Chitosan/Polyethylene Glycol Beads Crosslinked with Tripolyphosphate and Glutaraldehyde for Gastrointestinal Drug Delivery

This study reports on the preparation of chitosan (CS)/polyethylene glycol (PEG) hydrogel beads using sodium diclofenac (DFNa) as a model drug. Following the optimization of the polymer to drug ratio, the chitosan beads were modified by ionic crosslinking with sodium tripolyphosphate (TPP). The CS/PEG/DFNa beads obtained from a (w/w/w) ratio of 1/0.5/0.5 with crosslinking in 10% (w/v) TPP at pH 6.0 for 30 min yielded excellent DFNa encapsulation levels with over 90% loading efficiency. The dissolution profile of DFNa from CS/PEG/DFNa beads demonstrated that this formulation was able to maintain a prolonged drug release for approximately 8 h. Among the formulations tested, the CS/PEG/DFNa (1/0.5/1 (w/w/w)) beads crosslinked with a combination of TPP (10% (w/v) for 30 min) and glutaraldehyde (GD) (5% (w/v)) were able to provide minimal DFNa release in the gastric and duodenal simulated fluids (pH 1.2 and 6.8, respectively) allowing for a principally gradual drug release over 24 h in the intestinal (jejunum and ileum) simulated fluid (pH 7.4). Thus, overall the CS/PEG beads crosslinked with TPP and GD look to be a promising and novel alternative gastrointestinal drug release system.     

Exploring metal effects and synergistic interactions of ferric stimulation on azo-dye decolorization by new indigenous Acinetobacter guillouiae Ax-9 and Rahnella aquatilis DX2b

The first-attempt study deciphered metal-interacting effects on dye-decolorizing capabilities of indigenous bioelectricity-generating strains, Acinetobacter guillouiae Ax-9 and Rahnella aquatilis DX2b. Most of the metallic ions were inhibitory to color removal capabilities of these strains. However, with supplementation of 5 mM ferric chloride, specific decolorization rate (SDR) of Ax-9 increased by 55.48 % compared to Fe³⁺-free conditions. In contrast, SDR of DX2b decreased 75.35 % due to the inhibition of ferric chloride. On the other hand, ferric citrate could stimulate SDR of DX2b for 21.5 % at same dosage. Enzymatic assay indicated that Fe reductase activity was consistent with synergistic effects of ferric chloride on Ax-9, and ferric citrate on DX2b. Protein analysis via SDS-PAGE and identification of Tandem MS/MS afterwards showed that outer membrane protein (Omp) primarily deals with decolorization as a channeling regulation. Moreover, molecular modeling and bioinformatics data also provided detailed evidences to confirm the biological significance of Omp.     

Calcium hypochlorite for removing stalks on eggs of the green lacewing Chrysoperla genanigra (Neuroptera: Chrysopidae)

The release of lacewings eggs in the field requires the removal of their silk stalk, otherwise the release is impaired, as the eggs can become entangled in these pedicels. This study evaluated the effect of three concentrations of chlorine solution associated with two different exposure times for removing the stalks of eggs of Chrysoperla genanigra Freitas, and on the hatchability of the treated eggs. The concentrations used were 0.0 (control), 2.0, 4.0, and 6.0 % chlorine granules of hth^®—containing as major components calcium hypochlorite and sodium chloride—dissolved in distilled water. The times used for each concentration were 60 and 120 s. Fifty eggs were used for each treatment. After each treatment, the eggs were washed in running water and then dried with a directed air flow. The best results were obtained with the concentrations of 2 % for 120 s and 4 % for 60 and 120 s, removing 100 % of stalks without reducing hatchability. At 6 %, egg destruction started to occur. We conclude that calcium hypochlorite can be used for removing stalks of C. genanigra eggs with no reduction in hatchability.     

Characterization of Organogel Prepared from Rice Bran Oil with Cinnamic Acid

Cinnamic acid (CA) was added to rice bran oil (RBO) at concentrations range from 2 to 12% (w/w) to prepare functional fat-like organogel. The oil binding capacity and gel formation time of the obtained organogels were determined. The results showed that the oil binding capacity was increased and the gel formation time was significantly (p < 0.05) decreased with the increase in concentration of CA up to 10% (w/w). Based on these results, organogels prepared from RBO with 6, 8, and 10% CA (w/w) were chosen and subjected to further characterization analyses. The microscopy analysis showed different gel network structures and crystalline behavior for the organogels prepared with varying concentrations of CA. The rheological and thermal properties of the organogel were improved with the increase in CA concentration up to 10% (w/w). The X-ray diffractometer and Fourier Transform Infrared spectroscopy analyses indicated that the gel network formed based on crystalline of CA and physical entanglements among the organogel components. In addition, the organogel of RBO with 10% CA (w/w) showed a uniform and homogenized structure during storage at 5 °C for 28 days compared to the organogels prepared with 6 and 8%. Therefore, it can be suggested that cinnamic acid is a good organogelator for preparing functional fat-like organogel from rice bran oil.     

Effects of Partial Replacement of Gelatin in High Sugar Gels with Gellan on their Textural,Rhelogical, and Thermal Properties

Effects of partial replacement of gelatin in simulated gummy confections with either high acyl or deacylated gellan on their textural, rheological, and thermal properties were investigated. Atomic force microscopy (AFM) images of high acyl and deacylated gellan revealed that both gellan types formed finely stranded networks as a result from air-drying of dilute aqueous solutions, the strand thickness of which was approximately 0.5–1 nm. Simulated gummy confections containing 5.025–7.1 % w/w gelatin, 0–0.075 % w/w high acyl or deacylated gellan, and 73–75 % w/w corn syrup and sucrose combined were prepared and analyzed using texture profile analysis (TPA) and small amplitude oscillatory shear measurements. The principal component analysis (PCA) of textural attributes obtained from TPA identified a cluster in the first quadrant formed by samples containing 7.1 % w/w gelatin but no gellan and those containing 6.025 % w/w gelatin and 0.075 % w/w high acyl or deacylated gellan. All simulated gummy confections showed storage modulus (G′) values greater than loss modulus (G″) values at 0.1 rad/s, G″ increasing more steeply with increasing angular frequency, and G′-G″ crossovers within the examined angular frequency range (0.1–100 rad/s), typical of high solid biopolymer gels. Furthermore, increasing gellan concentration at a total concentration of the gelling agents (i.e., gelatin and gellan) of 6.1 % w/w increased the melting temperature. These results attest the feasibility of improving the heat stability of gummy confections by the partial replacement of gelatin with either high acyl or deacylated gellan with maintaining textures characteristics of gummy confections containing gelatin as the only gelling agent.     

Evaluation of the in vitro methods for micropropagation of Chondracanthus acicularis (Roth) Fredericq (Gigartinales, Rhodophyta): tissue culture and production of protoplasts

Tissue was cultured and protoplasts isolated from the carrageenophyte Chondracanthus acicularis with the aim of developing micropropagation as an alternative to harvesting raw material from natural beds. Both adventitious shoots and filamentous calluses were induced by tissue culture on medium solidified with 0.4–1 % (w/v) agar. Adventitious shoots were mainly produced from discoid bases while filamentous calluses were mainly induced from basal zones and sub-apical explants. A gradient of the regeneration ability was observed from the top to the bottom of the thallus. The discoid base was the most reactive explant and produced the highest number of adventitious shoots compared to basal zones and sub-apical explants, irrespective of the concentration of agar. Protoplasts were isolated enzymatically from the whole thallus using a combination of cellulase R-10 Onozuka, macerozyme R-10, and crude extract of the gland gut of algivorous molluscs. The highest mean yield of protoplasts (1.2?×?10⁶ protoplasts g^?1 fresh weight) was obtained after 16 h of digestion with an enzyme mixture containing 2 % (w/v) cellulase R-10, 1 % (w/v) macerozyme R-10 Onozuka, 4 % (v/v) crude extract of gut gland of Haliotis, 0.8 M mannitol, 50 mM sodium citrate, 0.3 % (w/v) bovine serum albumin. Depending on the conditions, mean protoplast yields ranged from 3.14?×?10⁵ to 1.2?×?10⁶ protoplasts g^?1 fresh weight. Different factors (storage duration, mannitol, sodium citrate, crude extract of the gland gut of algivorous molluscs) were tested to improve the yield of protoplasts but none has a significantly effect.     

Improvement in the docosahexaenoic acid production of <Emphasis Type="Italic">Schizochytrium</Emphasis> sp. S056 by replacement of sea salt

Schizochytrium is a marine microalga that requires high concentrations of sea salt for growth, although problems arise with significant amounts of chloride ions in the culture medium, which corrodes the fermenters. In this work, we evaluated that cell growth and docosahexaenoic acid (DHA) production can be improved when using 1 % (w/v) sodium sulfate instead of 2 % (w/v) sea salt in the culture medium for Schizochytrium sp. S056. In practice, the use of sodium sulfate as the sodium salt led to chloride ion levels in the medium that can be completely removed, thus avoiding fermenter corrosion during Schizochytrium sp. S056 growth, reducing cost and increasing DHA production, and simplifying the disposal of fermentation wastewater. Additionally, we demonstrated that the osmolality of growth media did not play a crucial role in the production of DHA. These findings may be significantly important to companies involved in production of PUFAs by marine microbes.     

Simultaneous wastewater treatment,electricity generation and biomass production by an immobilized photosynthetic algal microbial fuel cell

A photosynthetic algal microbial fuel cell (PAMFC) was constructed by the introduction of immobilized microalgae (Chlorella vulgaris) into the cathode chamber of microbial fuel cells to fulfill electricity generation, biomass production and wastewater treatment. The immobilization conditions, including the concentration of immobilized matrix, initial inoculation concentration and cross-linking time, were investigated both for the growth of C. vulgaris and power generation. It performed the best at 5 % sodium alginate and 2 % calcium chloride as immobilization matrix, initial inoculation concentration of 10⁶ cell/mL and cross-linking time of 4 h. Our findings indicated that C. vulgaris immobilization was an effective and promising approach to improve the performance of PAMFC, and after optimization the power density and Coulombic efficiency improved by 258 and 88.4 %, respectively. Important parameters such as temperature and light intensity were optimized on the performance. PAMFC could achieve a COD removal efficiency of 92.1 %, and simultaneously the maximum power density reached 2,572.8 mW/m³ and the Coulombic efficiency was 14.1 %, under the light intensity of 5,000 lux and temperature at 25 °C.     

Development of Transparent Curcumin Loaded Microemulsions by Phase Inversion Temperature (PIT) Method: Effect of Lipid Type and Physical State on Curcumin Stability

In this study, curcumin loaded transparent microemulsions obtained using the phase inversion temperature (PIT) method were developed. Different lipids (sunflower, peanut, castor, and extra virgin olive oil, tristearin, and tripalmitin) were tested as curcumin carrier in microemulsions. The obtained systems were analyzed for transparency, particle size, lipid crystal polymorphism, and curcumin stability at 20 °C up to 120 days. It was found that the maximum lipid content allowing transparent microemulsions (mean particle diameter of around 25 nm) to be obtained was greatly affected by the lipid characteristics. By using oils rich in long chain fatty acids, such as sunflower, peanut, and extra virgin olive oil, transparent microemulsions can be obtained with oil fractions up to 7.5 % (w/w). On the contrary, when fat containing crystals (e.g. tripalmitin or tristearin) was used, the maximum lipid loading capacity was reduced to 5 % (w/w). Castor oil, rich in polar groups, did not permit the formation of transparent microemulsions at any tested concentration (from 1 to 9 % w/w). The oil type also affected curcumin stability: curcumin degradation rate was lower in tristearin containing microemulsions than in those containing extra virgin olive oil. This result was attributed to the protective effect of solid lipid particles into lipid droplets.     

Enhancing lipid production of the marine diatom Chaetoceros gracilis: synergistic interactions of sodium chloride and silicon

Silicon deficiency is a lipid-promoting stress for many oleaginous diatoms. Literature reports suggest that reduced salinity in seawater, a primary component of which is sodium chloride, may inhibit metabolism of silicon in marine diatoms. We hypothesized that lowering sodium chloride below ocean levels may thus be effective in creating silicon stress and enhancing lipid productivity. We examined the interacting effects of silicon supply (0.05, 0.1, 0.2, and 0.8 mM) and sodium chloride concentration (50, 100, and 400 mM) on growth and lipid production in Chaetoceros gracilis. This was done in batch culture to facilitate the application of severe stress. Low levels of either sodium chloride or silicon resulted in at least 50 % increases in lipid content. The synergy of simultaneous, moderate sodium chloride and silicon stress resulted in lipid content up to 73 % of dry mass and lipid productivity of 1.7 g m^?2 day^?1; with a daily integrated photosynthetic photon flux of 17.3 mol photons m^?2 day^?1, the efficiency of lipid synthesis was thus 0.1 g mol^?1 of photons. Decreased silicon also resulted in a 5 % shift in lipid chain length from C18 to C16 fatty acids. We observed a strong sodium chloride/silicon interaction on total and ash-free dry mass densities that arose because low sodium chloride concentrations were inhibitory to growth, but the inhibition was overcome with excessive silicon supply. This observation suggests that low levels of sodium chloride may have affected metabolism of silicon. The findings of this study can be used to enhance lipid production in oleaginous marine diatoms.     

The ectomycorrhizae of <Emphasis Type="Italic">Lactarius rimosellus</Emphasis> and <Emphasis Type="Italic">Lactarius acatlanensis</Emphasis> with the endangered <Emphasis Type="Italic">Fagus grandifolia</Emphasis> var. <Emphasis Type="Italic">mexicana</Emphasis>

Gut bacterium Pantoea sp. is one of the predominant bacterial species in the larval gut of the diamondback moth, Plutella xylostella. The phenotypic characters of Pantoea sp. were investigated with BIOLOG phenotype MicroArray (PM) in this study. Totally 950 different metabolic phenotypes were tested using the PM plates 1–10. Results exhibited that Pantoea sp. was able to metabolize 37.37 % of the tested carbon sources, 91.32 % of nitrogen sources, 100 % of sulfur sources, and 98.31 % of phosphorus sources. Most informative utilization patterns for carbon sources of Pantoea sp. were organic acids and carbohydrates, and for nitrogen were various amino acids. The bacterium had 94 different biosynthetic pathways. It had a wide range of adaptabilities, and could still metabolize in osmolytes with up to 9 % sodium chloride, 6 % potassium chloride, 5 % sodium sulfate, 20 % ethylene glycol, 4 % sodium formate, 4 % urea, 5 % sodium lactate, 200 mmol/L sodium phosphate (pH 7.0), 100 mmol/L ammonium sulfate (pH 8.0), 100 mmol/L sodium nitrate, and 100 mmol/L sodium nitrite, respectively. It also exhibited active metabolism under pH values between 4.5 and 10. Pantoea sp. showed active decarboxylase activities while poor deaminase activities in the presence of various amino acids. The phenotypic characterization of Pantoea sp. increased our knowledge of the bacterium, in particular its interactions with insect hosts and the adaptability in gut environments, and showed us some possible approaches to controlling diamondback moth through decreasing Pantoea sp. density.     

Enhancing isomaltulose production by recombinant Escherichia coli producing sucrose isomerase: culture medium optimization containing agricultural wastes and cell immobilization

Isomaltulose is a structural isomer of sucrose commercially used in food industries. In this work, recombinant Escherichia coli producing sucrose isomerase (SIase) was used to convert sucrose into isomaltulose. To develop an economical industrial medium, untreated cane molasses (10.63 g l^?1), yeast extract (25.93 g l^?1), and corn steep liquor (10.45 g l^?1) were used as main culture compositions for SIase production. The relatively high SIase activity (14.50 ± 0.11 U mg DCW^?1) was obtained by the recombinant cells. To the best of our knowledge, this is the first investigation on SIase production by engineered E. coli using untreated cane molasses. The recombinant E. coli cells expressing the SIase gene were immobilized in calcium alginate gel in order to improve the efficiency of recycling. The immobilization was most effective with 2 % (w/v) sodium alginate and 3 % (w/v) calcium chloride. The optimal initial biomass for immobilization was 20 % (w/v, wet wt.), with a hardening time of 8 h for cell immobilization. The immobilized E. coli cells exhibited good stability for 30 batches with the productivity of 0.45 g isomaltulose g pellet^?1 h^?1. A continuous isomaltulose formation process using a column reactor remained stable for 40 days with 83 ± 2 % isomaltulose yield, which would be beneficial for economical production of isomaltulose.     

Carotenoid and lipid production by the autotrophic microalga Chlorella protothecoides under nutritional, salinity, and luminosity stress conditions

Today microalgae represent a viable alternative source for high-value products. The specie Chlorella protothecoides (Cp), heterotrophically grown, has been widely studied and provides a high amount of lutein and fatty acids (FA) and has a good profile for biodiesel production. This work studies carotenoid and FA production by autotrophic grown Cp. Cp was grown until the medium’s nitrogen was depleted, then diluted in NaCl solution, resulting in nutritional, luminosity, and salinity stresses. Different NaCl concentrations were tested (10, 20, 30 g/L) at two different dilutions. After dilution, a color shifting from green to orange-red was noticed, showing carotenoid production. The best production of both carotenoids and FA was attained with a 20 g/L NaCl solution. The total carotenoid content was 0.8 % w/w (canthaxanthin (23.3 %), echinenone (14.7 %), free astaxanthin (7.1 %), and lutein/zeaxanthin (4.1 %)). Furthermore, the total lipid content reached 43.4 % w/w, with a FA composition of C18:1 (33.64 %), C16:0 (23.30 %), C18:2 (11.53 %), and less than 12 % of C18:3, which is needed to fulfill the biodiesel quality specifications (EN 14214).     

Synthesis of poly(ε-caprolactone) by an immobilized lipase coated with ionic liquids in a solvent-free condition

Polycaprolactone (PCL) was synthesized by ring-opening polymerization of ε-caprolactone through two different enzymatic processes. The lipase from Candida antarctica B, immobilized on macroporous acrylic acid beads, was employed either untreated or coated with small amounts of ionic liquids (ILs). Monocationic ionic liquids, [C _n MIm][NTf₂] (n = 2, 6, 12), as well as a dicationic ionic liquid, ([C₄(C₆Im)₂][NTf₂]₂), were used to coat the immobilized lipase and also as the reaction medium. In both methods, the polarity, anion of the ILs concentration and viscosity strongly influenced the reaction. Coating the immobilized enzyme with ILs improved catalytic activity and less ILs was required to produce PCL with a higher molecular weight and reaction yield. At 60 °C and ILs/Novozyme-435 coating ratio of 3:1 (w/w) for 48 h, the highest M _w and reaction yield of PCL were 35,600 g/mol and 62 % in the case of [C₁₂MIm][NTf₂], while the M _w and reaction yield of PCL was 20,300 g/mol and 54 % with [C₁₂MIm][NTf₂] and catalyzed by untreated lipase.     

Cold-Set Gelation of Commercial Soy Protein Isolate: Effects of the Incorporation of Locust Bean Gum and Solid Lipid Microparticles on the Properties of Gels

This study aimed to evaluate the ability of commercial soy protein isolate (SPI) to form cold-set gels under different pHs (5–11), pre-heating temperatures (60 °C, 80 °C), CaCl₂ (0–15 mM) and SPI (5–15%, w/v) concentrations, and also select a formulation for the investigation of the effects of incorporating locust bean gum (LBG) (0–0.3%, w/v) and solid lipid microparticles (SLM) on gels rheological and microstructural properties. Gels were evaluated in terms of visual aspect, water-holding capacity, microstructure (using confocal laser scanning microscopy and cryo-scanning electronic microscopy) and rheological properties. SPI showed higher solubilities at pHs 7 (32.0%), 9 (51.6%) and 11 (100%). Self-supported gels were obtained under several conditions at alkaline pHs. At pH 7, only systems pre-heated to 80 °C with 15% (w/v) SPI and 10 or 15 mM CaCl₂ gave self-supported gels. At neutral pH, samples showed relative structural instability, which was minimized with LBG incorporation. Formulations G_SPI (pH 7, preheated to 80 °C, 15% (w/v) SPI, 10 mM CaCl₂) and G_MIX (pH 7, preheated to 80 °C, 15% (w/v) SPI, 0.2% (w/v) LBG, 15 mM CaCl₂) were selected for emulsion-filled gels (EFG) production. Power law parameters (K′, K″), calculated from frequency sweep results, revealed that non-filled G_MIX (K′: 472.1; K″: 77.6) was stronger than G_SPI (K′: 170.4; K″: 33.6). Besides, G_MIX showed microphase separation. SLM stabilized with Tween 80-Span 80 were active fillers in EFG, altering microstructures and increasing G’, G” and the Young’s modulus (1.8 to 2.1 kPa for G_SPI and 1.4 to 2.2 kPa for G_MIX).     

Tea stalks – a novel agro-residue for the production of tannase under solid state fermentation by Aspergillus niger JMU-TS528

A novel agro-residue, tea stalks, was tested for the production of tannase under solid-state fermentation (SSF) using Aspergillus niger JMU-TS528. Maximum yield of tannase was obtained when SSF was carried out at 28 °C, pH 6.0, liquid-to-solid ratio (v/w) 1.8, inoculum size 2 ml (1?×?10⁸ spores/ml), 5 % (w/v) ammonium chloride as nitrogen source and 5 % (w/v) lactose as additional carbon source. Under optimum conditions, tannase production reached 62 U/g dry substrate after 96 h of fermentation. Results from the study are promising for the economic utilization and value addition of tea stalks.     

A new method for fluorometric measurement of proline in grapes and wines

A method for the measurement of free proline in grapes and wines containing other amino acids is presented. The method included the removal of phenols and tryptophan by the pre-treatment of samples with activated carbon, the oxidation of proline to the corresponding primary amine with sodium hypochlorite, destruction of other amino acids by oxidation at pH 5.3 using a combination of hydrogen peroxide, ferric chloride, and sodium hypochlorite, and the formation of afluorescent product from the oxidized proline with o-phthalaldehyde in the presence of 2-mercaptoethanol and Brij 35 at pH 5.3. The proline in grapes and wines measured by this method agreed well with the values obtained by the method using an automatic amino acid analyzer with ninhydrin.     

Environmental Effects of Steam Explosion Pretreatment on Biogas from Maize—Case Study of a 500-kW Austrian Biogas Facility

Potential environmental impacts of biogas electricity from agricultural residues (maize stover) with steam explosion (SE) pretreatment were compared to a typical Austrian biogas system (maize silage) using the method of life cycle assessment. Besides the biogas plant, the system includes substrate production, a combined heat-and-power (CHP) unit, digestate management, and transportation. The stover scenario (including construction and operation of the SE unit) results in lower total climate change impacts than those of the typical biogas system (239 g CO₂-eq/kWh electricity vs. 287 g CO₂-eq/kWh electricity; 100-year global warming potential (GWP)), and this holds also for the other impact categories (e.g., cumulative energy demand, acidification, eutrophication). While uncertainties in other areas could change the results, based on the uncertainty information considered, the overall results for the two scenarios were significantly different. Methane slip emissions from the CHP exhaust account for the largest GWP share in both scenarios. Other large GWP contributions are from substrate production and grid electricity for plant operations. The findings were robust against worst-case assumptions about the energy requirements of the SE pretreatment.     

Production of fructose from highly concentrated date extracts using Saccharomyces cerevisiae

Large amounts of low-quality dates produced worldwide are wasted. Here, highly concentrated fructose syrups were produced via selective fermentation of date extracts with Saccharomyces cerevisiae. Syrups with 95.4–99.9 % (w/w) fructose yields were obtained from date extracts having an initial sugar range of 49–374 g/l without media supplementation; the corresponding ethanol yields were between 69 and 52 % (w/w). At 470 g initial sugars/l, fructose and ethanol yields were 84 and 47 % (w/w), respectively, and the product contained 62 % (w/w) fructose, which is higher than the widely available commercial 42 and 55 % (w/w) high fructose corn syrups. The commercial potential for conversion of waste dates to high-value products is thus demonstrated.