Application of F magnetic resonance to study the efficacy of fluorine labeled drugs in the three-dimensional cultured breast cancer cells

The cellular monitoring of tumor response to treatments is important for drug discovery and drug development in cancer therapy. We studied efficacy of Herceptin, a common breast cancer drug conjugated with a fluorine organic compound, perfluoro-15-crown-5-ether (PFCE) which easily forms biocompatible emulsions. Three new pharmaceutical forms of Herceptin, Herceptin/PFCE, Herceptin/PFCE/Lipoplex and Herceptin/PFCE/HydraLink were synthesized for the ex vivo study of their efficacy in breast cancer treatment. The emulsions were administered to 10⁹ cells mL⁻¹ of HER-2 positive human adenocarcinoma (MCF-7) cells and the same amount of human mammary epithelial cells (HMEC) cultured in three-dimensional (3D) geometry using hollow fiber bioreactor (HFB) device. Following drugs administration ex vivo, fluorine-19 magnetic resonance imaging (¹⁹F MRI) was applied for cells imaging to measure their viability and to study drug efficacy over 72 h. To ensure optimum drug tracking, HydraLink was used to provide stable binding affinity of emulsified Herceptin to receptor while cationic lipid (Lipofectamine) was used to enhance lipophilicity of the emulsions.After 72 h of treatment with Herceptin, Herceptin/PFCE, Herceptin/PFCE/Lipoplex and Herceptin/PFCE/HydraLink the viability of cells was 54 ± 2%, 49 ± 3%, 43 ± 5% and 42 ± 1%, respectively, as compared with control 93 ± 2%. The efficacy (EC₅₀) of Herceptin conjugated with emulsions was found to be 970 ± 13 μg mL⁻¹ for Herceptin/PFCE, 645 ± 11 μg mL⁻¹ for Herceptin/PFCE/Lipoplex, 678 ± 7 μg mL⁻¹ for Herceptin/PFCE/HydraLink and 1000 ± 3 μg mL⁻¹ for Herceptin. The results show that fluorine emulsions improved the efficacy of Herceptin and ¹⁹F signal intensity changes validated drug efficiency. The significant correlations between duration of treatments and MCF-7 cells viability were observed. While we studied breast cancer cells, the fluorine emulsions could be applied for treatment of other cancer cells overexpressing HER-2.     

γ-Cyclodextrin–polyurethane copolymer adsorbent for selective removal of endotoxin from DNA solution

Copolymer particles for removal of endotoxins (lipopolysaccharides, LPSs) were prepared by suspension copolymerization of γ-cyclodextrin (CyD) and 1,6-hexamethylenediisocyanate. The LPS-removing activity of the copolymer particles was compared with that of poly(ε-lysine)-immobilized Cellufine (cationic adsorbent) or polystyrene particles (hydrophobic adsorbent) by a batch method. When DNA was present in solution with LPSs under physiological conditions (pH 6.0, ionic strength of μ = 0.05–0.8), LPS-removing activity of the cationic or hydrophobic adsorbent was unsatisfactory because both the DNA and the LPSs were adsorbed onto each adsorbent. By contrast, the copolymer particles with γ-CyD cavity (CyD content: 14–20 mol%) could selectively remove LPSs from a DNA solution (50 μg ml⁻¹, pH 6.0, and μ = 0.05–0.2) containing LPSs (15 EU ml⁻¹) without the adsorption of DNA. The residual concentration of LPSs in the treated DNA solution was below 0.1 EU ml⁻¹, and the recovery of DNA was 99%.     

Primary hemocyte culture of Penaeus monodon as an in vitro model for white spot syndrome virus titration, viral and immune related gene expression and cytotoxicity assays

Immortal cell lines have not yet been reported from Penaeus monodon, which delimits the prospects of investigating the associated viral pathogens especially white spot syndrome virus (WSSV). In this context, a method of developing primary hemocyte culture from this crustacean has been standardized by employing modified double strength Leibovitz-15 (L-15) growth medium supplemented with 2% glucose, MEM vitamins (1×), tryptose phosphate broth (2.95 g l⁻¹), 20% FBS, N-phenylthiourea (0.2 mM), 0.06 μg ml⁻¹ chloramphenicol, 100 μg ml⁻¹ streptomycin and 100 IU ml⁻¹ penicillin and hemolymph drawn from shrimp grown under a bio-secured recirculating aquaculture system (RAS). In this medium the hemocytes remained viable up to 8 days. 5-Bromo-2′-deoxyuridine (BrdU) labeling assay revealed its incorporation in 22 ± 7% of cells at 24 h. Susceptibility of the cells to WSSV was confirmed by immunofluoresence assay using a monoclonal antibody against 28 kDa envelope protein of WSSV. A convenient method for determining virus titer as MTT₅₀/ml was standardized employing the primary hemocyte culture. Expression of viral genes and cellular immune genes were also investigated. The cell culture could be demonstrated for determining toxicity of a management chemical (benzalkonium chloride) by determining its IC₅₀. The primary hemocyte culture could serve as a model for WSSV titration and viral and cellular immune related gene expression and also for investigations on cytotoxicity of aquaculture drugs and chemicals.     

Sulfated polysaccharides from Laminaria angustata: Structural features and in vitro antiviral activities

Sulfated polysaccharides potently inhibit the infectivity of herpes simplex virus (HSV) in cultured cells. In this study, we have analyzed sulfated xylogalactofucan and alginic acid containing fractions generated from Laminaria angustata, a marine alga. The xylogalactofucan that has apparent molecular mass of 56 ± 5 kDa and unusually low sulfate content contains, inter alia, 1,3-, 1,4- and 1,2-linked fucopyranosyl residues. The algin (molecular mass: 32 ± 5 kDa) contains gulo- (55.5%) and mannuronic (44.5%) acid residues. Introduction of sulfate groups enhanced the macromolecules capability to inhibit the infection of cells by HSV-1. The 50% inhibitory concentration (IC₅₀) values of these macromolecules against HSV-1 were in the range of 0.2-25 μg ml⁻¹ and they lacked cytotoxicity at concentrations up to 1000 μg ml⁻¹. The sulfate content appeared to be an important hallmark of anti-HSV-1 activity. Our results suggest the feasibility of inhibiting HSV attachment to cells by direct interaction of polysaccharides with viral particles.     

Synergistic production of L-arabinose from arabinan by the combined use of thermostable endo- and exo-arabinanases from Caldicellulosiruptor saccharolyticus

The optimum conditions for the production of l-arabinose from debranched arabinan were determined to be pH 6.5, 75 °C, 20 g l⁻¹ debranched arabinan, 42 U ml⁻¹ endo-1,5-α-l-arabinanase, and 14 U ml⁻¹ α-l-arabinofuranosidase from Caldicellulosiruptor saccharolyticus and the conditions for sugar beet arabinan were pH 6.0, 75 °C, 20 g l⁻¹ sugar beet arabinan, 3 U ml⁻¹ endo-1,5-α-l-arabinanase, and 24 U ml⁻¹ α-l-arabinofuranosidase. Under the optimum conditions, 16 g l⁻¹l-arabinose was obtained from 20 g l⁻¹ debranched arabinan or sugar beet arabinan after 120 min, with a hydrolysis yield of 80% and a productivity of 8 g l⁻¹ h⁻¹. This is the first reported trial for the production of l-arabinose from the hemicellulose arabinan by the combined use of endo- and exo-arabinanases.     

Ethanol production from sweet sorghum juice using very high gravity technology: Effects of carbon and nitrogen supplementations

Ethanol production from sweet sorghum juice by Saccharomyces cerevisiae NP01 was investigated under very high gravity (VHG) fermentation and various carbon adjuncts and nitrogen sources. When sucrose was used as an adjunct, the sweet sorghum juice containing total sugar of 280 g l⁻¹, 3 g yeast extract l⁻¹ and 5 g peptone l⁻¹ gave the maximum ethanol production efficiency with concentration, productivity and yield of 120.68 ± 0.54 g l⁻¹, 2.01 ± 0.01 g l⁻¹ h⁻¹ and 0.51 ± 0.00 g g⁻¹, respectively. When sugarcane molasses was used as an adjunct, the juice under the same conditions gave the maximum ethanol concentration, productivity and yield with the values of 109.34 ± 0.78 g l⁻¹, 1.52 ± 0.01 g l⁻¹ h⁻¹ and 0.45 ± 0.01 g g⁻¹, respectively. In addition, ammonium sulphate was not suitable for use as a nitrogen supplement in the sweet sorghum juice for ethanol production since it caused the reduction in ethanol concentration and yield for approximately 14% when compared to those of the unsupplemented juices.     

Effects of the methane-inhibitors nitrate,nitroethane, lauric acid,Lauricidin and the Hawaiian marine algae Chaetoceros on ruminal fermentation in vitro

The effects of several methane-inhibitors on rumen fermentation were compared during three 24 h consecutive batch cultures of ruminal microbes in the presence of nonlimiting amounts of hydrogen. After the initial incubation series, methane production was reduced greater than 92% from that of non-treated controls (25.8 ± 8.1 μmol ml⁻¹ incubation fluid) in cultures treated with nitroethane, sodium laurate, Lauricidin^® or a finely-ground product of the marine algae, Chaetoceros (added at 1, 5, 5 and 10 mg ml⁻¹, respectively) but not in cultures treated with sodium nitrate (1 mg m1⁻¹). Methane production during two successive incubations was reduced greater than 98% from controls (22.5 ± 3.2 and 23.5 ± 7.9 μmol ml⁻¹, respectively) by all treatments. Reductions in amounts of volatile fatty acids and ammonia produced and amounts of hexose fermented, when observed, were most severe in sodium laurate-treated cultures. These results demonstrate that all tested compounds inhibited ruminal methane production in our in vitro system but their effects on fermentation differed.     

Butyric acid fermentation in a fibrous bed bioreactor with immobilized Clostridium tyrobutyricum from cane molasses

Butyrate fermentation by immobilized Clostridium tyrobutyricum was successfully carried out in a fibrous bed bioreactor using cane molasses. Batch fermentations were conducted to investigate the influence of pH on the metabolism of the strain, and the results showed that the fermentation gave a highest butyrate production of 26.2 g l⁻¹ with yield of 0.47 g g⁻¹ and reactor productivity up to 4.13 g l⁻¹ h⁻¹ at pH 6.0. When repeated-batch fermentation was carried out, long-term operation with high butyrate yield, volumetric productivity was achieved. Several cane molasses pretreatment techniques were investigated, and it was found that sulfuric acid treatment gave better results regarding butyrate concentration (34.6 ± 0.8 g l⁻¹), yield (0.58 ± 0.01 g g⁻¹), and sugar utilization (90.8 ± 0.9%). Also, fed-batch fermentation from cane molasses pretreated with sulfuric acid was performed to further increase the concentration of butyrate up to 55.2 g l⁻¹.     

Comparative pharmacokinetics of baicalin and wogonoside by liquid chromatography–mass spectrometry after oral administration of Xiaochaihu Tang and Radix scutellariae extract to rats

The aim of this study was to compare the pharmacokinetics of baicalin and wogonoside in rats following oral administration of Xiaochaihu Tang (Minor Radix Bupleuri Decoction) and Radix scutellariae extract. Thus, a specific LC–MS method was developed and validated for the determination of these flavonoids in the plasma of rats after oral administration Xiaochaihu Tang and Radix scutellariae extract. Chromatographic separation was performed on a Zorbax SB C₁₈ column (150 mm × 4.6 mm, i.d.: 5 μm) with 0.1% formic acid in water and acetonitrile by linear gradient elution. Baicalin, wogonoside and carbamazepine (internal standard, I.S.) were detected in select-ion-monitoring (SIM) mode with a positive electrospray ionization (ESI) interface. The following ions: m/z 447 for baicalin, m/z 461 for wogonoside and m/z 237 for the I.S. were used for quantitative determination. The calibration curves were linear over the concentration ranges from 0.1231 to 6.156 μg mL⁻¹ for baicalin and 0.08832 to 4.416 μg mL⁻¹ for wogonoside. The lower limit of detection (LLOD) based on a signal-to-noise ratio of 2 was 0.06155 μg mL⁻¹ for baicalin and 0.04416 μg mL⁻¹ for wogonoside. Intra-day and inter-day precisions (RSD%) were within 10% and accuracy (RE%) ranged from −6.4 to 4.4%. The extraction recovery at three QC concentrations ranged from 74.7 to 86.0% for baicalin and from 71.3 to 83.7% for wogonoside. The plasma concentrations of baicalin and wogonoside in rats at designated time periods after oral administration were successfully determined using the validated method, pharmacokinetic parameters were estimated by a non-compartment model. Following oral administration of Xiaochaihu Tang and Radix scutellariae extract, the t_1/2 of baicalin was 3.60 ± 0.90 and 5.64 ± 1.67, the C_max1 was 1.64 ± 0.99 and 5.66 ± 2.02, the t_max1 was 0.13 ± 0.05 and 0.20 ± 0.07, the C_max2 was 2.43 ± 0.46 and 3.18 ± 1.66, and the t_max2 were 6.40 ± 1.67 and 5.66 ± 2.02, respectively. Following oral administration of Xiaochaihu Tang and Radix scutellariae extract, the t_1/2 of wogonoside was 4.97 ± 1.68 and 7.71 ± 1.55, the C_max1 was 1.39 ± 0.83 and 1.45 ± 0.37, the t_max1 was 0.21 ± 0.20 and 0.17 ± 0.01, the C_max2 was 1.90 ± 0.55 and 1.42 ± 0.70, and the t_max2 was 5.60 ± 1.67 and 5.20 ± 1.79, respectively. A significant difference (p < 0.05) was observed for t_1/2, and the elimination of baicalin and wogonoside in Xiaochaihu Tang was increased.     

Cytoskeletal and developmental alterations in Ceratophyllum demersum induced by microcystin-LR, a cyanobacterial toxin

The aim of the present work was the investigation of microtubule organization related to developmental processes of Ceratophyllum demersum, a submergent aquatic macrophyte, as affected by microcystin-LR (MCY-LR), a cyanobacterial toxin. We studied the time- and dose-dependent effects of the cyanotoxin in a concentration range of 0.01-20 μg mL⁻¹ (0.01-20.1 μM) at exposure times of 2-16 d. At short term (4 d) of MCY-LR exposure we observed the inhibition of C. demersum shoot tip elongation. This phenomenon was already observed at 0.01 μg mL⁻¹ MCY-LR (reduction of shoot tip length to 56 ± 3.6% of controls) and correlated with the induction of cortical microtubule (CMT) reorientation from transverse to longitudinal known to induce radial expansion of meristematic cells instead of normal elongation. Concomitantly we detected the increase of the percentage of cells in early mitosis in shoot tip meristems, from 1.17 ± 0.2% for controls to 1.93 ± 0.3 at 0.01 μg mL⁻¹ MCY-LR and 6.19 ± 0.5 at 10 μg mL⁻¹ MCY-LR. Cyanotoxin exposure induced the inhibition of general shoot elongation that was more pronounced than inhibition of the increase of leaf whorl number or fresh weight in the treatment period and this was observable at as short as 2-4 d of 2.5 μg mL⁻¹ MCY-LR exposure. This observation further proved that the MCY-LR-induced inhibition of cell elongation is responsible mainly for growth inhibition in C. demersum. Concomitantly with developmental and growth changes MCY-LR decreased protein and chlorophyll content at 16 d of exposure: at 20 μg mL⁻¹ of cyanotoxin, protein content was reduced to 53.3 ± 2.8%, while total chlorophyll content to 46.53 ± 2.7% of controls. This is the first study showing that MCY-LR inhibits the growth of C. demersum through cytoskeletal reorganization. This plant proved to be a powerful model system for toxicological as well as plant cell biology studies.     

Determination of the major constituents in fruit of Arctium lappa L. by matrix solid-phase dispersion extraction coupled with HPLC separation and fluorescence detection

The arctiin and arctigenin in the fruit of Arctium lappa L. were extracted by matrix solid-phase dispersion (MSPD) and determined by high-performance liquid chromatography (HPLC) with fluorescence detection. The experimental conditions for the MSPD were optimized. Silica gel was selected as dispersion adsorbent and methanol as elution solvent. The calibration curve showed good relationship (r > 0.9998) in the concentration range of 0.010–5.0 μg mL⁻¹ for arctiin and 0.025–7.5 μg mL⁻¹ for arctigenin. The recoveries were between 74.4% and 100%. The proposed method consumed less sample, time and solvent compared with conventional methods, including ultrasonic and Soxhlet extraction.     

Enhanced ethanol production from sugarcane juice by galactose adaptation of a newly isolated thermotolerant strain of Pichia kudriavzevii

The thermotolerant yeast strain isolated from sugarcane juice through enrichment technique was identified as a strain of Pichiakudriavzevii (Issatchenkiaorientalis) through molecular characterization. The P. kudriavzevii cells adapted to galactose medium produced about 30% more ethanol from sugarcane juice than the non-adapted cells. The recycled cells could be used for four successive cycles without a significant drop in ethanol production. Fermentation in a laboratory fermenter with galactose adapted P. kudriavzevii cells at 40 °C resulted in an ethanol concentration and productivity of 71.9 g L⁻¹ and 4.0 g L⁻¹ h⁻¹, respectively from sugarcane juice composed of about 14% (w/v) sucrose, 2% (w/v) glucose and 1% (w/v) fructose. In addition to ethanol, 3.30 g L⁻¹ arabitol and 4.19 g L⁻¹ glycerol were also produced, whereas sorbitol and xylitol were not formed during fermentation. Use of galactose adapted P. kudriavzevii cells for ethanol production from sugarcane juice holds potential for scale-up studies.     

Cultivation of microalgae for oil production with a cultivation strategy of urea limitation

The microalgae, Chlorella sp., were cultivated in various culture modes to assess biomass and lipid productivity in this study. In the batch mode, the biomass concentrations and lipid content of Chlorella sp. cultivated in a medium containing 0.025–0.200 g L⁻¹ urea were 0.464–2.027 g L⁻¹ and 0.661–0.326 g g⁻¹, respectively. The maximum lipid productivity of 0.124 g d⁻¹ L⁻¹ occurred in a medium containing 0.100 g L⁻¹ urea. In the fed-batch cultivation, the highest lipid content was obtained by feeding 0.025 g L⁻¹ of urea during the stationary phase, but the lipid productivity was not significantly increased. However, a semi-continuous process was carried out by harvesting the culture and renewing urea at 0.025 g L⁻¹ each time when the cultivation achieved the early stationary phase. The maximum lipid productivity of 0.139 g d⁻¹ L⁻¹ in the semi-continuous culture was highest in comparison with those in the batch and fed-batch cultivations.     

Low molecular weight chitosans—Preparation with the aid of pronase,characterization and their bactericidal activity towards Bacillus cereus and Escherichia coli

The homogeneous low molecular weight chitosans (LMWC) of molecular weight 9.5–8.5 kDa, obtained by pronase catalyzed non-specific depolymerization (at pH 3.5, 37 °C) of chitosan showed lyses of Bacillus cereus and Escherichia coli more efficiently (100%) than native chitosan (< 50%). IR and ¹H-NMR data showed decrease in the degree of acetylation (14–19%) in LMWC compared to native chitosan (∼ 26%). Minimum inhibitory concentration of LMWC towards 10⁶ CFU ml^− 1 of B. cereus was 0.01% (w/v) compared to 0.03% for 10⁴ CFU ml^− 1 of E. coli. SEM revealed pore formation as well as permeabilization of the bacterial cells, as also evidenced by increased carbohydrate and protein contents as well as the cytoplasmic enzymes in the cell-free supernatants. N-terminal sequence analyses of the released proteins revealed them to be cytoplasmic/membrane proteins. Upon GLC, the supernatant showed characteristic fatty acid profiles in E. coli, thus subscribing to detachment of lipopolysaccharides into the medium, whereas that of B. cereus indicated release of surface lipids. The mechanism for the observed bactericidal activity of LMWC towards both Gram-positive and Gram-negative bacteria has been discussed.     

Assessment of olive-mill wastewater as a growth medium for lipase production by Candida cylindracea in bench-top reactor

Olive-mill wastewater (OMW) was investigated for its suitability to serve as a medium for lipase production by Candida cylindracea NRRL Y-17506. The OMW that best supported enzyme production was characterized by low COD and low total sugars content. In shake flask batch cultures, OMW supplementation with 2.4 g l⁻¹ NH₄Cl and 3 g l⁻¹ olive oil led to an enzyme activity of about 10 U ml⁻¹. The addition of glucose or malt extract and supplements containing organic N (e.g., peptone, yeast extract) either depressed or did not affect the enzyme production. Further experiments were then performed in a 3-l stirred tank reactor to assess the impact of medium pH and stirring speed on the yeast enzyme activity. The lipase activity was low (1.8 U ml⁻¹) when the pH was held constant at 6.5, significantly increased (18.7 U ml⁻¹) with uncontrolled pH and was maximum (20.4 U ml⁻¹) when the pH was let free to vary below 6.5. A stirring regime, that varied depending on the dissolved oxygen concentration in the medium, both prevented the occurrence of anoxic conditions during the exponential growth phase and enabled good lipase production (i.e., 21.6 U ml⁻¹) and mean volumetric productivity (i.e., 123.5 U l⁻¹ h⁻¹).     

Styrene removal from polluted air in one and two-liquid phase biotrickling filter: steady and transient-state performance and pressure drop control

A Sporothrix variecibatus-inoculated biotrickling filter (BTF) was examined for styrene removal, without and with the addition of silicone oil, at different empty bed residence times. The highest elimination capacities (ECs) were 172.8 (without silicone oil) and 670 g m⁻³ h⁻¹ (with silicone oil), respectively, corresponding to a 4-fold improvement in presence of oil. The addition of silicone oil formed a well-coalesced emulsion of fungi and silicone oil, resulting in filter-bed clogging. Clogging prevention strategies adopted were; (i) lowering the volume ratio of silicone oil from 10% to 2% (v/v), and (ii) periodic increase in trickling rate of the medium from 50 to 190 mL min⁻¹. During shock-load experiments, the BTF with silicone oil (2% v/v) could withstand high styrene loads, of up to 1900 g m⁻³ h⁻¹, when compared to the BTF without silicone oil (400 g m⁻³ h⁻¹).     

Toxicity of lead and zinc to developing mussel and sea urchin embryos: Critical tissue residues and effects of dissolved organic matter and salinity

Lead (Pb) EC50 values in the very sensitive early development phases (48–72 h post-fertilization) of the mussels Mytilus galloprovincialis and Mytilus trossolus and sea urchin Strongylocentrotus purpuratus in 100% sea water were: M. trossolus — 45 (95% C.I. = 22–72) μg L^− 1; M. galloprovincialis — 63 (36–94) μg L^− 1; S. purpuratus — 74 (50–101) μg L^− 1. Salinity thresholds for normal development varied: M. trossolus > 21 ppt; M. galloprovincialis > 28 ppt; S. purpuratus ≥ 30 ppt. Addition of two spectroscopically distinct dissolved organic matters (DOM) from fresh water (Nordic Reservoir) and sea water (Inshore) moderately decreased the toxicity of Pb to both mussels, but not in a concentration-dependent fashion, with only an approximate doubling of EC50 over the range of 1.4–11.2 mg C L^− 1. Independent Pb binding capacity determinations for DOC explained the lack of a relationship between DOM concentration and toxicity. Salinity had no effect on Pb toxicity down to 21 ppt in M. trossolus, and low salinity (21 ppt) did not enhance the protective effect of DOC. Both DOMs increased the toxicity of Pb in developing sea urchin embryos, in contrast to mussels. Relative to Pb, the organisms were 6–9 fold less sensitive to Zn on a molar basis in 100% seawater with the following Zn EC50s: M. trossolus — 135 (103–170) μg L^− 1; M. galloprovincialis — 172 (126–227) μg L^− 1, S. purpuratus — 151 (129–177) μg L^− 1. Nordic Reservoir and Inshore DOM (2–12 mg C L^− 1) had no significant effect on Zn toxicity to mussels, in accord with voltammetry data showing an absence of any strong ligand binding for Zn by DOMs. As with Pb, DOMs increased Zn toxicity to urchin larvae. Critical Tissue Residues (CTR) based on whole body concentrations of Pb and Zn were determined for M. galloprovincialis at 48 h and S. purpuratus at 72 h. The median lethal CTR values (LA50s), useful parameters for development of saltwater Biotic Ligand Models (BLMs), were approximately 4-fold higher on a molar basis for Zn than for Pb. The latter were not altered by DOM exposure, despite increased EC50 values, in accord with the tenets of the BLM.     

Facile and sensitive detection of protamine by enhanced room-temperature phosphorescence of Mn-doped ZnS quantum dots

Quantum dot (QD) nanohybrids provide an effective route to explore the new properties of materials and are increasingly used as highly valuable sensitive (bio) chemical probes. Interestingly, the room-temperature phosphorescence (RTP) of 3-mercaptopropionic acid (MPA)-capped Mn-doped ZnS QDs could be remarkably enhanced by the addition of protamine. Based on the above finding, a simple, sensitive, and selective method for rapid detection of protamine was successfully designed. With this method, protamine as a cationic peptide interacts electrostatically with MPA-capped Mn-doped ZnS QDs to form MPA-capped Mn-doped ZnS QD/protamine complexes, which leads to the aggregation of QDs and enhances the RTP intensity. Under the optimized conditions, the RTP intensity change was linearly proportional to the concentration of protamine in the range 0.2–3.0 μg ml⁻¹, and the limit of detection was 0.14 μg ml⁻¹. The proposed method was successfully applied to detect protamine in protamine sulfate injection and human serum samples with satisfactory results, and the recovery ranged from 96.5 to 105.6%.     

Nutritional value and intake of aquatic ferns (Azolla filiculoides Lam. and Salvinia molesta Mitchell.) in sows

Aquatic ferns (AFs) such as Azolla filiculoides and Salvinia molesta are grown on swine lagoons in the tropics and used in diets for pigs. The present work is aimed at evaluating their potential as feed ingredients for sows. When presented with ad libitum AFs, gilts weighing 110 ± 14 kg (mean ± SD), were able to ingest 9.1–9.7 kg fresh AF per day (from 597 to 630 g dry matter (DM) per day) and from 1240 to 1428 g DM per day when presented in a dry, ground form. A digestibility study was conducted, using sows weighing 213 ± 9 kg (mean ± SD), which were fed diets containing maize, soybean meal and 0, 150 or 300 g AF kg⁻¹ diet. The presence of AFs had a negative impact on the faecal digestibility of the crude protein, NDF and energy content of the whole diet (P<0.001) and on the ileal protein digestibility, especially with 300 g AFs kg⁻¹ diet. The level of AFs in the diet had no effect on stomach weight (P>0.05) but increased the weight of the rest of the gastrointestinal tract (P<0.001). The rate of AF fibre fermentation in the pig large intestine was measured using an in vitro gas test. The rates were much lower than tropical tree foliage, which can also be used in pig diets in the tropics. This could partly explain the low apparent digestibility of AFs in pigs. In conclusion, the inclusion level of AFs in rations for sows should be limited to 150 g AFs kg⁻¹ diet due to the low digestibility and energy density, as well as the negative impact on the digestibility of the whole diet.     

Techno-economic implications of improved high gravity corn mash fermentation

The performance of Saccharomyces cerevisiae MBG3964, a strain able to tolerate >18% v/v ethanol, was compared to leading industrial ethanol strain, Fermentis Ethanol Red, under high gravity corn mash fermentation conditions. Compared to the industrial ethanol strain, MBG3964 gave increased alcohol yield (140 g L⁻¹ vs. 126 g L⁻¹), lower residual sugar (4 g L⁻¹ vs. 32 g L⁻¹), and lower glycerol (11 g L⁻¹ vs. 12 g L⁻¹). After 72 h fermentation, MBG3964 showed about 40% viability, whereas the control yeast was only about 3% viable. Based on modelling, the higher ethanol tolerant yeast could increase the profitability of a corn-ethanol plant and help it remain viable through higher production, lower unit heating requirements and extra throughput. A typical 50 M gal y⁻¹ dry mill ethanol plant that sells dried distiller’s grain could potentially increase its profit by nearly $US3.4 M y⁻¹ due solely to the extra yield, and potentially another $US4.1 M y⁻¹ if extra throughput is possible.