Abstract  This study achieved resolution improvement for iodine speciation in the presence of an ion-pairing reagent by a pressure-driven capillary electrophoresis (CE) system. Addition of 0.01mM tetrabutyl ammonium hydroxide (TBAH) as the ion-pairing reagent into the electrophoretic buffer resulted in the complete separation of four iodine species (I(-), IO3(-), mono-iodothyrosine-MIT and di-iodothyrosine-DIT), because of the electrostatic interaction between TBAH and the negatively charged analytes. A +16kV separation voltage was applied along the separation capillary (50μm i.d., 80cm total and 60cm effective) with the inlet grounded. The detection wavelength was fixed at 210nm, and the pressure-driven flow rate was set at 0.12mLmin(-1) with an injected volume of 2μL. The optimal electrolyte consisted of 2mM borate, 2mM TBAH and 80% methanol with pH adjusted to 8.5. Baseline separation of iodine species was achieved within 7min. The detection limits for I(-), IO3(-), MIT and DIT were 0.052, 0.040, 0.032 and 0.025mgL(-1), respectively. The relative standard deviations of peak heights and areas were all below 3% for 5mgL(-1) and 5% for 1mgL(-1). Application of the proposed method was demonstrated by speciation analysis of iodine in two seaweed samples. The developed method offered satisfactory recoveries in the 91-99% range and good precisions (<5%). Good agreement between the determined values by the proposed CE method and the HPLC-ICP-MS method was also obtained. All results proved its great potential in routine analysis of iodine speciation in environmental, food and biological samples.

Abstract  An unprecedented stable isotope dilution assay for the genotoxic altertoxins along with exposure data of consumers is presented to enable a first risk assessment of these Alternaria toxins in foods. Altertoxins were produced as the most abundant Alternaria toxins in a modified Czapek-Dox medium with a low level of glucose as the carbon source and ammonium sulfate as the sole nitrogen source. Labeled altertoxins were synthesized in the same way using [(13)C6]glucose. Moreover, labeled alternariol, alternariol methyl ether, altenuene, and alternuisol were biosynthesized in another modified medium containing [(13)C6]glucose and sodium [(13)C2]acetate. A stable isotope dilution LC-MS/MS method was developed and used for food analysis. For altertoxin I, altertoxin II, alterperylenol, alternariol, and alternariol methyl ether, the limits of detection ranged from 0.09 to 0.53 μg kg(-1). The inter-/intra-day (n = 3 × 6) relative standard deviations of the method were below 13 %, and the recoveries ranged between 96 and 109 %. Among the various commercial food samples, some of the organic whole grains revealed low-level contamination with altertoxin I and alterperylenol, and paprika powder, which was heavily loaded with alternariol, alternariol methyl ether, and tentoxin, showed higher contamination level of altertoxin I and alterperylenol. Altertoxin II and III and stemphyltoxin III were not detectable. In addition, if the food was contaminated with altertoxins, it was likely to be co-contaminated with the other Alternaria toxins, but not necessarily vice versa. Maximum concentrations of altertoxin I and alterperylenol were detected in sorghum feed samples containing 43 and 58 μg kg(-1), respectively. This was significantly higher than that in the measured food samples.

Abstract  The purpose of this study was to find the effect of endurance training and thiamine supplementation on anti-fatigue during the exercise. Each nine students from K Women's University went through three cross-over treatments: placebo treatment, training treatment and thiamine treatment. Training treatment was performed with bicycle ergometer exercise for four weeks (five days per week). Each exercise was performed for an hour with intensity set at 70% (50rpm) of maximal oxygen uptake. Thiamine treatment group was given 10mg of thiamine tetrahydrofurfuryl disulfide per one kilogram for four weeks. The bicycle ergometer exercise was performed at 70% of maximal oxygen uptake in exercise intensity which 60 minutes of exercise was performed at 50rpm . Lactate concentration was significantly decreased during 15 to 30 minutes of exercise for those with training treatment and 15 to 60 minutes of exercise for those with thiamine treatment compared to placebo treatment group. Ammonia concentration was significantly decreased during 15 to 60 minutes of exercise and 15 to 30 minutes of recovery for those with training and thiamine treatment compared to placebo treatment. Resting blood thiamine concentrations of placebo treatment were significantly lower than training treatment. 60 minutes after the exercise, plasma thiamine concentration was significantly increased in all treatment group. To sum up the previous, thiamine intake during exercise positively benefits carbohydrate metabolism in a way that will decrease lactate concentration, ammonia concentration, and anti- fatigue by reducing the RPE. Therefore, we can consider thiamine intake to be utilized as similar benefits as endurance training.

Abstract  BACKGROUND/OBJECTIVE: A distinct suppressive effect of a whey protein (including glycomacropeptide)-enriched preload drink on subsequent food intake in comparison with a maltodextrin carbohydrate-enriched preload was demonstrated in an earlier companion study with the same female subjects; however, the potential mediators underlying the effect are unclear. The objective of this study was to investigate how the ingestion of a whey protein-enriched preload beverage affected postprandial plasma concentrations of several satiety-related gastrointestinal hormones and metabolites in comparison with a maltodextrin carbohydrate-enriched preload.

SUBJECTS/METHODS: Eighteen normal-weight women were studied in a single-blind, randomized block design. Blood samples were collected at various time intervals for 120 min after consumption of a test drink (300 ml, ~1300 kJ) enriched (45 g) with either maltodextrin carbohydrate or whey protein containing naturally present glycomacropeptide.

RESULTS: Plasma-active ghrelin concentrations decreased after both maltodextrin carbohydrate- and whey protein-enriched test drinks (P<0.05). The whey protein-enriched beverage led to increased plasma concentrations of cholecystokinin (CCK) at 60 and 75 min (P<0.05), glucagon-like peptide-1 (GLP-1) at 90 min (P<0.001), peptide tyrosine-tyrosine (PYY) at 90 and 120 min (P<0.01) and pancreatic polypeptide (PP) from 15 to 120 min (P<0.05) compared with maltodextrin carbohydrate. Total amino acid, urea and ammonia plasma concentrations were also higher after whey protein compared with maltodextrin carbohydrate ingestion (P<0.01).

CONCLUSIONS: Increased plasma concentrations of some gastrointestinal hormones related to satiety, particularly PP, and of amino acids and their metabolites, may have acted either singly or together to mediate the observed satiety response to whey protein.

Abstract  In this study, the effect of operational conditions on biofilm development and nitrification in moving-bed biofilm reactor (MBBR) was investigated. The reactor was operated in a continuously fed regime during 170 days and with theoretical hydraulic retention time of 7 h, respectively. The presence of chemical oxygen demand (COD) increased the time required to form stable nitrifying. Subsequent stepwise increase of influent COD caused an increment in total polysaccharide (PS) and protein (PN) content, which was accompanied by an attachment of the biofilm, as shown by atomic force microscope (AFM). PS and PN concentrations proved to be good indicators of biomass development and attachment in MBBR system. Reactor was operated and water quality was characterized before and after treatment. Parameters including pH, 5-day biochemical oxygen demand (BOD5), total suspended solids (TSS) (COD), PN, PS, and fecal bacteria in both raw and treated wastewater were monitored during the treatment. The removal rates of ammonium-nitrogen (NH4 (+)-N), BOD5, COD, and TSS are 95, 67.5, 69.2, and 73.33 %, respectively. The average bacterial reduction between the inlet and the outlet was of the order of 5 ± 1 logarithmic units for fecal coliforms. AFM showed that distinct biofilm and extracellular polymeric substances were formed in biofilm was thicker in the 70 days than in the 30 days. These results showed that the consumption rate for each substrate increased parabolically with biofilm thickness due to the increased amount of biomass Thus, MBBR can serve as a promising technology for wastewater treatment and can be scaled up for small communities in the developing countries.

Abstract  BACKGROUND:: Omeprazole (OME) is a proton pump inhibitor with a 58% bioavailability after a single oral dose. It is subject to marked inter-individual variations and significant drug-drug interactions. The authors developed a simple and rapid method based on liquid chromatography in tandem with mass spectrometry (LC/MS-MS) with solid phase extraction and isotope-labelled internal standard (IS) to monitor plasma levels of OME in pharmacokinetics and drug-drug interaction studies.

METHODS:: OME and its IS (OME-D3) were eluted with a Zorbax Extend C-18 rapid resolution column (4.6 mm x 50 mm, 3.5 μm) at 25°C, under isocratic conditions through a mobile phase consisting of 1 mM ammonium acetate, pH 8.5 (55%), and acetonitrile (45%). The flow rate was 0.8 mL/min, and the chromatogram run time was 1.2 min. OME was detected and quantified by LC-MS/MS with positive electrospray ionization, which operates in multiple-reaction monitoring mode.

RESULTS:: The method was linear in the range of 1.5-2000 ng/mL for OME. The validation assays for accuracy and precision, matrix effect, extraction recovery, and stability of the samples for OME did not deviate more than 20% for the lower limit of quantification and no more than 15% for other quality controls.

CONCLUSIONS:: These findings are consistent with the requirements of regulatory agencies. The method enables rapid quantification of OME concentrations and can be used in pharmacokinetic and drug-drug interaction studies.

Abstract  Currently, the mechanism(s) responsible for the regulation of urea transporter B (UT-B) expression levels in the epithelium of the rumen remain unclear. We hypothesized that rumen fermentation products affect ruminal UT-B expression. Therefore, the effects of short-chain fatty acids (SCFA), pH, ammonia, and urea on mRNA and protein levels of UT-B were assayed in primary rumen epithelial cell cultures and in rumen epithelium obtained from intact goats. In vitro, SCFA and acidic pH were found to synergetically stimulate both mRNA and protein expression of UT-B, whereas NH4Cl decreased mRNA and protein levels of UT-B at pH 6.8. Treatment with urea increased both levels at pH 7.4. When goats received a diet rich in nitrogen (N) and nonfiber carbohydrates (NFC), their rumen epithelium had higher levels of UT-B, and the rumen contained higher concentrations of SCFA and NH3-N with a lower pH. An increase in plasma urea-N concentration was also observed compared with the plasma of the goats that received a diet low in N and NFC. In a second feeding trial, goats that received a NFC-rich, but isonitrogenous, diet had higher mRNA and protein levels of UT-B, and higher levels of G protein-coupled receptor (GPR) 41 and GPR4, in their rumen epithelium. The ruminal concentrations of SCFA and NH3-N also increased, while a lower pH was detected. In contrast, the serum urea-N concentrations remained unchanged. These data indicate that ruminal SCFA and pH are key factors, via GPR4 and GPR41, in the dietary regulation of UT-B expression, and they have priority over changes in plasma urea.

Abstract  We investigated the impact of nutritional status on the physiological, metabolic and ion-osmoregulatory performance of European sea bass (Dicentrarchus labrax) when acclimated to seawater (32ppt), brackish water (20 and 10ppt) and hyposaline water (2.5ppt) for 2weeks. Following acclimation to different salinities, fish were either fed or fasted (unfed for 14days). Plasma osmolality, [Na(+)], [Cl(-)] and muscle water content were severely altered in fasted fish acclimated to 10 and 2.5ppt in comparison to normal seawater-acclimated fish, suggesting ion regulation and acid-base balance disturbances. In contrast to feed-deprived fish, fed fish were able to avoid osmotic perturbation more effectively. This was accompanied by an increase in Na(+)/K(+)-ATPase expression and activity, transitory activation of H(+)-ATPase (only at 2.5ppt) and down-regulation of Na(+)/K(+)/2Cl(-) gene expression. Ammonia excretion rate was inhibited to a larger extent in fasted fish acclimated to low salinities while fed fish were able to excrete efficiently. Consequently, the build-up of ammonia in the plasma of fed fish was relatively lower. Energy stores, especially glycogen and lipid, dropped in the fasted fish at low salinities and progression towards the anaerobic metabolic pathway became evident by an increase in plasma lactate level. Overall, the results indicate no osmotic stress in both feeding treatments within the salinity range of 32 to 20ppt. However, at lower salinities (10-2.5ppt) feed deprivation tends to reduce physiological, metabolic, ion-osmo-regulatory and molecular compensatory mechanisms and thus limits the fish's abilities to adapt to a hypo-osmotic environment.

Abstract  BackgroundMillions of human infections caused by arthropod-borne pathogens are initiated by the feeding of an infected mosquito on a vertebrate. However, interactions between the viruses and the mosquito vector, which facilitates successful infection and transmission of virus to a subsequent vertebrate host, are still not fully understood.FindingHere we describe early chikungunya virus (CHIKV) infectious events in cells derived from one of the most important CHIKV vectors, Aedes albopictus. We demonstrated that CHIKV infection of mosquito cells depended on acidification of the endosome as indicated by significant inhibition following prophylactic treatment with the lysosomotropic drugs chloroquine, ammonium chloride, and monensin, which is consistent with observations in mammalian cells. While all three agents inhibited CHIKV infection in C6/36 cells, ammonium chloride was less toxic to cells than the other agents.ConclusionThe observation of similar mechanisms for inhibition of CHIKV infection in mosquito and mammalian cell lines suggests that conserved entry pathways are utilized by CHIKV for vertebrate and invertebrate cell types.

Abstract  In drinking water, monochloramine may promote ammonia–oxidizing bacteria (AOB) growth because of concurrent ammonia presence. AOB use (i) ammonia monooxygenase for biological ammonia oxidation to hydroxylamine and (ii) hydroxylamine oxidoreductase for biological hydroxylamine oxidation to nitrite. In addition, monochloramine and hydroxylamine abiotically react, providing AOB a potential benefit by removing the disinfectant (monochloramine) and releasing growth substrate (ammonia). Alternatively and because biological hydroxylamine oxidation supplies the electrons (reductant) required for biological ammonia oxidation, the monochloramine/hydroxylamine abiotic reaction represents a possible inactivation mechanism by consuming hydroxylamine and inhibiting reductant generation. To investigate the abiotic monochloramine and hydroxylamine reaction's impact on AOB activity, the current study used batch experiments with Nitrosomonas europaea (AOB pure culture), ammonia, monochloramine, and hydroxylamine addition. To decipher whether hydroxylamine addition benefitted N. europaea activity by (i) removing monochloramine and releasing free ammonia or (ii) providing an additional effect (possibly the aforementioned reductant source), a previously developed cometabolism model was coupled with an abiotic monochloramine and hydroxylamine model for data interpretation. N. europaea maintained ammonia oxidizing activity when hydroxylamine was added before complete ammonia oxidation cessation. The impact could not be accounted for by monochloramine removal and free ammonia release alone and was concentration dependent for both monochloramine and hydroxylamine. In addition, a preferential negative impact occurred for ammonia versus hydroxylamine oxidation. These results suggest an additional benefit of exogenous hydroxylamine addition beyond monochloramine removal and free ammonia release, possibly providing reductant generation.

Abstract  Thermal hydrolysis pretreatment coupled with Thermophilic Anaerobic Digestion (TAD) for Waste Activated Sludge (WAS) treatment is a promising combination to improve biodegradation kinetics during stabilization. However, to date there is a limited knowledge of the anaerobic biomass composition and its impact on TAD process performances. In this study, the structure and dynamics of the microbial communities selected in two semi-continuous anaerobic digesters, fed with untreated and thermal pretreated sludge, were investigated. The systems were operated for 250 days at different organic loading rate. 16S rRNA gene clonal analysis and Fluorescence In Situ Hybridization (FISH) analyses allowed us to identify the majority of bacterial and archaeal populations. Proteolytic Coprothermobacter spp. and hydrogenotrophic Methanothermobacter spp. living in strict syntrophic association were found to dominate in TAD process. The establishment of a syntrophic proteolytic pathway was favoured by the high temperature of the process and enhanced by the thermal pretreatment of the feeding sludge. Proteolytic activity, alone or with thermal pretreatment, occurred during TAD as proven by increasing concentration of soluble ammonia and soluble COD (sCOD) during the process. However, the availability of a readily biodegradable substrate due to pretreatment allowed to significant sCOD removals (more than 55%) corresponding to higher biogas production in the reactor fed with thermal pretreated sludge. Microbial population dynamics analysed by FISH showed that Coprothermobacter and Methanothermobacter immediately established a stable syntrophic association in the reactor fed with pretreated sludge in line with the overall improved TAD performances observed under these conditions.

Abstract  Rectifying the accelerated chloramine decay after the onset of nitrification is a major challenge for water utilities that employ chloramine as a disinfectant. Recently, the evidence of soluble microbial products (SMPs) accelerating chloramine decay beyond traditionally known means was reported. After the onset of nitrification, with an intention to inactivate nitrifying bacteria and thus maintaining disinfectant residuals, breakpoint chlorination followed by re-chloramination is usually practiced by water utilities. However, what actually breakpoint chlorination does beyond known effects is not known, especially in light of the new finding of SMPs. In this study, experiments were conducted using severely nitrified chloraminated water samples (chloramine residuals <0.5 mg Cl2 L−1, nitrite residuals >0.1 mg N L−1 and an order of magnitude higher chloramine decay rate compared to normal decay) obtained from two laboratory scale systems operated by feeding natural organic matter (NOM) containing and NOM free waters. Results showed that the accelerated decay of chloramine as a result of SMPs can be eliminated by spiking higher free chlorine residuals (about 0.92 ± 0.03 to 1.16 ± 0.12 mg Cl2 L−1) than the stoichiometric requirement for breakpoint chlorination and nitrite oxidation. Further, accelerated initial chlorine decay showed chlorine preferentially reacts with nitrite and ammonia before destroying SMPs. This study, clearly demonstrated there is an additional demand from SMPs that needs to be satisfied to effectively recover disinfection residuals in subsequent re-chloramination.

Abstract  The whole-sediment Toxicity Identification Evaluation (TIE) approach is a useful technique that allows for the identification of the contaminants responsible for the toxicity of complex sediment samples. This study aimed to compare the effectiveness of this technique in identifying the causes of toxicity when the test organism used in the toxicity test is capable of ingesting sediment particles. Two forms of exposure were compared: whole-sediment (WS), which integrates dermic and dietary exposures; and sediment-water interface (SWI), which involves dermic exposure only. The combined analysis of the TIE experiments revealed that metals, ammonia and, at one station, organic compounds, were responsible for sediment toxicity. The integrated use of WS and SWI TIE manipulations provided a more complete overview of the causes of toxicity, and thus enabled a better comprehension of complex contamination situations and, consequently, a better ecological assessment.

Abstract  Prior investigation of an upflow anaerobic sludge blanket (UASB) reactor treating brewery wastes suggested that direct interspecies electron transfer (DIET) significantly contributed to interspecies electron transfer to methanogens. To investigate DIET in granules further, the electrical conductivity and bacterial community composition of granules in fourteen samples from four different UASB reactors treating brewery wastes were investigated. All of the UASB granules were electrically conductive whereas control granules from ANAMMOX (ANaerobic AMMonium OXidation) reactors and microbial granules from an aerobic bioreactor designed for phosphate removal were not. There was a moderate correlation (r=0.67) between the abundance of Geobacter species in the UASB granules and granule conductivity, suggesting that Geobacter contributed to granule conductivity. These results, coupled with previous studies, which have demonstrated that Geobacter species can donate electrons to methanogens that are typically predominant in anaerobic digesters, suggest that DIET may be a widespread phenomenon in UASB reactors treating brewery wastes.

Abstract  Ochratoxin A (OTA) is a widely spread nephrotoxic food contaminant mycotoxin. Unfortunately, attenuation or prevention of the toxic effects of OTA is still an unresolved problem. Molecular inclusion of OTA by cyclodextrins (CDs) results in complexes with low stability. In the human organism, OTA exists mostly in the dianionic state (OTA(2-)). Therefore, our major goal was to develop a chemically modified cyclodextrin which gives a more stable complex with OTA than the previously published derivatives and which shows stronger preference towards OTA(2-). In our fluorescence spectroscopic study we demonstrate that quaternary ammonium beta-cyclodextrin (QABCD) fulfils both of these requirements. The calculated stability constant of the QABCD-OTA(2-) complex was 28,840 M(-1) (about 200-fold higher than that of the β-CD-OTA(2-) complex). We hypothesize, that QABCD may be a suitable tool for the decontamination of different OTA-contaminated drinks; furthermore, for alleviation of the toxic effects of OTA, such complex formation may reduce its absorption from the intestine.

Abstract  MicroRNA-based therapeutic applications have fostered a growing interest in the development of microRNAs purification processes in order to obtain the final product with high purity degree, good quality and biologically active. The pre-miR-29 deficiency or overexpression has been associated to a number of clinically important diseases, and its therapeutic application can be considered. Monolithic columns emerged as a new class of chromatographic supports used in the plasmid DNA purification platforms, being an interesting alternative to the conventional particle-based columns. Thus, the current work describes, for the first time, a new affinity chromatography method that combines the high selectivity of agmatine ligands with the versatility of monoliths to specifically and efficiently purify pre-miR-29 from other small RNA species and Rhodovulum sulfidophilum impurities. The effect of different flow rates on pre-miR-29 separation was also evaluated. Moreover, breakthrough experiments were designed to study the effect of different RNA concentrations on the modified monolithic support binding capacity, being verified that the dynamic binding capacity for RNA molecules is dependent of the feed concentration. In order to achieve higher efficiency and selectivity, three different binding and elution strategies based on increased sodium chloride (1.75-3M) or arginine (100mM) and decreased ammonium sulfate (2.4-0M) stepwise gradients are described to purify pre-miR-29. As a matter of fact, by employing elution strategies using sodium chloride or arginine, an improvement in the final pre-miR-29 yields (97.33 and 94.88%, respectively) as well as purity (75.21 and 90.11%, respectively) were obtained. Moreover, the quality control analysis revealed that the level of impurities (proteins, endotoxins, sRNA) in the final pre-miR-29 sample was negligible. In fact, this new monolithic support arises as a powerful instrument on the microRNA purification to be used in further clinical applications, providing a more rapid and economical purification platform.

Abstract  This study investigated the effects of dietary nitrate addition on ruminal fermentation characteristics and microbial populations in goats. The involvement of Selenomonas ruminantium in nitrate and nitrite reduction in the rumen was also examined. As the result of nitrate feeding, the total concentration of ruminal volatile fatty acids decreased, whereas the acetate : propionate ratio and the concentrations of ammonia and lactate increased. Populations of methanogens, protozoa and fungi, as estimated by real-time PCR, were greatly decreased as a result of nitrate inclusion in the diet. There was modest or little impact of nitrate on the populations of prevailing species or genus of bacteria in the rumen, whereas Streptococcus bovis and S. ruminantium significantly increased. Both the activities of nitrate reductase (NaR) and nitrite reductase (NiR) per total mass of ruminal bacteria were increased by nitrate feeding. Quantification of the genes encoding NaR and NiR by real-time PCR with primers specific for S. ruminantium showed that these genes were increased by feeding nitrate, suggesting that the growth of nitrate- and nitrite-reducing S. ruminantium is stimulated by nitrate addition. Thus, S. ruminantium is likely to play a major role in nitrate and nitrite reduction in the rumen.

Abstract  BackgroundAdvances in the diagnosis and treatment of urea cycle disorders (UCDs) have led to a higher survival rate. The purpose of this study is to describe the characteristics of patients with urea cycle disorders in Spain.MethodsObservational, cross-sectional and multicenter study. Clinical, biochemical and genetic data were collected from patients with UCDs, treated in the metabolic diseases centers in Spain between February 2012 and February 2013, covering the entire Spanish population. Heterozygous mothers of patients with OTC deficiency were only included if they were on treatment due to being symptomatic or having biochemistry abnormalities.Results104 patients from 98 families were included. Ornithine transcarbamylase deficiency was the most frequent condition (64.4%) (61.2% female) followed by type 1 citrullinemia (21.1%) and argininosuccinic aciduria (9.6%). Only 13 patients (12.5%) were diagnosed in a pre-symptomatic state. 63% of the cases presented with type intoxication encephalopathy. The median ammonia level at onset was 298 ¿mol/L (169-615). The genotype of 75 patients is known, with 18 new mutations having been described. During the data collection period four patients died, three of them in the early days of life. The median current age is 9.96 years (5.29-18), with 25 patients over 18 years of age. Anthropometric data, expressed as median and z-score for the Spanish population is shown. 52.5% of the cases present neurological sequelae, which have been linked to the type of disease, neonatal onset, hepatic failure at diagnosis and ammonia values at diagnosis. 93 patients are following a protein restrictive diet, 0.84 g/kg/day (0.67-1.10), 50 are receiving essential amino acid supplements, 0.25 g/kg/day (0.20-0.45), 58 arginine, 156 mg/kg/day (109-305) and 45 citrulline, 150 mg/kg/day (105-199). 65 patients are being treated with drugs: 4 with sodium benzoate, 50 with sodium phenylbutyrate, 10 with both drugs and 1 with carglumic acid.ConclusionsStudies like this make it possible to analyze the frequency, natural history and clinical practices in the area of rare diseases, with the purpose of knowing the needs of the patients and thus planning their care.

Abstract  Systematic fundamental understanding of mass transport in osmosis-driven membrane processes is important for further development of this emerging technology. In this work, we investigate the role of membrane surface chemistry and charge on bidirectional solute diffusion in forward osmosis (FO). In particular, bidirectional diffusion of ammonium (NH4(+)) and sodium (Na(+)) is examined using FO membranes with different materials and surface charge characteristics. Using an ammonium bicarbonate (NH4HCO3) draw solution, we observe dramatically enhanced cation fluxes with sodium chloride feed solution compared to that with deionized water feed solution for thin-film composite (TFC) FO membrane. However, the bidirectional diffusion of cations does not change, regardless of the type of feed solution, for cellulose triacetate (CTA) FO membrane. We relate this phenomenon to the membrane fixed surface charge by employing different feed solution pH to foster different protonation conditions for the carboxyl groups on the TFC membrane surface. Membrane surface modification is also carried out with the TFC membrane using ethylenediamine to alter carboxyl groups into amine groups. The modified TFC membrane, with less negatively charged groups, exhibits a significant decrease in the bidirectional diffusion of cations under the same conditions employed with the pristine TFC membrane. Based on our experimental observations, we propose Donnan dialysis as a mechanism responsible for enhanced bidirectional diffusion of cations in TFC membranes.

Abstract  Robust start-up of the anaerobic ammonium oxidation (anammox) process from non-anammox-specific seeding material was achieved by using an inoculation with sludge-treating industrial [Formula: see text]-, organics- and N-rich yeast factory wastewater. N-rich reject water was treated at 20°C, which is significantly lower than optimum treatment temperature. Increasing the frequency of biomass fluidization (from 1-2 times per day to 4-5 times per day) through feeding the reactor with higher flow rate resulted in an improved total nitrogen removal rate (from 100 to 500 g m(-3)d(-1)) and increased anammox bacteria activity. As a result of polymerase chain reaction (PCR) tests, uncultured planctomycetes clone 07260064(4)-2-M13-_A01 (GenBank: JX852965) was identified from the biomass taken from the reactor. The presence of anammox bacteria after cultivation in the reactor was confirmed by quantitative PCR (qPCR); an increase in quantity up to ∼2×10(6) copies g VSS(-1) during operation could be seen in qPCR. Statistical modelling of chemical parameters revealed the roles of several optimized parameters needed for a stable process.

Abstract  Increased availability of rapidly fermentable carbohydrates and a great proportion of corn-derived CP in the diet may result in a degradable intake protein (DIP) deficit. Therefore, ruminal DIP deficit may result from high dietary inclusion of processed corn grain and small to moderate inclusion of corn distillers grains (DG). Two experiments were conducted to evaluate the effect of increasing dietary DIP concentration through the inclusion of urea on feedlot cattle performance, carcass characteristics, ruminal fermentation, total tract digestibility, and purine derivatives-to-creatinine (PDC) index. In Exp. 1, 42 steers (428 ± 5 kg initial BW) were assigned randomly to 1 of 3 diets containing (DM basis) 0 (control [CON]), 0.4 (low urea [LU]), or 0.6% urea (high urea [HU]) to provide 6.4, 7.5, or 8.0% dietary DIP, respectively, and 12% high-moisture corn (HMC), 20% corn dried DG with solubles (DDGS), 10% ryegrass haylage, 2.9% dry supplement, and dry-rolled corn (DRC). Steers were fed ad libitum once daily using a Calan gate system. Carcass-adjusted final BW and DMI were similar among treatments (P ≥ 0.58). Carcass-adjusted ADG was greater (P ≤ 0.04) for the HU diet compared with the LU and CON diets and was similar (P = 0.73) between the LU and CON diets. Carcass-adjusted G:F was greater (P = 0.03) for the HU diet compared with the LU diet, tended (P = 0.09) to be greater compared with the CON diet, and was similar (P = 0.61) between the LU and CON diets. Carcass characteristics were similar (P ≥ 0.34) among treatments. In Exp. 2, 4 ruminally cannulated steers (347 ± 18 kg initial BW) were randomly assigned to a replicated 2 × 2 Latin square design. Steers were fed the same CON or HU diet used in Exp. 1 ad libitum once daily. Differences in the PDC index were used as indicators of differences in microbial CP synthesis. Ruminal pH, OM intake, and starch and CP digestibility were not affected by treatment (P ≥ 0.13). Digestibility of OM and NDF and ruminal concentration of ammonia-N and total VFA were greater (P ≤ 0.04) for the HU diet compared with the CON diet. The PDC index was similar (P = 0.81) between treatments at 2 h before feed delivery: 4% lower and 14% greater for the HU diet compared with the CON diet at 4 and 10 h after feed delivery, respectively (P < 0.01). These results suggest that, due to limited DIP supplied by a DRC- and HMC-based feedlot diet containing 20% DDGS, urea supplementation resulted in improved ruminal fermentation and feed digestibility, which may explain the concurrently improved cattle performance.

Abstract  OBJECTIVE: To study immunization procedures and preparation methods of specific IgY antibodies (IgY-Hp, IgY-IB) produced by hens immunized with Helicobacter pylori (Hp) bacterial antigen and recombinant Hp specific antigen IB, detect the inhibition effects on Hp growth and Hp urease activity, and study the effects of oral administration for treating Hp infection.

METHODS: By using recombinant cholera toxin subunit B (rCTB) as an adjuvant, hens received intramuscular injection immunization for continuous 7 times at an interval of 14 days. Then, the eggs were collected; IgY was purified.

RESULTS: On day 49 after hens were immunized, levels of two antibodies all reached 1:12800; after they were purified by Ammonium sulfate precipitation, their purity was over 80%. IgY-Hp could inhibit Hp growth and inhibit Hp urease activity; although in vitro, IgY-IB could not inhibit Hp growth but could inhibit Hp urease activity. The experiments in vivo found that when IgY-Hp or IgY-IB with sucralfate dual oral therapy was used to treat Hp infected mouse model, the cure rate all could reach 83.3%.

CONCLUSION: According to immunization procedure, high titer specific IgY antibody (1:12800) can be obtained in 49 days and its titer remains stable. Oral administration of the specific IgY antibodies in Hp infected mice can reach a cure rate of 83.3%, and the antibodies are expected to become new drugs and therapeutic methods of targeted therapy against Hp infection.

Abstract  Several factors affecting erythritol production from glycerol by Yarrowia lipolytica Wratislavia K1 strain were examined in batch fermentations. Ammonium sulfate, monopotassium phosphate, and sodium chloride were identified as critical medium components that determine the ratio of polyols produced. The central composite rotatable experimental design was used to optimize medium composition for erythritol production. The concentrations of ammonium sulfate, monopotassium phosphate, and sodium chloride in the optimized medium were 2.25, 0.22, and 26.4 g L(-1), respectively. The C:N ratio was found as 81:1. In the optimized medium with 100 g L(-1) of glycerol the Wratislavia K1 strain produced 46.9 g L(-1) of erythritol, which corresponded to a 0.47 g g(-1) yield and a productivity of 0.85 g L(-1) hr(-1). In the fed-batch mode and medium with the total concentration of glycerol at 300 g L(-1) and C:N ratio at 81:1, 132 g L(-1) of erythritol was produced with 0.44 g g(-1) yield and a productivity of 1.01 g L(-1) hr(-1.)

Abstract  In this study, a fast UHPLC-MS/MS method was developed and validated for the determination of a novel potent carvone Schiff base of isoniazid (CSB-INH) in rat plasma using carbamazepine as an internal standard (IS). After a single-step protein precipitation by acetonitrile, CSB-INH and IS were separated on an Acquity BEH(TM) C18 column (50 × 2.1 mm, 1.7 µm) under an isocratic mobile phase, consisting of acetonitrile: 10 mM ammonium acetate (95:5, v/v), at a flow rate of 0.3 mL/min. Quantification was performed on a triple quadrupole tandem mass spectrometer in multiple reactions monitoring mode by using positive electrospray ionization source. The precursor to product ion transitions were set at m/z 270.08 → 79.93 for CSB-INH and m/z 237.00 → 178.97 for IS. The proposed method was validated in compliance with US Food and Drug Administration and European Medicines Agency guidelines for bioanalytical method validation. The method was found to be linear in the range of 0.35-2500 ng/mL (r(2)  ≥ 0.997) with a lower limit of quantification of 0.35 ng/mL. The intra- and inter-day precision values were ≤12.0% whereas accuracy values ranged from 92.3 to 108.7%. In addition, other validation results were within the acceptance criteria and the method was successfully applied in a pharmacokinetic study of CSB-INH in rats. Copyright © 2014 John Wiley & Sons, Ltd.

Abstract  Phenylketonuria (PKU) was the first inherited metabolic disease in which dietary treatment was found to prevent the disease's clinical features. Treatment of phenylketonuria remains difficult due to progressive decrease in adherence to diet and the presence of neurocognitive defects despite therapy. This review aims to summarize the current literature on new treatment strategies. Additions to treatment include new, more palatable foods based on glycomacropeptide that contains very limited amount of aromatic amino acids, the administration of large neutral amino acids to prevent phenylalanine entry into the brain or tetrahydropterina cofactor capable of increasing residual activity of phenylalanine hydroxylase. Moreover, human trials have recently been performed with subcutaneous administration of phenylalanine ammonia-lyase, and further efforts are underway to develop an oral therapy containing phenylanine ammonia-lyase. Gene therapy also seems to be a promising approach in the near future.