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The low-cost production of cellulolytic complexes that present high action at mild conditions is one of the major bottlenecks for the economic viability of the production of cellulosic ethanol. The influence of agro-industrial residues was assessed to enhance endoglucanase production by Aspergillus niger 426 grown in solid state fermentation. The highest percentage of lignin degradation was found on soybean hulls (56%) followed by sugarcane bagasse (36%) and rice straw (8.5%). The cellulose degradation, around 90%, was observed on soybean hulls and sugarcane bagasse, but only 50% on rice straw, and maximum production of endoglucanase (112.34 ± 0.984 U mL-1) was observed for soybean hulls. The best Experimental Mixture Design condition was under cultivation of 2.5 g of sugarcane bagasse, 2.3 g of rice straw and 5.2 g of soybean hulls, leading to a maximum activity of 138.92 ± 0.02 U mL-1. The statistical methodology enabled an increase of over 20% in the production of endoglucanase using agro-industrial waste. These data demonstrate that A. niger 426 is a potential source of cellulases which can be obtained by solid state fermentation using agro-industrial waste.

The development of novel antioxidant compounds with high efficacy and low toxicity is of utmost importance in the medicine and food industries. Moreover, with increasing concerns about the safety of synthetic components, scientists are beginning to search for natural sources of antioxidants, especially essential oils (EOs). The combination of EOs may produce a higher scavenging profile than a single oil due to better chemical diversity in the mixture. Therefore, this exploratory study aims to assess the antioxidant activity of three EOs extracted from Cymbopogon flexuosus , Carum carvi , and Acorus calamus in individual and combined forms using the augmented-simplex design methodology. The in vitro antioxidant assays were performed using DPPH and ABTS radical scavenging approaches. The results of the Chromatography Gas-Mass spectrometry (CG-MS) characterization showed that citral (29.62%) and niral (27.32%) are the main components for C. flexuosus , while d -carvone (62.09%) and d -limonene (29.58%) are the most dominant substances in C. carvi . By contrast, β-asarone (69.11%) was identified as the principal component of A. calamus (30.2%). The individual EO exhibits variable scavenging activities against ABTS and DPPH radicals. These effects were enhanced through the mixture of the three EOs. The optimal antioxidant formulation consisted of 20% C. flexuosus , 53% C. carvi , and 27% A. calamus for DPPHIC50. Whereas 17% C. flexuosus , 43% C. carvi , and 40% A. calamus is the best combination leading to the highest scavenging activity against ABTS radical. These findings suggest a new research avenue for EOs combinations to be developed as novel natural formulations useful in food and biopharmaceutical products.

Kojic monooleate (KMO) is an ester derived from a fungal metabolite of kojic acid with monounsaturated fatty acid, oleic acid, which contains tyrosinase inhibitor to treat skin disorders such as hyperpigmentation. In this study, KMO was formulated in an oil-in-water nanoemulsion as a carrier for better penetration into the skin. The nanoemulsion was prepared by using high and low energy emulsification technique. D-optimal mixture experimental design was generated as a tool for optimizing the composition of nanoemulsions suitable for topical delivery systems. Effects of formulation variables including KMO (2.0%-10.0% w/w), mixture of castor oil (CO):lemon essential oil (LO; 9:1) (1.0%-5.0% w/w), Tween 80 (1.0%-4.0% w/w), xanthan gum (0.5%-1.5% w/w), and deionized water (78.8%-94.8% w/w), on droplet size as a response were determined. Analysis of variance showed that the fitness of the quadratic polynomial fits the experimental data with -value (2,479.87), a low -value ( <0.0001), and a nonsignificant lack of fit. The optimized formulation of KMO-enriched nanoemulsion with desirable criteria was KMO (10.0% w/w), Tween 80 (3.19% w/w), CO:LO (3.74% w/w), xanthan gum (0.70% w/w), and deionized water (81.68% w/w). This optimum formulation showed good agreement between the actual droplet size (110.01 nm) and the predicted droplet size (111.73 nm) with a residual standard error <2.0%. The optimized formulation with pH values (6.28) showed high conductivity (1,492.00 µScm ) and remained stable under accelerated stability study during storage at 4°C, 25°C, and 45°C for 90 days, centrifugal force as well as freeze-thaw cycles. Rheology measurement justified that the optimized formulation was more elastic (shear thinning and pseudo-plastic properties) rather than demonstrating viscous characteristics. In vitro cytotoxicity of the optimized KMO formulation and KMO oil showed that IC (50% inhibition of cell viability) value was >100 µg/mL. The survival rate of 3T3 cell on KMO formulation (54.76%) was found to be higher compared to KMO oil (53.37%) without any toxicity sign. This proved that the KMO formulation was less toxic and can be applied for cosmeceutical applications.

The health risks associated with synthetic preservatives have intensified the search for natural antimicrobial alternatives. Crocus sativus L. (saffron) generates abundant by-products, such as tepals and leaves, which are rich in bioactive compounds with demonstrated antimicrobial potential. Compared to other natural alternatives, saffron by-products offer distinct advantages, including a unique combination of phenolic compounds (e.g., ellagic acid, rutin) and carotenoids (e.g., crocin) that act synergistically against both Gram-positive and Gram-negative bacteria, as well as fungi. Additionally, these by-products represent a sustainable solution, with approximately 63 kg of agricultural waste generated per kg of saffron spice. This study optimized the antibacterial and antifungal efficacy of saffron extracts using a simplex centroid mixture design. Phytochemical analysis using high-performance liquid chromatography with diode-array detection (HPLC–DAD) identified key antimicrobial compounds, including ellagic acid (68.43% in leaves, 50.31% in tepals) and crocin (9.59% in stigmas). Antimicrobial assays against Staphylococcus aureus , Escherichia coli , Candida albicans , and Geotrichum candidum revealed that stigma extracts exhibited superior antibacterial activity (MIC = 25 mg/mL for S. aureus and E. coli ), while tepal and leaf extracts showed promising antifungal effects (MIC = 12.5 mg/mL for G. candidum ). The mixture design approach uncovered synergistic interactions, with an equimolar combination of stigma, tepal, and leaf extracts (33:33:33) demonstrating the strongest antibacterial activity (MIC = 25 mg/mL) and a ternary mixture (34% stigma, 30% leaf, 36% tepal) achieving the lowest antifungal MIC (6.25 mg/mL). These findings highlight saffron by-products as highly effective and sustainable natural antimicrobials, providing a cost-efficient (40–60% reduction compared to conventional extracts) and multi-functional alternative to synthetic preservatives. Their dual functionality (antimicrobial + natural coloring) and agricultural waste origin make them particularly valuable for industrial applications in food preservation, pharmaceuticals, and biopharmaceuticals. The integration of statistical modeling maximizes their potential, meeting the growing demand for safe, natural antimicrobial solutions with clear competitive advantages

The optimization of existing natural antioxidants that are highly effective is crucial for advancements in medicine and the food industry. Due to growing concerns regarding the safety of synthetic antioxidants, researchers are increasingly focusing on natural sources, particularly essential oils (EOs). Combining EOs might enhance antioxidant activity due to increased chemical diversity. This study investigates, for the first time, the antioxidant properties of EOs from Lavandula dentata, Rosmarinus officinalis, and Myrtus communis, both individually and in combination, using the augmented-simplex design methodology. The in vitro evaluation of the antioxidant activity was performed using DPPH and ABTS radical scavenging assays. Chromatography gas-mass spectrometry (CG-MS) revealed that 1,8-cineol (37.27%) and pinocarveol (12.67%) are the primary components of L. dentata; verbenone (16.90%), camphor (15.00%), and camphene (11.03%) are predominant in R. officinalis; while cineol (43.32%) is the main component of M. communis. The EOs showed varying scavenging activities against ABTS and DPPH radicals, with DPPH assay values ranging from 194.10 ± 3.01 to 541.19 ± 3.72 µg/mL and ABTS assay values ranging from 134.07 ± 1.70 to 663.42 ± 2.99 µg/mL. These activities were enhanced when the EOs were combined. The optimal antioxidant blend for DPPHIC50 consisted of 20% L. dentata, 50% R. officinalis, and 30% M. communis. For the highest ABTS radical scavenging activity, the best combination was 18% L. dentata, 43% R. officinalis, and 40% M. communis. These results highlight the potential of EO combinations as new natural formulations for use in cosmeceutical, food, and pharmaceutical sectors.

This research aimed to examine the interplay among carotenoids, total polyphenols, and indole alkaloids for the discovery of new bioactive compounds using “simplex-centroid” designs and response surface methodology (RSM). The antioxidant (AOA) and anti-inflammatory (AIA) activities of mixtures containing different proportions of each compound were evaluated by measuring the percentage inhibition of oxidation and spontaneous inflammation. The results showed that the highest AOAs were obtained with different mixtures of polyphenols, indole alkaloids, and carotenoids. The DPPH test revealed that a mixture of 16.94% polyphenols and 83.05% indole alkaloids produced the best results, whereas, for the ABTS test, 100% polyphenols gave the best results. The FRAP assay showed that the best-performing mixture was composed of 85.96% carotenoids and 14.03% polyphenols, with an IC 50 of 8.85 ± 1.17 μg/mL. In contrast, the most potent AIA was obtained with 100% carotenoids. Model validation tests were performed and showed that the experimentally observed values for AOA and AIA were not significantly different from the values estimated by the fitted models. The results also showed that the RSM model can be used to estimate the behavior of mixed compounds in the development of new functional ingredients or the discovery of new bioactive compounds with pharmacological properties. Finally, the results also suggested that there is a good correlation between several AOA and AIA assays, which makes natural substances combining these biological activities particularly interesting. Graphical abstract

The synergistic anticancer effect of docetaxel (DTX) and curcumin (CCM) has emerged as an attractive therapeutic candidate for lung cancer treatment. However, the lack of optimal bioavailability because of high toxicity, low stability, and poor solubility has limited their clinical success. Given this, an aerosolized nanoemulsion system for pulmonary delivery is recommended to mitigate these drawbacks. In this study, DTX- and CCM-loaded nanoemulsions were optimized using the D-optimal mixture experimental design (MED). The effect of nanoemulsion compositions towards two response variables, namely, particle size and aerosol size, was studied. The optimized formulations for both DTX- and CCM-loaded nanoemulsions were determined, and their physicochemical and aerodynamic properties were evaluated as well. The MED models achieved the optimum formulation for DTX- and CCM-loaded nanoemulsions containing a 6.0 wt% mixture of palm kernel oil ester (PKOE) and safflower seed oils (1:1), 2.5 wt% of lecithin, 2.0 wt% mixture of Tween 85 and Span 85 (9:1), and 2.5 wt% of glycerol in the aqueous phase. The actual values of the optimized formulations were in line with the predicted values obtained from the MED, and they exhibited desirable attributes of physicochemical and aerodynamic properties for inhalation therapy. Thus, the optimized formulations have potential use as a drug delivery system for a pulmonary application.

Background/Objectives: The rise of antibiotic-resistant pathogens has become a global health crisis, necessitating the development of alternative antimicrobial strategies. This study aimed to optimize the antibacterial effects of essential oils (EOs) from Thymus satureioides, Lavandula angustifolia, and Origanum majorana, enhancing their efficacy through optimized mixtures. Methods: This study utilized a simplex–centroid design to optimize the mixture ratios of EOs for maximal antibacterial and antioxidant effectiveness. The chemical profiles of the EOs were analyzed using gas chromatography-mass spectrometry (GC-MS). The antibacterial activity was assessed against Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa using minimum inhibitory concentration (MIC) tests, while antioxidant activity was evaluated through DPPH (2,2-diphenyl-1-picrylhydrazyl), and ABTS (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) assays. Results: The optimized essential oil mixtures demonstrated potent antibacterial activity, with MIC values of 0.097% (v/v) for E. coli, 0.058% (v/v) for S. aureus, and 0.250% (v/v) for P. aeruginosa. The mixture ratios achieving these results included 76% T. satureioides, and 24% O. majorana for E. coli, and varying proportions for other strains. Additionally, L. angustifolia essential oil exhibited the strongest antioxidant activity, with IC50 values of 84.36 µg/mL (DPPH), and 139.61 µg/mL (ABTS), surpassing both the other EOs and standard antioxidants like BHT and ascorbic acid in the ABTS assay. Conclusions: The study successfully demonstrates that optimized mixtures of EOs can serve as effective natural antibacterial agents. The findings highlight a novel approach to enhance the applications of essential oils, suggesting their potential use in food preservation and biopharmaceutical formulations. This optimization strategy offers a promising avenue to combat antibiotic resistance and enhance food safety using natural products.

Psoriasis is a chronic autoimmune disease that cannot be cured. It can however be controlled by various forms of treatment, including topical, systemic agents, and phototherapy. Topical treatment is the first-line treatment and favored by most physicians, as this form of therapy has more patient compliance. Introducing a nanoemulsion for transporting cyclosporine as an anti-inflammatory drug to an itchy site of skin disease would enhance the effectiveness of topical treatment for psoriasis. The addition of nutmeg and virgin coconut-oil mixture, with their unique properties, could improve cyclosporine loading and solubility. A high-shear homogenizer was used in formulating a cyclosporine-loaded nanoemulsion. A D-optimal mixture experimental design was used in the optimization of nanoemulsion compositions, in order to understand the relationships behind the effect of independent variables (oil, surfactant, xanthan gum, and water content) on physicochemical response (particle size and polydispersity index) and rheological response (viscosity and -value). Investigation of these variables suggests two optimized formulations with specific oil (15% and 20%), surfactant (15%), xanthan gum (0.75%), and water content (67.55% and 62.55%), which possessed intended responses and good stability against separation over 3 months' storage at different temperatures. Optimized nanoemulsions of pH 4.5 were further studied with all types of stability analysis: physical stability, coalescence-rate analysis, Ostwald ripening, and freeze-thaw cycles. In vitro release proved the efficacy of nanosize emulsions in carrying cyclosporine across rat skin and a synthetic membrane that best fit the Korsmeyer-Peppas kinetic model. In vivo skin analysis towards healthy volunteers showed a significant improvement in the stratum corneum in skin hydration.

Hydrochlorothiazide (HCTZ)/losartan potassium (LOS-K) was used as a model drug to prepare compound tablets through the investigation of the compression and mechanical properties of mixed powders to determine the formulation and preparation factors, followed by D-optimal mixture experimental design to optimize the final parameters. The type and amount of lactose monohydrate (SuperTab®14SD, 19.53–26.91%), microcrystalline cellulose (MCC PH102, 32.86–43.31%), pre-gelatinized starch (Starch-1500, 10.96–15.91%), and magnesium stearate (0.7%) were determined according to the compressive work, stress relaxation curves, and Py value. Then, the compression mechanism of the mixed powder was investigated by the Kawakita equation, Shapiro equation, and Heckel analysis, and the mixed powder was classified as a Class-II powder. The compaction pressure (150–300 MPa) and tableting speed (1200–2400 Tab/h) were recommended. A D-optimal mixture experimental design was utilized to select the optimal formulation (No 1, 26.027% lactose monohydrate, 32.811% MCC PH102, and 15.462% pregelatinized starch) according to the drug dissolution rate, using Hyzaar® tablets as a control. Following oral administration in beagle dogs, there were no significant differences in bioavailability between the No. 1 tablet and the Hyzaar® tablet in HCTZ, losartan carboxylic acid (E-3174), and LOS-K (F < F0.05). Thus, formulation and preparation factors were determined according to the combination of the compression and mechanical properties of the mixed powder and quality of tablets, which was demonstrated to be a feasible method in direct powder compression.