Fractionation Of Extract Research Articles

Chronic endometritis. Local prooxidant, antioxidant and immunometabolic disorders. An open-label, prospective, randomized study

Aim. To determine local features of lipoperoxidation processes, antioxidant defense, state of matrix metalloproteinases in chronic endometritis. Materials and methods. Eighty women aged 29.33±6.71 (min-max [24–36]) with histologically and immunohistochemically confirmed chronic endometritis and a history of reproductive losses were examined (group 1). The control group consisted of 30 patients without chronic endometritis confirmed by the results of a histochemical study and no history of reproductive losses, suffering from infertility of tubal-peritoneal origin at the age of 30.11±6.9 years (min-max [24–37]; group 2). All 130 patients underwent transvaginal echography of the pelvic organs, and histological and immunohistochemical examination of endometrial samples was performed. The activity of lipid peroxidation factors was analyzed: primary, secondary and final products of lipid peroxidation in heptane and isopropanolol fractions of uterine cavity aspirate, enzymes of the antioxidant system: superoxide dismutase, catalase, glutathione peroxidase, deoxyribonuclease I (DNase), by spectrophotometric method, remodeling factors – matrix metalloproteinases MMP-2, -9 by flow fluorimetry. Results. In patients of group 1, according to transvaginal echography of the pelvic organs, M-echo was 6.29±1.22 mm, in group 2 – 9.10±1.15 mm. In chronic endometritis, disruption of pro-oxidant and activity of antioxidant factors in the endometrium in women with chronic endometritis and a history of reproductive losses. In the heptane fraction of the phenolic extract, the concentration of primary products of lipid peroxidation (diene conjugates) was 12.98% higher than in the control, the content of secondary products of lipid peroxidation (ketodienes) was higher by 15.45%, conjugated trienes by 18.67% higher than the values in the control group, which indicates increased lipid peroxidation of endometrial cell membranes during CE. In the isopropanol phase, the products of lipid peroxidation of phospholipid acyl residues are detected. In the isopropanol fraction of the lipid extract, the amount of diene conjugates was higher, ketodienes by 24.32% and conjugated trienes by 26.14%. An increase in the activity of the enzymes superoxide dismutase by 23.98%, catalase by 14.34%, glutathione peroxidase by 12.39%, and DNase by 2.9 times was recorded compared to the indicators of group 2. Conclusion. Chronic endometritis is accompanied by changes in the system of pro-oxidant and antioxidant factors, the content of tissue remodeling products. The pathogenetic changes underlying oxidative stress in the endometrium may be increased leukocyte infiltration, imbalance of primary, secondary and end products of lipid peroxidation, enzymes superoxide dismutase, catalase, glutathione peroxidase, deoxyribonuclease I (DNase), expression of matrix metalloproteinases-2, -9.

Preparative Fractionation of Brazilian Red Propolis Extract Using Step-Gradient Counter-Current Chromatography.

Propolis is a resinous bee product with a very complex composition, which is dependent upon the plant sources that bees visit. Due to the promising antimicrobial activities of red Brazilian propolis, it is paramount to identify the compounds responsible for it, which, in most of the cases, are not commercially available. The aim of this study was to develop a quick and clean preparative-scale methodology for preparing fractions of red propolis directly from a complex crude ethanol extract by combining the extractive capacity of counter-current chromatography (CCC) with preparative HPLC. The CCC method development included step gradient elution for the removal of waxes (which can bind to and block HPLC columns), sample injection in a single solvent to improve stationary phase stability, and a change in the mobile phase flow pattern, resulting in the loading of 2.5 g of the Brazilian red propolis crude extract on a 912.5 mL Midi CCC column. Three compounds were subsequently isolated from the concentrated fractions by preparative HPLC and identified by NMR and high-resolution MS: red pigment, retusapurpurin A; the isoflavan 3(R)-7-O-methylvestitol; and the prenylated benzophenone isomers xanthochymol/isoxanthochymol. These compounds are markers of red propolis that contribute to its therapeutic properties, and the amount isolated allows for further biological activities testing and for their use as chromatographic standards.

Phytochemical profiling and therapeutic potential of Baliospermum montanum: A promising inhibitor of COX-2/15-LOX enzymes and NO production in LPS-stimulated RAW 264.7 macrophages

IntroductionBaliospermum montanum (BM) is traditionally used to treat various ailments such as inflammation, rheumatoid arthritis, pain, etc. The root part of BM is well documented for its anti-inflammatory activity. However, the present study evaluates the multitargeted anti-inflammatory activity of the less explored BM leaf part. This is the first study investigating the COX-2, 15-LOX dual inhibiting potential and NO production inhibition of BM leaf extracts, fractions and compounds. MethodsThe BM leaf's water, ethanol, and hydroethanolic extracts were evaluated for anti-inflammatory activity by assessing the inhibition of COX-2, 15-LOX, and NO production. Additionally, the antioxidant potential of the extracts was determined through DPPH, ABTS, and FRAP assays. The most active ethanol extract was fractionated into 12 fractions and tested for anti-inflammatory, antioxidant, phenol and flavonoid content. Compounds were isolated from the most active fractions and identified using NMR techniques. The isolated compounds were checked for their anti-inflammatory activity. In addition, HPLC analysis of ethanol extract, fractions, and isolated compounds was carried out. ResultsThe ethanol extract exhibited higher inhibition of COX-2, 15-LOX enzymes and NO production (IC50 35.75 ± 1.95 μg/mL) in LPS-treated RAW 264.7 cells. It also showed higher antioxidant activity. Following the fractionation of the ethanol extract using different solvents, only the ethyl acetate fractions demonstrated inhibition of COX-2, 15-LOX enzymes, and NO production. The ethyl acetate and acetone fractions are excellent sources of antioxidants, phenolic and flavonoid compounds. An effort to find the active principles from these ethyl acetate fractions resulted in the isolation and identification of four potent anti-inflammatory agents, namely, (-)-epiafzelechin, afzelechin-(4α→8)-afzelechin, afzelechin-(4α→8)-epiafzelechin and β-sitosterol-d-glycoside. The first three compounds were reported for the first time from this plant and afzelechin-(4α→8)-afzelechin was found to be a potent inhibitor of the COX-2/15 LOX enzyme as well as NO production (IC50 = 5.9 ± 1.09 µM). ConclusionBased on these findings, we conclude that ethanol extract, fractions, and isolated compounds exert anti-inflammatory activity through multiple therapeutic mechanisms. The present study scientifically supports the traditional use of plant BM for treating inflammation and other related disorders.

Evaluation of the merit of ethanolic extract of Annona reticulata as an anti-cancer agent in human colon cancer cell lines (HCT-116)

Background Colon cancer is the third most common cancer type worldwide. Novel alternative therapeutic anti-cancer drugs against colon cancer with less toxicity are to be explored . This study was aimed to explore the anti-proliferative and anti-migratory activity of various fractions of Annona reticulata ethanolic leaf extract on human colon cancer cell lines (HCT-116) and to explore the potential molecular targets from the most potent plant extract fraction. Methods After obtaining ethical clearance from the institutional ethics committee, the extract and fractions were prepared and a preliminary analysis of the phytochemical was done qualitatively. Total phenolic and flavonoids were determined. Ethanolic leaf extract and its fractions were subjected to cytotoxicity analysis using the sulforhodamine B assay and the most promising fraction which showed the highest viability was selected to study anti-migratory activity. The anti-migratory effect was studied using a scratch wound healing assay. Gas chromatography-mass spectrometry (GC-MS) was done to identify the major phytocompounds present in the fraction. The major five phytocompounds identified from the GC-MS were subjected to bioinformatics analysis. Result Among the four fractions, the petroleum ether fraction exhibited the highest anti-proliferative activity. The migration of colon cancer cells was significantly inhibited by the extract and petroleum ether fraction. The major phytocompounds identified from GC-MS were phytol (13.03%), 2,6-bis (3,4-methylenedioxyphenyl)-3,7-dioxabicyclo (3.3.0) octane (11.95%), gamma.-sitosterol (10.45%), alpha.-tocopherol-beta.-D-mannoside (7.50%) and 3-amino-4-piperonyl-5-pyrazolone (5.84%). The bioinformatics analysis of these phytochemicals showed a high potential to affect the levels of key proteins driving colon cancer progression, inhibiting the enzymes and proteins overexpressed in cancer. Conclusion The outcome of this study endorses the potential of phytochemicals of the petroleum ether fraction of ethanolic leaf extract of Annona reticulata for the development of a new chemotherapeutic agent in the treatment of colon cancer.

Novel therapeutic activities of dragon blood from palm tree Daemonorops draco for the treatment of chronic diabetic wounds

BackgroundThe clinical efficacy of Jinchuang Ointment, a traditional Chinese medicine (TCM), in treating chronic non-healing diabetic wounds has been demonstrated over the past decades. Both in vitro and in vivo angiogenic activities have been reported for its herbal ingredients, including dragon blood from the palm tree Daemonorops draco and catechu from Uncaria gambir Roxb. Additionally, crude extracts of dragon blood have exhibited hypoglycemic effects not only in animal studies but also in cell-based in vitro assays.ResultsOur findings indicate that crude dragon blood extract promotes the differentiation of myoblasts into myotubes. Partially purified fractions of dragon blood crude extract significantly enhance the expression of muscle cell differentiation-related genes such as myoG, myoD, and myoHC. Our results also demonstrate that crude extracts of dragon blood can inhibit platelet-derived growth factor-induced PAI-1 expression in primary rat vascular smooth muscle cells, thereby favoring changes in hemostasis towards fibrinolysis. Consistent with previous reports, reduced expression of plasminogen activator inhibitor 1 (PAI-1) accelerates wound healing. However, further separation resulted in a significant loss of both activities, indicating the involvement of more than one compound in these processes. Stem cells play a crucial role in muscle injury repair. Neither dragon blood nor catechu alone stimulated the proliferation of human telomerase reverse transcriptase (hTERT)-immortalized and umbilical cord mesenchymal stem cells. Interestingly, the proliferation of both types of stem cells was observed when crude extracts of dragon blood and catechu were present together in the stem cell growth medium.ConclusionsDragon blood from D. draco offers multifaceted therapeutic benefits for treating chronic nonhealing diabetic wounds from various perspectives. Most drugs in Western medicine consist of small molecules with defined ingredients. However, this is not the case in TCM, as the activities of dragon blood reported in this study. Surprisingly, the activities documented here align with descriptions in ancient Chinese medical texts dating back to A.D. 1625.

Larvicidal activity of liverworts’ (Marchantia paleacea) ethyl acetate fraction against Athalia proxima

Abstract Indonesia is an agrarian country with extensive agricultural land and a variety of crop types. One of the challenges faced by farmers when cultivating crops is pest infestation. The larval stage of Athalia proxima attacks cruciferous plants, leading to a 65-80% decrease in productivity. Continuous use of synthetic larvicides has negative impacts such as pest resistance, harm to natural plant enemies, and environmental pollution. This research aims to investigate the larvicidal activity of the ethyl acetate fraction of ethanol extract from liverworts (Marchantia paleacea) against A. proxima. The stages of compound isolation included extraction and fractionation. Compound groups in the fraction were identified through phytochemical screening. The larvicidal activity testing method used was the spray method. The concentrations of liverwort ethyl acetate fraction used were 0.4%, 0.6%, 0.8%, and 1%. Distilled water was used as a negative control. Larval mortality was observed 24 hours after application and analyzed using probit analysis to determine the LC50 value. Phytochemical screening results indicated that the ethyl acetate fraction tested positive for alkaloids, phenolics, flavonoids, terpenoids, and steroids. The LC50 value of the ethyl acetate fraction of the ethanol extract from M. paleacea was 0.33%. Therefore, the ethyl acetate fraction of M. paleacea exhibited high larvicidal activity against A. proxima.

Bioassay-guided fractionation of Verbascum thapsus extract and its combination with polyvinyl alcohol in the form electrospun nanofibrous membrane for efficient wound dressing application

Verbascum thapsus (V. thpsus), family Scrophulariaceae, has considerable importance in traditional medicine worldwide because of its antioxidant and anti-inflammatory activities. V. thpsus was used in traditional medicines as a useful drug for lung disease, sore throat, wound healing, and treatment of whooping cough. The aim of this study was to extract of V. thpsus bioactive fraction using antibacterial assay guided fractionation methodology and develop a system based on electrospun nanofibrous membrane (NFM) that can be effective by releasing the extract of V. thapsus for antibacterial and wound healing applications. For this purpose, the fractionation of total extract was done using Liquid-Liquid extraction method. The selected fraction based on its anti-bacterial activity was then subjected to the silica gel column chromatography for further purification. Since electrospinning is an economical and relatively simple method to produce continuous and uniform nanofibers, and due to its high specific surface area, adjustable pore size, and flexibility, special attention has been paid to loaded the most effective fraction on PVA nanofibers for applications such as wound dressings. The obtained result showed that, the purified V. Thapsus extract has a concentration-dependent antimicrobial activity against Escherichia coli and Staphylococcus aureus. The phytochemically analyses of bioactive fraction by High-performance liquid chromatography (HPLC) proved the presence of 6 phenolic acids, 4-hydroxybenzoic acid, chlorogenic acid, caffeic acid, p-coumaric acid, ferulic acid, and flavonoid, rutin, as the major compounds. Also, physicochemical characterization of PVA-selected extract loaded electrospun nanofibrous membranes (NFM) were analyzed by scanning electron microscope (SEM), Fourier transform infrared spectrometer (FT-IR). MTT and hemolysis assays were done to affirm the biocompatibility of fabricated scaffolds. Release profile of extract loaded- NFM showed continues release of extract from mat during 90h. Moreover, the capability of these NFM in wound healing application was evaluated in-vitro and in-vivo. The cell viability test (MTT), cell adhesion images, antioxidant, antibacterial, hemolysis assays and in-vitro and in-vivo wound healing assays confirmed that fabricated NFM containing 5 % butanolic extract were the most biocompatible scaffold for wound dressing applications and accelerates the rate of wound closure. The obtained outcomes confirmed that V. thpsus/PVA NFM can be considered as promising scaffold for potential wound dressings.

Evaluation of Flavonoid-rich Fraction of Portulaca Grandiflora Aerial Part Extract in Atherogenic Diet-induced Atherosclerosis.

Portulaca grandiflora is a tiny, upright herb that contains a variety of chemical components, including alkaloids, glycosides, mucilage, proteins, tannins, flavonoids, saponins, polysaccharides, and triterpenoids possessing properties that may help with atherosclerosis. The reported pharmacological properties of Portulaca grandiflora are antioxidant, antidiabetic, antiasthmatic, antibacterial, antiulcer and anti-inflammatory properties. The yield of methanol extract is higher than that of ethanol and acetone, and its phytoconstituents, like flavonoids and polyphenols, and has potent antioxidant properties. In order to determine the effectiveness of Portulaca grandiflora methanol extract fraction against high-fat diet (HFD)-induced hyperlipidemia, hemodynamic change, antioxidant levels, and vascular dysfunction in rats, a study was carried out on a flavonoid-rich methanol extract fraction of the aerial part of Portulaca grandiflora Hook. This method involves a study of 30 days involving male Wistar rats (240-250 g) (n=5) that were fed with an Ath diet. Study groups were divided into (i) The Control Group, (ii) the Diseases Control Group, (iii) Disease + Standard drug (Atorvastatin 20mg/kg, orally, (iv) Disease + Test Extract dose 1 (Portulaca grandiflora 200 mg/kg orally), and (v) Disease + Test Extract dose 2 (Portulaca grandiflora 400 mg/kg orally). Both the test drug Portulaca grandiflora and the standard drug Atorvastatin were given orally for 30 days. At the end of the study, blood samples were taken to measure the serum lipid profile, alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, and levels of oxidative tissue stress. Hemodynamic parameters and aortic staining were performed. Portulaca grandiflora treatment improved the lipid profile and considerably reduced oxidative stress levels. Aortic staining examination revealed a marked reduction in atherosclerotic lesions. These results revealed that Portulaca grandiflora is an effective treatment approach in preventing atherosclerotic lesion progression, which is attributed to its protection against oxidative stress and various enzymatic activities in the Atherogenic model.

Anticancer Activity of Sargassum fluitans Extracts in Different Cancer Cells.

The arrival of large quantities of Sargassum in the Mexican Caribbean Sea has generated major environmental, health and economic problems. Although Sargassum has been used in the generation of some commercial products, few studies have described its possible applications as a source of compounds with anticancer activity. This study aimed to evaluate the antiproliferative effects of different Sargassum extracts on various cancer cell lines. Furthermore, LC/QTOF-MS was used to identify the compounds related to the antiproliferative effect. First, determination of the seaweed was performed, and dichloromethane, chloroform and methanol extracts were obtained. The extracts were evaluated for their antiproliferative effects by MTT in breast (MDAMB- 231 and MCF-7), prostate (DU-145), lung (A549) and cervical (SiHa) cancer cell lines. Finally, LC/QTOFMS identified the compounds related to the antiproliferative effect. The authentication showed Sargassum fluitans as the predominant species. The extracts of dichloromethane and chloroform showed an antiproliferative effect. Interestingly, the fractionation of the chloroform extract showed two fractions (FC1 and FC2) with antiproliferative activity in MDA-MB-231, SiHa and A549 cancer cell lines. On the other hand, three fractions of dichloromethane extract (FD1, FD4 and FD5) also showed antiproliferative effects in the MDA-MB-231, MCF-7, SiHa and DU-145 cancer cell lines. Furthermore, LC/QTOF-MS revealed the presence of eight major compounds in FC2. Three compounds with evidence of anticancer activity were identified (D-linalool-3-glucoside, (3R,4S,6E,10Z)-3,4,7,11-tetramethyl-6,10-tridecadienal and alpha-tocotrienol). These findings showed that Sargassum fluitans extracts are a possible source of therapeutic agents against cancer and could act as scaffolds for new drug discovery.

Antioxidant and Anti-Inflammatory Activities of the Extract and Bioaccessible Fraction of Mango Peel in Muffins.

Mango peel is a production waste and can cause environmental problems, but its nutritional value consists of bioactive compounds that could be beneficial for human health. The aim of this study is to determine the bioactive compounds, antioxidant and anti-inflammatory activities of mango peels and their use in muffins. The content of polyphenols, carotenoids and total phenols as well as the antioxidant activity of mango peel extract were evaluated. The anti-inflammatory activity of the extract was investigated using Caco-2 cell assay. The mango peel powder was then incorporated into muffin formulations. The sensory properties of these fortified muffins were evaluated. The total phenolic content, antioxidant activity and anti-inflammatory properties of the muffin extracts and their bioaccessible fractions were also analysed. The mango peel contained quercetin, phenolic compounds, α-carotene, β-carotene and lutein, which have antioxidant potential. In Caco-2 cells exposed to induced inflammation, the mango peel powder extract (γ=10, 50 and 100 µg/mL) attenuated the production of reactive oxygen species (ROS), tumour necrosis factor-alpha (TNF-α) and interleukin-8 (IL-8), while maintaining cell viability. Muffins supplemented with 5 % mango peel powder showed good sensory properties, but not as good as those of plain muffins without mango peel powder. The total phenolic content and antioxidant activities in both the extract and the bioaccessible fraction of the supplemented muffins were higher than those observed in the standard muffins. Moreover, the extract and bioaccessible fraction of the supplemented muffins resulted in a higher reduction of ROS, IL-8 and TNF-α production in Caco-2 cells than those obtained from the standard muffins. This study is the first to investigate the protective effects of mango peel and muffins supplemented with mango peel powder against IL-1β-induced oxidative damage in Caco-2 cells. The results confirm that both mango peel and the supplemented muffins inhibited the production of inflammatory markers, including ROS and cytokines. These findings suggest that mango peel could be a valuable component of functional food formulations including dietary supplements.

The Age of Buried Carbon Changes the Greenhouse Gas Budget of a Dam

AbstractDams are a globally relevant source of greenhouse gases (GHG), which impairs their function as a source of green energy. High burial rates of organic carbon (OC) in dam sediments may partly or fully offset the emissions. We argue that only the burial of carbon fixed in the timespan of dam operation changes the GHG balance. Here, we took sediment cores from a temperate dam. We analyzed radiocarbon age and OC molecular composition by laser desorption ionization mass spectrometry in the bulk OC and in four chemical extract fractions. The bulk samples contained modern OC, fixed after 1950. However, the extracted OC was of different ages (modern to 1900 years BP). Fractions with OC measured as old (>960 years BP) accounted for 57% of total sediment OC. Correlations of molecular composition with extract age suggest that these older fractions contained insignificant amounts of modern OC. We conclude that a substantial proportion of buried carbon did not originate from the contemporary atmosphere and cannot be offset against recent GHG emissions.

Efficacy of Albizia malacophylla (A.Rich.) Walp. (Leguminosae) methanol (80%) leaf extract and solvent fractions against Plasmodium berghei-induced malaria in mice model

Ethnopharmacological relevanceNovel drugs are needed to address the issue of malarial infection resistance; natural items can be a different source of these medications. Albizia malacophylla (A. Rich.) Walp. (Leguminosae) is listed as one of the antimalarial medicinal plants in Ethiopian folk medicine. However, there are no reports regarding the biological activity or phytochemistry of the plant. Aim of the studyThus, this study aimed to evaluate the A. malacophylla crude extract and solvent fractions' in vivo antimalarial activity utilizing 4-day suppressive, preventative, and curative tests in mice infected with P. berghei. Materials and methodsThe parasite Plasmodium berghei, which causes rodent malaria, was used to infect healthy male Swiss Albino mice, weighing 23–28 g and aged 6–8 weeks. Solvent fractions such as methanol, water, and chloroform were given in addition to an 80% methanolic extract at 100, 200, and 400 mg/kg doses. A Conventional test such as parasitemia, survival time, body weight, temperature, and packed cell capacity were employed to ascertain factors such as the suppressive, curative, and preventive tests. ResultsEvery test substance dramatically reduced the number of parasites in every experiment. Crude extract (with the highest percentage suppression of 67.78%) performs better antimalarial effect than the methanol fraction, which is the most efficient solvent fraction with a percentage suppression of 55.74%. With a suppression value of 64.83% parasitemia level, the therapeutic effects of 80% methanolic crude extract were greater than its curative and preventative effects in a four-day suppressive test. The survival period (17 days) was longer with the hydroalcoholic crude extract dose of 400 mg/kg than with other doses of the materials under investigation. ConclusionsThe results of this investigation validate the antimalarial characteristics of A. malacophylla leaf extract. The crude extract prevented weight loss, a decline in temperature, and a reduction in PCV. The results demonstrate that the plant has a promising antimalarial effect against P. berghei, hence supporting the traditional use of the plant. Therefore, it could serve as a foundation for the development of new antimalarial drugs.

Further Polycyclic Quinones of Micromonospora sp.

The crude acetone extract of a marine Micromonospora sp. strain associated with Eudistoma vannnamei was fractioned with hexane and ethyl acetate. The crude extract and both soluble fractions were assayed against several bacteria strains. The new polycyclic quinones 12-hydroxy-9-propyltetracene-6,1-dione (1), 5,12-dihydroxy-4-methoxy-9-propyltetracene-5,12-dione (2), and 4,6-dihydroxy-3-methoxycarbonyl- methyl-6a-(oxobutyl)-5,12-anthraquinone (3), along with the known 4,6-dihydroxy-3-methoxycarbonyl-methyl-6a-(oxo-3-methyl-butyl)-5,12-anthraquinone (4) and 4,6-dihydroxy-3-methoxycarbonyl-methyl-6a-(oxopentyl)-5,12-anthraquinone (5) were isolated from the hexane-soluble fraction, while from the active ethyl acetate fraction were isolated the known 4,6,11-trihydroxy-9-propyltetracene-5,12-dione (6), 4-methoxy-9-propyltetracene-6,11-dione (7), 7,8,9,10-tetrahydro-9-hydroxy-4-methoxy-9-propyltetracene-6,11-dione (8), and 10β-carbomethoxy-7,8,9,10-tetrahydro-4,6,7α,9α,11-pentahydroxy-9-propyltetracene-5,12-dione (9). The structures of the new compounds were established by interpretation of HRMS and NMR techniques. A study of molecular docking was performed with the compounds from the active ethyl acetate fraction to correlate tentatively with the antimicrobial activity. Molecular docking, RMSD, RMSF, and MM-GBSA evaluations were performed to investigate the inhibitory activity of 6-8 against the protein PDB-codex 1MWT, being considered a promising target for studying drug development responsible for inhibiting replication of Staphylococcus aureus. Penicillin G was used as the standard inhibitory. Anthracyclinones 6-8 were the best hydrolase inhibitor with affinity energy -8.1 to -7.9 kcal/mol compared to penicillin G, which presented -6.9 kcal/mol. Both 8 and 7 present potent inhibitory effects against hydrolase through molecular dynamics simulation and exhibit favorable drug-like properties, promising new hydrolase blockers to fight bacterial infections from Staphylococcus aureus.

ANTIOXIDANT ACTIVITY TEST OF SUNGKAI LEAF (Peronema canescens Jack) ETHANOL EXTRACT, N-HEXANE AND ETHYL ACETATE FRACTION

Research on antioxidant activity test of ethanol extract, n-hexane fraction and ethyl acetate of Sungkai leaves (Peronema canescens Jack) has been carried out. Sungkai plant is a traditional medicinal plant that is widely used for traditional medicine. Sungkai leaf secondary metabolites (Peronema canescens Jack) contain alkaloids, flavonoids, saponins, phenols, terpenoids. Sungkai leaves (Peronema canescens Jack) were extracted using maceration method with 70% ethanol solvent and fractionated using n-hexane and ethyl acetate solvents. Testing the antioxidant activity using the DPPH method (2,2-diphenyl-1-pikhrylhydrazil) obtained IC50 (Inhibition Concentration) of ethanol extract, ethyl acetate and n-hexane fractions, respectively 5.6356 µg/mL, 46.4707 µg/mL, and 153.7020 µg/mL.

Elucidating the mechanism of hepatotoxicity in Euodia rutaecarpa: insights from QSAR toxicity prediction and metabolomics

Objective: Euodia rutaecarpa, (Wu Zhu Yu) a Chinese medicine clinically used to treat gastrointestinal disorders, has been widely employed. However, Euodia rutaecarpa is regarded as a small toxic traditional Chinese medicine in the Chinese Pharmacopoeia and other herbal works. Using toxicity predictions combined with in vitro and in vivo studies, this study aimed to identify the toxic components and toxic target organs of Euodia rutaecarpa, and explore its toxic mechanism from a metabolic perspective. Methods: The toxic target organs of Euodia rutaecarpa were identified through in vitro and in vivo studies. In vitro toxicity screening was performed by alkaloid enrichment and isolation. The potential toxicity of compounds was predicted by Absorption, Distribution, Metabolism, Excretion, and Toxicity Predictor (ADMET Predictor) based on Quantitative Structure–Activity Relationship (QSAR) construction. In addition, the study integrated the serum metabolomic analysis after the administration of potentially toxic components to clarify the effect of potentially toxic substances on metabolism in mice. Results: Comparing the acute toxicity in mice of different extraction methods and before and after processing, it was evident that Euodia rutaecarpa alcoholic extract had the highest toxicity, and the target organ of Euodia rutaecarpa toxicity was the liver. The alkaloid fraction of alcoholic extract of Euodia showed strong cytotoxicity. The potential toxicity of Euodia rutaecarpa was calculated and predicted by ADMET Predictor, and alkaloids are suspected to be responsible for the toxicity of Euodia rutaecarpa. Evodiamine significantly reduced the number of cells and increased the mitochondrial membrane potential in vitro. Different metabolites were significantly identified by serum metabolomics, of which bile acid metabolism and steroid hormone biosynthesis are the key pathways of hepatotoxicity. Conclusions: Clarify the scientific significance of clinical use of processed products by comparing the acute toxicity of different extraction methods before and after processing. Combining the toxicity prediction based on QSAR with the toxicity screening in vitro and in vivo, the potential toxic target organs and toxic components of Euodia rutaecarpa can be identified. Through metabolomics, we preliminarily revealed that the hepatotoxicity of Euodia rutaecarpa may be related to bile acid metabolism and steroid hormone biosynthesis. This study lays the foundation for elucidating the mechanism of Euodia rutaecarpa and evaluating its safety and quality.

<i>In vitro</i> influence on fibrinolysis parameters of aqueous extracts of mushrooms growing on woody plants

Introduction. Various compounds of mushroom extracts are of interest as a source of new active pharmaceutical substances with antithrombotic activity.Aim. The aim of the work is to determine the potential antithrombotic activity in vitro of aqueous extracts of xylotrophic mushrooms: Lentinula edodes, Pholiota squarrosa, Flammulina velutipes (Curtis) Signer, Kuehneromyces mutabilis, Armillaria cepistipes, Armillaria borealis, Hypholoma capnoides, Lentinellus cochleatus, Léccinum aurantíacum, Pleurotus ostreatus.Materials and methods. The study of fibrinolytic activity of water extracts of mushrooms in vitro was carried out. The effect of test objects on plasminogen activation, on the activity of enzyms of the fibrinolysis system α2-antiplasmin, on the time of euglobulin clot lysis and on the concentration of fibrinogen was evaluated. For the analysis we used reagent kits of SPD «RENAM» and LLC Company «Тechnology-Standard» (Russia).Results and discussion. Pronounced antithrombotic activity is demonstrated by an aqueous extract of L. edodes and fractions of aqueous extract. Extracts of L. edodes affect all studied parameters of fibrinolysis to varying degrees. Caps and stems of P. squarrosa, K. mutabilis, and F. velutipes aqueous extracts and fractions of aqueous extracts also demonstrate antithrombotic (fibrinolytic) activity in vitro, having a statistically significant effect on various parameters of fibrinolysis. A less pronounced effect on fibrinolysis parameters in comparison with the above samples is observed when using an aqueous extract of L. cochleatus marked. A. cepistipes, A. borealis, H. capnoides, L. aurantiacum, P. ostreatus fruiting bodies aqueous extracts do not affect the studied parameters of fibrinolysis and the concentration of fibrinogen.Conclusion. The data obtained make it possible to recommend extracts from L. edodes, F. velutipes, P. squarrosa and K. mutabilis for further study, for example, in order to develop functional food products for cardiovascular diseases or to create antithrombotic drugs.

Therapeutic effect of the solvent fraction of hexane leaf extract of Tapinanthus bangwensis (Engl. & K. Krause) (Loranthaceae) in alloxan-induced pathology in diabetic rats

Plant-based products are gradually replacing pharmaceuticals in treating ailments, including diabetes, due to their safety, cost-effectiveness, potency, and availability. Therefore, the current study looked into the therapeutic effect of the solvent fraction of hexane leaf extract of Tapinanthus bangwensis (HEXETACF) (Loranthaceae) in alloxan-induced pathology in diabetic rats. The biochemical parameters were estimated using analytical grade kits via spectrophotometric method. The laboratory rats were distributed into group W (five rats on feed and water), group X (seven rats + 150 mg/kg alloxan solution only), group Y (seven rats + 150 mg/kg alloxan solution + 200 mg/kg BW silymarin for 21 days), and group Z (seven rats + 150 mg/kg alloxan solution + 250 mg/kg BW HEXETACF for 21 days). The results showed that HEXETACF and silymarin (SILY) reduced blood glucose concentration by 33.77% and 34.80%, respectively, after the 21st day of treatment (p < 0.05). Additionally, alkaline phosphatase (ALP) and alanine aminotransferase (ALT) activity in SILY and HEXETACF were significantly decreased compared to the diabetic group (p < 0.05), but no significant decrease in aspartate aminotransferase (AST) activity was observed between the test samples and the diabetic group. Furthermore, the test samples lowered malondialdehyde (MDA) levels, by improving glutathione, superoxide dismutase (SOD), and catalase (CAT) activity. The HEXETACF and SILY significantly decreased triglyceride levels (TG) compared to the diabetic group at p < 0.05. They also reduced low-density lipoprotein (LDL) and cholesterol levels and increased the high-density lipoprotein levels compared to the diabetic group. Additionally, no significant decrease in serum electrolytes (Na+, K+, and Cl-), urea, and creatinine (including albumin and total protein) values was observed in HEXETACF and SILY, while hematological indices increased compared to the diabetic group. Histology results revealed that the test samples had normalized glomeruli, β-islet cells, and hepatocytes. However, a trace of mild congestion was noticed in the STDG. But edemic blood congestion was observed in the diabetic group. In conclusion, the current result demonstrated that HEXETACF may be a promising antidiabetic agent that could replace mSILY.

Cytotoxicity of Fractions of Dracontium spruceanum Chloroform Extract in MDA-MB-231 and MCF-7 Breast Cancer Cell Lines

Background: Breast cancer is a major health issue responsible for numerous deaths worldwide. Medicinal plants are rich in compounds with potential anticancer properties. Objectives: This study aimed to assess the cytotoxicity of fractions from the Dracontium spruceanum chloroform extract on MDA-MB-231 and MCF-7 breast cancer cell lines. Methods: Eleven fractions were screened for cytotoxic effects on cell viability using the MTT assay. Subsequently, the two most cytotoxic fractions were selected, and the half-maximal inhibitory concentration (IC50) was calculated using two-fold dilution concentrations ranging from 100 to 1.56 µg/mL. Results: Fractions F43 and F50 showed the highest cytotoxicity at 100 µg/mL. The IC50 of F43 was 270.8 ± 18.07 µg/mL and 133.0 ± 17.99 µg/mL for MDA-MB-231, and 252.2 ± 21.94 µg/mL and 144.3 ± 23.14 µg/mL for MCF-7 cells at 24 and 48 hours, respectively. For F50, IC50 values were 107.2 ± 2.97 µg/mL and 126.1 ± 14.77 µg/mL for MDA-MB-231, and 236.4 ± 58.7 µg/mL and 104.5 ± 10.54 µg/mL for MCF-7 at corresponding times. Conclusions: Dracontium spruceanum fractions demonstrated moderate cytotoxicity on MDA-MB-231 and MCF-7 breast cancer cell lines, indicating their anticancer potential. Further research is required to identify the metabolites responsible for this activity and to elucidate the molecular mechanisms involved.

Fractionation and Identification of Phenolic Compounds from Ethylacetate Fraction of Ageratum Conyzoides Methanolic Leaf Extract

Natural phenols are significant functional substances that are commonly present in plants and provide advantages for human health. Phenolics can be utilized for a variety of therapeutic applications, such as antioxidant, anti-inflammatory, antidiabetic, and antibacterial activities. This work aimed to fractionate and identify the phenolic compounds present in ethylacetate fractions of the methanolic leaf extracts of A. conyzoides. Fractionation and purification of the ethylacetate fraction using column chromatography and TLC, followed by FTIR and LC-MS for identification, were employed in this work. While the FTIR spectroscopy was used to identify the fractions with –OH functional group, LCMS was used to identify the phenolics. After fractionation and purification, six ethylacetate samples were obtained, each with different Rf values, of which three showed a pronounced presence of OH functional groups in phenolics. Subsequently, four phenolic compounds were identified: furocoumaric acid, liquiritin, isorhamnetin, and syringin. This research has demonstrated that ethylacetate fraction of A. conyzoides methanol leaf extract is a good source of the phenolic compounds: Furocoumaric acid, Liquiritin, Isorhamnetin, and syringin.

Toxicity of Bioactive Compounds of Strychnos pseudoquina (Loganiaceae) in Spodoptera frugiperda (Noctuidae)

The objective of this study was to assess the toxicity of leaf and bark extracts of Strycns pseudoquina and their fractions, as well as flavonoids isolated from its bark, in a population of Spodoptera frugiperda. Crude bark and leaf extracts, as well as fractions of these extracts, were obtained using a rotary evaporator and different solvents; the ethyl acetate fraction was obtained, and flavonoid compounds were isolated, consisting mostly of 3-O-methylquercetin and minimally of strychnobiflavone. The crude extracts, extract fractions, and isolated flavonoids were applied separately to maize leaves, which were offered to S. frugiperda larvae; then, biological parameters of this insect population were evaluated. Toxic effects of topical applications of the isolated flavonoids on the larvae were also evaluated. The results highlighted the larvicidal effects of the crude bark extract and its fractions and the significant toxicity of this crude extract (LC50 = 0.048%), which was six times more toxic to larvae than its ethyl acetate fraction (LC50 = 0.288%). The investigated flavonoids showed no significant larvicidal effect. The pronounced larvicidal effect of S. pseudoquina bark crude extract and its fractions on S. frugiperda indicates that this plant presents an insecticidal potential to be explored in integrated pest management programs.