Microchip for sustained drug delivery by diffusion through microchannels

To enable sustained drug delivery, we prepared microchips of simple structure for drug release based on diffusion through microchannels. The microchips were fabricated with poly(methyl methacrylate), embedded with one or more microwells and microchannels of controlled length. The channels were filled with biocompatible polymer, poly(ethylene glycol), to serve as a drug diffusion barrier. The wells served as drug reservoirs and were filled with a fine powder of a model compound, fluorescein. Three different drug delivery microchip designs were prepared, each equipped with a channel of 1, 4, or 8 mm length. Drug release from these devices all exhibited a delay followed by sustained release over time. As the channel length increased from 1 to 8 mm, the onset time and duration of drug release also increased from 0.5 to 7 day and from 11 days to 28, respectively. We also prepared microchips equipped with multiple microwells, each connected to a channel of different length. In this way, a chip with channels of 1, 4, and 8 mm length exhibited a continuous drug release from 0.5 to 35 days. A future study is in progress to develop the microchips made of biodegradable materials. Therefore, we conclude that a microchip embedded with multiple sets of microwells and microchannels of different length can be designed to enable sustained drug release for controlled and prolonged periods of time.     

Powdered edible locust enhances nitrogen excretion and presents a low-energy value in growing rats

Edible insects are increasingly recommended as novel sustainable protein sources, but the nutritional properties of edible insects have not been well studied. We investigated whether locust powder can be used as a nutritionally functional food resource with a low energy value using a rat model. Twenty-five male Wistar rats (4-week-old) were fed a basal diet (12 g daily) to which a fixed amount of locust powder (0, 0.5, 1.0, or 2.0 g) was added for 20 days (L0, L0.5, L1.0, and L2.0 groups, respectively). In the sucrose standard group, rats received 12 g of the basal diet daily and 2.0 g of sucrose daily for 20 days (S2.0 group). Body-weight gain and the nutritional composition of the carcasses and feces were determined to estimate the available energy value of locust powder. The L0.5 group had the lowest carcass fat content and energy accumulation, but these values were increased by locust powder in a dose-dependent manner. The net energy value of locust powder was estimated to be 2.78 kcal/g, which was expected to be lower than the calculated theoretical value (4.25 kcal/g) and that of sucrose (3.94 kcal/g). Fecal nitrogen excretion was increased by dietary locust powder in a dose-dependent manner (correlation coefficient, R = 0.98), and the carcass nitrogen percentage was not changed, regardless of the dietary content of the locust powder, indicating an increased excretion of proteins or other non-protein nitrogen compounds derived from the locust powder. These findings suggest that locust powder can be used as a novel food material with a low energy value for humans.     

Ammonia emissions and biodegradation of organic carbon during sewage sludge composting with different extra carbon sources

Ammonia emissions during composting result in the reduced value of agronomic production and can also pollute the air. To evaluate the influence of various carbon sources on ammonia emissions, six composting experiments were carried out with different amendments of carbon sources (glucose, sucrose and straw powder). The cumulative ammonia volatilizations were reduced from 3.11 g/kg (R6) to 2.46 g/kg (R1), 2.17 g/kg (R2), 2.23 g/kg (R4) and 1.93 g/kg (R5). Compared to the control, no significant difference of ammonia emissions and carbon degradation was observed for the mixture of R3 (3.15 g/kg), which was amended with straw powder alone. The co-addition of sucrose and straw powder led to the lowest ammonia emissions. According to these results, a higher C/N ratio did not necessarily indicate an effective solution for reducing ammonia emissions, and not all readily available carbon compounds were helpful in reducing ammonia emissions. The addition of sucrose promoted the decomposition of organic carbon during the intensive stage of ammonia emissions, and the combination of straw and sucrose prolonged this promotion. Thus, the co-addition of sucrose and straw powder made it possible to reduce ammonia emissions drastically by nitrogen immobilization.     

5-Fluorouracil-Loaded BSA Nanoparticles: Formulation Optimization and In Vitro Release Study

Over the past few decades, there has been considerable interest in developing protein nanoparticles as drug delivery devices. The underlying rationale is their exceptional characteristics, namely biodegradability and nonantigenicity. Herein, phase separation method was used to prepare 5-fluorouracil-loaded bovine serum albumin (BSA) nanoparticles. Drug release was tracked by continuous flow dialysis technique. Effect of process variables on loading efficiency of 5-fluorouracil was investigated and optimized through Taguchi’s M16 design with the amount of entrapped drug as response. Optimum condition was found to be 2 mg/mL of 5-fluorouracil, 3.7 mL of added ethanol, 176 μL of glutaraldehyde, drug–protein incubation time of 30 min, and pH of 8.4 for 200 mg of BSA in 2 mL drug solution. pH had the most noticeable effect on the amount of entrapped drug, but glutaraldehyde had the least. Mean diameter and zeta potential of fabricated nanoparticles under these conditions were 210 nm and −31.7 mV, respectively. Drug-loaded BSA nanoparticles suspension maintained constant release of drug for 20 h under experimental conditions, so this colloidal drug carrier is capable of releasing drug in a sustained manner.     

Drug-Cyclodextrin-Vesicles Dual Carrier Approach for Skin Targeting of Anti-acne Agent

In the present study attempt was made for preparation of isotretinoin-hydroxypropyl β cyclodextrin (HP-β-CD) inclusion complex and encapsulate this complex in elastic liposomes to study the effect of dual carrier approach on skin targeting of isotretinoin. The isotretinoin HP-β-CD complex was prepared by freeze-drying method and characterized by IR spectroscopy. The drug and drug-CD complex loaded elastic liposomal formulation were prepared and characterized in vitro, ex-vivo and in vivo for shape, size, entrapment efficiency, no. of vesicles per cubic mm, in vitro skin permeation and deposition study, photodegradation and skin toxicity assay. The transdermal flux for different vesicular formulations was observed between 10.5 ± 0.5 to 13.9 ± 1.6 μg/cm²/h. This is about 15-21 folds higher than that obtained from drug solution (0.7 ± 0.1 μg/cm²/h) and 4-5 folds higher than obtained with drug-CD complex solution (2.7 ± 0.1 μg/cm²/h). The amount of drug deposit was found to increase significantly (p < 0.05) by cyclodextrin complexation (30.1 ± 0.1 μg). The encapsulation of this complex in elastic liposomal formulation further increases its skin deposition (262.2 ± 21 μg). The results of skin irritation study using Draize test also showed the significant reduction in skin irritation potential of isotretinoin elastic liposomal formulation in comparison to free drug. The results of the present study demonstrated that isotretinoin elastic liposomal formulation possesses great potential for skin targeting, prolonging drug release, reduction of photodegradation, reducing skin irritation and improving topical delivery of isotretinoin.     

Control of Transdermal Permeation of Hydrocortisone Acetate from Hydrophilic and Lipophilic Formulations

The purpose of this research was the preparation of four formulations containing hydrocortisone acetate (HCA) for topical application, including two aqueous systems (hydrophilic microemulsion and aqueous gel) and two systems with dominant hydrophobicity (hydrophobic microemulsion and ointment). The formulations were tested for the release and permeation of HCA across an animal membrane. The release of HCA was found comparable for the four systems. The two microemulsions promote permeation across an ex-vivo membrane, examined by means of a Franz cell. Hydrophobic microemulsion guarantees the highest solubility (2,370 μg/ml) and flux (133 μg/cm².h) of the drug, since it contains almost 40% Transcutol, a permeation enhancer. Gel and ointment provide lower solubility and flux, being the values, related to the ointment, the lowest ones (562 μg/ml and 0.4 μg/cm².h). Experimental results allow the conclusion that gel and ointment can be suitable when it is desirable to minimize absorption of topically applied HCA as to keep the drug restricted to the diseased area and prevent side effects of the systemic presence of HCA.     

Combined efficacy of Vigna radiata (L.) R. Wilczek and Amorphophallus paeoniifolius (Dennst.) Nicolson on serum lipids in albino rats

Coronary Artery Disease (CAD) is a major killer disease throughout the world. Dyslipidemia is a major contributor to the risk of CAD. Several dietary articles traditionally used in India and other South Asian countries reduced dyslipidemia. The present study was undertaken to evaluate the combined effect of Mung bean (Vigna radiata) and Elephant foot yam (Amorphophallus paeoniifolius) on serum lipids and atherogenic indices in albino rats and to compare it with a standard drug Cholestyramine. Thirty healthy albino rats of both sexes (150–200 g) were randomized to 5 groups of 6 animals each. The grouping were done based on the following criteria: Group I: Normal Control Group, Group II: (Standard Group): Cholestyramine resin 5 mg/kg bw, Group III: (Half Dose Group): Drug powder at 540 mg/kg bw, Group IV: (Effective Dose Group): Drug powder at 1080 mg/kg bw, and Group V: (Double Dose Group): Drug powder at 2160 mg/kg bw. Lipid profile was estimated at the beginning and after 30 days of treatment. The Effective and Double doses of the drug reduced Total cholesterol along with levels of Triglycerides, Low density lipoprotein and Very low density lipoprotein levels significantly (p < 0.01) along with a significant (p < 0.01) increase in high density lipoproteins (HDL) in rats. There was also significant (p < 0.01) improvement in atherogenic indices like Castelli Risk Index I, Non HDL C/HDL, Castelli risk Index II, TG/HDL, Atherogenic coefficient and Atherogenic Index of Plasma. The combination of powdered sprouted mung bean and yam powder have excellent lipid lowering potential.     

Formulation and characterization of Aloe vera gel and tomato powder containing cream

Herbal cosmetics are a valuable gift of nature and their demand is increasing in the world market. They have good activity with no side effects as compared to synthetic cosmetics. Herbal cosmetics are beauty products and because of natural ingredients, they possess desirable physiological activities. The present research work aimed to formulate and characterize cost-effective herbal nourishment cream containing Aloe vera gel and tomato powder. Four formulations i.e. F1, F2, F3, and F4 were prepared with varying concentrations of Aloe vera gel and tomato powder, while the composition of the cream base was kept the same. The oil phase of the cream base consisted of liquid paraffin (5.0 ml), stearic acid (4.8 g) and coconut oil (3.0 ml). The aqueous phase consisted of triethanolamine (1.5 ml), glycerin (5.0 ml), citric acid (0.4 g), methylparaben (0.1 g), water (q.s.) and key ingredients. The concentration of Aloe vera gel was 2.0 ml, 4.0 ml, 6.0 ml, 8.0 ml and the concentration of tomato powder was 0.2 g, 0.4 g, 0.6 g, 0.8 g in F1, F2, F3, and F4 respectively. The characterization of the formulated creams was carried out by standard methods, such as homogeneity test, smear type, irritancy test, pH test, spreadability test and acid value test. The stability test was carried out at different temperature conditions at 40 °C, below 5 °C and room temperature. All creams were homogeneous, non-irritant and easily removable. The pH of creams was in the range of 7.3–7.6 which is safe for human skin. Spreadability was in the range of 9–13. Acid values of the prepared creams were 5.6, 5.6, 6.4 and 7.2 respectively. Stability was examined by two parameters, homogeneity and pH change. The economical herbal creams prepared by using fewer chemicals are good for skin with moisturizing properties. The efficiency of creams can be improved by increasing the concentration of Aloe vera gel and tomato powder.     

Comparison between three different LCIA methods for aquatic ecotoxicity and a product environmental risk assessment

Background and Objective  In the OMNIITOX project 11 partners have the common objective to improve environmental management tools for the assessment of (eco)toxicological impacts. The detergent case study aims at: i) comparing three Procter &c Gamble laundry detergent forms (Regular Powder-RP, Compact Powder-CP and Compact Liquid-CL) regarding their potential impacts on aquatic ecotoxicity, ii) providing insights into the differences between various Life Cycle Impact Assessment (LCIA) methods with respect to data needs and results and iii) comparing the results from Life Cycle Assessment (LCA) with results from an Environmental Risk Assessment (ERA). Material and Methods  The LCIA has been conducted with EDIP97 (chronic aquatic ecotoxicity) [1], USES-LCA (freshwater and marine water aquatic ecotoxicity, sometimes referred to as CML2001) [2, 3] and IMPACT 2002 (covering freshwater aquatic ecotoxicity) [4]. The comparative product ERA is based on the EU Ecolabel approach for detergents [5] and EUSES [6], which is based on the Technical Guidance Document (TGD) of the EU on Environmental Risk Assessment (ERA) of chemicals [7]. Apart from the Eco-label approach, all calculations are based on the same set of physico-chemical and toxicological effect data to enable a better comparison of the methodological differences. For the same reason, the system boundaries were kept the same in all cases, focusing on emissions into water at the disposal stage. Results and Discussion  Significant differences between the LCIA methods with respect to data needs and results were identified. Most LCIA methods for freshwater ecotoxicity and the ERA see the compact and regular powders as similar, followed by compact liquid. IMPACT 2002 (for freshwater) suggests the liquid is equally as good as the compact powder, while the regular powder comes out worse by a factor of 2. USES-LCA for marine water shows a very different picture seeing the compact liquid as the clear winner over the powders, with the regular powder the least favourable option. Even the LCIA methods which result in die same product ranking, e.g. EDIP97 chronic aquatic ecotoxicity and USES-LCA freshwater ecotoxicity, significantly differ in terms of most contributing substances. Whereas, according to IMPACT 2002 and USES-LCA marine water, results are entirely dominated by inorganic substances, the other LCIA methods and the ERA assign a key role to surfactants. Deviating results are mainly due to differences in the fate and exposure modelling and, to a lesser extent, to differences in the toxicological effect calculations. Only IMPACT 2002 calculates the effects based on a mean value approach, whereas all other LCIA methods and the ERA tend to prefer a PNEC-based approach. In a comparative context like LCA the OMNIITOX project has taken the decision for a combined mean and PNEC-based approach, as it better represents the ‘average’ toxicity while still taking into account more sensitive species. However, the main reason for deviating results remains in the calculation of the residence time of emissions in the water compartments. Conclusion and Outlook  The situation that different LCIA methods result in different answers to the question concerning which detergent type is to be preferred regarding the impact category aquatic ecotoxicity is not satisfactory, unless explicit reasons for the differences are identifiable. This can hamper practical decision support, as LCA practitioners usually will not be in a position to choose the ’right’ LCIA method for their specific case. This puts a challenge to the entire OMNIITOX project to develop a method, which finds common ground regarding fate, exposure and effect modelling to overcome the current situa-tion of diverging results and to reflect most realistic conditions.     

Bioadhesive Microspheres for Bioavailability Enhancement of Raloxifene Hydrochloride: Formulation and Pharmacokinetic Evaluation

Raloxifene hydrochloride (R-HCl), a BCS class II drug, remains a mainstay in the prevention and pharmacologic therapy of osteoporosis. Its absolute bioavailability, however, is 2% due to poor solubility and extensive first pass metabolism. The present study describes two simultaneous approaches to improve its bioavailability, complexation of R-HCl with cyclodextrin(s), and formulation of mucoadhesive microspheres of the complex using different proportions of carbopol and HPMC. Microspheres were pale yellow in color, free-flowing, spherical, and porous in outline. The particle size ranged between 3 and 15 μm, and entrapment efficiency was found to be within 81.63% to 87.73%. A significant improvement in the solubility of R-HCl was observed, and it differed with the combination of excipients used. X-ray diffraction and differential scanning calorimetry studies revealed that enhancement in drug solubility was resulted due to a change in its crystallinity within the formulation. Microspheres possessed remarkable mucoadhesion and offered controlled drug release, lasting up to 24 h. They produced a sharp plasma concentration–time profile of R-HCl within 30 min post-administration to Wistar rats. [AUC]_0–24 h was found to be 1,722.34 ng h/ml, and it differed significantly to that of pure drug powder (318.28 ng h/ml). More than fivefold increase in AUC and more than twofold increase in MRT were observed. FT-IR studies evidenced no interaction among drug and excipients. The results of this study showed that mucoadhesive microspheres could be a viable approach to improve the pharmacokinetic profile of R-HCl.     

Production of biodiesel from acid waste lard

The objective of the present work was: (i) to enable biodiesel production from acid waste lard; (ii) to study the esterification reaction as possible pre-treatment at different temperatures, catalyst amount and reaction times; (iii) to evaluate biodiesel quality according to EN 14214 after basic transesterification of the pre-treated fat; and (iv) to predict the impact of using such waste as raw material in mixture with soybean oil. Temperature and catalyst amount were the most important reaction conditions which mostly affected biodiesel quality, namely viscosity and purity. The selected pre-treatment conditions were 65 °C, 2.0 wt% H₂SO₄ and 5 h, which allowed obtaining a product with a viscosity of 4.81 mm² s⁻¹ and a purity of 99.6 wt%. The proposed pre-treatment was effective to enable acid wastes as single raw materials for biodiesel production with acceptable quality; however, low yields were obtained (65 wt%). Alkali transesterification of a mixture of waste lard and soybean oil resulted in a product with a purity of 99.8 wt% and a yield of 77.8 wt%, showing that blending might be an interesting alternative to recycle such wastes. Also, because in addition to using conventional and relatively economical processes, some biodiesel properties depending on the raw material composition (such as the iodine value) might even be improved.     

Development of doxorubicin loading platform based albumin-sporopollenin as drug carrier

Albumin is thought as an drug carrier for doxorubicin (DOX). The binding of doxorubicin to albumin was studied on the surface of sporopolleninin (SP) to produce a new drug system based natural materials. Human serum albumin (HSA) was immobilized on SPIONs in 20 mM Tris buffer, 7.4 of pH. Data showed that binding amount of HSA has been found to be as 285.53 µg to the 25 mg of Sporopolleninin which also bounded 319.76 µM of DOX. Binding of protein and drug to Sp were clarified by SEM, EDX and FT-IR analysis.     

Dissolution improvement of high drug-loaded solid dispersion

This study focused on an investigation of a high drug-loaded solid dispersion system consisting of drug, carrier, and surfactant. Solid dispersions of a water-insoluble ofloxacin (OFX) with polyethylene glycol (PEG) of different molecular weights, namely binary solid dispersion systems, were prepared at drug to carrier not less than 5∶5. Polysorbate 80, a nonionic surfactant, was incorporated into the binary solid dispersion systems as the third component to obtain the ternary solid dispersion systems. The powder x-ray diffraction and differential scanning calorimetric studies indicated that crystalline OFX existed in the solid dispersions with high drug loading. However, a decreased crystallinity of the solid dispersions obtained revealed that a portion of OFX was in an amorphous state. The results indicated a remarkably improved dissolution of drug from the ternary solid dispersion systems when compared with the binary solid dispersion systems. This was because of polysorbate 80, which improved wettability and solubilized the non-molecularly dispersed or crystalline fraction of OFX.     

Micropropagation and hairy root culture of Ophiorrhiza alata Craib for camptothecin production

An efficient system was developed for the in vitro micropropagation and hairy root culture of Ophiorrhiza alata Craib for camptothecin (CPT) production. Shoot multiplication on leaf and node explants from germinated seeds of O. alata was successful on half-strength Murashige and Skoog medium supplemented with varying amounts of kinetin and α-naphthaleneacetic acid. Node explants grown in vitro were successfully infected by Agrobacterium rhizogenes TISTR 1450 for the establishment of hairy root culture. The amount of CPT in various parts of O. alata was analyzed by HPLC. The accumulation of CPT in transformed hairy roots was twice that in soil-grown plants (785 ± 52 and 388 ± 32 μg/g dry wt, respectively). In the presence of a polystyrene resin (Diaion HP-20) that absorbed CPT, the CPT content in the culture media increased sevenfold compared with controls (1,036 and 151 μg per 250 ml medium, respectively). These results enable the feasible production of CPT of O. alata by means of a cell culture strategy. These measures can help safeguard the plant from extinction.     

The Transdermal Patches for Site-Specific Delivery of Letrozole: A New Option for Breast Cancer Therapy

The aim of this work was to evaluate capability of site-specific delivery of a transdermal patch through determination of letrozole in local tissues disposition in female mice. After transdermal administration, the letrozole levels in skin, muscle, and plasma were 10.4–49.3 μg/g, 1.64–6.89 μg/g, and 0.35–1.64 μg/mL, respectively. However, after the mice received letrozole suspension, the drug concentration of plasma and muscle were 0.20–4.80 μg/mL and 0.15–2.38 μg/g. There was even no drug determined in skin through all experiments. Compared with oral administration, the transdermal patch for site-specific delivery of letrozole could produce high drug concentrations in skin and muscle and meanwhile obtain low drug level in plasma. These findings show that letrozole transdermal patch is an appropriate delivery system for application to the breast tumor region for site-specific drug delivery to obtain a high local drug concentration and low circulating drug concentrations avoiding the risk of systemic side effects.     

Effect of Plasticizer on Release Kinetics of Diclofenac Sodium Pellets Coated with Eudragit RS 30 D

The present study was designed to investigate the effect of two plasticizers, i.e., triethyl citrate (TEC) and polyethylene glycol 6000 (PEG 6000) on the in vitro release kinetics of diclofenac sodium from sustained-release pellets. Ammonio methacrylate copolymer type B (Eudragit RS 30 D) is used as the release-retarding polymer. Both plasticizers were used at 10% and 15% (w/w) of Eudragit RS 30 D. Pellets were prepared by powder layering technology and coated with Eudragit RS 30 D by air suspension technique. Thermal properties of drug and drug-loaded beads were studied using differential scanning calorimeter (DSC). DSC thermogram represented the identity of raw materials and exhibited no interaction or complexation between the active and excipients used in the pelletization process. Dissolution study was performed by using USP apparatus 1. No significant difference was observed among the physical properties of the coated pellets of different batches. When dissolution was performed as pure drug, about 8.22% and 90% drug was dissolved at 2 h in 0.1 N HCl and at 30 min in buffer (pH 6.8), respectively. From all formulations, the release of drug in acid media was very negligible (maximum 1.8 ± 0.08% at 2 h) but in buffer only 12% and 30% drug was released at 10 h from coated pellets containing TEC and PEG 6000, respectively, indicating that Eudragit RS 30 D significantly retards the drug release rate and that drug release was varied according to the type and amount of plasticizers used. The amount of TEC in coating formulation significantly effected drug release (p < 0.001), but the effect of PEG 6000 was not significant. Formulations containing PEG 6000 released more drug (98.35 ± 2.35%) than TEC (68.01 ± 1.04%) after 24 h. Different kinetic models like zero order, first order, and Higuchi were used for fitting drug release pattern. Zero order model fitted best for diclofenac release in all formulations. Drug release mechanism was derived with Korsmeyer equation.     

Effects of Saccharomyces cerevisiae supplementation on immune response,hematological parameters,body composition and disease resistance in rainbow trout,Oncorhynchus mykiss (Walbaum, 1792)

Natural substances are now generally preferred over chemical and synthetic compounds for the growth and immune enhancement of aquatic organisms. The aim of this study was to evaluate the effect of Saccharomyces cerevisiae extract and hydrolyzed powder on immunity, hematological parameters and body composition in rainbow trout, Oncorhynchus mykiss. Six hundred rainbow trout (50 ± 5 g mean weight) were acclimated to laboratory conditions and then randomly divided into four groups of triplicate tanks. The first group was fed with a commercial diet (control) without supplementation. The second and third groups were given a diet supplemented with 1% of yeast extract and hydrolyzed powder, respectively. The fourth group was also fed with a basal diet supplemented with 0.5% of both substances. Fish were cultured in 300‐L polyethylene tanks for 60 days; immune and hematological parameters, fillet composition and disease resistance were analyzed at days 0, 30 and 60. Results showed that a combination of Saccharomyces cerevisiae extract and hydrolyzed powder could improve the immunity and alter hematological parameters of the rainbow trout compared to the control. Mortality rates of fish fed yeast extract and hydrolyzed powder were also lower than in fish fed the control diet after challenging with Yersinia ruckeri. There were no significant changes in rainbow trout fillet composition compared to the control. It can be concluded that fish diet supplementation with a mixture of yeast extract hydrolyzed powder is preferable compared to each one used alone.     

Optimization of PEGylation Conditions for BSA Nanoparticles Using Response Surface Methodology

Chemical coupling of polyethylene glycol (PEG) to proteins or particles (PEGylation), prolongs their circulation half-life by greater than 50-fold, reduces their immunogenicity, and also promotes their accumulation in tumors due to enhanced permeability and retention effect. Herein, phase separation method was used to prepare bovine serum albumin (BSA) nanoparticles. PEGylation of BSA nanoparticles was performed by SPA activated mPEG through their free amino groups. Effect of process variables on PEGylation efficiency of BSA nanoparticles was investigated and optimized through response surface methodology with the amount of free amino groups as response. Optimum conditions was found to be 32.5 g/l of PEG concentration, PEG-nanoparticle incubation time of 10 min, incubation temperature of 27°C, and pH of 7 for 5 mg of BSA nanoparticles in 1 mL phosphate buffer. Analysis of data showed that PEG concentration had the most noticeable effect on the amount of PEGylated amino groups, but pH had the least. Mean diameter and zeta potential of PEGylated nanoparticles under these conditions were 217 nm and −14 mV, respectively. In conclusion, PEGylated nanoparticles demonstrated reduction of the negative surface charge compared to the non modified particles with the zeta potential of −31.7 mV. Drug release from PEGylated nanoparticles was almost slower than non-PEGylated ones, probably due to existence of a PEG layer around PEGylated particles which makes an extra resistance in opposition to drug diffusion.     

Rational orthologous pathway and biochemical process engineering for adipic acid production using Pseudomonas taiwanensis VLB120

Microbial bioprocessing based on orthologous pathways constitutes a promising approach to replace traditional greenhouse gas- and energy-intensive production processes, e.g., for adipic acid (AA). We report the construction of a Pseudomonas taiwanensis strain able to efficiently convert cyclohexane to AA. For this purpose, a recently developed 6-hydroxyhexanoic acid (6HA) synthesis pathway was amended with alcohol and aldehyde dehydrogenases, for which different expression systems were tested. Thereby, genes originating from Acidovorax sp. CHX100 and the XylS/Pm regulatory system proved most efficient for the conversion of 6HA to AA as well as the overall cascade enabling an AA formation activity of up to 48.6 ± 0.2 U g_CDW⁻¹. The optimization of biotransformation conditions enabled 96% conversion of 10 mM cyclohexane with 100% AA yield. During recombinant gene expression, the avoidance of glucose limitation was found to be crucial to enable stable AA formation. The biotransformation was then scaled from shaking flask to a 1 L bioreactor scale, at which a maximal activity of 22.6 ± 0.2 U g_CDW⁻¹ and an AA titer of 10.2 g L⁻¹ were achieved. The principal feasibility of product isolation was shown by the purification of 3.4 g AA to a purity of 96.1%. This study presents the efficient bioconversion of cyclohexane to AA by means of a single strain and thereby sets the basis for an environmentally benign production of AA and related polymers such as nylon 6,6.     

Increased yield and CO2 sequestration potential with the C4 cereal Sorghum bicolor cultivated in basaltic rock dust‐amended agricultural soil

Land‐based enhanced rock weathering (ERW) is a biogeochemical carbon dioxide removal (CDR) strategy aiming to accelerate natural geological processes of carbon sequestration through application of crushed silicate rocks, such as basalt, to croplands and forested landscapes. However, the efficacy of the approach when undertaken with basalt, and its potential co‐benefits for agriculture, require experimental and field evaluation. Here we report that amending a UK clay‐loam agricultural soil with a high loading (10 kg/m²) of relatively coarse‐grained crushed basalt significantly increased the yield (21 ± 9.4%, SE) of the important C₄ cereal Sorghum bicolor under controlled environmental conditions, without accumulation of potentially toxic trace elements in the seeds. Yield increases resulted from the basalt treatment after 120 days without P‐ and K‐fertilizer addition. Shoot silicon concentrations also increased significantly (26 ± 5.4%, SE), with potential benefits for crop resistance to biotic and abiotic stress. Elemental budgets indicate substantial release of base cations important for inorganic carbon removal and their accumulation mainly in the soil exchangeable pools. Geochemical reactive transport modelling, constrained by elemental budgets, indicated CO₂ sequestration rates of 2–4 t CO₂/ha, 1–5 years after a single application of basaltic rock dust, including via newly formed soil carbonate minerals whose long‐term fate requires assessment through field trials. This represents an approximately fourfold increase in carbon capture compared to control plant–soil systems without basalt. Our results build support for ERW deployment as a CDR technique compatible with spreading basalt powder on acidic loamy soils common across millions of hectares of western European and North American agriculture.