Diatom taxa and assemblages for establishing nutrient criteria of lakes with anthropogenic hydrologic alteration

Stressor-response models offer guidance for concentration-based nutrient criteria in lakes under human intervention. Diatom-based statistics from biological responses were incorporated to derive taxon-specific and community-level change points (thresholds) of phosphorous and nitrogen in 77 Yangtze floodplain lakes. Diatom metrics relating with conductivity were adopted as response variables, since conductivity explained the maximum variation (38.1%) in diatom assemblages via Bootstrapped regression trees. Nonparametric change-point analysis and Threshold Indicator Taxa ANalysis showed threshold responses of diatom community structure at 0.05–0.08 mg TP/L in connected lakes and 0.02–0.04 mg TP/L in isolated lakes. Distinct community change points of sensitive diatoms occurred at 0.96–1.63 mg TN/L in connected lakes and 0.52–0.63 mg TN/L in isolated lakes. Diatom community structures of tolerant taxa were substantially altered beyond 0.22–0.23 mg/L in connected lakes and 0.52–0.69 mg NO_x/L in isolated lakes. Hydrological river-lake connectivity differed significantly in ecological nutrient criteria with more TN/TP criteria and less NO_x criteria in connected lakes. Given the ecological significance and biological integrity, diatom-based statistics can provide more reliable change points (thresholds) for nutrient criteria than Chl a-nutrient relationships.     

Effects of rapid temperature fluctuations prior to breeding on reproductive efficiency in replacement gilts

Rapidly cooling pigs after heat stress (HS) results in a pathophysiological condition, and because rapid temperature fluctuations may be associated with reduced reproductive success in sows, it lends itself to the hypothesis that these conditions may be linked. Objectives were to determine the effects of rapid cooling on thermal response and future reproductive success in pigs. Thirty-six replacement gilts (137.8±0.9 kg BW) were estrus synchronized and then 14.1±0.4 d after estrus confirmation, pigs were exposed to thermoneutral conditions (TN; n=12; 19.7±0.9°C) for 6 h, or HS (36.3±0.5°C) for 3 h, followed by 3 h of rapid cooling (HSRC; n=12; immediate TN exposure and water dousing) or gradual cooling (HSGC; n=12; gradual decrease to TN conditions) repeated over 2 d. Vaginal (T_V) and gastrointestinal tract temperatures (T_GI) were obtained every 15 min, and blood was collected on d 1 and d 2 during the HS and recovery periods at 180 and 60 min, respectively. Pigs were bred 8.3±0.8 d after thermal treatments over 2 d. Reproductive tracts were collected and total fetus number and viability were recorded 28.0±0.8 d after insemination. HS increased T_V and T_GI (P=0.01; 0.98 °C) in HSRC and HSGC compared to TN pigs. During recovery, T_V was reduced from 15 to 105 min (P=0.01; 0.33 °C) in HSRC compared to HSGC pigs, but no overall differences in T_GI were detected (P<0.05; 39.67 °C). Rapid cooling increased (P<0.05) TNFα compared to HSGC and TN pigs during recovery-d 1 (55.2%), HS-d 2 (35.1%), and recovery-d 2 (64.9%). Viable fetuses tended to be reduced (P=0.08; 10.5%) and moribund fetuses tended to be increased (P=0.09; 159.3%) in HSRC compared to HSGC and TN pigs. In summary, rapid cooling prior to breeding may contribute to reduced fetal viability and reproductive success in pigs.     

The role of plants in the removal of nutrients at a constructed wetland treating agricultural (dairy) wastewater,Ontario, Canada

The South Nation River Watershed, in eastern Ontario, Canada, is an agricultural watershed impacted by excess nutrient loading primarily from agricultural activities. A constructed wetland for the treatment of agricultural wastewater from a 150-cow dairy operation in this watershed was monitored in its eighth operating season to evaluate the proportion of total nitrogen (TN) (approximated by total Kjeldahl nitrogen (TKN) due to low NO₃⁻) and total phosphorus (TP) removal that could be attributed to storage in Typha latifolia L. and Typha angustifolia L., which dominate this system. Nutrient loading rates were high, with 16.2 kg ha⁻¹ d⁻¹ N and 3.4 kg ha⁻¹ d⁻¹ P entering the wetland and loading the first wetland cell. Plant uptake accounted for 0.7% of TKN removal when the vegetated free water surface cells were considered together. However, separately, in the second wetland cell with lower N and P loading rates, plants accounted for 9% of TKN, 21% of NH₄⁺ and 5% of TP removal. Plant uptake was significant to overall removal given wetland age and nutrient loading. Nutrient storage during the growing season at this constructed wetland helped reduce the nutrient load entering the watershed, already stressed by intensive local agriculture.     

Estimation of the long-term nutrient budget and thresholds of regime shift for a large shallow lake in China

In this study, we apply an integrated empirical and mechanism approach to estimate a comprehensive long-term (1953–2012) total nitrogen (TN) and total phosphorus (TP) loading budget for the eutrophic Lake Chaohu in China. This budget is subsequently validated, firstly, by comparing with the available measured data in several years, and secondly, by model simulations for long-term nutrient dynamics using both Vollenweider (VW) model and dynamic nonlinear (DyN) model. Results show that the estimated nutrient budget is applicable for further evaluations. Surprisingly, nutrient loading from non-point sources (85% for TN and 77% for TP on average) is higher than expectation, suggesting the importance of nutrient flux from the soil in the basin. In addition, DyN model performs relatively better than VW model, which is attributed to both the additional sediment recycling process and the parameters adjusted by the Bayesian-based Markov Chain Monte Carlo (MCMC) method. DyN model further shows that the TP loading thresholds from the clear to turbid state (631.8 ± 290.16 t y⁻¹) and from the turbid to clear state (546.0 ± 319.80 t y⁻¹) are significantly different (p < 0.01). Nevertheless, the uncertainty ranges of the thresholds are largely overlapped, which is consistent with the results that the eutrophication of Lake Chaohu is more likely to be reversible (74.12%) than hysteretic (25.53%). The ecosystem of Lake Chaohu shifted from the clear to turbid state during late 1970s. For managers, approximately two-thirds of the current TP loading must be reduced for a shift back with substantial improvement in water quality. Because in practice the reduction of loading from a non-point source is very difficult and costly, additional methods beyond nutrient reduction, such as water level regulation, should be considered for the lake restoration.     

Thymidine esters as substrate analogue inhibitors of angiogenic enzyme thymidine phosphorylase in vitro

Thymidine phosphorylase (TP) catalyzes the cleavage of thymidine into thymine and 2-deoxy-α-d-ribose-1-phosphate. Elevated activity of TP prevents apoptosis, and induces angiogenesis which ultimately leads to tumor growth and metastasis. Critical role of TP in cancer progression makes it a valid target in anti-cancer research. Discovery of small molecules as TP inhibitors is vigorously pursued in cancer therapy. In the present study, we functionalized thymidine as benzoyl ester to synthesize compounds 3–16. In vitro evaluation of thymidine esters for their thymidine phosphorylase inhibition activity was subsequently carried out. Compounds 4, 10, 14, and 15 showed good activities with lower IC₅₀ values than the standard, 7-deazaxanthine (IC₅₀ = 41.0 ± 1.63 μM). Among them, compound 14 showed five folds higher activity (IC₅₀ = 7.5 ± 0.8 μM), while 4 (IC₅₀ = 18.5 ± 1.0 μM) and 10 (IC₅₀ = 18.8 ± 1.2 μM) showed two folds higher activity than the standard. Compound 15 showed slightly better activity (IC₅₀ = 33.3 ± 1.5 μM) to the standard. Potent compounds were further subjected to kinetic and molecular docking studies to identify their mode of inhibition, and to study their interactions with the protein at atomic level, respectively. All active compounds were non-cytotoxic to mouse fibroblast 3T3 cell line. These results identify thymidine esters as substrate analogue (substrate-like) inhibitors of angiogenic enzyme thymidine phosphorylase for further studies.     

Characteristics of nitrous oxide (N2O) emission from intermittently aerated sequencing batch reactors (IASBRs) treating slaughterhouse wastewater at low temperature

This study investigated the characteristics of nitrous oxide (N₂O) emission from intermittently aerated sequencing batch reactors (IASBRs) treating high strength slaughterhouse wastewater at 11 °C, where partial nitrification followed by denitrification (PND) was achieved. N₂O generation and emission was examined at three aeration rates of 0.4, 0.6, and 0.8 L air/min in three IASBRs (SBR1, SBR2, and SBR3, respectively). The slaughterhouse wastewater contained chemical oxygen demand (COD) of 6057 ± 172.6 mg/L, total nitrogen (TN) of 576 ± 15.1 mg/L, total phosphorus (TP) of 52 ± 2.7 mg/L and suspended solids (SS) of 1843 ± 280.5 g/L. In the pseudo-steady state, the amount of N₂O emission was up to 5.7–11.0% of incoming TN. The aeration rate negatively affected N₂O emission and the ratio of N₂O emission to incoming TN was reduced by 48.2% when the aeration rate was increased from 0.4 to 0.8 L air/min. Results showed that more N₂O was generated in non-aeration periods than in aeration periods. Lower DO concentrations enhanced N₂O generation in the aeration periods (probably via nitrifier denitrification) while low DO concentrations (lower than 0.2 mg/L) did not affect N₂O generation in the non-aeration periods (probably via heterotrophic denitrification). When PHB was utilized as the organic substrate for denitrification, there was a high N₂O generation potential. It was estimated that 1.8 mg N₂O-N was generated accompanying per mg PHB consumed.     

Paracellular absorption in laboratory mice: Molecule size-dependent but low capacity

Water-soluble nutrients are absorbed by the small intestine via transcellular and paracellular processes. The capacity for paracellular absorption seems lower in nonfliers than in fliers, although that conclusion rests largely on a comparison of relatively larger nonflying mammals (> 155 g) and relatively smaller flying birds (< 155 g). We report on paracellular absorption in laboratory mice, the smallest nonflying mammal species studied to date. Using a standard pharmacokinetic technique, we measured the extent of absorption (fractional absorption = f) of inert carbohydrate probes: l-arabinose (M_r = 150.13 Da) and cellobiose (342.3) that are absorbed exclusively by the paracellular route, and 3-O-methyl d-glucose (3OMD-glucose) (M_r = 194) absorbed both paracellularly and transcellularly. f was measured accurately in urine collection trials of 5–10 h duration. Absorption of 3OMD-glucose by mice was essentially complete (f = 0.95 ± 0.07) and much higher than that for l-arabinose (f = 0.21 ± 0.02), indicating that in mice, like other nonflying mammals, > 80% of glucose is absorbed by mediated process(es) rather than the passive, paracellular route. As in all other vertebrates, absorption of cellobiose (f = 0.13 ± 0.02) was even lower than that for l-arabinose, suggesting an equivalent molecular size cut-off for flying and nonflying animals and thus a comparable effective TJ aperture. An important ecological implication is that smaller water-soluble plant secondary metabolites that have been shown to be absorbed by the paracellular path in cell culture, such as phenolics and alkaloids, might be absorbed in substantial amounts by bats and small birds relative to nonflying mammals such as mice.     

Effects of urban and non-urban land cover on nitrogen and phosphorus runoff to Chesapeake Bay

The aim of this study was to determine the effects of catchment and riparian stream buffer-wide urban and non-urban land cover/land use (LC/LU) on total nitrogen (TN) and total phosphorus (TP) runoff to the Chesapeake Bay. The effects of the composition and configuration of LC/LU patches were explored in particular. A hybrid-statistical-process model, the SPAtially Referenced Regression On Watershed attributes (SPARROW), was calibrated with year 1997 watershed-wide, average annual TN and TP discharges to Chesapeake Bay. Two variables were predicted: (1) yield per unit watershed area and (2) mass delivered to the upper estuary. The 166,534 km² watershed was divided into 2339 catchments averaging 71 km². LC/LU was described using 16 classes applied to both the catchments and also to riparian stream buffers alone. Seven distinct landscape metrics were evaluated. In all, 167 (TN) and 168 (TP) LC/LU class metric combinations were tested in each model calibration run. Runs were made with LC/LU in six fixed riparian buffer widths (31, 62, 125, 250, 500, and 1000 meters (m)) and entire catchments. The significance of the non-point source type (land cover, manure and fertilizer application, and atmospheric deposition) and factors affecting land-to-water delivery (physiographic province and natural or artificial land surfaces) was assessed. The model with a 31 m riparian stream buffer width accounted for the highest variance of mean annual TN (r² = 0.9366) and TP (r² = 0.7503) yield (mass for a specified time normalized by drainage area). TN and TP loadings (mass for a specified time) entering the Chesapeake Bay were estimated to be 1.449 × 10⁸ and 5.367 × 10⁶ kg/yr, respectively. Five of the 167 TN and three of the 168 TP landscape metrics were shown to be significant (p-value ≤ 0.05) either for non-point sources or land-to-water delivery variables. This is the first demonstration of the significance of riparian LC/LU and landscape metrics on water quality simulation in a watershed as large as the Chesapeake Bay. Land cover metrics can therefore be expected to improve the precision of estimated TN and TP annual loadings to the Chesapeake Bay and may also suggest changes in land management that may be beneficial in control of nutrient runoff to the Chesapeake Bay and similar watersheds elsewhere.     

The threshold responses of phytoplankton community to nutrient gradient in a shallow eutrophic Chinese lake

Excessive nutrient loads resulted in cascading trophic effects and ecosystem responses. Aims of this study were to determine if the thresholds in nutrient gradient related to phytoplankton community composition could be identified in eutrophic lake, and further to analyze the change of phytoplankton assemblage along the nutrient concentration based on Threshold Indicator Taxa ANalysis (TITAN). The results presented the significant community thresholds estimate for negative taxa declining at 1.650 mg/L TN and 131.5 μg/L TP, as well as simultaneously increasing for positive taxa at 1.665 mg/L TN and 151.5 μg/L TP along nutrient enrichment gradient. However, there was unremarkable change point determined for TN:TP ratios in Lake Dianchi. Elevated TN and TP altered the phytoplankton assemblage, even may induce the fade of algal blooms across the threshold in the hypertrophic lake. The findings could provide implications for deeply deciphering abrupt transitions for phytoplankton assemblage and developing nutrient tactics to protect the lake ecosystems.     

Terpenoids with cytotoxic activity from the branches and leaves of Pyrus pashia

Twenty terpenoids, including a new triterpenoid (1) and a new monoterpenoid (20), were isolated from the branches and leaves of Pyrus pashia. The structures of two new compounds were determined to be 2α, 3β, 27-trihydroxyolean-12-en-28-oic acid (1) and (4α)-3-(5,5-dimethyltetrahydrofuranyl)-1-buten-3-ol 3-O-β-d-glucopyranoside (20) on the basis of spectroscopic analysis (IR, HRESIMS, 1D and 2D NMR) and chemical method. Some of the isolated compounds were evaluated for their cytotoxic activity against a panel of human cancer cell lines by MTT assay, using cisplatin as a positive control. Compound 14 exhibited cytotoxic activities against A549 (IC₅₀ = 19.18 ± 4.26 μM), Hela (IC₅₀ = 12.56 ± 3.89 μM), SGC7901 (IC₅₀ = 10.48 ± 1.95 μM) and NHI-1975 (IC₅₀ = 7.38 ± 2.31 μM) cell lines as well as compound 12 displayed cytotoxic activities against A549 (IC₅₀ = 14.71 ± 1.47 μM) and Hela (IC₅₀ = 12.22 ± 1.88 μM) cell lines.     

Repeated fed-batch cultivation of Arthrospira (Spirulina) platensis using urea as nitrogen source

Arthrospira (Spirulina) platensis (Nordstedt) Gomont was autotrophically cultivated for biomass production in repeated fed-batch process using urea as nitrogen source, with the aim of making large-scale production easier, increasing cell productivity and then reducing the production costs. It was investigated the influence of the ratio of renewed volume to total volume (R), the urea feeding time (t_f) and the number of successive repeated fed-batch cycles on the maximum cell concentration (X_m), cell productivity (P_x), nitrogen-to-cell conversion yield (Y_x/n), maximum specific growth rate (μ_m) and protein content of dry biomass. The experimental results demonstrated that R = 0.80 and t_f = 6 d were the best cultivation conditions, being able to simultaneously ensure, throughout the three fed-batch cycles, the highest average values of three of the five responses (X_m = 2101 ± 113 mg L^?1, P_x = 219 ± 13 mg L^?1 d^?1 and Y_x/n = 10.3 ± 0.8 g g^?1).     

A nutrient biotic index (NBI) for use with benthic macroinvertebrate communities

Aquatic macroinvertebrates have been among the principal biological communities used for freshwater monitoring and assessment for several decades, but macroinvertebrate biomonitoring has not incorporated nutrient measures into assessment strategies. Two nutrient biotic indices were developed for benthic macroinvertebrate communities, one for total phosphorus (NBI-P), and one for nitrate (NBI-N). Weighted averaging was used to assess the distributions of 164 macroinvertebrate taxa across TP and NO₃⁻ gradients and to establish nutrient optima and subsequent nutrient tolerance values. Both the NBI-P and NBI-N were correlated with increasing mean TP and NO₃⁻ values (r = 0.68 and r = 0.57, respectively, p < 0.0001). A three-tiered scale of eutrophication for TP and NO₃⁻ (oligotrophic: ≤0.0175 mg/l TP, ≤0.24 mg/l NO₃⁻, mesotrophic: >0.0175 to ≤0.065 mg/l TP, >0.24 to ≤0.98 mg/l NO₃⁻, eutrophic: >0.065 mg/l TP, >0.98 mg/l NO₃⁻) was also established through cluster analysis of invertebrate communities using Bray–Curtis (quantitative) similarity. Significant differences (p < 0.0001) were detected between median NBI-P and NBI-N scores among the three trophic states. Therefore, the nutrient biotic indices (NBIs) appear to accurately reflect changes in stream trophic state. Multimetric water quality assessments were also used to identify thresholds of impairment among the three trophic states. Hodges-Lehman estimation indicated that the greatest change in assessment results occurred between the mesotrophic and eutrophic states. The eutrophic state also represented the highest percentage of overall impairment. Therefore, the suggested threshold for nutrient impairment is the boundary between mesotrophic and eutrophic (0.065 mg/l TP and 0.98 mg/l NO₃⁻). The corresponding NBI-P score (6.1) and NBI-N score (6.0) for this threshold incorporate predictive capabilities into the NBIs. The NBI and index score thresholds of impairment will provide monitoring programs with a robust measure of stream nutrient status and serve as a useful tool in enforcing regional nutrient criteria.     

Synthesis of pyrimidine-2,4,6-trione derivatives: Anti-oxidant,anti-cancer, α-glucosidase, β-glucuronidase inhibition and their molecular docking studies

This paper describes a facile protocol, efficient, and environmentally benign for the synthesis a series of barbiturate acid substituted at C5 position 3a–o. The desired compounds subjected in vitro for different set of bioassays including against anti-oxidant (DPPH and super oxide scavenger assays), anti-cancer, α-glucosidase and β-glucuronidase inhibitions. Compound 3m (IC₅₀ = 22.9 ± 0.5 μM) found to be potent α-glucosidase enzyme inhibitors and showed more activity than standard acarbose (IC₅₀ = 841 ± 1.73 μM). Compound 3f (IC₅₀ = 86.9 ± 4.33 μM) found to be moderate β-Glucuronidase enzyme inhibitors and showed activity comparatively less than the standard d-saccharic acid 1,4-lactone (IC₅₀ = 45.75 ± 2.16 μM). Furthermore, in sillico investigation was carried out to investigate bonding mode of barbiturate acid derivatives.     

Regional nutrient thresholds in wadeable streams of New York State protective of aquatic life

Human influence on the landscape has caused nutrients in surface waters to increase to the point where their presence has substantially altered biological communities. Because this is a nationally recognized problem, the United States Environmental Protection Agency (USEPA) tasked each state, tribe, and territory to adopt numeric nutrient criteria. Here we integrate the concept of ecological thresholds with the derivation of effects-based numeric nutrient criteria. Acceptable levels of risk exceeding predefined biocriteria were determined using conditional probability and nonparametric changepoint analysis. We show how certain community metrics exhibit threshold responses to nutrients. Using these thresholds, we suggest nutrient values protective of aquatic life and characterize community composition. Nutrient criteria were suggested for two aggregations of USEPA's nutrient ecoregions in New York State an upland pristine forested region (Ecoregions VIII and XI) and a nutrient-enriched lowland region (Ecoregions VII and XIV). Of 11 biological community metrics evaluated, 5 had a strong response to nutrients (NBI-P, NBI-N, HBI, TRI, and DMA). Maximum probabilities of exceeding the biological impairment thresholds established for these metrics ranged from 81% to 100%. Changepoint analysis conducted on probability outcomes of these metrics resulted in nutrient thresholds at or above USEPA nutrient guidance values, depending on ecoregion and nutrient variable (Ecoregion VIII/XI: 15 μg/L TP, 472 μg/L TN, 150 μg/L NO₃-N, Ecoregion VII/XIV: 17 μg/L TP, 1133 μg/L TN, 356 μg/L NO₃-N). Results of taxonomic similarity percentages (SIMPER) and species contributions indicate that several orders of macroinvertebrates and diatoms exhibit significant shifts in their percent of contributions to sample similarity in response to changes in nutrient concentrations.     

Nonparametric approaches for estimating regional lake nutrient thresholds

Nonparametric approaches including a classification and regression tree (CART), a nonparametric changepoint analysis (nCPA) and a Bayesian hierarchical modeling (BHM) method were developed to determine ecoregional nutrient response thresholds. A CART analysis revealed that hierarchical structure was important for predicting Chl a concentrations from total nitrogen (TN) and total phosphorus (TP). The nCPA and BHM methods confirmed the CART results for each node in the tree, and the 90% confidence interval for each threshold was calculated to quantify uncertainty. The CART, nCPA, and BHM methods suggested that the nutrient criteria differed significantly within certain nutrient ecoregions and that numerical nutrient criteria of 0.0150–0.222 mg/L TP and 0.300–1.766 mg/L TN may control Chl a concentrations in the various lake ecoregions. The results of this analysis suggest that the integration of CART, nCPA and BHM might be useful for determining nutrient thresholds.     

Lowering of oxidative stress improves endothelial function in healthy subjects with habitually low intake of fruit and vegetables: A randomized controlled trial of antioxidant- and polyphenol-rich blackcurrant juice

Inadequate intake of the recommended five-a-day fruit and vegetable portions might contribute to increased cardiovascular disease risk. We assessed the effects of dietary intake of a blackcurrant juice drink, rich in vitamin C and polyphenols, on oxidative stress and vascular function. This was a double-blind, placebo-controlled, parallel group study of 66 healthy adults who habitually consume <2 portions of fruit and vegetables per day. Participants were randomly allocated to consume 250 ml of placebo (flavored water) or low or high blackcurrant juice drink four times a day for 6 weeks. Flow-mediated dilation (FMD) and plasma concentrations of F₂-isoprostanes and vitamin C were measured. In the high blackcurrant juice drink group FMD increased significantly (5.8±3.1 to 6.9±3.1%, P=0.022) compared with the placebo group (6.0±2.2 to 5.1±2.4%). Plasma vitamin C concentration increased significantly in the low (38.6±17.6 to 49.4±21.0 µmol/L, P<0.001) and high (34.6±20.4 to 73.8±23.3 µmol/L, P<0.001) blackcurrant juice drink groups compared with the placebo group (38.1±21.0 to 29.0±17.6 µmol/L). F₂-isoprostane concentrations were significantly lower in the high blackcurrant juice drink group (225±64 pg/ml) compared with the low blackcurrant juice drink (257±69 pg/ml, P=0.002) and placebo group (254±59 pg/ml, P=0.003). At follow-up, changes in plasma vitamin C correlated significantly with changes in FMD (r=0.308, P=0.044). Consumption of blackcurrant juice drink high in vitamin C and polyphenols can decrease oxidative stress and improve vascular health in individuals with habitually low dietary fruit and vegetable intake.     

Structure and water permeability of fully hydrated diphytanoylPC

Diphytanoylphosphatidylcholine (DPhyPC) is a branched chain lipid often used for model membrane studies, including peptide/lipid interactions, ion channels and lipid rafts. This work reports results of volume measurements, water permeability measurements P_f, X-ray scattering from oriented samples, and X-ray and neutron scattering from unilamellar vesicles at T = 30 °C. We measured the volume/lipid V_L = 1426 ± 1 Å³. The area/lipid was found to be 80.5 ± 1.5 Å² when both X-ray and neutron data were combined with the SDP model analysis (Ku?erka, N., Nagle, J.F., Sachs, J.N., Feller, S.E., Pencer, J., Jackson, A., Katsaras, J., 2008. Lipid bilayer structure determined by the simultaneous analysis of neutron and X-ray scattering data. Biophys. J. 95, 2356–2367); this is substantially larger than the area of DOPC which has the largest area of the common linear chain lipids. P_f was measured to be (7.0 ± 1.0) × 10^?3 cm/s; this is considerably smaller than predicted by the recently proposed 3-slab model (Nagle, J.F., Mathai, J.C., Zeidel, M.L., Tristram-Nagle, S., 2008. Theory of passive permeability through lipid bilayers. J. Gen. Physiol. 131, 77–85). This disagreement can be understood if there is a diminished diffusion coefficient in the hydrocarbon core of DPhyPC and that is supported by previous molecular dynamics simulations (Shinoda, W., Mikami, M., Baba, T., Hato, M., 2004. Molecular dynamics study on the effects of chain branching on the physical properties of lipid bilayers. 2. Permeability. J. Phys. Chem. B 108, 9346–9356). While the DPhyPC head–head thickness (D_HH = 36.4 Å), and Hamaker parameter (H = 4.5 × 10^?21 J) were similar to the linear chain lipid DOPC, the bending modulus (K_C = 5.2 ± 0.5 × 10^?21 J) was 30% smaller. Our results suggest that, from the biophysical perspective, DPhyPC belongs to a different family of lipids than phosphatidylcholines that have linear chain hydrocarbon chains.     

Synthesis of alpha amylase inhibitors based on privileged indole scaffold

Twenty five derivatives of indole carbohydrazide (1–25) had been synthesized. These compounds were characterized using ¹H NMR and EI-MS, and further evaluated for their α-amylase inhibitory potential. The analogs (1–25) showed varying degree of α-amylase inhibitory potential.ranging between 9.28 and 599.0 µM when compared with standard acarbose having IC₅₀ value 8.78 ± 0.16 µM. Six analogs, 25 (IC₅₀ = 9.28 ± 0.153 µM), 22 (IC₅₀ = 9.79 ± 0.43 µM), 4 (IC₅₀ = 11.08 ± 0.357 µM), 1 (IC₅₀ = 12.65 ± 0.169 µM), 8 (IC₅₀ = 21.37 ± 0.07 µM) and 14 (IC₅₀ = 43.21 ± 0.14 µM) showed potent α-amylase inhibition as compared to the standard acarbose (IC₅₀ = 8.78 ± 0.16 µM). All other analogs displayed good to moderate inhibitory potential. Structure-activity relationship was established through the interaction of the active compounds with enzyme active site with the help of docking studies.     

Optimisation and scale-up of α-glucuronidase production by recombinant Saccharomyces cerevisiae in aerobic fed-batch culture with constant growth rate

α-Glucuronidase (EC 3.2.1.139) of family GH 115 from Scheffersomyces stipitis is a valuable enzyme for the modification of water-soluble xylan into insoluble biopolymers, due to its unique ability to act on polymeric xylans. The influence of growth rate on the production of α-glucuronidase by recombinant Saccharomyces cerevisiae MH1000pbk10D-glu in glucose-limited fed-batch culture was studied at 14 and 100 L scale. At and below the critical specific growth rate (μ_crit) of 0.12 h⁻¹ at 14 L scale, the biomass yield coefficient (Y_x/s) remained constant at 0.4 g g⁻¹ with no ethanol production, whereas ethanol yields relative to biomass (k_eth/x) of up to 0.54 g g⁻¹ and a steady decrease in Y_x/s were observed at μ > 0.12 h⁻¹. Production of α-glucuronidase was growth associated at a product yield (k_α-glu/x) of 0.45 mg g⁻¹, with the highest biomass (37.35 g/L) and α-glucuronidase (14.03 mg/L) concentrations, were recorded during fed-batch culture at or near to μ_crit. Scale-up with constant k_La from 14 to 100 L resulted in ethanol concentrations of up to 2.5 g/L at μ = 0.12 h⁻¹. At this scale, α-glucuronidase yield could be maximised at growth rates below μ_crit, to prevent localised high glucose concentration pockets at the feed entry zone that would induce oxido-reductive metabolism. This is the first report where recombinant production of α-glucuronidase (EC 3.2.1.139) by S. cerevisiae was optimised for application at pilot scale.     

Differences in preovulatory follicle dynamics and timing of preovulatory LH surge affect fertility of maiden sheep reared in semi-arid extensive conditions

In the current study follicular dynamics, pituitary function, ovulatory response and luteal activity of 30 maiden Barbarine sheep were analyzed according to oestrus occurrence and lambing outcome after oestrus synchronisation with cloprostenol. Animals were retrospectively classified in three groups named as O? (n = 7, ewes not displaying oestrus), O+L? (n = 7, ewes showing oestrus but failing to lamb) and O+L+ (n = 16; ewes showing oestrus and lambing thereafter). All the sheep ovulated and daily transrectal ultrasonographies revealed that preovulatory follicles were present at cloprostenol injection in all the animals. In sheep O+L+ and O+L?, 50% and 57% of the ovulatory follicles were the largest follicles at cloprostenol treatment (mean size of 4.1 ± 0.26 mm and 4.3 ± 0.74 mm, respectively). In O? ewes, the same percentage was higher (86%, P < 0.05 when compared to group O+L+; mean size of 4.0 ± 0.46 mm). The number of large follicles and the final diameter of the ovulatory follicles at oestrous tended thereafter to be higher in group O+L+ (1.4 ± 0.1 and 6.4 ± 0.2) than in groups O+L? (1 ± 0.2 and 5.7 ± 0.36) and O? (0.9 ± 0.2 and 5.9 ± 0.5, respectively). Conversely, the number of medium follicles at oestrus detection was higher in the group O+L? (2.1 ± 0.3, P < 0.05) than in the other two groups (1 ± 0.2 and 1 ± 0.3 for O+L+ and O? respectively). Timing of preovulatory LH surge was earlier for ewes O? (24.0 ± 4.75, P < 0.05) than for sheep O+L+ and O+L? (37.9 ± 2.45 h and 38.0 ± 4.75 h, respectively) and 94% of O+L+ ewes had a LH surge between 16 h and 64 h after cloprostenol injection compared to 57% in O+L? and O? groups (P < 0.05). Thus, maiden Barbarine sheep failing to display oestrus or conceive showed alterations in their follicular dynamics and, thereafter, pituitary function and ovulatory response.