Evaluation of phosphorus distribution and bioavailability in sediments of a subtropical wetland reserve in southeast China

The phosphorus (P) fractions and bioavailable P in the sediments from the Quanzhou Bay Estuarine Wetland Nature Reserve were investigated using chemical extraction methods for the first time to study the distribution and bioavailability of P in the reserve sediments. A hypothesis was presented suggesting that the bioavailable P in the sediments could be evaluated using the P fractions. The total phosphorus (TP), inorganic phosphorus (IP), organic phosphorus (OP), non-apatite phosphorus (NAIP), and apatite phosphorus (AP) contents in the sediments were in the ranges of 303.87–761.59 mg kg⁻¹, 201.22–577.66 mg kg⁻¹, 75.83–179.16 mg kg⁻¹, 28.86–277.90 mg kg⁻¹, and 127.36–289.94 mg kg⁻¹, respectively. The water soluble phosphorus (WSP), readily desorbable phosphorus (RDP), algal available phosphorus (AAP), and NaHCO₃ extractable phosphorus (Olsen-P) contents in the sediments were in the ranges of 0.58–357.17 mg kg⁻¹, 80.77–586.75 mg kg⁻¹, 1.09–24.12 mg kg⁻¹, and 54.96–676.82 mg kg⁻¹, respectively. The correlation analysis results showed that the NAIP was the major component of the bioavailable P and that the impact of the AP on the bioavailable phosphorus may be minimal. Due to the low TP content in the sediments of the Quanzhou Bay Estuarine Wetland Nature Reserve, the potential pollution risks of P in the sediments may not be very high. The results also show that the bioavailable P concentrations in the sediments of the Quanzhou Bay Estuarine Wetland Nature Reserve could not be evaluated by measuring the P fractions and that the hypothesis was untenable.     

Lipid biomarker evidence for determining the origin and distribution of organic matter in surface sediments of Lake Taihu,Eastern China

Lipid biomarkers from surface sediments of Lake Taihu (Eastern China) were analyzed in order to determine the origin and spatial distribution of sediment organic matter (OM), which is necessary to understand the regional carbon cycles and design environmental management strategies for lake systems. The results indicated significant heterogeneity in the distribution of autochthonous (algae-, photosynthetic bacteria- and macrophyte-based) and allochthonous (terrestrial plant-based) OM in sediments across the lake. Allochthonous OM inputs, indicated by long-chain n-alkane and long-chain n-alkanol biomarkers, generally declined in abundance from northwestern (Zhushan Bay and Meiliang Bay) to southeastern (East Bays) parts of the lake, suggesting a critical influence of hydrology, and in particular of inflowing rivers, which mainly enter the lake from the west and drain from the east. Autochthonous OM, on the other hand, appeared to reflect variations in overall nutrient status and habit type across the lake. Cyanobacterial OM inputs, identified by short-chain n-alkanes, were most abundant in sediment from the most severely polluted zones in Lake Taihu, namely Zhushan Bay and Meiliang Bay. OM derived from diatoms, indicated by brassicasterol and highly branched isoprenoids (HBIs), was most abundant in sediments from the East Bays, a clear-water zone with relatively low levels of nutrient input. Macrophyte OM input, indicated by the middle-chain n-alkanes and Paq ((n-C23 + n-C25)/(n-C23 + n-C25 + n-C29 + n-C31)), was only identified in sediments from the East Bays. The lowest recorded inputs for both autochthonous and allochthonous OM were in sediments from open areas with significant sediment resuspension, including Gonghu Bay, Central and Western Region. This finding might reflect degradation mineralization of OM in the water column during sediment resuspension.     

Effect of suplemental nitrogen source and feeding frequency on nutrient supply to lambs fed a kikuyu grass (Pennisetum clandestinum) hay-based diet

Eight castrated male lambs (35 ± 4 kg live weight), fed a basal diet of kikuyu grass hay, were used in a replicated 4 × 4 Latin Square experiment with a 2 × 2 factorial arrangement of treatments to evaluate the effect of supplemental feeding frequency and source of rumen degradable N on intake, digestibility, ruminal fermentation, and microbial protein yield. Treatments were supplementation with cassava meal plus calcium caseinate or cassava meal plus urea offered at a rate of 7 g/kg live weight daily in one or two meals per day. Lambs were fed twice daily in such manner to allow ad libitum comsumption of forage. There was significant feeding frequency by N source interaction on variables of intake. In general, intake of feed components was higher (P ≤ 0.05) by lambs offered the caseinate-supplement twice daily over intake observed in lambs given the others diet treatments. Digestibility of feed components was neither affected by supplemental N source (DM, P = 0.541; OM, P = 0.585; NDF, P = 0.828) nor by feeding frequency (DM, P = 0.122; OM, P = 0.175; NDF, P = 0.591). Urinary excretion of N increased (P ≤ 0.05) in lambs supplemented twice daily whereas N retention was similar for all treatments (N source, P = 0.748; feeding frequency, P = 0.418). Microbial protein entering into the small intestine was affected by the interaction between feeding frequency and N source such as an increasing (P < 0.10) in this variable was observed when lambs received the caseinate but not the urea supplement twice daily. Efficiency of microbial protein synthesis, however, was not affected by treatments (N source, P = 0.588; feeding frequency, P = 0.334). Rumen pH averaged 6.70 and it was neither affected by N source (P = 0.827) nor by feeding frequency (P = 0.740). Ruminal concentration of ammonia N was not affected by feeding frequency (P = 0.144) while it increased (P < 0.05) when urea rather than caseinate was the supplemental N source (mean of 7.61 mg/dl vs. 6.00 mg/dl). Concentration of sugars in rumen fluid was higher (P ≤ 0.05) in lambs supplemented once a day compared to twice daily (mean of 49.4 mg/dl vs. 34.4 mg/dl) for both N sources. A significant (P ≤ 0.05) N source by feeding frequency interaction effect was observed for ruminal concentrations of α-amino N compounds. In urea treatment α-amino N concentration increased (P ≤ 0.05) in lambs receiving the supplement twice daily compared to once a day (mean of 4.59 mg/dl vs. 3.70 mg/dl) while in caseinate treatment it was higher (P ≤ 0.05) in lambs offered the supplement in one meal per day compared to twice daily (mean of 5.29 mg/dl vs. 4.07 mg/dl). In conclusion, for ruminants fed a tropical grass-based diet, starch-rich supplement containing non-protein N as N source may be offered only once a day whereas the supply of nutrients may be improved if degradable true protein is included as N source and supplement is offered in two meals per day.     

Phosphorus retention capacity of agricultural headwater ditch sediments under alkaline condition in purple soils area,China

Phosphorus (P) retention by headwater ditch sediments adsorption plays a pivotal ecological role in P buffering in freshwater ecosystems. Previous studies focused on headwater ditch sediment adsorption and its P retention capacity in acid conditions, but little information is available for headwater ditches under alkaline condition. In this study, adsorption behavior of phosphorus was investigated in headwater ditch sediments under alkaline condition using a batch equilibrium technique, thus determining phosphorus retention capacity of headwater ditch sediments collected at 11 sites at base-flow on 2 March 2006 in purple soils area of China. Results showed that headwater ditch sediments had elevated phosphorus sorption maximum (S_max) values (122.72–293.23 mg P kg^?1) and P binding energy (K) values (1.64–8.65 L mg^?1), while they had low equilibrium phosphorus concentration (EPC₀) (0.001–0.108 mg L^?1) and degree of phosphorus saturation (DSP) (1.93–10.19%). Analysis of EPC₀ and soluble P concentration indicated that sediments acted as a sink for P across all headwater ditches. Therefore, there were high intrinsic P retention capacities of headwater ditch sediments. Positive correlations of both K and S_max with oxalate-extractable Fe (r of 0.93 and 0.81, p < 0.05) and total carbon (TC) (r of 0.89 and 0.74, p < 0.05) were found, thus suggesting that organic matter and amorphous or poorly crystalline Fe would play dominant roles in P adsorption in the headwater ditch sediments under alkaline condition. Since neither S_max nor K were correlated with CCE (CaCO₃) (r of 0.15 and ?0.06, p > 0.05), the high-energy sorptive surfaces of Fe oxides were more important than CaCO₃ in P sorption of sediment under alkaline condition. At the same time, these poor correlations between CCE and K and S_max imply a non-linear relationship between P retention and the content of carbonate. The negative correlations of both K and S_max with pH (r of–0.73, and–0.58, p < 0.05) revealed that an increase in pH would not improve sediment retention capacity under alkaline conditions.     

Short-time response in root morphology of Vallisneria natans to sediment type and water-column nutrient

Rooted submerged macrophytes can absorb significant amounts of nutrients from both sediment and water. We investigated root morphology of Vallisneria natans in mesocosm plastic bins, in response to three types of sediment (sandy loam, clay, and a 50:50 (v/v) mixture of the two sediments) and two levels of water-column nutrient (well water and nutrient medium). Compared to the plants grown in the clay or mixed sediments, root diameter decreased (0.39–0.41 versus 0.36–0.37 mm) but total root length per plant increased (0.87–1.27 versus 1.14–1.62 m) when grown in sandy loam. Increase of nutrient availability in water column led to decreased specific root length (306–339 versus 258–281 m g⁻¹). However, both sediment type and water-column nutrient had no impacts on root number (ranged from 19 to 24 number of roots per plant). Root weight ratio, root:leaf mass ratio and root:leaf length ratio generally decreased with enhanced nutrient availability in sediment or water. Plant growth was affected by sediment type alone (P < 0.05), rather than water-column nutrient (P > 0.05). However, plant N and P contents were significantly impacted by both sediment type (P ≤ 0.001) and water-column nutrient (P < 0.05). Increase of nutrient availability in sediment or water led to increased plant N (ranged from 2.47 to 4.77 mg g⁻¹) and P concentrations (ranged from 42.8 to 62.0 mg g⁻¹). These results indicate that considerable variation in root morphology of V. natans exists in response to the fertility of the sediment it is rooted in.     

Quantitative analysis of RU38486 (mifepristone) by HPLC triple quadrupole mass spectrometry

A sensitive liquid chromatography–mass spectrometric method was validated for the quantification of RU38486 (mifepristone) in human and murine plasma. The analyte and internal standard (alfaxolone) were extracted by liquid–liquid extraction with diethyl ether, resolved on a C18 column using gradient elution with methanol and ammonium acetate and detected after positive electrospray ionization (m/z 430 → 372; m/z 333 → 297, respectively). Quantification was linear over the range 0.5–500 ng (r² > 0.997), precise and accurate (intra-assay RSD ≤ 7.2%, RME ≤ 8.2%; inter-assay RSD ≤ 15.7% RME ≤ 10.2%). The limit of quantification (LOQ) was 50 pg injected on column, permitting reproducible analysis of RU38486 in small volumes of plasma.     

RANKL/OPG in primary cultures of osteoblasts from post-menopausal women. Differences between osteoporotic hip fractures and osteoarthritis

The OPG/RANKL/RANK system is important in the balance between bone formation and resorption.We used primary human osteoblasts (hOBs) cells to examine the impact of 17-β-estradiol (E2) or/and 1,25-dihydroxyvitamin D (1,25D) in OPG/RANKL system in 28 post-menopausal (PM) women; (a) with hip fracture (OP) or (b) with osteoarthritis (OA). The hOB from OP patients proliferated slower during the first stage, than the OA women (31.5 ± 2.6 and 21.4 ± 1.3 days, respectively, p < 0.05). The OP group secreted significantly higher OPG protein levels than the OA women (10.1 ± 2.6 and 4.4 ± 0.8 pmol/L, respectively, p < 0.05). The 1,25D and 1,25D+E2 induce an increase in RANKL and RANKL/OPG mRNA expression in OP patients above 200% (p < 0.05).HOBs from the osteoporotic hip initially proliferate slower but after reaching the first cellular confluence, the proliferation rate is equal in both groups. Furthermore, hOBs from hips with OP present a higher protein secretion of OPG, and higher RANKL and RANKL/OPG expression ratio in response to 1,25D and 1,25D+E2, than hOBs from OA women. All this could suggest that the greater bone loss that characterizes OP patients can be mediated due to differences in the secretion and expression of the RANKL/OPG system in response to different stimuli.     

Fecal nitrogen to estimate intake and digestibility in grazing ruminants

This study aimed at evaluating forage intake and digestibility in ruminants using fecal nitrogen content, as well as validating a non-linear model to estimate digestibility in ruminants. A total of 58 conventional metabolism trials, carried out with sheep fed 27 forages (offered pure or in mixture) used in Rio Grande do Sul (RS) during the period 1969–1989 was analyzed. OM intake and OM digestibility (OMD) results were regressed linearly against fecal N, and OMD was also estimated from fecal crude protein (N × 6.25) content by a non-linear regression model. Fecal nitrogen excretion estimated forage intake in sheep with an R² = 0.73, whereas a low R² value of 0.36 was observed for OMD estimates. The equation obtained using the non-linear model was OMD = 0.7326 ? 0.3598 exp [(?0.9052 CP (g/kg OM))/100]. The parameters a (0.7326) and b (0.3598) estimated by the equation for all forages were significant (P<0.00001) and there was no effect of type of forage (P=0.38). The mean prediction error (MSPE), was 0.2379, indicating that the equation fit well to the data. The difference between estimated and observed organic matter digestibility was mainly caused by random variation (0.9765). The results indicated that the equation using the non-linear model developed with all forages can be used with enough precision to estimate the OM digestibility of forage consumed by sheep in Rio Grande do Sul.     

Release of ferulic acid from corn cobs by alkaline hydrolysis

The process of corn cobs alkaline hydrolysis to produce solutions with high hydroxy-cinnamic acids content was investigated. In particular the attention was focused on the solubilisation of ferulic acid (FA) and related compounds, mainly p-coumaric acid (p-CA). Although these compounds have applications as antioxidants, the purpose of this work was to obtain FA solutions that can be used as feedstock for the biotechnological production of vanillin in future studies. The effects of different concentrations of NaOH (0.2 ≤ C_a ≤ 2.0N) and solid/liquid ratios (0.028 ≤ S/L ≤ 0.168 g/g) on the solubilisation of FA versus time have been investigated at room temperature. Optimal hydrolysis conditions (C_a = 0.5N, S/L = 0.084 g/g after 6 h) ensured the production of hydrolysates with relatively high contents of both FA (1171 ± 34 mg/L) and p-coumaric acid (2156 ± 64 mg/L), which can be used in future studies for the microbial transformation into vanillin.     

Effects of phosphorus and nitrogen amendments on the growth of Egeria najas

The effect of nutrient addition on the growth of E. najas was evaluated in a dose response experiment using sand amended with phosphorus (P) and nitrogen (N), and in enrichment trials with N and P amendments to natural sediments. Plants, water and sediment came from lagoons of the Upper Paraná River Floodplain and from Itaipu Reservoir (Brazil). Relative growth rates (RGRs) of E. najas shoots, based on dry mass (DM), varied from 0.03 to 0.060 d⁻¹ for both nutrients. Root:shoot biomass ratios were related to sediment exchangeable P (r = −0.419; P = 0.03) and N (r = −0.54; P = 0.006), however root RGR was not related to sediment nutrient concentrations. When natural sediments were amended with N and P, neither shoot nor root RGRs differed among treatments for substrata from either the reservoir or the floodplain lagoons (P > 0.05). Comparison of nutrient concentrations measured in natural sediments collected from several sites in both the Upper Paraná River Floodplain (range 49–213 μg P g⁻¹ DM; 36–373 μg N g⁻¹ DM) and Itaipu Reservoir (range 43–402 μg P g⁻¹ DM; 7.9–238 μg N g⁻¹ DM) showed that sediment N and P from these systems usually exceeded minimum requirements necessary for E. najas growth, as measured in the dose response experiment. Together, these results indicate that E. najas, at least in early stages of development, responds to sediment nutrient amendments and relies upon bottom sediments to meet its N and P requirements and that for at least two Brazilian ecosystems, growth of this species is not limited by insufficient sediment N or P. Thus, reducing N and P in water is not enough to control E. najas growth in short time periods in these ecosystems.     

Stimuli responsive polymorphism of C12NO/DOPE/DNA complexes: Effect of pH,temperature and composition

N,N-dimethyldodecylamine-N-oxide (C₁₂NO) is a surfactant that may exist either in a neutral or cationic protonated form depending on the pH of aqueous solutions. Using small angle X-ray diffraction (SAXD) we observe the rich structural polymorphism of pH responsive complexes prepared due to DNA interaction with C₁₂NO/dioleoylphosphatidylethanolamine (DOPE) vesicles and discuss it in view of utilizing the surfactant for the gene delivery vector of a pH sensitive system. In neutral solutions, the DNA uptake is low, and a lamellar L_α phase formed by C₁₂NO/DOPE is prevailing in the complexes at 0.2 ≤ C₁₂NO/DOPE < 0.6 mol/mol. A maximum of ~ 30% of the total DNA volume in the sample is bound in a condensed lamellar phase L_α^C at C₁₂NO/DOPE = 1 mol/mol and pH 7.2. In acidic conditions, a condensed inverted hexagonal phase H_II^C was observed at C₁₂NO/DOPE = 0.2 mol/mol. Commensurate lattice parameters, a_HC ≈ d_LC, were detected at 0.3 ≤ C₁₂NO/DOPE ≤ 0.4 mol/mol and pH = 4.9–6.4 suggesting that L_α^C and H_II^C phases were epitaxially related. While at the same composition but pH ~ 7, the mixture forms a cubic phase (Pn3m) when the complexes were heated to 80 °C and cooled down to 20 °C. Finally, a large portion of the surfactant (C₁₂NO/DOPE > 0.5) stabilizes the L_α^C phase in C₁₂NO/DOPE/DNA complexes and the distance between DNA strands (d_DNA) is modulated by the pH value. Both the composition and pH affect the DNA binding in the complexes reaching up to ~ 95% of the DNA total amount at acidic conditions.     

Expanded bed adsorption of an alkaline lipase from Pseudomona cepacia

An extracellular lipase was isolated from Pseudomona cepacia by expanded bed adsorption on an Amberlite 410 ion-exchange resin. Enzyme characterization and hydrodynamic study of a chromatography column were done. Enzyme purification was done at three condition of expanded bed height (H): at one and half (6 cm), at two (8 cm) and at three (12 cm) times the fixed bed height (H₀ = 4 cm). The results showed that the experimental data was fitted to the Richardson and Zaki equation, and the comparison between the experimental and calculated terminal velocities showed low relative error. In enzyme purification for better condition, a purification factor of about 80 times was found at 6 cm of expanded bed height, or 1.5 times of expansion degree. Purified lipase had an optimal pH and a temperature of 8 and 37 °C, respectively.     

Phosphorus sorption by sediments in a subtropical constructed wetland receiving stormwater runoff

This study investigated the potential of using a mixture of fishpond bund material, completely decomposed granite and river sand as substrate in a constructed wetland for phosphorus removal. Core samples were collected from the newly constructed Hong Kong Wetland Park (HKWP) receiving influent stormwater from a nearby new town, and batch incubation experiments were conducted to determine the P sorption characteristics of sediments. The HKWP sediments adsorbed the majority of available P in the initial 20 min of incubation, with a first-order rate constant of 1.01–2.11 h^?1. Sediments in the reedbeds and freshwater marshes possessed a great capacity for P adsorption with the high Langmuir sorption maxima (478–858 mg kg^?1) and Freundlich adsorption constants (417–672 L kg^?1) obtained, attributable to the high amorphous iron and aluminium concentrations compared to other constructed wetlands. Moreover, sediment equilibrium P concentrations were generally low (4.6–23.6 μg L^?1), facilitating a net P adsorption by sediments under moderate P loadings. Yet, the amount of P adsorbed by the HKWP sediments was limited by the low ambient porewater P concentrations and there was even a risk of P desorption when sediments in the freshwater marshes were resuspended into the water column. While substrates in the HKWP demonstrated a great potential for P adsorption, consideration should also be given to P loadings in influent water to fully utilize the P sorption capacity of sediments and enhance the P removal efficiency of constructed wetlands.     

Differential regulation of extracellular matrix constituents in myocardial remodeling with and without heart failure following pressure overload

Patients with aortic stenosis develop various degrees of myocardial hypertrophy and heart failure (HF) despite comparable transvalvular gradients. An important element in the transition from compensated hypertrophy to HF is dilatation of the left ventricle (LV). The molecular pathology associated with LV dilatation and development of HF is not known. Thus, we examined potential differences in the regulation of myocardial extracellular matrix (ECM) constituents in mice with hypertrophy only (ABnonHF) and with HF (ABHF) as response to comparable pressure overload. The ascending aorta was banded, or left loose in sham-operated mice. Increased lung weight and left atrial diameter indicating pulmonary congestion were used to identify ABHF mice. Cardiac function and geometry were evaluated by echocardiography. Despite comparable pressure gradients and cardiac output, ABHF had reduced fractional shortening (23%), reduced systolic (28%) and diastolic (32%) tissue velocity and increased LV internal dimension in diastole (10%) and systole (17%) (LVIDd/s) compared to ABnonHF (p ≤ 0.05). Microarray analyses identified 120 differently regulated genes related to ECM in ABHF compared to ABnonHF (p ≤ 0.05). Interestingly, in ABHF, we found a 24% (p ≤ 0.05) reduction of the LV collagen VIII protein levels despite increased levels of LV total collagen by 23% (p ≤ 0.05). LV collagen VIII correlated negatively with LVIDd (R = 0.55, p = 0.03) and LVIDs (R = 0.72, p = 0.002). As this protein may function as a “sealant” binding collagen fibrils together, reduction of collagen VIII could potentially contribute to LV dilatation and development of HF.     

Effect of supplementation with pelleted citrus pulp on digestibility and intake in beef cattle fed a tropical grass-based diet (Cynodon nlemfuensis)

Citrus pulp is an important by-product for sub-tropical and tropical ruminant animal production. In this study, three steers (average body weight = 324 ± 16 kg) were randomly assigned to three levels of pelleted citrus pulp (PCP) supplementation (0, 1.25, and 2.5 kg animal⁻¹ d⁻¹; as-fed) in a 3 × 3 Latin square design to evaluate its effects on forage intake, digestion, and ruminal pH. The basal diet was stargrass (Cynodon nlemfuensis) harvested and chopped every day and fed fresh. Supplementation with increasing amounts of PCP tended (P≤0.10) to result in a linear increase in digestibility of total diet dry matter (DM) and organic matter (OM), but no effects were noticed for digestibility of forage DM or total diet neutral detergent fiber. Forage DM intake decreased linearly (P=0.03) with increasing PCP supplementation, although the decrease tended (quadratic; P=0.08) to be of greater magnitude at the highest level of supplementation. Both a linear increase (P<0.01) and a quadratic trend (greatest increase with first level of supplementation; P=0.09) were also observed for intake of total digestible OM. Average ruminal pH was between 6.6 and 7.2 and was not affected (P=0.29) by supplementation treatment. Although supplementation with PCP depressed forage consumption somewhat, little effect on forage digestion was observed. The provision of digestible OM in the form of supplement was greater than that lost via depressed forage consumption, resulting in an overall increase in energy supply. Our results suggest that high levels of citrus pulp to beef cattle can lower forage intake, but increase total energy intake. High levels of citrus pulp supplementation could be beneficial in combination with forages high in rumen dagradable protein. Systems using grasses with higher ruminally degradable protein content than we used, may benefit from this extra supply of energy which should be tested in a further experiment.     

Response in root morphology and nutrient contents of Myriophyllum spicatum to sediment type

Sediment may play an important role during the submerged macrophyte decline in the eutrophication progress. In order to investigate the response in root morphology and nutrient contents of submerged macrophytes Myriophyllum spicatum to sediment, five sediment types were treated and used (five types of sediment were used in the experiment: treatment 1 was nature sediment + sand, a 50:50 (v/v) mixture, treatment 2 was the studied sediment only, treatment 3 was sediment + nitrogen (N, NH₄Cl 400 mg kg^?1), treatment 4 was sediment + phosphorus (P, NaH₂PO₄ 300 mg kg^?1); treatment 5 was sediment + phosphorus (P, NaH₂PO₄ 600 mg kg^?1)). The results show that the root N content was only significantly affected by adding N in sediments and P was elevated by adding N and P. The root mass and its percentage increased at first, the peak values were reached at 35 d, and then decreased. The root growth was restrained by adding sand and N in sediments, root senescence process was delayed at the later experimental time by adding P in sediments. The increase of root volume showed a similar trend to that of root growth, except for plant with P addition where root volume remained high after 35 d. The root volume decreased while the main root number increased significantly by adding sand in sediments. The mean root length and main root diameter were reduced by adding P in sediments. The compatible sediment nutrient condition is necessary to restore submerged macrophytes in a degraded shallow lake ecosystem, and the effect of sediment on the root morphology and nutrient content is one of the important aspects restricting the restoration of submerged macrophytes.     

Using liquid chromatography–tandem mass spectrometry to quantify monohydroxylated metabolites of polycyclic aromatic hydrocarbons in urine

We present an assay which employs enzyme digestion and solid phase extraction followed by liquid chromatography–tandem mass spectrometry to simultaneously quantify 16 hydroxylated polycyclic aromatic hydrocarbons (OHPAHs) in 3-ml samples of urine. The analytes consisted of 2-, 3-, and 4-ring OHPAHs, namely, 1- and 2-hydroxynaphthalene (1- and 2-OHNAP), 2-hydroxyfluorine (2-OHFLU), 1-, 2-, 3-, 4-, and 9-hydroxyphenanthrene (1-, 2-, 3-, 4-, and 9-OHPHE), 1-hydroxypyrene (1-OHPYR), 1- and 2-hydroxybenzo(a)anthracene (1- and 2-OHBAA), 3- and 6-hydroxychrysene (3- and 6-OHCHR) and 3-, 7-, and 9-hydroxybenzo(a)pyrene (3-, 7-, and 9-OHBAP). The method was validated using urine samples from steel workers and control subjects. The coefficients of variation of the method for the particular analytes were between 7% and 27% and the limits of quantitation were between 0.002 and 0.010 μg/l urine. The 2- and 3-ring OHPAHs were easily quantified in all subjects. However, 1-OHPYR was the only representative of the 4- and 5-ring metabolites that could be quantified. Pairwise correlations showed that all OHPAHs were highly correlated with each other (0.553 ≤ r ≤ 0.910) and with 1-OHPYR (0.614 ≤ r ≤ 0.910), the metabolite most widely accepted as a short-term biomarker of exposure to PAHs. The analyte, 2-OHNAP exhibited the lowest pairwise correlations with the other OHPAHs (0.542 ≤ r ≤ 0.628), presumably due to confounding by smoking. Metabolites of phenanthrene, an abundant PAH and the smallest to possess a bay region, are promising OHPAHs for characterizing both exposures to PAHs and the various metabolic pathways.     

Cotyledonary responses to maternal selenium and dietary restriction may influence alterations in fetal weight and fetal liver glycogen in sheep

To examine the effects of maternal supranutritional selenium (Se) and nutrient restriction during mid and late gestation on placental characteristics and fetal liver glycogen, ewes received either adequate Se (ASe) or high Se (HSe) prior to breeding. On d 64 of gestation, ASe and HSe ewes remained at 100% of requirements (controls; CON) or were restricted (RES; 60% of requirements). On d 135 of gestation, fetal weight (P ≤ 0.08) was greatest in both HSe and CON ewes. Placentome number, mass, and caruncular and cotyledonary weight were not different (P ≥ 0.17) among treatments. Fetal mass:placental mass ratio was less (P = 0.06) in RES compared to CON ewes. Compared to ASe, HSe exhibited increased (P ≤ 0.08) cellular proliferation and DNA concentration and decreased (P = 0.07) cellular size in cotyledonary tissue. Nutritional restriction decreased (P ≤ 0.08) cotyledonary protein concentration and cellular size. VEGF receptor 1 (Flt) mRNA in cotyledonary tissue was greater in HSe compared with ASe ewes (P = 0.06) and in RES compared with CON ewes (P = 0.08). There was no effect of diet on caruncular growth variables (P ≥ 0.13) or on placental vascularity (P ≥ 0.11). Progesterone was greater (P ≤ 0.08) in ASe–RES ewes compared to all groups at d 90 and ASe–CON and HSe–CON at d 104. Although fetal glucose and cortisol concentrations were not affected by diet, fetal liver glycogen was greater (P = 0.04) in ASe–RES compared to ASe–CON and HSe–RES ewes with HSe–CON being intermediate. Both Se and nutritional plane may impact placental function and fetal growth, as fetal weight and liver glycogen are altered despite similar placental vascularity measurements.     

Simultaneous saccharification and fermentation of kraft pulp by recombinant Escherichia coli for phenyllactic acid production

Simultaneous saccharification and fermentation (SSF) of renewable cellulose for the production of 3-phenyllactic acid (PhLA) by recombinant Escherichia coli was investigated. Kraft pulp recovered from biomass fractionation processes was used as a model cellulosic feedstock and was hydrolyzed using 10–50 filter paper unit (FPU) g⁻¹ kraft pulp of a commercial cellulase mixture, which increased the glucose yield from 21% to 72% in an enzyme dose-dependent manner. PhLA fermentation of the hydrolyzed kraft pulp by a recombinant E. coli strain expressing phenylpyruvate reductase from Wickerhamia fluorescens TK1 produced 1.9 mM PhLA. The PhLA yield obtained using separate hydrolysis and fermentation was enhanced from 5.8% to 42% by process integration into SSF of kraft pulp (20 g L⁻¹) in a complex medium (pH 7.0) at 37 °C. The PhLA yield was negatively correlated with the initial glucose concentration, with a five-fold higher PhLA yield observed in culture medium containing 10 g L⁻¹ glucose compared to 100 g L⁻¹. Taken together, these results suggest that the PhLA yield from cellulose in kraft pulp can be improved by SSF under glucose-limited conditions.     

Bioavailability and extraction of heavy metals from contaminated soil by Atriplex halimus

Pot experiments were performed to evaluate the phytoremediation capacity of plants of Atriplex halimus grown in contaminated mine soils and to investigate the effects of organic amendments on the metal bioavailability and uptake of these metals by plants. Soil samples collected from abandoned mine sites north of Madrid (Spain) were mixed with 0, 30 and 60 Mg ha⁻¹ of two organic amendments, with different pH and nutrients content: pine-bark compost and horse- and sheep-manure compost. The increasing soil organic matter content and pH by the application of manure amendment reduced metal bioavailability in soil stabilising them. The proportion of Cu in the most bioavailable fractions (sum of the water-soluble, exchangeable, acid-soluble and Fe–Mn oxides fractions) decreased with the addition of 60 Mg ha⁻¹ of manure from 62% to 52% in one of the soils studied and from 50% to 30% in the other. This amendment also reduced Zn proportion in water-soluble and exchangeable fractions from 17% to 13% in one of the soils. Manure decreased metal concentrations in shoots of A. halimus, from 97 to 35 mg kg⁻¹ of Cu, from 211 to 98 mg kg⁻¹ of Zn and from 1.4 to 0.6 mg kg⁻¹ of Cd. In these treatments there was a higher plant growth due to the lower metal toxicity and the improvement of nutrients content in soil. This higher growth resulted in a higher total metal accumulation in plant biomass and therefore in a greater amount of metals removed from soil, so manure could be useful for phytoextraction purposes. This amendment increased metal accumulation in shoots from 37 to 138 mg pot⁻¹ of Cu, from 299 to 445 mg pot⁻¹ of Zn and from 1.8 to 3.7 mg pot⁻¹ of Cd. Pine bark amendment did not significantly alter metal availability and its uptake by plants. Plants of A. halimus managed to reduce total Zn concentration in one of the soils from 146 to 130 mg kg⁻¹, but its phytoextraction capacity was insufficient to remediate contaminated soils in the short-to-medium term. However, A. halimus could be, in combination with manure amendment, appropriate for the phytostabilization of metals in mine soils.