de l'efficacité de la dialyse, de la modélisation et de la médecine factuelle. L'article conclut que le Kt/V de Gotch, le Kt/V de Lysaght et le Kt/V standardisé sont inadéquats pour la mesure de l'efficacité de la dialyse. En effet, une mesure idéale de l'efficacité de la dialyse serait proportionnelle aux toxines éliminées par les reins, adaptable à la modalité de dialyse, facile à utiliser (largement dénuée de calculs) et représentative de la pathologie/physiologie du patient.

Life cycle assessment of lignocellulosic ethanol: a review of key factors and methods affecting calculated GHG emissions and energy use.

Lignocellulosic ethanol has potential for lower life cycle greenhouse gas emissions compared to gasoline and conventional grain-based ethanol. Ethanol production 'pathways' need to meet economic and environmental goals. Numerous life cycle assessments of lignocellulosic ethanol have been published over the last 15 years, but gaps remain in understanding life cycle performance due to insufficient data, and model and methodological issues. We highlight key aspects of these issues, drawing on literature and a case study of corn stover ethanol. Challenges include the complexity of feedstock/ecosystems and market-mediated aspects and the short history of commercial lignocellulosic ethanol facilities, which collectively have led to uncertainty in GHG emissions estimates, and to debates on LCA methods and the role of uncertainty in decision making.

Life cycle air emissions impacts and ownership costs of light-duty vehicles using natural gas as a primary energy source.

This paper aims to comprehensively distinguish among the merits of different vehicles using a common primary energy source. In this study, we consider compressed natural gas (CNG) use directly in conventional vehicles (CV) and hybrid electric vehicles (HEV), and natural gas-derived electricity (NG-e) use in plug-in battery electric vehicles (BEV). This study evaluates the incremental life cycle air emissions (climate change and human health) impacts and life cycle ownership costs of non-plug-in (CV and HEV) and plug-in light-duty vehicles. Replacing a gasoline CV with a CNG CV, or a CNG CV with a CNG HEV, can provide life cycle air emissions impact benefits without increasing ownership costs; however, the NG-e BEV will likely increase costs (90% confidence interval: $1000 to $31 000 incremental cost per vehicle lifetime). Furthermore, eliminating HEV tailpipe emissions via plug-in vehicles has an insignificant incremental benefit, due to high uncertainties, with emissions cost benefits between -$1000 and $2000. Vehicle criteria air contaminants are a relatively minor contributor to life cycle air emissions impacts because of strict vehicle emissions standards. Therefore, policies should focus on adoption of plug-in vehicles in nonattainment regions, because CNG vehicles are likely more cost-effective at providing overall life cycle air emissions impact benefits.

Ethanol or bioelectricity? Life cycle assessment of lignocellulosic bioenergy use in light-duty vehicles.

Our study evaluates life cycle energy use and GHG emissions of lignocellulosic ethanol and bioelectricity use in U.S. light-duty vehicles. The well-to-pump, pump-to-wheel, and vehicle cycle stages are modeled. All ethanol (E85) and bioelectricity pathways have similar life cycle fossil energy use (~ 100 MJ/100 vehicle kilometers traveled (VKT)) and net GHG emissions (~5 kg CO2eq./100 VKT), considerably lower (65-85%) than those of reference gasoline and U.S. grid-electricity pathways. E85 use in a hybrid vehicle and bioelectricity use in a fully electric vehicle also have similar life cycle biomass and total energy use (~ 350 and ~450 MJ/100 VKT, respectively); differences in well-to-pump and pump-to-wheel efficiencies can largely offset each other. Our energy use and net GHG emissions results contrast with findings in literature, which report better performance on these metrics for bioelectricity compared to ethanol. The primary source of differences in the studies is related to our development of pathways with comparable vehicle characteristics. Ethanol or vehicle electrification can reduce petroleum use, while bioelectricity may displace nonpetroleum energy sources. Regional characteristics may create conditions under which either ethanol or bioelectricity may be the superior option; however, neither has a clear advantage in terms of GHG emissions or energy use.

Electricity production from anaerobic digestion of household organic waste in Ontario: techno-economic and GHG emission analyses.

The first Feed-in-Tariff (FiT) program in North America was recently implemented in Ontario, Canada to stimulate the generation of electricity from renewable sources. The life cycle greenhouse gas (GHG) emissions and economics of electricity generation through anaerobic digestion (AD) of household source-separated organic waste (HSSOW) are investigated within the FiT program. AD can potentially provide considerable GHG emission reductions (up to 1 t CO(2)eq/t HSSOW) at relatively low to moderate cost (-$35 to 160/t CO(2)eq) by displacing fossil electricity and preventing the emission of landfill gas. It is a cost-effective GHG mitigation option compared to some other FiT technologies (e.g., wind, solar photovoltaic) and provides unique additional benefits (waste diversion, nutrient recycling). The combination of electricity sales at a premium rate, savings in waste management costs, and economies of scale allow AD facilities processing >30,000 t/yr to be cost-competitive against landfilling. However, the FiT does not sufficiently support smaller-scale facilities that are needed as a transition to larger, more economically viable facilities. Refocusing of the FiT program and waste policies are needed to support the adoption of AD of HSSOW, which has not yet been developed in the Province, while more costly technologies (e.g., photovoltaic) have been deployed.

Comparison of the safety and pharmacokinetics of ST-246® after i.v. infusion or oral administration in mice, rabbits and monkeys.

BACKGROUND: ST-246® is an antiviral, orally bioavailable small molecule in clinical development for treatment of orthopoxvirus infections. An intravenous (i.v.) formulation may be required for some hospitalized patients who are unable to take oral medication. An i.v. formulation has been evaluated in three species previously used in evaluation of both efficacy and toxicology of the oral formulation. METHODOLOGY/PRINCIPAL FINDINGS: The pharmacokinetics of ST-246 after i.v. infusions in mice, rabbits and nonhuman primates (NHP) were compared to those obtained after oral administration. Ten minute i.v. infusions of ST-246 at doses of 3, 10, 30, and 75 mg/kg in mice produced peak plasma concentrations ranging from 16.9 to 238 µg/mL. Elimination appeared predominately first-order and exposure dose-proportional up to 30 mg/kg. Short i.v. infusions (5 to 15 minutes) in rabbits resulted in rapid distribution followed by slower elimination. Intravenous infusions in NHP were conducted at doses of 1 to 30 mg/kg. The length of single infusions in NHP ranged from 4 to 6 hours. The pharmacokinetics and tolerability for the two highest doses were evaluated when administered as two equivalent 4 hour infusions initiated 12 hours apart. Terminal elimination half-lives in all species for oral and i.v. infusions were similar. Dose-limiting central nervous system effects were identified in all three species and appeared related to high C(max) plasma concentrations. These effects were eliminated using slower i.v. infusions. CONCLUSIONS/SIGNIFICANCE: Pharmacokinetic profiles after i.v. infusion compared to those observed after oral administration demonstrated the necessity of longer i.v. infusions to (1) mimic the plasma exposure observed after oral administration and (2) avoid C(max) associated toxicity. Shorter infusions at higher doses in NHP resulted in decreased clearance, suggesting saturated distribution or elimination. Elimination half-lives in all species were similar between oral and i.v. administration. The administration of ST-246 was well tolerated as a slow i.v. infusion.

A non-dimensional analysis of hemodialysis.

BACKGROUND: Non-dimensional analysis is a powerful approach that can be applied to multivariate problems to better understand their behaviour and interpret complex interactions of variables. It is has not been rigorously applied to the parameters that define renal dialysis treatments and may provide insight into the planning of hemodialysis treatments. METHODS: Buckingham's non-dimensional approach was applied to the parameters that define hemodialysis treatments. Non-dimensional groups were derived with knowledge of a mass transfer model and independent of it. Using a mass transfer model, the derived non-dimensional groups were plotted to develop an understanding of key relationships governing hemodialysis and toxin profiles in patients with end-stage renal disease. RESULTS: Three non-dimensional groups are sufficient to describe hemodialysis, if there is no residual renal function (RRF). The non-dimensional groups found represent (1) the number of half-lives that characterize the mass transfer, (2) the toxin concentration divided by the rise in toxin concentration without dialysis for the cycle time (the inverse of the dialysis frequency), and (3) the ratio of dialysis time to the cycle time. If there is RRF, one additional non-dimensional group is needed (the ratio between cycle time and intradialytic elimination rate constant). Alternate non-dimensional groups can be derived from the four unique groups. CONCLUSIONS: Physical interpretation of the non-dimensional groups allows for greater insight into the parameters that determine dialysis effectiveness. This technique can be applied to any toxin and facilitates a greater understanding of dialysis treatment options. Quantitative measures of dialysis adequacy should be based on dimensional variables.

Thermoinactivation mechanism of glucose isomerase.

In this article, the mechanisms of thermoinactivation of glucose isomerase (GI) from Streptomyces rubiginosus (in soluble and immobilized forms) were investigated, particularly the contributions of thiol oxidation of the enzyme's cysteine residue and a "Maillard-like" reaction between the enzyme and sugars in high fructose corn syrup (HFCS). Soluble GI (SGI) was successfully immobilized on silica gel (13.5 microm particle size), with an activity yield between 20 and 40%. The immobilized GI (IGI) has high enzyme retention on the support during the glucose isomerization process. In batch reactors, SGI (half-life =145 h) was more stable than IGI (half-life =27 h) at 60 degrees C in HFCS, whereas at 80 degrees C, IGI (half-life =12 h) was more stable than SGI (half-life =5.2 h). IGI was subject to thiol oxidation at 60 degrees C, which contributed to the enzyme's deactivation. IGI was subject to thiol oxidation at 80 degrees C, but this did not contribute to the deactivation of the enzyme. SGI did not undergo thiol oxidation at 60 degrees C, but at 80 degrees C SGI underwent severe precipitation and thiol oxidation, which caused the enzyme to deactivate. Experimental results show that immobilization suppresses the destabilizing effect of thiol oxidation on GI. A "Maillard-like" reaction between SGI and the sugars also caused SGI thermoinactivation at 60, 70, and 80 degrees C, but had minimal effect on IGI. At 60 and 80 degrees C, IGI had higher thermostability in continuous reactors than in batch reactors, possibly because of reduced contact with deleterious compounds in HFCS.

Properties and performance of glucoamylases for fuel ethanol production.

Studies were conducted on maltodextrin saccharification and on simultaneous saccharification and fermentation (SSF) with various commercial glucoamylases. In kinetics studies, none of the glucoamylases were able to completely convert maltodextrin into glucose. Typically, about 85% conversion was obtained, and glucose yields were about 75%. Typically, the kinetics were biphasic, with 1 h of rapid conversion, then a significant reduction in rate. Data were consistent with strong product inhibition and/or enzyme inactivation. Some glucoamylases followed first-order kinetics, initially slower at dextrin conversion, but eventually achieving comparable conversion and glucose concentrations. Most of the glucoamylases were more active at 55 degrees C than at 35 degrees C, but pH had little effect on activity. Screening studies in an SSF system demonstrated little difference between the glucoamylases, with a few exceptions. Subsequent targeted studies showed clear differences in performance, depending on the fermentation temperature and yeast used, suggesting that these are key parameters that would guide the selection of a glucoamylase.

Absence of adverse effects in cynomolgus macaques treated with CNTO 95, a fully human anti-alphav integrin monoclonal antibody, despite widespread tissue binding.

PURPOSE: CNTO 95 is a fully human anti-alphav integrin monoclonal antibody that inhibits macaque and rodent angiogenesis and inhibits human tumor growth in rodents. The purpose of these studies was to evaluate the preclinical safety of long-term administration of CNTO 95 in cynomolgus macaques. EXPERIMENTAL DESIGN: The in vitro binding profiles of CNTO 95 to human and macaque tissues and the in vivo binding to macaque tissues was evaluated by immunohistochemistry. The preclinical safety of CNTO 95 (10 and 50 mg/kg, i.v.) was evaluated in macaques treated once per week for up to 6 months. Safety was evaluated by clinical observations, ophthalmic and physical examinations (including heart rate, blood pressure, and electrocardiogram), clinical pathology (including coagulation parameters), and comprehensive anatomic pathology. The effect of CNTO 95 (50 mg/kg, i.v.) on incisional wound healing was evaluated in macaques. RESULTS: The tissue binding studies showed that CNTO 95 bound with mild to moderate intensity to macaque and human endothelial cells, epithelial cells, and vascular smooth muscle cells in most normal tissues examined. CNTO 95 showed strong to intense staining to the positive control tissue, human placenta. Despite the widespread binding to normal tissues, treatment of cynomolgus macaques with CNTO 95 produced no signs of toxicity and no histopathologic changes in any of the tissues examined (including ovaries and bone growth plates). CNTO 95 did not impair wound healing. CONCLUSION: These studies show that CNTO 95 is safe and, unlike some other angiogenesis inhibitors, does not seem to inhibit normal physiologic angiogenesis.

Characterization and performance of immobilized amylase and cellulase.

The performance of cellulase and amylase immobilized on siliceous supports was investigated. Enzyme uptake onto the support depended on the enzyme source and immobilization conditions. For amylase, the uptake ranged between 20 and 60%, and for cellulase, 7-10%. Immobilized amylase performance was assessed by batch kinetics in 100-300 g/L of corn flour at 65 degrees C. Depending on the substrate and enzyme loading, between 40 and 60% starch conversion was obtained. Immobilized amylase was more stable than soluble amylase. Enzyme samples were preincubated in a water bath at various temperatures, then tested for activity. At 105 degrees C, soluble amylase lost approximately 55% of its activity, compared with approximately 30% loss for immobilized amylase. The performance of immobilized cellulase was evaluated from batch kinetics in 10 g/L of substrate (shredded wastepaper) at 55 degrees C. Significant hydrolysis of the wastepaper was also observed, indicating that immobilization does not preclude access to and hydrolysis of insoluble cellulose.

The aryl hydrocarbon receptor mediates degradation of estrogen receptor alpha through activation of proteasomes.

2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and other aryl hydrocarbon receptor (AhR) ligands suppress 17beta-estradiol (E)-induced responses in the rodent uterus and mammary tumors and in human breast cancer cells. Treatment of ZR-75, T47D, and MCF-7 human breast cancer cells with TCDD induces proteasome-dependent degradation of endogenous estrogen receptor alpha (ERalpha). The proteasome inhibitors MG132, PSI, and PSII inhibit the proteasome-dependent effects induced by TCDD, whereas the protease inhibitors EST, calpain inhibitor II, and chloroquine do not affect this response. ERalpha levels in the mouse uterus and breast cancer cells were significantly lower after cotreatment with E plus TCDD than after treatment with E or TCDD alone, and our results indicate that AhR-mediated inhibition of E-induced transactivation is mainly due to limiting levels of ERalpha in cells cotreated with E plus TCDD. TCDD alone or in combination with E increases formation of ubiquitinated forms of ERalpha, and both coimmunoprecipitation and mammalian two-hybrid assays demonstrate that TCDD induces interaction of the AhR with ERalpha in the presence or absence of E. In contrast, E does not induce AhR-ERalpha interactions. Thus, inhibitory AhR-ERalpha cross talk is linked to a novel pathway for degradation of ERalpha in which TCDD initially induces formation of a nuclear AhR complex which coordinately recruits ERalpha and the proteasome complex, resulting in degradation of both receptors.

Degradation of phenol using tyrosinase immobilized on siliceous supports.

The degradation of phenol by tyrosinase immobilized on chemically modified sodium aluminosilicate (NaA), calcium aluminosilicate (CaA), and silica gel was studied. Phenol conversion by immobilized tyrosinase ranged between approximately 15% and 60%, depending upon the initial phenol concentration, pH, and enzyme loading. Tyrosinase immobilized on CaA and on NaA could be re-used repeatedly without any decrease in performance. However, in studies at pH 8.0, significant enzyme inhibition was observed, since phenol conversion was rapid for approximately 20 min, then reached a plateau. The inhibition was reversible; activity was restored upon placing the immobilized enzyme in fresh substrate. Reducing the pH to 6.8 from 8.0 led to higher conversion of phenol, and decreased the inhibition of the immobilized enzyme.

Hypoxia induces proteasome-dependent degradation of estrogen receptor alpha in ZR-75 breast cancer cells.

Regulation of estrogen receptor alpha (ERalpha) plays an important role in hormone responsiveness and growth of ER-positive breast cancer cells and tumors. ZR-75 breast cancer cells were grown under conditions of normoxia (21% O(2)) or hypoxia (1% O(2) or cobaltous chloride), and hypoxia significantly increased hypoxia-inducible factor 1alpha protein within 3 h after treatment, whereas ERalpha protein levels were dramatically decreased within 6-12 h, and this response was blocked by the proteasome inhibitor MG-132. In contrast, hypoxia induced only minimal decreases in cellular Sp1 protein and did not affect ERalpha mRNA; however, hypoxic conditions decreased basal and 17beta-estradiol-induced pS2 gene expression (mRNA levels) and estrogen response element-dependent reporter gene activity in ZR-75 cells. Although 17beta-estradiol and hypoxia induce proteasome-dependent degradation of ERalpha, their effects on transactivation are different, and this may have implications for clinical treatment of mammary tumors.

Cooperative coactivation of estrogen receptor alpha in ZR-75 human breast cancer cells by SNURF and TATA-binding protein.

SNURF is a small RING finger protein that binds the zinc finger region of steroid hormone receptors and enhances Sp1- and androgen receptor-mediated transcription in COS and CV-1 cells. In this study, we show that SNURF coactivates both wild-type estrogen receptor alpha (ERalpha) (4-fold)- and HE19 (ERalpha deletion of activation function 1 (AF1)) (210-fold)-mediated activation of an estrogen-responsive element promoter in ZR-75 cells. In mammalian two-hybrid assays in ZR-75 cells SNURF interactions were estrogen (E2)-dependent and were not observed with the antiestrogen ICI 182,780. ERalpha interacted with multiple regions of SNURF; SNURF interactions with ERalpha were dependent on AF2, and D538N, E542Q, and D545N mutations in helix 12 abrogated both SNURF-ERalpha binding and coactivation. Moreover, peptide fusion proteins that inhibit interactions between helix 12 of ERalpha with LXXLL box-containing proteins also blocked ERalpha coactivation by SNURF. However, cotransfection of SNURF with prototypical steroid receptor coactivators 1, 2, and 3 that contain LXXLL box motifs did not enhance E2 responsiveness, whereas TATA-binding protein (TBP) and SNURF cooperatively coactivated ERalpha-mediated transactivation. The results are consistent with a unique model for cooperative coactivation of ERalpha that requires ligand binding, repositioning of helix 12, recruitment of TBP, and interaction with SNURF, which binds both ERalpha and TBP.