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1.
ACS Omega ; 9(23): 25189-25202, 2024 Jun 11.
Article in English | MEDLINE | ID: mdl-38882090

ABSTRACT

Consumer demand for plant-based alternatives drives innovation in nondairy beverages. This study explores the development of a novel sesame milk with enhanced functionality using an artificial neural network (ANN) and milk permeate integration. An ANN model effectively optimized water-based sesame milk (WSM) extraction, maximizing total solids (T.S.) recovery. The ANN model's predicted T.S. yield (99.65%) closely matched the actual value (95.18%), demonstrating its potential for optimizing high-yield production. Furthermore, milk permeate was incorporated (5:1 ratio) to create permeate-based sesame milk (PSM), which supported the growth of lactic acid bacteria, suggesting its potential as a growth medium for future probiotic applications. PSM also displayed superior nutritional value and sensory characteristics compared to WSM. These findings highlight the promise of ANN-powered optimization and milk permeate integration for creating innovative sesame milk alternatives with enhanced probiotic viability and sensory appeal. Future research should focus on ANN optimization of alternative-based-plant milk, including permeate-based sesame milk production, the health benefits of LAB fermentation, and consumer preferences for flavors and textures. Optimizing fermentation and LAB selection remain key for commercial success.

2.
Heliyon ; 10(7): e28359, 2024 Apr 15.
Article in English | MEDLINE | ID: mdl-38560145

ABSTRACT

Due to increasing concerns about environmental impact and toxicity, developing green and sustainable methods for nanoparticle synthesis is attracting significant interest. This work reports the successful green synthesis of silver (Ag), silver-titanium dioxide (Ag@TiO2), and silver-selenium dioxide (Ag@SeO2) nanoparticles (NPs) using Beta vulgaris L. extract. Characterization by XRD, SEM, TEM, and EDX confirmed the successful formation of uniformly distributed spherical NPs with controlled size (25 ± 4.9 nm) and desired elemental composition. All synthesized NPs and the B. vulgaris extract exhibited potent free radical scavenging activity, indicating significant antioxidant potential. However, Ag@SeO2 displayed lower hemocompatibility compared to other NPs, while Ag@SeO2 and the extract demonstrated reduced inflammation in a carrageenan-induced paw edema animal model. Interestingly, Ag@TiO2 and Ag@SeO2 exhibited strong antifungal activity against Rhizoctonia solani and Sclerotia sclerotium, as evidenced by TEM and FTIR analyses. Generally, the findings suggest that B. vulgaris-derived NPs possess diverse biological activities with potential applications in various fields such as medicine and agriculture. Ag@TiO2 and Ag@SeO2, in particular, warrant further investigation for their potential as novel bioactive agents.

3.
RSC Adv ; 14(10): 7088-7111, 2024 Feb 21.
Article in English | MEDLINE | ID: mdl-38414990

ABSTRACT

This study bio-synthesized Ag@SeO2 bmNPs successfully, using turmeric ethanol extract, and characterized them using various techniques. The FT-IR analysis reveals the involvement of these plant-derived compounds, especially phenolics, in the reduction process by acting as electron donors and stabilizing/capping agents. Zeta potential analysis showed a slight negative surface charge for the stability of Ag@SeO2 NPs, where TEM revealed spherical nanoparticles with an average size of 20 nm. The XRD confirmed crystallinity and a core-shell structure, and EDX identified elements consistent with Ag@SeO2 and a 3 : 1 Ag/Se atomic ratio. Further, SEM supported the spherical shape and uniform size. These findings highlight the successful biosynthesis of Ag@SeO2 bmNPs with promising properties for diverse applications. Moreover, the Box-Behnken design (BBD) and artificial neural network (ANN) model were engaged to optimize Ag@SeO2 bmNP biosynthesis. BBD identified significant influences of pH, bioconversion temperature, time, and turmeric concentration on bmNP yield, with adjusted R2 and predictive R2 being 0.9075 and 0.8829, respectively. However, its limitations were revealed by a significant lack of fit. ANN modeling with a 3-5-7-1 topology showed superior predictive accuracy and identified optimal conditions for maximizing yield (pH 9.83, 51.7 °C, 1.0 h, 3.71 mg mL-1 turmeric). Validation experiments confirmed the model's reliability. Turmeric extract exhibited significantly higher amounts of phenolics, and flavonoids compared to the bmNPs, suggesting its potential for strong antioxidant activity. Both turmeric extract and bmNPs displayed antioxidant activity in ABTS and DPPH assays, with turmeric extract being the most potent due to its curcuminoid content. The potential activity of Ag@SeO2 bmNPs against S. aureus, K. pneumonia, E. coli, and B. cereus was investigated, with inhibition zones ranging from 22 to 32 mm. The MIC values of tested NPs towards pathogenic bacteria ranged from 165.625 and 331.25 µg mL-1.

4.
Heliyon ; 9(9): e20063, 2023 Sep.
Article in English | MEDLINE | ID: mdl-37809880

ABSTRACT

Biodiesel is a sustainable, and renewable alternative to fossil fuels that can be produced from various biological sources with the aid of lipases. This study developed a simple and novel fungal system for lipase biosynthesis to be used for catalyzing the oily residuals into biodiesel, employing the artificial neural network (ANN), and semi-solid-state fermentation (SSSF). Nigella sativa was selected among agro-industrial oily residuals as a substrate for lipase biosynthesis by Aspergillus flavipes MH47297. The effect of cultural humidity (X1), the surfactant; Brij 35 (X2), and inoculum density (X3) on lipase biosynthesis were researched based on the matrix of Box-Behnken design (BBD). The ANN together with a new fungal candidate and SSSF were then applied for the first time to model the biosynthesis process of lipase. The optimum predicted cultural conditions varied according to the model. The optimum predicted conditions were estimated separately by BBD (X1 = 5.8 ml water/g, X2 = 46.6 µl/g, and X3 = 62156610 spore/g) and ANN (X1 = 5.4 ml water/g, X2 = 54.2 µl/g, and X3 = 100000000 spore/g) models. Based on the modeling process, the response of lipase was calculated to be 214.95 (BBD) and 217.72 U (ANN), which revealed high consistency with the experimental lipase yield (209.13 ± 3.27 U for BBD, and 218 ± 2.01 U for ANN). Despite both models showing high accuracy, ANN was more accurate and surpassed the BBD model. Gas chromatography analysis showed that lipase successfully converted corn oil to biodiesel (29.5 mg/l).

5.
Polymers (Basel) ; 15(18)2023 Sep 14.
Article in English | MEDLINE | ID: mdl-37765609

ABSTRACT

Melanin as a natural polymer is found in all living organisms, and plays an important role in protecting the body from harmful UV rays from the sun. The efficiency of fungal biomass (Aureobasidium pullulans) and its extracellular melanin as Cr(VI) biosorbents was comparatively considered. The efficiency of Cr(VI) biosorption by the two sorbents used was augmented up to 240 min. The maximum sorption capacities were 485.747 (fungus biomass) and 595.974 (melanin) mg/g. The practical data were merely fitted to both Langmuir and Freundlich isotherms. The kinetics of the biosorption process obeyed the pseudo-first-order. Melanin was superior in Cr(VI) sorption than fungal biomass. Furthermore, four independent variables (contact time, initial concentration of Cr(VI), biosorbent dosage, and pH,) were modeled by the two decision trees (DTs). Conversely, to equilibrium isotherms and kinetic studies, DT of fungal biomass had lower errors compared to DT of melanin. Lately, the DTs improved the efficacy of the Cr(VI) removal process, thus introducing complementary and alternative solutions to equilibrium isotherms and kinetic studies. The Cr(VI) biosorption onto the biosorbents was confirmed and elucidated through FTIR, SEM, and EDX investigations. Conclusively, this is the first report study attaining the biosorption of Cr(VI) by biomass of A. pullulans and its extracellular melanin among equilibrium isotherms, kinetic study, and algorithmic decision tree modeling.

6.
Microorganisms ; 11(8)2023 Jul 31.
Article in English | MEDLINE | ID: mdl-37630517

ABSTRACT

The high prevalence of fungal resistance to antifungal drugs necessitates finding new antifungal combinations to boost the antifungal bioactivity of these agents. Hence, the aim of the present investigation was to greenly synthesize zinc oxide nanoparticles (ZnO-NPs) using an aqueous leaf extract of Salvia officinalis and investigate their antifungal activity and synergistic efficiency with common antifungal agents. The biofabricated ZnO-NPs were characterized to detect their physicochemical properties. A disk diffusion assay was employed to investigate the antifungal effectiveness of the greenly synthesized ZnO-NPs and evaluate their synergistic patterns with common antifungal agents. The Candida tropicalis strain was detected to be the most susceptible strain to ZnO-NPs at both tested concentrations of 50 and 100 µg/disk, demonstrating relative suppressive zones of 19.68 ± 0.32 and 23.17 ± 0.45 mm, respectively. The minimum inhibitory concentration (MIC) of ZnO-NPs against the C. tropicalis strain was 40 µg/mL, whereas the minimum fungicidal concentration (MFC) was found to be 80 µg/mL. The highest synergistic efficiency of the biogenic ZnO-NPs with terbinafine antifungal agent was detected against the C. glabrata strain, whereas the highest synergistic efficiency was detected with fluconazole against the C. albicans strain, demonstrating relative increases in fold of inhibition area (IFA) values of 6.82 and 1.63, respectively. Moreover, potential synergistic efficiency was detected with the nystatin antifungal agent against the C. tropicalis strain with a relative IFA value of 1.06. The scanning electron microscopy (SEM) analysis affirmed the morphological deformations of candidal cells treated with the biosynthesized ZnO-NPs as the formation of abnormal infoldings of the cell wall and membranes and also the formation of pores in the cell wall and membranes, which might lead to the leakage of intracellular constituents. In conclusion, the potential synergistic efficiency of the biogenic ZnO-NPs with terbinafine, nystatin, and fluconazole against the tested candidal strains highlights the potential application of these combinations in formulating novel antifungal agents of high antimicrobial efficiency. The biogenic ZnO nanoparticles and antifungal drugs exhibit powerful synergistic efficiency, which highlights their prospective use in the formulation of efficient antimicrobial medications, including mouthwash, ointments, lotions, and creams for effective candidiasis treatment.

7.
Sci Rep ; 13(1): 13545, 2023 08 19.
Article in English | MEDLINE | ID: mdl-37598271

ABSTRACT

The effect of three independent variables (i.e., tyrosine, sucrose, and incubation time) on melanin production by Aureobasidium pullulans AKW was unraveled by two distinctive approaches: response surface methodology (i.e. Box Behnken design (BBD)) and artificial neural network (ANN) in this study for the first time ever using a simple medium. Regarding BBD, sucrose and incubation intervals did impose a significant influence on the output (melanin levels), however, tyrosine did not. The validation process exhibited a high consistency of BBD and ANN paradigms with the experimental melanin production. Concerning ANN, the predicted values of melanin were highly comparable to the experimental values, with minor errors competing with BBD. Highly comparable experimental values of melanin were achieved upon using BBD (9.295 ± 0.556 g/L) and ANN (10.192 ± 0.782 g/L). ANN accurately predicted melanin production and showed more improvement in melanin production by about 9.7% higher than BBD. The purified melanin structure was verified by scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDX), X-ray diffraction pattern (XRD), and thermogravimetric analysis (TGA). The results verified the hierarchical architecture of the particles as small compasses by SEM analysis, inter-layer spacing in the XRD analysis, maximal atomic % for carbon, and oxygen atoms in the EDX analysis, and the great thermal stability in the TGA analysis of the purified melanin. Interestingly, the current novel endophytic strain was tyrosine-independent, and the uniquely applied ANN paradigm was more efficient in modeling the melanin production with appreciate amount on a simple medium in a relatively short time (168 h), suggesting additional optimization studies for further maximization of melanin production.


Subject(s)
Melanins , Tyrosine , Neural Networks, Computer , Sucrose
8.
Microorganisms ; 11(4)2023 Apr 04.
Article in English | MEDLINE | ID: mdl-37110368

ABSTRACT

Nosocomial bacterial and fungal infections are one of the main causes of high morbidity and mortality worldwide, owing to the high prevalence of multidrug-resistant microbial strains. Hence, the study aims to synthesize, characterize, and investigate the antifungal and antibacterial activity of silver nanoparticles (AgNPs) fabricated using Camellia sinensis leaves against nosocomial pathogens. The biogenic AgNPs revealed a small particle diameter of 35.761 ± 3.18 nm based on transmission electron microscope (TEM) graphs and a negative surface charge of -14.1 mV, revealing the repulsive forces between nanoparticles, which in turn indicated their colloidal stability. The disk diffusion assay confirmed that Escherichia coli was the most susceptible bacterial strain to the biogenic AgNPs (200 g/disk), while the lowest sensitive strain was found to be the Acinetobacter baumannii strain with relative inhibition zones of 36.14 ± 0.67 and 21.04 ± 0.19 mm, respectively. On the other hand, the biogenic AgNPs (200 µg/disk) exposed antifungal efficacy against Candida albicans strain with a relative inhibition zone of 18.16 ± 0.14 mm in diameter. The biogenic AgNPs exposed synergistic activity with both tigecycline and clotrimazole against A. baumannii and C. albicans, respectively. In conclusion, the biogenic AgNPs demonstrated distinct physicochemical properties and potential synergistic bioactivity with tigecycline, linezolid, and clotrimazole against gram-negative, gram-positive, and fungal strains, respectively. This is paving the way for the development of effective antimicrobial combinations for the effective management of nosocomial pathogens in intensive care units (ICUs) and health care settings.

9.
Microorganisms ; 11(3)2023 Mar 02.
Article in English | MEDLINE | ID: mdl-36985218

ABSTRACT

The drug resistance of bacterial pathogens causes considerable morbidity and death globally, hence there is a crucial necessity for the development of effective antibacterial medicines to address the antibacterial resistance issue. The bioprepared zinc oxide nanoparticles (ZnO-NPs) were prepared utilizing the flower extract of Hibiscus sabdariffa and then characterized using different physicochemical techniques. The antibacterial effectiveness of the bioprepared ZnO-NPs and their synergism with fosfomycin were evaluated using disk diffusion assay against the concerned pathogens. Transmission electron microscopy (TEM) investigation of the bioprepared ZnO-NPs showed that their average particle size was 18.93 ± 2.65 nm. Escherichia coli expressed the highest sensitivity to the bioinspired ZnO-NPs with a suppressive zone of 22.54 ± 1.26 nm at a concentration of 50 µg/disk, whereas the maximum synergistic effect of the bioinspired ZnO-NPs with fosfomycin was noticed against Klebsiella pneumoniae strain with synergism ratio of 100.29%. In conclusion, the bioinspired ZnO-NPs demonstrated significant antibacterial and synergistic efficacy with fosfomycin against the concerned nosocomial bacterial pathogens, highlighting the potential of using the ZnO NPs-fosfomycin combination for effective control of nosocomial infections in intensive care units (ICUs) and health care settings. Furthermore, the biogenic ZnO-NPs' potential antibacterial action against food pathogens such as Salmonella typhimurium and E. coli indicates their potential usage in food packaging applications.

10.
Microorganisms ; 10(9)2022 Sep 10.
Article in English | MEDLINE | ID: mdl-36144416

ABSTRACT

Ralstonia solanacearum is one of the globally significant plant pathogens that infect a wide host range of economically important plants. A study was conducted to evaluate the hypothesis that an avirulent strain of R. solanacearum can act as a biocontrol mediator for managing potato bacterial wilt. Virulent R. solanacearum was isolated and identified (GenBank accession number; OP180100). The avirulent strain was obtained from the virulent strain through storage for 3 weeks until the development of deep red colonies. The virulent strain had higher lytic activity than the avirulent strain. Tubers' treatments by the avirulent strain of R. solanacearum, (supernatant, boiled supernatant, and dead cells) significantly reduced plant disease rating and increased the growth, physiological activities, and biomass of potato compared to the untreated, infected control. The major components detected by GC-MS in the supernatant revealed 10.86% palmitic acid (virulent), and 18.03% 1,3-dioxolane, 2,4,5-trimethyl- (avirulent), whereas the major component in the boiled supernatant was 2-hydroxy-gamma-butyrolactone in the virulent (21.17%) and avirulent (27.78%) strains. This is the first research that assessed the influence of boiled supernatant and dead cells of virulent and avirulent R.solanacearum strains in controlling bacterial wilt disease. Additional work is encouraged for further elucidation of such a topic.

11.
Environ Res ; 212(Pt C): 113421, 2022 09.
Article in English | MEDLINE | ID: mdl-35568233

ABSTRACT

The current study aimed to isolate biodegradable soil fungi capable of metabolizing diazinon. The collected soil samples were investigated for diazinon pollution to detect the pesticide level in the polluted soil samples. Food poisoning techniques were utilized to preliminary investigate the biodegradation efficiency of the isolated fungal strains to diazinon pesticide using solid and liquid medium and also to detect their tolerance to different concentrations. GC-MS analysis of control and treated flasks were achieved to determine the diazinon residues for confirmation of the biodegradation efficiency. The total diazinon residues in the collected soil samples was found to be 0.106 mg/kg. Out of thirteen fungal strains isolated form diazinon polluted soils, six strains were potentially active in diazinon biodegradation. Food poisoning technique showed that A. niger, B. antennata, F. graminearum, P. digitatum, R. stolonifer and T. viride strains recorded fungal growth diameters of 65.2 ± 0.18, 57.5 ± 0.41, 47.2 ± 0.36, 56.5 ± 0.27, 85.0 ± 0.01, 85.0 ± 0.06 mm respectively in the treated group which were non significantly different compared to that of control (P > 0.05), indicating the high efficiency of these strains in diazinon degradation compared to the other isolated strains. GC-MS analysis revealed that B. antennata was the most efficient strain in diazinon degradation recording 32.24 ± 0.15 ppm concentration after 10 days incubation. Linear regression analysis confirmed that B. antennata was the most effective biodegradable strain recording the highest diazinon dissipation (83.88%) with the lowest T1/2 value of 5.96 days while T. viride, A. niger, R. stolonifer and F. graminearum exhibited a high biodegradable activities reducing diazinon to 80.26%, 78.22%, 77.36% and 75.43% respectively after 10 days incubation. In conclusion, these tolerant fungi could be considered as promising, eco-friendly and biodegradable fungi for the efficient and potential removal of hazardous diazinon from polluted soil.


Subject(s)
Foodborne Diseases , Pesticides , Soil Pollutants , Biodegradation, Environmental , Diazinon/analysis , Diazinon/chemistry , Diazinon/metabolism , Fungi , Pesticides/analysis , Soil/chemistry , Soil Microbiology , Soil Pollutants/analysis
12.
Front Microbiol ; 13: 772417, 2022.
Article in English | MEDLINE | ID: mdl-35401430

ABSTRACT

Plant residuals comprise the natural habitat of the plant pathogen; therefore, attention is currently focusing on biological-based bioprocessing of biomass residuals into benefit substances. The current study focused on the biodegradation of peanut plant residual (PNR) into citric acid (CA) through a mathematical modeling strategy. Novel endophytic Trichoderma longibrachiatum WKA55 (GenBank accession number: MZ014020.1), having lytic (cellulase, protease, and polygalacturonase) activity, and tricalcium phosphate (TCP) solubilization ability were isolated from peanut seeds and used during the fermentation process. As reported by HPLC, the maximum CA (5505.1 µg/g PNR) was obtained after 9 days in the presence of 15.49 mg TCP, and 15.68 mg glucose. GC-MS analysis showed other bioactive metabolites in the filtrate of the fermented PNR. Practically, the crude product (40%) fully inhibited (100%) the growth and spore germination of three mycotoxinogenic fungi. On peanuts, it improved the seed germination (91%), seedling features, and vigor index (70.45%) with a reduction of abnormal seedlings (9.33%). The current study presents the fundamentals for large-scale production in the industry for the sustainable development of PNR biomass as a natural source of bioactive metabolites, and safe consumption of lignocellulosic-proteinaceous biomass, as well. T. longibrachiatum WKA55 was also introduced as a novel CA producer specified on PNR. Application of the resulting metabolite is encouraged on a large scale.

14.
Plants (Basel) ; 10(2)2021 Feb 23.
Article in English | MEDLINE | ID: mdl-33672364

ABSTRACT

During preservation, Jerusalem artichoke (JA) tubers are subjected to deterioration by mold fungi under storage, which signifies a serious problem. A new blue mold (Penicillium polonium) was recorded for the first time on JA tubers. Penicillium mold was isolated, identified (morphologically, and molecularly), and deposited in GenBank; (MW041259). The fungus has a multi-lytic capacity, facilitated by various enzymes capable of severely destroying the tuber components. An economic oil-based procedure was applied for preserving and retaining the nutritive value of JA tubers under storage conditions. Caraway and clove essential oils, at a concentration of 2%, were selected based on their strong antifungal actions. JA tubers were treated with individual oils under storage, kept between peat moss layers, and stored at room temperature. Tubers treated with both oils exhibited lower blue mold severity, sprouting and weight loss, and higher levels of carbohydrates, inulin, and protein contents accompanied by increased levels of defense-related phytochemicals (total phenols, peroxidase, and polyphenol oxidase). Caraway was superior, but the results endorse the use of both essential oils for the preservation of JA tubers at room temperature, as an economic and eco-safe storage technique against the new blue mold.

15.
Article in English | MEDLINE | ID: mdl-31885644

ABSTRACT

Organophosphorus pesticides (OPs) are widely used for agricultural and housekeeping purposes. Exposure to OPs is associated with the progression of several health issues. Antioxidant agents may be powerful candidates to minimise adverse reactions caused by OPs. The aim of the present study was to evaluate the nephroprotective effects of red beetroot extract (RBR) against chlorpyrifos- (CPF-) induced renal impairments. CPF induced kidney dysfunction, as demonstrated by changes in serum creatinine and urea levels. Moreover, CPF exposure induced oxidative stress in the kidneys as determined by increased malondialdehyde and nitric oxide levels, decreased glutathione content, decreased catalase, superoxide dismutase, glutathione peroxidase, and glutathione reductase activities, and decreased nuclear factor (erythroid-derived 2)-like-2 factor expression. In addition, CPF induced inflammation in renal tissue as evidenced by increased release of tumor necrosis factor-alpha and interleukin-1ß and upregulation of inducible nitric oxide synthase. Furthermore, CPF promoted cell death as demonstrated by decreased Bcl-2 and increased Bax and caspase-3 levels. Treatment with RBR one hour prior to CPF treatment blocked the effects observed in response to CPF alone. Our results suggest that RBR could be used to alleviate CPF-induced nephrotoxicity through antioxidant, anti-inflammatory, and antiapoptotic activities.

16.
Biomolecules ; 9(7)2019 07 07.
Article in English | MEDLINE | ID: mdl-31284640

ABSTRACT

Exposure to organophosphorus insecticides causes several health problems to animals and humans. Red beetroot (RBR) is rich in antioxidant ingredients and possesses a promising hepatoprotective activity. This study evaluated the potential of RBR extract to prevent chlorpyrifos (CPF)-induced liver injury, with an emphasis on oxidative stress, inflammation and apoptosis. Rats received 10 mg/kg CPF and were treated with 300 mg/kg RBR extract for 28 days. CPF caused liver injury evidenced by elevated serum levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and bilirubin, along with several histological alterations. Hepatic lipid peroxidation (LPO) and nitric oxide (NO) levels, as well as inducible nitric oxide synthase (iNOS) and pro-inflammatory cytokines were increased in CPF-intoxicated rats. RBR prevented CPF-induced histological alterations, and ameliorated liver function, LPO, NO, iNOS and pro-inflammatory cytokines. RBR boosted glutathione and antioxidant enzymes, and increased Nrf2 expression. In addition, RBR diminished Bax and caspase-3, and increased Bcl-2 expression. In conclusion, RBR prevented CPF-induced liver injury via attenuation of oxidative stress, inflammation and apoptosis. RBR enhanced antioxidant defenses, suggesting that it could be used as a potential therapeutic intervention to minimize CPF hepatotoxicity.


Subject(s)
Beta vulgaris/chemistry , Chemical and Drug Induced Liver Injury/drug therapy , Chlorpyrifos/antagonists & inhibitors , Inflammation/drug therapy , Plant Extracts/pharmacology , Animals , Antioxidants/analysis , Antioxidants/metabolism , Apoptosis/drug effects , Biomarkers/analysis , Biomarkers/metabolism , Chemical and Drug Induced Liver Injury/metabolism , Chlorpyrifos/chemistry , Inflammation/chemically induced , Inflammation/metabolism , Molecular Structure , Oxidative Stress/drug effects , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Plant Roots/chemistry , Rats , Rats, Wistar
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