Suboptimal Temperature Acclimation Affects Kennedy Pathway Gene Expression, Lipidome and Metabolite Profile of Nannochloropsis salina during PUFA Enriched TAG Synthesis.

In humans, dietary polyunsaturated fatty acids (PUFAs) are involved in therapeutic processes such as prevention and treatment of cardiovascular diseases, neuropsychiatric disorders, and dementia. We examined the physiology, PUFA accumulation and glycerol lipid biosynthesis in the marine microalga Nannochloropsis salina in response to constant suboptimal temperature (<20 °C). As expected, N. salina exhibited significantly reduced growth rate and photosynthetic activity compared to optimal cultivation temperature. Total fatty acid contents were not significantly elevated at reduced temperatures. Cultures grown at 5 °C had the highest quantity of eicosapentanoic acid (EPA) (C20:5n3) and the lowest growth rate. Additionally, we monitored broadband lipid composition to model the occurrence of metabolic alteration and remodeling for various lipid pools. We focused on triacylglycerol (TAG) with elevated PUFA content. TAGs with EPA at all three acyl positions were higher at a cultivation temperature of 15 °C. Furthermore, monogalactosyldiacylglycerol and digalactosyldiacylglycerol, which are polar lipids associated with chloroplast membranes, decreased with reduced cultivation temperatures. Moreover, gene expression analysis of key genes involved in Kennedy pathway for de novo TAG biosynthesis revealed bimodal variations in transcript level amongst the temperature treatments. Collectively, these results show that Nannochloropsis salina is a promising source of PUFA containing lipids.

Heme-independent Redox Sensing by the Heme-Nitric Oxide/Oxygen-binding Protein (H-NOX) from Vibrio cholerae.

Heme nitric oxide/oxygen (H-NOX)-binding proteins act as nitric oxide (NO) sensors among various bacterial species. In several cases, they act to mediate communal behavior such as biofilm formation, quorum sensing, and motility by influencing the activity of downstream signaling proteins such as histidine kinases (HisKa) in a NO-dependent manner. An H-NOX/HisKa regulatory circuit was recently identified in Vibrio cholerae, and the H-NOX protein has been spectroscopically characterized. However, the influence of the H-NOX protein on HisKa autophosphorylation has not been evaluated. This process may be important for persistence and pathogenicity in this organism. Here, we have expressed and purified the V. cholerae HisKa (HnoK) and H-NOX in its heme-bound (holo) and heme-free (apo) forms. Autophosphorylation assays of HnoK in the presence of H-NOX show that the holoprotein in the Fe(II)-NO and Fe(III) forms is a potent inhibitor of HnoK. Activity of the Fe(III) form and aerobic instability of the Fe(II) form suggested that Vibrio cholerae H-NOX may act as a sensor of the redox state as well as NO. Remarkably, the apoprotein also showed robust HnoK inhibition that was dependent on the oxidation of cysteine residues to form disulfide bonds at a highly conserved zinc site. The importance of cysteine in this process was confirmed by mutagenesis, which also showed that holo Fe(III), but not Fe(II)-NO, H-NOX relied heavily upon cysteine for activation. These results highlight a heme-independent mechanism for activation of V. cholerae H-NOX that implicates this protein as a dual redox/NO sensor.

The regulation of photosynthetic structure and function during nitrogen deprivation in Chlamydomonas reinhardtii.

The accumulation of carbon storage compounds by many unicellular algae after nutrient deprivation occurs despite declines in their photosynthetic apparatus. To understand the regulation and roles of photosynthesis during this potentially bioenergetically valuable process, we analyzed photosynthetic structure and function after nitrogen deprivation in the model alga Chlamydomonas reinhardtii. Transcriptomic, proteomic, metabolite, and lipid profiling and microscopic time course data were combined with multiple measures of photosynthetic function. Levels of transcripts and proteins of photosystems I and II and most antenna genes fell with differing trajectories; thylakoid membrane lipid levels decreased, while their proportions remained similar and thylakoid membrane organization appeared to be preserved. Cellular chlorophyll (Chl) content decreased more than 2-fold within 24 h, and we conclude from transcript protein and (13)C labeling rates that Chl synthesis was down-regulated both pre- and posttranslationally and that Chl levels fell because of a rapid cessation in synthesis and dilution by cellular growth rather than because of degradation. Photosynthetically driven oxygen production and the efficiency of photosystem II as well as P700(+) reduction and electrochromic shift kinetics all decreased over the time course, without evidence of substantial energy overflow. The results also indicate that linear electron flow fell approximately 15% more than cyclic flow over the first 24 h. Comparing Calvin-Benson cycle transcript and enzyme levels with changes in photosynthetic (13)CO2 incorporation rates also pointed to a coordinated multilevel down-regulation of photosynthetic fluxes during starch synthesis before the induction of high triacylglycerol accumulation rates.

Quantitative and qualitative evaluation of kernel anthocyanins from southwestern United States blue corn.

BACKGROUND: Anthocyanin-rich blue corn is an emerging specialty crop in the USA. The antioxidant properties of blue corn offer health benefits in the human diet. The objectives of this study were to identify, characterize and quantify the anthocyanins from blue corn. Hypotheses tested were that total anthocyanin content was similar among southwestern US accessions and that it would vary across locations. It was also examined whether different anthocyanin components were unique to certain genotypes. RESULTS: Across all locations and accessions, an average of 0.43 g kg(-1) total anthocyanin content (TAC) was observed. Accessions Santa Clara Blue and Ohio Blue displayed the highest TAC. The TAC of accession Flor del Rio was lower by nearly a factor of six. A total of five anthocyanin components were identified. Cyanidin 3-glucoside was the most abundant, followed by pelargonidin and peonidin 3-glucoside. Succinyl and disuccinyl glycosidic forms of cyanidin were also identified. Cyanidin 3-disuccinylglucoside was newly identified as a novel form of anthocyanin. CONCLUSION: Quantitative and qualitative anthocyanin expression was determined to be relatively stable across multiple southwestern environments. Increased expression of red and purple pigmentation in accession Flor del Rio appeared to be associated more with reduced TAC and cyanidin 3-glucoside than with elevated pelargonidin per se. A previously unreported anthocyanin component in blue corn, cyanidin 3-disuccinylglucoside, is present in southwestern landraces. © 2016 Society of Chemical Industry.

Occurrence and behavior of uranium and thorium series radionuclides in the Permian shale hydraulic fracturing wastes.

Over the last decade, there has been a rapid growth in the use of hydraulic fracturing (fracking) to recover unconventional oil and gas in the Permian Basin of southeastern New Mexico (NM) and western Texas. Fracking generates enormous quantities of wastes that contain technologically enhanced naturally occurring radioactive materials (TENORM), which poses risks to human health and the environment because of the relatively high doses of radioactivity. However, very little is known about the chemical composition and radioactivity levels of Permian Basin fracking wastes. Here, we report chemical as well as radiochemical compositions of hydraulic fracking wastes from the Permian Basin. Radium, the major TENORM of interest in unconventional drilling wastes, varied from 19.1 ± 1.2 to 35.9 ± 3.2 Bq/L for 226Ra, 10.3 ± 0.5 to 21.5 ± 1.2 Bq/L for 228Ra, and 2.0 ± 0.05 to 3.7 ± 0.07 Bq/L for 224Ra. In addition to elevated concentrations of radium, these wastewaters also contain elevated concentrations of dissolved salts and divalent cations such as Na+ (31,856-43,000 mg/L), Ca2+ (668-4123 mg/L), Mg2+ (202-2430 mg/L), K+ (148-780 mg/L), Sr2+ (101-260 mg/L), Cl- (5160-66,700 mg/L), SO42- (291-1980 mg/L), Br- (315-596 mg/L), SiO2 (20-32 mg/L), and high total dissolved solid (TDS) of 5000-173,000 mg/L compared to background waters. These elevated levels are of radiological significance and represent a major source of Ra in the environment. The recent discovery of large deposits of recoverable oil and gas in the Permian Basin will lead to more fracking, TENORM generation, and radium releases to the environment. This paper evaluates the potential radiation risks associated with TENORM wastes generated by the oil and gas recovery industry in the Permian Basin.

δ-Aminolevulinic acid dehydratase single nucleotide polymorphism 2 and peptide transporter 2*2 haplotype may differentially mediate lead exposure in male children.

Child low-level lead (Pb) exposure is an unresolved public health problem and an unaddressed child health disparity. Particularly in cases of low-level exposure, source removal can be impossible to accomplish, and the only practical strategy for reducing risk may be primary prevention. Genetic biomarkers of increased neurotoxic risk could help to identify small subgroups of children for early intervention. Previous studies have suggested that, by way of a distinct mechanism, δ-aminolevulinic acid dehydratase single nucleotide polymorphism 2 (ALAD(2)) and/or peptide transporter 2*2 haplotype (hPEPT2*2) increase Pb blood burden in children. Studies have not yet examined whether sex mediates the effects of genotype on blood Pb burden. Also, previous studies have not included blood iron (Fe) level in their analyses. Blood and cheek cell samples were obtained from 306 minority children, ages 5.1 to 12.9 years. (208)Pb and (56)Fe levels were determined with inductively coupled plasma-mass spectrometry. General linear model analyses were used to examine differences in Pb blood burden by genotype and sex while controlling for blood Fe level. The sample geometric mean Pb level was 2.75 µg/dl. Pb blood burden was differentially higher in ALAD(2) heterozygous boys and hPEPT2*2 homozygous boys. These results suggest that the effect of ALAD(2) and hPEPT2*2 on Pb blood burden may be sexually dimorphic. ALAD(2) and hPEPT2*2 may be novel biomarkers of health and mental health risks in male children exposed to low levels of Pb.

Potable-quality water recovery from primary effluent through a coupled algal-osmosis membrane system.

A coupled algal-osmosis membrane treatment system was studied for recovering potable-quality water from municipal primary effluent. The core components of the system included a mixotrophic algal process for removal of biochemical oxygen demand (BOD) and nutrients, followed by a hybrid forward osmosis (FO)-reverse osmosis (RO) system for separation of biomass from the algal effluent and production of potable-quality water. Field experiments demonstrated consistent performance of the algal system to meet surface discharge standards for BOD and nutrients within a fed-batch processing time of 2-3 days. The hybrid FO-RO system reached water productivity of 1.57 L/m2-h in FO using seawater as draw solution; and permeate flux of 3.50 L/m2-h in brackish water RO (BWRO) and 2.07 L/m2-h in seawater RO (SWRO) at 2068 KPa. The coupled algal-membrane system achieved complete removal of ammonia, fluoride, and phosphate; over 90% removal of calcium, sulfate, and organic carbon; and 86-89% removal of potassium and magnesium. Broadband characterization using high resolution mass spectrometry revealed extensive removal of organic compounds, particularly wastewater surfactants upon algal treatment. This study demonstrated long-term performance of the FO system at water recovery of 90% and with membrane cleaning by NaOH solution.

Bio-crude oil from hydrothermal liquefaction of wastewater microalgae in a pilot-scale continuous flow reactor.

To explore the feasibility of scaling up hydrothermal liquefaction (HTL) of algal biomass, a pilot-scale continuous flow reactor (CFR) was operated to produce bio-crude oil from algal biomass cultivated in urban wastewater. The CFR system ran algal slurry (5 wt.% solids loading) at 350 °C and 17 MPa for 4 h without any clogging issues. Bio-crude oil chemistry was characterized by high-resolution Fourier transform mass spectroscopy (FT-MS), proton nuclear magnetic resonance spectroscopy (1H NMR), bomb calorimetry, and elemental analysis. Bio-crude oil yield of 28.1 wt% was obtained with higher heating values of 38-39 MJ/kg. The quality of light bio-crude oil produced from the CFR system was comparable in terms of molecular structures to bio-crude oil produced in a batch reactor.

Hydrothermal liquefaction of Galdieria sulphuraria grown on municipal wastewater.

Two strains of Galdieria sulphuraria algae, 5587.1 and SOOS, were grown on municipal wastewater to develop energy-positive treatment systems. Hydrothermal liquefaction (HTL) of 5-10 wt% algal biomass solids was conducted at 310-350 °C for 5-60 min to produce bio-crude oil. HTL product yields and energy recovery were compared to those from previous studies using G. sulphuraria grown on a modified Cyanidium medium. Total bio-crude oil yields were lower (11.2-23.0 wt%) and char yields were higher (22.6-36.4 wt%) for HTL of algae grown on actual wastewater compared with that grown on media (31.4 wt% and 4.8 wt%, respectively), indicating a potential limitation for using yields from media-based studies. High-resolution mass spectroscopy of bio-crude oil provides new insights into differences in composition based on growth media. Energy recovery in total bio-crude oil and char at 350 °C was 17-28% and 14-19%, respectively, for the 5587.1 strain, and 23-27% and 14-25%, respectively, for the SOOS strain.

Bimolecular Cross-Metathesis of a Tetrasubstituted Alkene with Allylic Sulfones.

Exquisite control of catalytic metathesis reactivity is possible through ligand-based variation of ruthenium carbene complexes. Sterically hindered alkenes, however, remain a generally recalcitrant class of substrates for intermolecular cross-metathesis. Allylic chalcogenides (sulfides and selenides) have emerged as "privileged" substrates that exhibit enhanced turnover rates with the commercially available second-generation ruthenium catalyst. Increased turnover rates are advantageous when competing catalyst degradation is limiting, although specific mechanisms have not been defined. Herein, we describe facile cross-metathesis of allylic sulfone reagents with sterically hindered isoprenoid alkene substrates. Furthermore, we demonstrate the first example of intermolecular cross-metathesis of ruthenium carbenes with a tetrasubstituted alkene. Computational analysis by combined coupled cluster/DFT calculations exposes a favorable energetic profile for metallacyclobutane formation from chelating ruthenium ß-chalcogenide carbene intermediates. These results establish allylic sulfones as privileged reagents for a substrate-based strategy of cross-metathesis derivatization.

Adsorption and photocatalytic oxidation of ibuprofen using nanocomposites of TiO2 nanofibers combined with BN nanosheets: Degradation products and mechanisms.

This study investigated the adsorption and photocatalytic activity of titanium dioxide (TiO2)-boron nitride (BN) nanocomposites for the removal of contaminants of emerging concern in water using ibuprofen as a model compound. TiO2 nanofibers wrapped by BN nanosheets were synthesized by electrospinning method. Characterization of the nanocomposite photocatalysts indicated that the BN nanosheets improved the light absorbance and reduced the recombination of the photoexcited charge carriers (e- and h+). The photocatalytic oxidation products and mechanisms of ibuprofen by the TiO2-BN catalysts were elucidated using a multiple analysis approach by high performance liquid chromatography, ultraviolet absorbance, dissolved organic carbon, fluorescence excitation-emission matrices, and electrospray ionization-liquid chromatography-tandem mass spectrometry. The experimental results revealed that the photocatalytic oxidation by the TiO2-BN nanocomposites is a multi-step process and the interactions between ibuprofen molecules and the TiO2-BN nanocomposites govern the adsorption process. The increasing BN nanosheet content in the TiO2 nanofibers facilitated the breakdown of ibuprofen degradation intermediates (hydroxyibuprofen, carboxyibuprofen, and oxypropyl ibuprofen). Kinetic modeling indicated both adsorption and photocatalytic oxidation of ibuprofen by the TiO2-BN nanocomposites followed the first-order kinetic model. The photocatalytic oxidation rate increased with the increasing BN content in the nanocomposite catalysts, which was attributed to the enhanced light absorption capacity and the separation efficiency of the photoexcited electron (e-)-hole (h+) pairs. Multiple photocatalytic cycles were conducted to investigate the reusability and regeneration of the nanofibers for ibuprofen degradation.

Automated liquid injection field desorption/ionization for Fourier transform ion cyclotron resonance mass spectrometry.

We describe automation of liquid injection field desorption/ionization (LIFDI) for reproducible sample application, improved spectral quality, and high-throughput analyses. A commercial autosampler provides reproducible and unattended sample application. A custom-built field desorption (FD) controller allows data station or front panel control of source parameters including high-voltage limit/ramp rate, emitter heating current limit/ramp rate, and feedback control of emitter heating current based on ion current measurement. Automated LIFDI facilitates ensemble averaging of hundreds of Fourier transform ion cyclotron resonance mass spectra for increased dynamic range, mass accuracy, and S/N ratio relative to single-application FD experiments, as shown here for a South American crude oil. This configuration can be adapted to any mass spectrometer with an LIFDI probe.

High-performance mass spectrometry: Fourier transform ion cyclotron resonance at 14.5 Tesla.

We describe the design and current performance of a 14.5 T hybrid linear quadrupole ion trap Fourier transform ion cyclotron resonance mass spectrometer. Ion masses are routinely determined at 4-fold better mass accuracy and 2-fold higher resolving power than similar 7 T systems at the same scan rate. The combination of high magnetic field and strict control of the number of trapped ions results in external calibration broadband mass accuracy typically less than 300 ppb rms, and a resolving power of 200,000 (m/Delta m50% at m/z 400) is achieved at greater than 1 mass spectrum per second. Novel ion storage optics and methodology increase the maximum number of ions that can be delivered to the FTICR cell, thereby improving dynamic range for tandem mass spectrometry and complex mixture applications.

Co-liquefaction of mixed culture microalgal strains under sub-critical water conditions.

We report the co-liquefaction performance of unicellular, red alga Cyanidioschyzon merolae and Galdieria sulphuraria under sub-critical water conditions within a stainless-steel batch reactor under different temperatures (150-300°C), residence time (15-60min), and Cyanidioschyzon merolae to Galdieria sulphuraria mass loading (0-100%). Individual liquefaction of C. merolae and G. sulphuraria at 300°C achieved maximum biocrude oil yield of 18.9 and 14.0%, respectively. The yield of biocrude oil increased to 25.5%, suggesting a positive synergistic effect during the co-liquefaction of 80-20mass loading of C. merolae to G. sulphuraria. The biocrude oils were analyzed by FT-ICR MS which showed that co-liquefaction did not significantly affect the distribution of product compounds compared to individual oils. The co-liquefied biocrude and biochar have a higher-heating-value of 35.28 and 7.96MJ/kg. Ultimate and proximate analysis were performed on algae biomass, biocrude and biochar.

Stabilization and prolonged reactivity of aqueous-phase ozone with cyclodextrin.

Recalcitrant organic groundwater contaminants, such as 1,4-dioxane, may require strong oxidants for complete mineralization. However, their efficacy for in-situ chemical oxidation (ISCO) is limited by oxidant decay and reactivity. Hydroxypropyl-ß-cyclodextrin (HPßCD) was examined for its ability to stabilize aqueous-phase ozone (O3) and prolong oxidation potential through inclusion complex formation. Partial transformation of HPßCD by O3 was observed. However, HPßCD proved to be sufficiently recalcitrant, because it was only partially degraded in the presence of O3. The formation of a HPßCD:O3 clathrate complex was observed, which stabilized decay of O3. The presence of HPßCD increased the O3 half-life linearly with increasing HPßCD:O3 molar ratio. The O3 half-life in solutions increased by as much as 40-fold relative to HPßCD-free O3 solutions. Observed O3 release from HPßCD and indigo oxidation confirmed that the formation of the inclusion complex is reversible. This proof-of-concept study demonstrates that HPßCD can complex O3 while preserving its reactivity. These results suggest that the use of clathrate stabilizers, such as HPßCD, can support the development of a facilitated-transport enabled ISCO for the O3 treatment of groundwater contaminated with recalcitrant compounds.

Hydrothermal liquefaction of Cyanidioschyzon merolae and the influence of catalysts on products.

This work investigates the hydrothermal liquefaction (HTL) of Cyanidioschyzon merolae algal species under various reaction temperatures and catalysts. Liquefaction of microalgae was performed with 10% solid loading for 30min at temperatures of 180-300°C to study the influences of two base and two acid catalysts on HTL product fractions. Maximum biocrude oil yield of 16.98% was obtained at 300°C with no catalyst. The biocrude oil yield increased to 22.67% when KOH was introduced into the reaction mixture as a catalyst. The algal biocrude and biochar has a higher heating values (HHV) of 32.22MJkg-1 and 20.78MJkg-1 respectively when no catalyst was used. Gas chromatography time of flight mass spectrometry (GC/TOFMS) was employed to analyze the biocrude oil composition, and elemental analysis was performed on the algae, biocrude and biochar samples. Analysis of the HTL aqueous phase revealed the presence of valuable products.

Synthesis of C59Hx and C58Hx fullerenes stabilized by hydrogen.

Prolonged hydrogenation of C(60) molecules by reaction with H(2) at elevated temperature and pressure results in fragmentation and collapse of the fullerene cage structure. However, fragments can be preserved by immediate termination of dangling bonds by hydrogen. Here we demonstrate that not only fullerene fragments but also hydrogenated fragmented fullerenes (e.g., C(58)H(40) and C(59)H(40)) can be synthesized in bulk amount by high-temperature hydrogenation of C(60). We confirm successful synthesis of these species by matrix-assisted laser desorption ionization time-of-flight mass spectrometry and complete speciation of the resultant complex fullerene mixtures by high-resolution field desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry.

Composition of hydrofullerene mixtures produced by C(60) reaction with hydrogen gas revealed by high-resolution mass spectrometry.

Complex solid hydrofulleride mixtures were synthesized by prolonged hydrogenation of C(60) at 120 bar hydrogen pressure, 673 K temperature, and different reaction periods. The high degree of hydrogenation was confirmed by infrared spectroscopy and X-ray diffraction. The identity of hydrogenation products was determined by high-resolution field desorption/ionization Fourier transform ion cyclotron resonance mass spectrometry. Despite partial gas-phase fragmentation of hydrofullerene ions during mass analysis, the data suggest that the synthesized mixtures consist of mostly C(58-60)H(x) hydrofullerenes. Increasing the duration of hydrogenation results in synthesis of C(59)H(x) and C(58)H(x) as major products. Possible hydrofullerene fragmentation pathways during both material synthesis and mass spectrometric analysis are discussed. Gas-phase fragmentation in the mass spectrometer arises from hydrofullerene ions C(60)H(x)(+) with x > 40 and C(59)H(44)(+) with drastically decreased molecular stability relative to the known C(60)H(36).

δ-Aminolevulinic acid dehydratase single nucleotide polymorphism 2 (ALAD2) and peptide transporter 2*2 haplotype (hPEPT2*2) differently influence neurobehavior in low-level lead exposed children.

Delta-aminolevulinic acid dehydratase single nucleotide polymorphism 2 (ALAD2) and peptide transporter haplotype 2*2 (hPEPT2*2) through different pathways can increase brain levels of delta-aminolevulinic acid and are associated with higher blood lead burden in young children. Past child and adult findings regarding ALAD2 and neurobehavior have been inconsistent, and the possible association of hPEPT2*2 and neurobehavior has not yet been examined. Mean blood lead level (BLL), genotype, and neurobehavioral function (fine motor dexterity, working memory, visual attention and short-term memory) were assessed in 206 males and 215 females ages 5.1-11.8years. Ninety-six percent of children had BLLs<5.0µg/dl. After adjusting for covariates (sex, age and mother's level of education) and sibling exclusion (N=252), generalized linear mixed model analyses showed opposite effects for the ALAD2 and hPEPT2*2 genetic variants. Significant effects for ALAD2 were observed only as interactions with BLL and the results suggested that ALAD2 was neuroprotective. As BLL increased, ALAD2 was associated with enhanced visual attention and enhanced working memory (fewer commission errors). Independent of BLL, hPEPT2*2 predicted poorer motor dexterity and poorer working memory (more commission errors). BLL alone predicted poorer working memory from increased omission errors. The findings provided further substantiation that (independent of the genetic variants examined) lowest-level lead exposure disrupted early neurobehavioral function, and suggested that common genetic variants alter the neurotoxic potential of low-level lead. ALAD2 and hPEPT2*2 may be valuable markers of risk, and indicate novel mechanisms of lead-induced neurotoxicity. Longitudinal studies are needed to examine long-term influences of these genetic variants on neurobehavior.

BABA and Phytophthora nicotianae Induce Resistance to Phytophthora capsici in Chile Pepper (Capsicum annuum).

Induced resistance in plants is a systemic response to certain microorganisms or chemicals that enhances basal defense responses during subsequent plant infection by pathogens. Inoculation of chile pepper with zoospores of non-host Phytophthora nicotianae or the chemical elicitor beta-aminobutyric acid (BABA) significantly inhibited foliar blight caused by Phytophthora capsici. Tissue extract analyses by GC/MS identified conserved change in certain metabolite concentrations following P. nicotianae or BABA treatment. Induced chile pepper plants had reduced concentrations of sucrose and TCA cycle intermediates and increased concentrations of specific hexose-phosphates, hexose-disaccharides and amino acids. Galactose, which increased significantly in induced chile pepper plants, was shown to inhibit growth of P. capsici in a plate assay.