CTAB Protocol for Obtaining High-Quality Total RNA from Hibiscus rosa-sinensis and Other Related Plants of the Family Malvaceae.

Malvaceae family, also known as the Mallow family, is a family of flowering plants containing Hibiscus rosa-sinensis and other plants of high medicinal value. This study focuses on the challenges associated with high-quality RNA extraction from Hibiscus rosa-sinensis and its related plants characterized by high levels of mucilage and phenolic compounds in their tissues. High mucilage and secondary metabolite content pose obstacles in obtaining high-quality RNA, negatively impacting downstream applications, such as gene expression analysis. Our research aimed to develop an efficient RNA extraction method tailored to the unique characteristics of Malvaceae family plants especially Hibiscus rosa-sinensis. Through the substitution of NaCl with KCl, a crucial component of the CTAB buffer, our methodology successfully addressed the challenges posed by high mucilage and phenolic compound levels. This modification led to a significant reduction in sample viscosity, which is because of the high mucilage in these plants. Our modified CTAB extraction method yielded significantly more RNA with higher purity than the conventional CTAB methods alone. The extracted RNA was largely intact, as indicated by 28S/18S ratios and RIN values, yielding high-quality RNA with improved purity suggested by the 260/280 and 260/230 ratios. The proposed approach not only serves as a solution to the specific challenges encountered in Hibiscus rosa-sinensis but also holds promise for broader applications across different plants within the family.

Simultaneous production of biofuel, and removal of heavy metals using marine alga Turbinaria turbinata as a feedstock in NEOM Region, Tabuk.

Depletion of fossil fuel and pollution by heavy metals are two major global issues. The cell wall of algae consists of polymers of polysaccharides such as cellulose, hemicellulose, alginate, starch, and many others that are readily hydrolyzed to monosaccharides and hence are amenable to fermentation into bioethanol. Moreover, algae contain lipids that may undergo trans-esterification to biodiesel, and can be absorbed by heavy metals. In this study, extraction of lipids from Turbinaria turbinata (common brown alga) from the beach of Sharma, NEOM, Tabuk, Saudi Arabia by different solvents hexane, methanol, and hexane: methanol (1:1), and trans-esterification was performed to obtain biodiesel and investigated by GC.MS. The alga residue after fats extractions by different solvents was used in bioremediation synthetic wastewater containing 50 ppm of As-3, Co+2, Cu+2, Fe+2, Mn+2, and Zn+2. The residue of defatted alga was hydrolyzed by 2% H2SO4 and then fermented to obtain bioethanol. The combination of hexane: methanol (1:1) gave the greatest amount of petroleum hydrocarbons, which contain Tetradecane, 5-methyl, Octacosane, Pentatriacontane, and a small amount of Cyclotrisiloxane, Hexamethyl. The most effective removal % was obtained with alga residue defatted by hexane: methanol (1:1), and methanol, 100% removal of As-3, 83% Co+2, 95% Cu+2, 97.25% Fe+2, Mn+2 79.69%, Zn+2 90.15% with 2 g alga /L at 3 hours. The lowest value of sugar was obtained with hexane: methanol residue but gave the highest bioethanol efficiency. Thus, it is possible to use Turbinaria turbinata, or brown alga as a feedstock to produce bio-diesel, and bioethanol, and to remove heavy metals from wastewater, which may have a great economic and environmental significance.

Biogenic Nanoparticles Silver and Copper and Their Composites Derived from Marine Alga Ulva lactuca: Insight into the Characterizations, Antibacterial Activity, and Anti-Biofilm Formation.

Bacterial pathogens cause pain and death, add significantly to the expense of healthcare globally, and pose a serious concern in many aspects of daily life. Additionally, they raise significant issues in other industries, including pharmaceuticals, clothing, and food packaging. Due to their unique properties, a great deal of attention has been given to biogenic metal nanoparticles, nanocomposites, and their applications against pathogenic bacteria. This study is focused on biogenic silver and copper nanoparticles and their composites (UL/Ag2 O-NPS, Ul/CuO-NPs, and Ul/Ag/Cu-NCMs) produced by the marine green alga Ulva lactuca. The characterization of biogenic nanoparticles UL/Ag2 O-NPS and Ul/CuO-NPs and their composites Ul/Ag/Cu-NCMs has been accomplished by FT-IR, SEM, TEM, EDS, XRD, and the zeta potential. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) experiments were conducted to prove antibacterial activity against both Gram-positive and Gram-negative bacteria and anti-biofilm. The FTIR spectroscopy results indicate the exiting band at 1633 cm-1, which represents N-H stretching in nanocomposites, with a small shift in both copper and silver nanoparticles, which is responsible for the bio-reduction of nanoparticles. The TEM image reveals that the Ul/Ag/Cu-NCMs were hexagonal, and the size distribution ranged from 10 to 35 nm. Meanwhile, Ul/CuO-NPs are rod-shaped, whereas UL/Ag2 O-NPS are spherical. The EDX analysis shows that Cu metal was present in a high weight percentage over Ag in the case of bio-Ag/Cu-NCMs. The X-ray diffraction denotes that Ul/Ag/Cu-NCMs, UL/CuO-NPs, and UL/Ag2 O-NPS were crystalline. The results predicted by the zeta potential demonstrate that Ul/Ag/Cu-NCMs were more stable than Ul/CuO-NPs. The antibacterial activity of UL/Ag2 O-NPS, Ul/Ag/Cu-NCMs, and UL/CuO-NPs was studied against eleven Gram-negative and Gram-positive multidrug-resistant bacterial species. The maximum inhibition zones were obtained with UL/Ag2 O-NPS, followed by Ul/Ag/Cu-NCMs and Ul/CuO-NPs in all the tested bacteria. The maximum anti-biofilm percentage formed by E. coli KY856933 was obtained with UL/Ag2 O-NPS. These findings suggest that the synthesized nanoparticles might be a great alternative for use as an antibacterial agent against different multidrug-resistant bacterial strains.

Pseudomonas alcaliphila NEWG-2 as biosorbent agent for methylene blue dye: optimization, equilibrium isotherms, and kinetic processes.

In comparison to physicochemical and chemical methods, microbial dye biosorption is regarded as an eco-effective and economically viable alternative and is a widely applied method due to its high efficiency and compatibility with the environment. Therefore, the idea of this study is to clarify to what extent the viable cells and the dry biomass of Pseudomonas alcaliphila NEWG-2 can improve the biosorption of methylene blue (MB) from a synthetic wastewater sample. The array of Taguchi paradigm has been conducted to ascertain five variables affecting the biosorption of MB by broth forms of P. alcaliphila NEWG. The data of MB biosorption were familiar to the predicted ones, indicating the precision of the Taguchi model's prediction. The maximum biosorption of MB (87.14%) was achieved at pH 8, after 60 h, in a medium containing 15 mg/ml MB, 2.5% glucose, and 2% peptone, with sorting the highest signal-to-noise ratio (38.80). FTIR spectra detected various functional groups (primary alcohol, α, ß-unsaturated ester, symmetric NH2 bending, and strong C-O stretching) on the bacterial cell wall that participated in the biosorption of MB. Furthermore, the spectacular MB biosorption ability was validated by equilibrium isotherms and kinetic studies (the dry biomass form), which were derived from the Langmuir model (qmax = 68.827 mg/g). The equilibrium time was achieved in about 60 min, with 70.5% of MB removal. The biosorption kinetic profile might be adequately represented by pseudo-second order and Elovich models. The changes in the bacterial cells before and after the biosorption of MB were characterized using a scanning electron microscope. As realized from the aforementioned data, the bacterium is a talented, effective, eco-friendly, and low-cost bio-sorbent for the decolorization and remedy of an industrial effluent containing MB from an aqueous environment. The current outcomes in the biosorption of MB molecules promote the use of the bacterial strain as viable cells and/or dry biomass in ecosystem restoration, environmental cleanup, and bioremediation studies.

Dental Composites with Magnesium Doped Zinc Oxide Nanoparticles Prevent Secondary Caries in the Alloxan-Induced Diabetic Model.

Antibacterial restorative materials against caries-causing bacteria are highly preferred among high-risk patients, such as the elderly, and patients with metabolic diseases such as diabetes. This study aimed to enhance the antibacterial potential of resin composite with Magnesium-doped Zinc oxide (Mg-doped ZnO) nanoparticles (NPs) and to look for their effectiveness in the alloxan-induced diabetic model. Hexagonal Mg-doped ZnO NPs (22.3 nm diameter) were synthesized by co-precipitation method and characterized through ultraviolet-visible (UV-Vis), Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), scanning electron microscopy (SEM), and energy dispersive spectroscopy (EDS) analysis. The Mg-doped ZnO NPs (1, 2.5 and 5% w/w) were then evaluated for antibacterial activity using a closed system in vitro biofilm model. Significant enhancement in the antibacterial properties was observed in composites with 1% Mg-doped ZnO compared to composites with bare ZnO reinforced NPs (Streptococcus mutans, p = 0.0005; Enterococcus faecalis, p = 0.0074, Saliva microcosm, p < 0.0001; Diabetic Saliva microcosm, p < 0.0001). At 1−2.5% Mg-doped ZnO NPs concentration, compressive strength and biocompatibility of composites were not affected. The pH buffering effect was also achieved at these concentrations, hence not allowing optimal conditions for the anaerobic bacteria to grow. Furthermore, composites with Mg-doped ZnO prevented secondary caries formation in the secondary caries model of alloxan-induced diabetes. Therefore, Mg-doped ZnO NPs are highly recommended as an antibacterial agent for resin composites to avoid biofilm and subsequent secondary caries formation in high-risk patients.

Effectiveness of Se/ZnO NPs in Enhancing the Antibacterial Activity of Resin-Based Dental Composites.

Biofilm formation in the resin-composite interface is a major challenge for resin-based dental composites. Using doped z nanoparticles (NPs) to enhance the antibacterial properties of resin composites can be an effective approach to prevent this. The present study focused on the effectiveness of Selenium-doped ZnO (Se/ZnO) NPs as an antibacterial nanofiller in resin composites and their impact on their mechanical properties. Pristine and Se/ZnO NPs were synthesized by the mechanochemical method and confirmed through UV-Vis Spectroscopy, FTIR (Fourier Transform Infrared) analysis, X-ray Diffraction (XRD) crystallography, Scanning Electron Microscopy (SEM), Energy Dispersive Spectroscopy (EDS), and Zeta analysis. The resin composites were then modified by varying concentrations of pristine and Se/ZnO NPs. A single species (S. mutans and E. faecalis) and a saliva microcosm model were utilized for antibacterial analysis. Hemolytic assay and compressive strength tests were also performed to test the modified composite resin's cytotoxicity and mechanical strength. When incorporated into composite resin, 1% Se/ZnO NPs showed higher antibacterial activity, biocompatibility, and higher mechanical strength when compared to composites with 1% ZnO NPs. The Se/ZnO NPs has been explored for the first time as an efficient antibacterial nanofiller for resin composites and showed effectiveness at lower concentrations, and hence can be an effective candidate in preventing secondary caries by limiting biofilm formation.

Innovative binary sorption of Cobalt(II) and methylene blue by Sargassum latifolium using Taguchi and hybrid artificial neural network paradigms.

The present investigation has been designed by Taguchi and hybrid artificial neural network (ANN) paradigms to improve and optimize the binary sorption of Cobalt(II) and methylene blue (MB) from an aqueous solution, depending on modifying physicochemical conditions to generate an appropriate constitution for a highly efficient biosorption by the alga; Sargassum latifolium. Concerning Taguchi's design, the predicted values of the two responses were comparable to actual ones. The biosorption of Cobalt(II) ions was more efficient than MB, the supreme biosorption of Cobalt(II) was verified in run L21 (93.28%), with the highest S/N ratio being 39.40. The highest biosorption of MB was reached in run L22 (74.04%), with a S/N ratio of 37.39. The R2 and adjusted R2 were in reasonable values, indicating the validity of the model. The hybrid ANN model has exclusively emerged herein to optimize the biosorption of both Cobalt(II) and MB simultaneously, therefore, the ANN model was better than the Taguchi design. The predicted values of Cobalt(II) and MB biosorption were more obedience to the ANN model. The SEM analysis of the surface of S. latifolium showed mosaic form with massive particles, as crosslinking of biomolecules of the algal surface in the presence of Cobalt(II) and MB. Viewing FTIR analysis showed active groups e.g., hydroxyl, α, ß-unsaturated ester, α, ß-unsaturated ketone, N-O, and aromatic amine. To the best of our knowledge, there are no reports deeming the binary sorption of Cobalt(II) and MB ions by S. latifolium during Taguchi orthogonal arrays and hybrid ANN.

Characterizations of Alpha-Cellulose and Microcrystalline Cellulose Isolated from Cocoa Pod Husk as a Potential Pharmaceutical Excipient.

Cellulose is a non-toxic, bio-degradable, and renewable biopolymer which is abundantly available in nature. The most common source of commercial microcrystalline cellulose is fibrous wood pulp. Cellulose and its derivatives have found wide commercial applications in the pharmaceutical, cosmetic, food, paper, textile, and engineering industries. This study aims to isolate and characterize cellulose forms from cocoa pod husk (CPH) and to assess its mechanical and disintegration properties as a direct compression excipient in metronidazole tablets. Two isolated cellulose types (i.e., cocoa alpha-cellulose (CAC) and cocoa microcrystalline cellulose (C-MCC)) were compared with avicel (AV). CAC and C-MCC were characterized for their physicochemical properties using Scanning Electron Microscopy (SEM), FTIR spectroscopy, Differential Scanning Calorimetry (DSC), and X-Ray Powder Diffraction (XRD). Metronidazole tablets were produced by direct compression with cellulose. The mechanical and disintegration properties of the tablets were evaluated. CAC and C-MCC yield was 42.3% w/w and 38.25% w/w, respectively. Particle diameters were significantly different with CAC (282.22 µm) > C-MCC (161.32 µm) > AV (72.51 µm). CAC and C-MCC had a better flow than AV. SEM revealed the fibrous nature of the cellulose. FTIR and XRD analysis confirmed the presence of cellulose with crystallinity index of 69.26%, 43.83%, and 26.32% for AV, C-MCC, and CAC, respectively. C-MCC and AV are more crystalline and thermally stable at high temperatures compared to CAC. The mechanical and disintegration properties of C-MCC and AV tablets complied with pharmacopeia specifications. Taken together, C-MCC isolated from CPH displayed some fundamental characteristics suitable for use as a pharmaceutical excipient and displayed better properties compared to that of AV.

Assessment of Antioxidant and Anticancer Activities of Microgreen Alga Chlorella vulgaris and Its Blend with Different Vitamins.

There is a very vital antioxidant extracted from microgreen alga. Chlorella vulgaris has major advantages and requires high yield worldwide. Some microalgae require vitamins for their growth promotion. This study was held to determine the impact of different vitamins including Thiamine (B1), Riboflavin (B2), Pyridoxine (B6), and Ascorbic acid (c) at concentrations of 0.02, 0.04, 0.06, and 0.08 mg/L of each. Each vitamin was added to the BG11 growth medium to determine the effect on growth, total carbohydrate, total protein, pigments content, antioxidant activities of Chlorella vulgaris. Moreover, antitumor effects of methanol extract of C. vulgaris without and with the supplement of thiamine against Human prostate cancer (PC-3), Hepatocellular carcinoma (HEPG-2), Colorectal carcinoma (HCT-116) and Epitheliod Carcinoma (Hela) was estimated in vitro. C. vulgaris supplemented with various vitamins showed a significant increase in biomass, pigment content, total protein, and total carbohydrates in comparison to the control. Thiamine was the best vitamin influencing as an antioxidant. C. vulgaris supplemented with thiamine had high antitumor effects in vitro. So, it's necessary to add vitamins to BG11 media for enhancement of the growth and metabolites.

Characterization and Anticancer Activity of Biosynthesized Au/Cellulose Nanocomposite from Chlorella vulgaris.

Therapeutic selectivity is a critical issue in cancer therapy. As a result of its adjustable physicochemical characteristics, the Au/cellulose nanocomposite currently holds a lot of potential for solving this challenge. This work was designed to prepare a Au/cellulose nanocomposite with enhanced anticancer activity through the regulation of the mitogen-activated protein kinases (MAPK) signaling pathway. Nanocellulose, nanogold (AuNPs), and a Au/cellulose nanocomposite were biosynthesized from microgreen alga Chlorella vulgaris. Using UV-Vis absorption spectroscopy, transmission electron microscope (TEM), zeta potential analyzer, and Fourier transform infrared spectroscopy (FTIR), the synthesized nanoparticles were confirmed and characterized. In human alveolar basal epithelial cells (A549 cells), the selectivity and anticancer activity of the produced nanoparticles were evaluated. The cytotoxicity results revealed that the inhibitory concentration (IC50) of the Au/cellulose nanocomposite against A549 cancer lung cells was 4.67 ± 0.17 µg/µL compared to 182.75 ± 6.45 µg/µL in the case of HEL299 normal lung fibroblasts. It was found that treatment with nanocellulose and the Au/cellulose nanocomposite significantly increased (p < 0.05) the relative expression of tumor suppressor 53 (p53) in comparison to control cells. They also significantly (p < 0.05) decreased the relative expression of the Raf-1 gene. These findings indicate that nanocellulose and the Au/cellulose nanocomposite regulate cell cycles mostly via the motivation of p53 gene expression and reduction of Raf-1 gene expression.