Microalgae are considered a rich source of high-value metabolites with an array of nutraceutical and pharmaceutical applications. Different strategies have been developed for cultivating microalgae at large-scale photobioreactors but high cost and low productivity are the major hurdles. Optimizing the composition of media for the cultivation of microalgae to induce biomass production and high-value metabolite accumulation has been considered as an important factor for sustainable product development. In this study, the effect of plant growth regulators together with basal microalgal cultivation medium on biomass, total lipid, and EPA production was studied using the Plackett-Burman model and Response surface methodology. The traditional one-factor-at-a-time optimization approach is laborious, time-consuming, and requires more experiments which makes the process and analysis more difficult. The Designed PB model was found to be significant for biomass (396 mg/L), lipid (254 mg/L), and EPA (5.6%) production with a P value < 0.05. The major objective of this study is to formulate a medium for EPA production without compromising the growth properties. Further, we had formulated a new media using RSM to achieve the goal and the significant variables selected were NaNO3, NaH2PO4, and IAA and was found to be significant with 16.72% EPA production with a biomass production of 893 mg/L with a P value < 0.05. The formulated medium can be used in large-scale cultivation systems which can enhance biomass production as well as the omega 3 fatty acid production in marine microalgae Nannochloropsis oceanica.
UNASSIGNED: The online version contains supplementary material available at 10.1007/s43393-021-00069-1.

To enhance the ribonucleic acid (RNA) productivity for industrial applications, this study employed strain screening and medium optimization to improve the content of RNA in Cyberlindnera jadinii. A rapid screening method, combining atmospheric and room temperature plasma mutagenesis, 48-deep-well plates fermentation, and microplate reader detection, was developed. A mutant strain named WB15 with high RNA content was successfully obtained, exhibiting the RNA content of 156 ± 4.5 mg/g DCW, 1.4 times of the starting strain CCTCC AY 92020. Furthermore, Plackett-Burman design and response surface methodology were employed to identify three significant factors (yeast extract, soybean peptone, and KH2PO4) affecting the RNA content. By utilizing the optimal medium composed of 13.43 g/L yeast extract, 12.12 g/L soybean peptone and 2.78 g/L KH2PO4, the RNA content of WB15 further increased to 184 ± 4.9 mg/g DCW. Additionally, the mutant strain WB15 exhibited a greater cellular width compared to AY 92020, along with increased growth rate and single-cell RNA content by 22% and 48.9%, respectively. Perturbations in ribosome assembly, specifically a reduction in the ratio of ribosomal proteins to ribosomal RNA of the large subunit, might indirectly contribute to the higher RNA content in the WB15 strain. Overall, the combination of rapid screening with fermentation medium optimization proved to be an effective approach for improving the RNA content of C. jadinii, thus facilitating the industrial production of RNA.

A novel simple, specific, sensitive, accurate and precise reversed phase high performance liquid chromatographic method (RP-HPLC/UV) was developed and validated for the simultaneous estimation of Glycopyrronium bromide (GLY), Indacaterol acetate (IND) and Mometasone furoate (MOF) in pure form, in laboratory prepared mixtures and in pharmaceutical dosage form. Experimental design methodology was applied by using Plackett-Burman and face-centered composite designs to achieve the best resolution with minimum experimental trials. The designed model was statistically analyzed, graphically presented by surface plots and the relationships between coefficients of the derived polynomial equations were interpreted. Chromatographic separation was achieved on Inertsil ODS C18 column (250 ×4.6 mm, 5 µm) at ambient temperature using a mobile phase composed of methanol: 0.1% glacial acetic acid (pH4) in a gradient elution at a flow rate 1 mL /min. UV detection was carried out at 233 nm. Response was found to be linear in the concentration range of 20-120 µg /mL with regression coefficient (r2 = 0.999) for GLY, 50-300 µg /mL with regression coefficient (r2 = 0.9995) for IND and 50-300 µg /mL with regression coefficient (r2 = 0.9998) for MOF. The method was validated as per ICH guidelines and satisfactory results were achieved. The method was successfully applied for the analysis of the cited drugs in their fixed dose combination (FDC) pharmaceutical formulation. Statistical comparison between the results obtained by the proposed method and the reference methods for GLY, IND and MOF showed no significant difference. The developed method could be implemented in quality control aspects of the cited drugs. Four green metrics were used to evaluate the new RP-HPLC/UV method\'s greenness and compare it to other published techniques.

The production of biopolymers from waste resources is a growing trend, especially in high-population countries like Egypt. Beta-glucan (β-glucan) belongs to natural polysaccharides that are derived from plant and microbial origins. In this study, following increasing demands for β-glucan owing to its bioactive properties, a statistical model to enhance microbial β-glucan production was evaluated for its usefulness to the food and pharmaceutical industries. In addition, a trial to convert β-glucan polymer to nanostructure form was done to increase its bioactivity.
Ingredients of low-cost media based on agro-industrial wastes were described using Plackett-Burman and central composite design of response surface methodology for optimizing yeast β-glucan. Minerals and vitamin concentrations significantly influenced β-glucan yield for Kluyveromyces lactis and nitrogen and phosphate sources for Meyerozyma guilliermondii. The maximum predicted yields of β-glucan recovered from K. lactis and M. guilliermondii after optimizing the medium ingredients were 407 and 1188 mg/100 ml; respectively. For the first time, yeast β-glucan nanoparticles (βGN) were synthesized from the β-glucan polymer using N-dimethylformamide as a stabilizer and characterized using UV-vis spectroscopy, transmission electron microscope (TEM), dynamic light scattering (DLS) and Fourier transform infrared spectroscopy (FT-IR). The average size of βGN was about 300 nm as determined by DLS. The quantitative variation of functional groups between β-glucan polymer and βGN was evaluated by FT-IR for explaining the difference in their biological activity against Normal Homo sapiens-Hela contaminant and Hepatic cancer cell lines.
Enriching the low-cost media based on agro-industrial wastes with nutritional ingredients improves the yield of yeast β-glucan. The present study succeeds to form β-glucan nanoparticles by a simple method.

A fast and straightforward reversed-phase dispersive liquid-liquid microextraction (RP-DLLME) using a deep eutectic solvent (DES) procedure to determine free tryptophan in vegetable oils was developed. The influence of eight variables affecting the RP-DLLME efficiency has been studied by a multivariate approach. A Plackett-Burman design for screening the most influential variables followed by a central composite response surface methodology led to an optimum RP-DLLME setup for a 1 g oil sample: 9 mL hexane as the diluting solvent, vortex extraction with 0.45 mL of DES (choline chloride-urea) at 40 °C, without addition of salt, and centrifugation at 6000 rpm for 4.0 min. The reconstituted extract was directly injected into a high-performance liquid chromatography (HPLC) system working in the diode array mode. At the studied concentration levels, the obtained method detection limits (MDL) was 11 mg/kg, linearity in matrix-matched standards was R2 ≥ 0.997, relative standard deviations (RSD) was 7.8%, and average recovery was 93%. The combined use of the recently developed DES -based RP-DLLME and HPLC provides an innovative, efficient, cost-effective, and more sustainable method for the extraction and quantification of free tryptophan in oily food matrices. The method was employed to analyze cold-pressed oils from nine vegetables (Brazil nut, almond, cashew, hazelnut, peanut, pumpkin, sesame, sunflower, and walnut) for the first time. The results showed that free tryptophan was present in the range of 11-38 mg/100 g. This article is important for its contributions to the field of food analysis, and for its development of a new and efficient method for the determination of free tryptophan in complex matrices, which has the potential to be applied to other analytes and sample types.

The present work proposes a method to characterize, calibrate, and compare, any 2D SLAM algorithm, providing strong statistical evidence, based on descriptive and inferential statistics to bring confidence levels about overall behavior of the algorithms and their comparisons. This work focuses on characterize, calibrate, and compare Cartographer, Gmapping, HECTOR-SLAM, KARTO-SLAM, and RTAB-Map SLAM algorithms. There were four metrics in place: pose error, map accuracy, CPU usage, and memory usage; from these four metrics, to characterize them, Plackett-Burman and factorial experiments were performed, and enhancement after characterization and calibration was granted using hypothesis tests, in addition to the central limit theorem.

Cryopreservation of sperm is a routine technology in many livestock species, but not in swine. Frozen sperm must result in acceptable conception rates and produce 11 to 12 piglets/litter to be competitive with traditional cooled semen. The development of an extender that results in high post-thaw sperm quality and acceptable litter size requires the identification of factors that markedly affect post-thaw semen quality. The present study aims to first identify factors in boar sperm cryopreservation that significantly affect post-thaw sperm quality using an efficient, cost-effective, and relatively rapid approach. The Plackett-Burman experimental design is ideal for the screening of factors at their extreme, greatly reducing the amount of time and resources needed for a follow-up, full factorial design. Using commercial semen, a 9-factor, 12-run Plackett-Burman design was used on 10 boars split between 12 treatments. Through this method, glycerol concentration, cooling rate, antioxidant supplementation with GameteGuard (Membrane Protective Technologies, Inc. Fort Collins, CO), and straw size were identified as highly influential factors that affect post-thaw sperm quality. Extender type, starting osmolality, sodium dodecyl sulfate addition, and stepwise addition of glycerol were also influential for some but not all post-thaw sperm parameters (P < 0.05). Equilibration time in the straws before freezing was determined to have no impact on post-thaw sperm quality parameters. Using the Plackett-Burman design, it can be concluded that four of the nine factors warrant detailed investigation in full factorial experiments in the development of boar sperm cryopreservation extenders.
Freezing of sperm is a routine in many livestock species, but not in pigs, because it results in lower conception in small litter sizes. This study aims to identify factors in pig sperm freezing protocols that affect sperm quality using an efficient and relatively rapid approach. The Plackett–Burman experimental design is one such method used to rapidly screen factors and was the method used for this study. Using commercial pig semen, freezing factors were tested to determine their impact on sperm quality after freezing. Through this method, glycerol concentration (prevents cell damage), cooling rate (speed used to cool sperm to refrigerator temperature), antioxidants, and straw size (straws in which sperm are packaged for freezing) were identified as highly influential factors that affect sperm health. Extender type (the chemical base used to freeze sperm), starting osmolality (the concentration of salts and sugars in a solution), sodium dodecyl sulfate addition (detergent), and stepwise addition of glycerol (to prevent sperm damage) were also influential for some but not all sperm health measures tested. Using the Plackett–Burman design, it can be concluded that four of the nine factors warrant detailed investigation in the development of boar sperm freezing extenders.

Blackcurrant (Ribes nigrum L.) is a fruit rich in vitamins, fatty acids, minerals, essential oils and phenolic compounds, including anthocyanins. In the present work, two anthocyanin extraction methods from blackcurrant samples based on Ultrasound-Assisted Extraction (UAE) and Enzyme-Assisted Extraction (EAE) have been developed. A Plackett-Burman design with seven variables has been preliminary used for both UAE and EAE in order to determine the most influential variables in each methodology. After that, a Box-Behnken design was employed to optimize the extraction methods. The composition of the extraction solvent (% EtOH in water) has been the most influential variable for both UAE and EAE. The optimal extraction times have been 5 min for UAE and 10 min for EAE. No differences have been observed in anthocyanin extraction with both methodologies. Both methods have been applied to blackcurrant-derived products and proven their suitability for quality control analysis.

Cyanobacteria comprise a good natural resource of a potential variety of neuro-chemicals, including acetylcholinesterase inhibitors essential for Alzheimer\'s disease treatment. Accordingly, eight different cyanobacterial species were isolated, identified, and evaluated on their growth on different standard nutrient media. It was found that the modified Navicula medium supported the highest growth of the test cyanobacteria. The effects of methylene chloride/methanol crude extracts of the test cyanobacteria on acetylcholinesterase activity were examined and compared. Anabaena variabilis (KU696637.1) crude extract recorded the highest acetylcholinesterase inhibition (62 ± 1.3%). Navicula medium chemical components were optimized through a Plackett-Burman factorial design. The biomass of Anabaena variabilis increased significantly when grown on the optimized medium compared to that of control. The chemical analysis of the fractions derived from Anabaena variabilis showed the presence of two compounds in significant amounts: the flavonoid 5,7-dihydroxy-2-phenyl-4H-chrome-4-one and the alkaloid 4-phenyl-2-(pyridin-3-yl) quinazoline. Molecular docking studies revealed that both compounds interact with the allosteric binding site of acetylcholinesterase at the periphery with π-π stackings with Tyr341 and Trp286 with good, predicted partition coefficient. The compounds obtained from this study open the door for promising drug candidates to treat Alzheimer\'s disease for their better pharmacodynamics and pharmacokinetic properties.

Rhodotorula yeasts which are known as carotenogenic yeasts have a great industrial value due to their ability to produce carotenoids. In particular, the isolated yeast Rhodotorula sp. (strain ATL72) has been reported to be a promising producer of high concentrations of carotenoids. A combination of central composite design (CCD) and Plackett-Burman (PB) design was used to optimize carotenoids produced by this yeast. The optimum production of carotenoids was completed when the yeast was grown in a production medium composed of 3.7 g/L malt extract, 7.7 g/L fructose, 9 g/L urea, 35 g/L NaCl, and 1 g/L yeast extract at 27.5 °C, pH 6.7, and 180 rpm. Two batch runs in 1 L and 7 L bioreactors were conducted which increased the productivity of carotenoid concentration from 21.5 mg/L after 98 h of incubation at the level of the shake flask to 229.9 mg/L after 47 h of incubation at the level of 7 L bioreactor. The carotenoid pigment was extracted in dimethylsulfoxide (DMSO), acetone, petroleum ether, and sodium chloride, and subsequently identified and characterized using UV-visible scanning, thin layer chromatography, and gas chromatography/mass spectrometry.