Crude Fraction Research Articles

Phytochemical analysis, in vitro and in silico effects from Alstonia boonei De Wild stem bark on selected digestive enzymes and adipogenesis in 3T3-L1 preadipocytes

BackgroundObesity is a global health issue arising from the unhealthy accumulation of fat. Medicinal plants such as Alstonia boonei stem bark has been reported to possess body weight reducing effect in obese rats. Thus, this study sought to investigate the in vitro and in silico effects of fractions from Alstonia boonei stem bark on selected obesity-related digestive enzymes and adipogenesis in 3T3-L1 preadipocytes.MethodTwo fractions were prepared from A. boonei: crude alkaloid fraction (CAF) and crude saponin fraction (CSF), and their phytochemical compounds were profiled using Liquid chromatography with tandem mass spectrometry (LCMS/MS). The fractions were assayed for inhibitory activity against lipase, α-amylase and α-glucosidase, likewise their antiadipogenic effect in 3T3-L1 adipocytes. The binding properties with the 3 enzymes were also assessed using in silico tools.ResultsEleven alkaloids and six saponin phytochemical compounds were identified in the CAF and CSF using LCMS/MS. The CAF and CSF revealed good inhibitory activity against pancreatic lipase enzyme, but weak and good activity against amylase respectively while only CSF had inhibitory activity against α-glucosidase. Both fractions showed antiadipogenic effect in the clearance of adipocytes and reduction of lipid content in 3T3-L1 adipocytes. The LCMS/MS identified compounds (41) from both fractions demonstrated good binding properties with the 3 enzymes, with at least the top ten compounds having higher binding energies than the reference inhibitors (acarbose and orlistat). The best two docked compounds to the three enzymes were firmly anchored in the substrate binding pockets of the enzymes. In a similar binding pattern as the reference acarbose, Estradiol-17-phenylpropionate (-11.0 kcal/mol) and 3α-O-trans-Feruloyl-2 α -hydroxy-12-ursen-28-oic acid (-10.0 kcal/mol) interacted with Asp197 a catalytic nucleophile of pancreatic amylase. Estradiol-17-phenylpropionate (-10.8 kcal/mol) and 10-Hydroxyyohimbine (-10.4 kcal/mol) interacted with the catalytic triad (Ser152-Asp176-His263) of pancreatic lipase while Estradiol-17-phenylpropionate (-10.1 kcal/mol) and 10-Hydroxyyohimbine (-9.9 kcal/mol) interacted with Asp616 and Asp518 the acid/base and nucleophilic residues of modelled α-glucosidase.ConclusionThe antiobesity effect of A. boonei was displayed by both the alkaloid and saponin fractions of the plant via inhibition of pancreatic lipase and adipogenesis.

Boiling point modeling for petroleum speciation data

An empirical model is presented for generating the boiling curve of a petroleum sample based on parameters that are experimentally available using typical speciation techniques. The approach estimates the boiling point distribution of the detected components and allows a comparison with the simulated distillation profile of the sample. The model is shown to predict the boiling behavior of the speciation-derived average components in crude oil distillation fractions, which match the measured average distillation temperature of the fractions. The model is then applied to generate boiling distributions, based on mass spectrometric and gas chromatographic speciation data, which match the measured simulated-distillation curves with less than 20 °C difference. The model is also applied to the isolated fraction of saturated compounds, where the modeled boiling profile matches well with the measured distillation profile of the entire sample. This confirms that the model, although built using Fourier transform ion cyclotron resonance mass spectrometry data of aromatic compounds, extends to data obtained using comprehensive 2-dimensional gas chromatography, and also describes the boiling behavior of saturated compounds speciated by field desorption time of flight mass spectrometry. Consequently, the modeled boiling distribution enables an assessment of how well a speciation measurement describes the sample, and provides an estimation of lower and higher-boiling portions that have been omitted in the data set.

Antispasmodic Effect of Alstonia boonei De Wild. and Its Constituents: Ex Vivo and In Silico Approaches.

Alstonia boonei, belonging to the family Apocynaceae, is one of the best-known medicinal plants in Africa and Asia. Stem back preparations are traditionally used as muscle relaxants. This study investigated the antispasmodic properties of Alstonia boonei Stem back and its constituents. The freeze-dried aqueous Stem back extract of A. boonei, as well as dichloromethane (DCM), ethyl acetate, and aqueous fractions, were evaluated for their antispasmodic effect via the ex vivo method. Two compounds were isolated from the DCM fraction using chromatographic techniques, and their antispasmodic activity was evaluated. An in silico study was conducted by evaluating the interaction of isolated compounds with human PPARgamma-LBD and human carbonic anhydrase isozyme. The Stem back crude extract, DCM, ethyl acetate, and aqueous fractions showed antispasmodic activity on high-potassium-induced (K+ 80 mM) contractions on isolated rat ileum with IC50 values of 0.03 ± 0.20, 0.02 ± 0.05, 0.03 ± 0.14, and 0.90 ± 0.06 mg/mL, respectively. The isolated compounds from the DCM fraction were β-amyrin and boonein, with only boonein exhibiting antispasmodic activity on both high-potassium-induced (IC50 = 0.09 ± 0.01 µg/mL) and spontaneous (0.29 ± 0.05 µg/mL) contractions. However, β-amyrin had a stronger interaction with the two proteins during the simulation. The isolated compounds boonein and β-amyrin could serve as starting materials for the development of antispasmodic drugs.

FTIR characterization, phytochemical, antibacterial, and antioxidant properties of Pterocarpus Osun stembark and leaf extracts

Antimicrobial resistance threatens the prevention and treatment of an ever-increasing range of infections caused by microorganisms. It becomes necessary therefore, to determine compounds from natural sources that can be used to develop novel medicines with potent antimicrobial properties. Although, the stem bark of Pterocarpus osunhas long been used to treat diarrhea, dysentery, and other gastrointestinal problems, there are no published pharmacological reports to authenticate these uses. The leaf and stem bark of Pterocarpus osunwere extracted exhaustively and successively with n-hexane, ethyl acetate and methanol. The antioxidant capacities of the extracts were measured using the 2,2-Diphenyl-1- PicrylHydrazyl (DPPH) scavenging technique and IC50 calculated. The extracts were screened for secondary metabolites and antibacterial activities. The result of the DPPH scavenging activities of the crude fractions compared favorably with that of the ascorbic acid used as standard control. 85.0% activity was recorded at concentration of 62.5μg/mL for the methanol stem bark fraction and, the least activity of 62.7% for ethyl acetate leaf extract at a concentration of 1000μg/mL. Terpenes, saponins, steroids, alkaloids, tannins, and flavonoids were detected in the extracts. Four of the eleven human pathogens used for antimicrobial screening were sensitive, four others were resistant. Zones of inhibition measured ranged between 18 and 29mm. Minimum inhibitory concentration and minimal bacteria/fungicidal concentrations were measured between 1.25 to 5.00mg/mL. The FTIR spectra of the powdered samples and extracts’ fractions reported.

Natural products from Rhynchostylis retusa (Orchidaceae), their chemophenetic significance and bioactivity

Rhynchostylis retusa (L.) Blume is traditionally used for the treatment of pain, inflammation, and skin diseases. The study aims the bioassay-guided isolation of chemical constituents from R. retusa using human breast cancer cell line MDA-MB-231 and murine RAW 264.7 macrophages. The cell viability assay revealed that crude extract, hexane, and chloroform fractions inhibited the viability of MDA-MB-231 and RAW 264.7 macrophages. The hexane fraction led to the isolation of four fatty acids; 1-octene (1), (9Z,12Z,15Z)-octadeca-9,12,15-trienoic acid (2), methyl oleate (3), methyl linolenate (4); and one steroid, stigmasterol (5). The isolation and purification of chemical constituents from the most potent chloroform fraction yielded two phenylpropanoids, dihydroconiferyl dihydro-p-coumarate (6) and retusiusine B (7); one bis-nor-lignan, 3,3′-dimethoxy-4,4′-dihydroxy-9,9′-bis-nor-lignan (8) and one bibenzyl, gigantol (9); and vanillin (10). This is the first report of compounds 1–10 from R. retusa (L.) Blume and the genus Rhynchostylis. 3,3′-dimethoxy-4,4′-dihydroxy-9,9′-bis-nor-lignan (8) is new from natural sources. Gigantol (9) is a bioactive compound of R. retusa as it showed an inhibitory effect on the viability of MDA-MB-231 cells and RAW 264.7 macrophages. The bioactivity of R. retusa is possibly due to the presence of gigantol (9) in the plant. Thus, the study corroborates the traditional uses of R. retusa for the treatment of pain and inflammation. The chemophenetic significance of the isolated compounds is also discussed in this study.

Comparative <i>in-vitro</i> antioxidant studies of crude methanol extract and four solvent fractions of red cultivar <i>Allium cepa</i> l. bulbs

Oxidative stress-induced disease conditions are on the increase and there is need for natural sources of antioxidant for relieving stress. The crude extract of Allium cepa L. has been well published as a potential natural source of antioxidant. But there is inadequate information on the antioxidant activities of fractionated extracts of the plant. The in-vitro antioxidant capacity of crude extract of red cultivar A. cepa bulbs and its four solvent fractions were studied and compared using Nitric oxide radical inhibitory assay (NORIA), FRAP and DPPH antioxidant models. The results of the three models revealed that the crude methanol extract exhibited very good antioxidant activities. However, with the aid of DPPH, the crude methanol extract, ethyl acetate and methanol fractions exhibited excellent antioxidant activities in comparison with ascorbic acid. Serial extraction did not make any of the fractionated extracts better than the crude extract of the red cultivar A. cepa bulbs. These results also showed that red cultivar A. cepa bulbs is a natural source of antioxidants and could serve as therapeutic agent in the prevention or slowing down of oxidative stress. Further studies are currently underway to identify the active component responsible for the observed antioxidant properties.

Verifying the Optimal Extraction Method and Content of Red Okra Extracts and Crude Pectin Using Response Surface Methodology (RSM)

Recently, many studies have reported the properties and functionality of okra pectin. However, these studies are about green okra pods, and pectin in red okra pods, stems, and roots has not been reported. Therefore, this study aimed to optimize the red okra extraction method using response surface methodology (RSM) analysis and evaluate the effects of extraction time, temperature, solvent ratio, and pH on the extract yield and crude pectin content. Based on RSM analysis, 4.35 h, 98.04 °C, 23.34 solvent ratio, and pH 3.36 are the optimal parameters for extracting crude pectin from red okra, and the crude pectin content was predicted to be 40.83%. When red okra was extracted under these extraction conditions (4 h, 100 °C, 23 solvent ratio, and pH 3), the extraction yield was 45.26%, and the crude pectin content was 38.42%, which was similar to the yield obtained under the conditions derived from the RSM analysis. In addition, the pH control using hydrochloric acid was replaced with citric acid, and the changes in extract yield and crude pectin content were compared. When citric acid was used, the extract yield was 49.15% (8.6% increase), and the crude pectin content was 42.76 ± 2.56% (11.3% increase); compared to when hydrochloric acid was used, the yield increased. Finally, the standardization of red okra raw materials was determined by analyzing the extraction yield and crude pectin content by part, harvest time, and size of red okra using the established extraction method using citric acid. As a result, it was confirmed that the extraction yield obtained from the established extraction method was reached from the extraction of red okra fruits up to 12 cm in size that were harvested between July and November. Additionally, compared to the non-pectin fraction, the crude pectin fraction isolated from red okra pod extract showed significantly higher total phenolic content (TPC) and total flavonoid content (TFC). These findings, reported for the first time, may contribute to the development of processes to purify red okra pectin, functional evaluation studies of pectin, and potential applications of red okra extract in various industries and research.

EVALUACIÓN DE UNA UNIDAD HPHT ACOPLADA CON SONDA NIR PARA DETERMINAR EL INICIO DE LA PRECIPITACIÓN DE ASFALTENOS BAJO DIFERENTES PRESIONES

Asphaltenes are characterized as the crude oil fraction with the highest molar mass and polarity, presetting mainly (poly) aromatic groups. The flocculation and deposition of asphaltenes causes large losses to the oil industry. Understanding the phase behavior of asphaltenes under conditions closer to those found in reservoirs is important. Therefore, LMCP/UFRJ started operating a high-pressure high-temperature (HPHT) unit coupled to a near-infrared spectrometer probe that can use different flocculants. This work describes the development of a procedure as well as the validation of the results obtained from this unit. Due to the complex composition of crude oil, model systems (MS) were prepared with asphaltenes extracted with n-pentane (C5I) and n-heptane (C7I). The experiments were carried out at atmospheric pressure, titrated with n-heptane, and at 100 and 300 bar titrated with propane. As expected, C7I asphaltenes were more unstable, presenting lower precipitation onset than C5I asphaltenes under ambient conditions and higher pressures. However, for both MS, the stability increased with rising pressure when using propane as a solvent. The proposed method to evaluate asphaltenes precipitation onset was effective for MS in toluene and dead crude oil, and is a promising alternative for investigation of different types of crude oil.

Antioxidant Activity, Cytotoxicity, and Identification of Secondary Metabolites of Kigelia africana from Waterpark Platinum Riau

Antioxidants can prevent reactive oxygen-associated diseases, which trigger carcinogenesis, cardiovascular disease, and premature aging. India and Africa have mostly practiced sausage trees (Kigelia africana) as traditional medicine, whereas Indonesia is still limited. The research aims to determine the phytochemistry and bioactivity of the n-hexane fractions from crude methanol extract of the leaves and bark of the sausage tree as antioxidants and their toxicity. The sausage tree is derived from Waterpark Platinum Riau. The sausage trees were macerated with methanol and fractionated by n-hexane and ethyl acetate. All samples were tested for their antioxidant to 1,1-diphenyl-2-picrylhydrazyl (DPPH) and cytotoxicity to Artemia salina Leach larvae. Phytochemical results of methanol crude extract, n-hexane, and ethyl acetate fractions of leaves and sausage bark showed the presence of all secondary metabolites except alkaloids in the n-hexane fraction. Antioxidant and cytotoxic activity of stem and leaf bark ethyl acetate fraction was stronger than crude methanol extract and n-hexane fraction. Based on LC-MS/MS data, the secondary metabolite components that have contributed strongly antioxidant activity of this study are flavonoid compounds such as kaempferol and the derivatives, lignans (cubebin), and steroids (pregnant).

Prediction and Rationalization of Abundant C-N-H Molecules in Different Environments.

The extreme chemical diversity of CmNnHk molecules is at the same time very important (central in organic chemistry) and difficult to rationalize in the sense that some molecules are abundant and easy to synthesize, while others are rare and difficult to make. Using the recently developed criteria of molecular "magicity", combined with evolutionary structure prediction and quantum-chemical calculations, we study these molecules in a wide range of compositions (0 ≤ m ≤ 13, 0 ≤ n ≤ 4, and 0 ≤ k ≤ 14). "Magic" molecules are defined as those that are lower in energy than any isochemical mixture of molecules with the nearest compositions. The predicted "magic" molecules are in good agreement with compounds found in versatile environments (interstellar and circumstellar media, Titan's lower atmosphere, and crude oil fractions) and in experimental chemistry. This work shows the predictive power of our approach, capable of predicting and explaining stable molecules in complex systems.

Использование бактериальной инокуляции для улучшения вегетативного роста и нутрицевтических качеств китайской капусты в условиях регулируемой агроэкосистемы

Abstract. In recent years, plant inoculation with beneficial plant growth-promoting microorganisms PGPR bacteria in agriculture has been steadily increasing, and they are expected to partially replace chemical fertilizers, pesticides and other growth regulators in the future, providing an environmentally friendly solution for sustainable farming practices. Consequently, the purpose of this study was to investigate the bacterial inoculation of Bacillus cereus to improve the vegetative growth and nutraceutical qualities of Chinese cabbage pak-choi plants grown by hydroponics under regulated agroecosystem conditions. Objects of the study are the plants of Chinese cabbage pak-choy (Brassica campestris ssp. cheninsis L.). Methods. When conducting research, we measured biometric indicators, determined the indicators of crude, dry matter, dietary fiber and mass fraction of ash of pak-choy plants according to GOST. The contents of vitamins, cations, anions and amino acids in plant products were estimated by capillary electrophoresis using “Kapel’-205” device (Russia). The obtained results were processed statistically by means of analysis of variance. Scientific novelty. The efficiency of root inoculation of Bacillus cereus bacteria in the cultivation of pak-choy plants by hydroponics in closed agro-ecosystems has been determined. The dependence of the improvement of quality indicators in the biomass of bacterial inoculation of pak-choi plants has been established. Results. As a result of studies, it was found that the inoculation of B. cereus had a positive effect on the development of the assimilating apparatus of pak-choi plants. The formation of the main morpho-biometric parameters (crude, dry weight, shoot length and number of leaves) of plants was not significantly affected by bacterial inoculation. All obtained values were at the control level. Pak-choi plants inoculated with bacteria showed a significant increase in concentration of mineral elements (K, S and P), amino acids (proline, phenylalanine, valine, histidine and lysine) and vitamins (ascorbic acid and choline).

Evaluation of in vitro Antioxidant, Antimicrobial and Cytotoxic Activities of Crude Ethanol Extract and Fractions of Achillea sintenisii Hub. Mor.

Objective: The Achillea species have been used to treat various ailments due to its anti-inflammatory, hemostatic, spasmolytic and cholagogue effects in the Turkish traditional medicine. However, there is no biological activity studies on some Achillea species except for the well-knowns. This work aimed to determine the antioxidant, antimicrobial and cytotoxic activities of the crude ethanolic extracts and fractions of Achillea sintenisii using in vitro methods. 
 Methods: The antioxidant activity was investigated by DPPH (1,1´-diphenyl- 2-picrylhydrazyl), ABTS (2,2´-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging, total phenol and total flavonoid content, and iron chelating methods. Antimicrobial activity evaluated by micro-plate dilution method against five test organisms. Cytotoxicity was determined by MTT method using MCF-7 breast cancer cell line and PC-3 prostate cancer cell line. Apoptosis was also measured by AO/EB staining. 
 Results: The n-Hexane fractions showed the highest antimicrobial and cytotoxic activities, respectively. Administration of the extracts on the cancer cells showed a concentration dependent inhibition on cell proliferation. The anti-proliferation effect could be via apoptosis and associated with the cell death. 
 Conclusion: The results showed that the extracts demonstrated antioxidant, antimicrobial, and cytotoxic activity, also supports the claims of traditional usage.

Insecticidal Activity of Tannins from Selected Brown Macroalgae against the Cotton Leafhopper Amrasca devastans.

Seaweeds, also known as marine macroalgae, are renewable biological resources that are found worldwide and possess a wide variety of secondary metabolites, including tannins. Drifted brown seaweed (DBSW) is particularly rich in tannins and is regarded as biological trash. The cotton leaf hopper Amrasca devastans (Distant) has caused both quantitative and qualitative losses in cotton production. Drifted brown seaweeds (DBSWs) were used in this study to extract, qualitatively profile, and quantify the levels of total tannins, condensed tannins, hydrolyzable tannins, and phlorotannins in the seaweeds; test their insecticidal activity; and determine the mechanism of action. The largest amount of tannin extract was found in Sargassum wightii Greville (20.62%) using the Soxhlet method (SM). Significantly higher amounts of hydrolyzable tannins (p = 0.005), soluble phlorotannins (p = 0.005), total tannins in the SM (p = 0.003), and total tannins in the cold percolation method (p = 0.005) were recorded in S. wightii. However, high levels of condensed tannins (CTAs) were observed in Turbinaria ornata (Turner) J. Agardh (p = 0.004). A. devastans nymphs and adults were examined for oral toxicity (OT) and contact toxicity (CT) against DBSW tannin crude extract and column chromatographic fractions 1 (Rf = 0.86) and 2 (Rf = 0.88). Stoechospermum polypodioides (J.V. Lamouroux) J. Agardh crude tannin was highly effective against A. devastans using the OT method (LC50, 0.044%) when compared with the standard gallic acid (LC50, 0.044%) and tannic acid (LC50, 0.122%). Similarly, S. wightii fraction 2 (LC50, 0.007%) showed a greater insecticidal effect against A. devastans adults in OT than gallic acid (LC50, 0.034%) and tannic acid (LC50, 0.022%). The mechanism of action results show that A. devastans adults treated with crude tannin of T. ornata had significantly decreased amylase, protease (p = 0.005), and invertase (p = 0.003) levels when compared with the detoxification enzymes. The levels of glycosidase, lactate dehydrogenase, esterase, lipase, invertase, and acid phosphate activities (p = 0.005) of S. wightii were reduced when compared with those of the Vijayneem and chemical pesticide Monocrotophos. In adult insects treated with LC50 concentrations of S. wightii tannin fraction 1, the total body protein (9.00 µg/µL) was significantly reduced (OT, LC50-0.019%). The SDS-PAGE analysis results also show that S. wightii tannin fraction 1 (OT and CT), fraction 2 (OT), and S. polypodioides fraction 2 (CT) had a significant effect on the total body portion level, appearance, and disappearance of some proteins and polypeptides. This study shows that the selected brown macroalgae can be utilized for the safer management of cotton leaf hoppers.

Assessment of asphaltene and resin fractions in crude oil using laser-induced fluorescence spectroscopy based on modified Beer-Lambert (LIFS-MBL)

Crude oil is one of the most significant petrogenic sources of polycyclic aromatic compounds (PACs). These substances play an essential role in the pollution of the marine environment. Therefore, the rapid identification of this pollutant source and its fractions is vital. For this purpose, a fast and on-site method of laser-induced fluorescence spectroscopy based on modified Beer-Lambert (LIFS-MBL) is proposed here using solvent densitometry. Three optical parameters of the self-quenching (K), the extinction (α), and the peak concentration (Cp) are experimentally extracted from MBL graphs. Note that the parameters above are known to be unique characteristics of various crude oils. The corresponding compounds are generally classified into saturate, aromatic, resin, and asphaltene fractions, abbreviated as SARA. Differentiation among these fractions is achieved using the LIFS-MBL method by selecting the optimal excitation wavelength at 405 nm. This line effectively rules out the light aromatic rings and focuses on heavy fractions. The correlation of optical parameters with heavy oil fractions is verified according to analysis of variance. Statistical relations are proposed to calculate crude oil fractions values. The values of light fractions including saturate and aromatic components can also be determined by the heavy fractions. In this method, the test time is notably reduced from four days using the standard methods to less than half an hour according to the presented LIFS-MBL technique.

Assessment of the Environmental Impact of Solid Oil Materials Based on Pyrolysis Oil.

One method of managing used car tires is decomposition by thermochemical conversion methods. By conducting the process at temperatures of 450-750 °C, three fractions are obtained from tires: oil, gas, and solid. The liquid product of the pyrolysis of used car tires is pyrolysis oil, which consists of aromatic, polyaromatic, and aliphatic hydrocarbons. Unconventional building materials were obtained from tire pyrolysis oil and the environmental impact was evaluated. Blocks made from pyrolysis oil showed mechanical strength of up to about 1700 N. No heavy metals or polycyclic aromatic hydrocarbons, which were found in the crude heavy-PO fraction, were detected in the filtrates after incubation of the block obtained from the heavy-PO fraction at 240 °C. The highest inhibition of Sorghum saccharatum shoot (74.4%) and root (57.5%) growth was observed for solid materials from the medium-PO fraction obtained at 240 °C. The most favorable values of the parameters for the process of obtaining blocks based on post-PO were an annealing temperature of 180 °C, time of 20 h, and mass ratio of catalyst to catalyzed oil of 0.045.

Purifying waxes through selective extraction of triacylglycerols from a bioethanol production side-stream using supercritical carbon dioxide

Agro-industrial sourced bioethanol fermentation produces a lipid slurry as a co-product. Corn/sorghum-based (3:2, w/w) post-fermentation lipid slurry was fractionated into crude oil and wax-rich fractions using supercritical carbon dioxide (SC-CO2). The process was optimized for temperature (35–75 °C), pressure (8–40 MPa), and time (2–6 h) for two responses (crude oil and wax yields) via response surface methodology (Box-Behnken design). The optimum conditions were determined as 35 °C, 40 MPa, and 4.1 h for maximum crude oil extraction (47.5 ± 1.4%, w/w) with minimum wax content (1.0 ± 0.2%, w/w). The crude oil fraction had a melting point of 17 °C, whereas the wax-rich fraction exhibited a melting point of 60 °C. The wax-rich fraction was composed of waxy compounds, wax esters, and aldehyde dimers. Overall, this study optimized SC-CO2 extraction conditions to selectively extract triacylglycerols from a bioethanol lipid slurry to purify waxes for food and non-food applications.

In vitro antioxidant, antibacterial, cytotoxic, and epigenetic screening of crude extract and fractions of the marine sponge Neopetrosia exigua from Mauritius waters

The marine sponge Neopetrosia exigua is known as a goldmine of novel compounds, yet its pharmacological activities remain poorly characterised. Herein, this study investigates the bioactivities of N. exigua collected from Mauritius waters. The crude extract (dichloromethane: methanol), hexane, ethyl acetate and aqueous fractions obtained from N. exigua were subjected to in vitro antioxidant assays. Their antibacterial activities were evaluated using the broth microdilution method to determine the minimum inhibitory concentration (MIC). The cytotoxic and epigenetic activities were further screened using the MTT assay and a cell-based image system that measures de-repression of a silenced Green Fluorescent Protein (GFP) reporter gene, respectively. Higher antioxidant activity was recorded for the ethyl acetate fraction as demonstrated by its significant ferric reducing antioxidant power, radical scavenging, and metal chelating activities relative to control (p < 0.05). The best antibacterial profile was presented by the ethyl acetate fraction against Cutibacterium acnes (MIC: 0.039 mg/ml), Streptococcus mutans (MIC: 0.078 mg/ml) and Mycobacterium smegmatis (MIC: 0.313 mg/ml). Similarly, the fraction displayed significant cytotoxicity against the human liposarcoma SW872 cells with IC50 value of 44.34 ± 2.64 µg/ml and GFP re-activation capacity of 43.79 ± 3.19% (p < 0.05). This work conveys interesting data on the antioxidant, antimicrobial, and anticancer properties of N. exigua. In particular, this study indicates the promising potential of N. exigua as a reservoir of epigenetically active agents that can modulate transcription of silenced genes involved in carcinogenesis. Hence, further investigations to isolate the active constituents is actively warranted.

Invitro Anticoccidial Effect of Fractions of Chromolaena Odorata Extract on Sporulating Eimeria Tenalla Oocysts

Coccidiosis is an important cause of digestive system disorders in birds, especially the poultry. The aim of this study was to evaluate the in-vitro anticoccidial effects of the different fractions of Chromolaena odorata extracts on sporulated Eimeria tenella oocysts. Sporulation inhibition bioassay was used to evaluate the activity of crude methanol fraction of Chromolaena odorata extracts; n-hexane, ethyl acetate, and methanol fractions of Chromolaena odorata on sporulation of Eimeria tenella, which was used to cause coccidiosis. in the birds following oral inoculation of experimental broiler birds with oocysts. Post-mortem examination of dead birds showed lesions consistent with Eimeria tenella. The fractions of the Chromolaena odorata were examined for the oocysticidal activities. Amprolium an anticoccidial drug was used as control. The fractions were used in 4 doses: 0 mg/ml; 0.1 mg/ml; 0.2 mg/ml and 0.4 mg/ml. The control drug was used in corresponding doses. The highest oocysticidal activity of tested plant extracts and fractions was recorded. The n-hexane fraction of Chromolaena odorata gave the highest oocysticidal activity. This oocysticidal activity was noticed at lower concentrations but activity was found to be dose-dependent. The highest oocysticidal activity for the n-hexane fraction of Chromolaena odorata was at 65.67±2.08%. Amprolium showed no oocysticidal activity at all concentrations. However, other fractions of the Chromolaena odorata showed different levels of oocysticidal activities. The methanol fractions were secondary to the n-hexane while the ethyl acetate was the least. In conclusion, this study provides the basis for using Chromolaena odorata, especially the n-hexane fraction as preventive agents against coccidiosis and suggests their use as a feed additive in the control of coccidiosis.

Oliveria decumbens Vent. (Apiaceae): Biological screening and chemical compositions

Ethnopharmacological relevanceOliveria decumbens Vent. (Apiaceae), a single aromatic species in Iran, is traditionally used for healing inflammation, gastrointestinal disorders, and infections. Aim of the studyRegarding the importance of O. decumbens in traditional medicine, we aimed to set out the plant's biological screening and analyze the chemical components of the active fractions. Materials and methodsAir-dried O. decumbens aerial parts were macerated by ethanol:water (70:30). Using a liquid-liquid extraction (LLE) technique, n-hexane, dichloromethane (DCM), ethyl acetate (EtOAc), n-butanol (n-BuOH), and water were successively used to fractionate the crude extract into different portions. Various biological activities were performed on the crude extract, fractions, and some experiments on pure compounds. The bioassays were as follows: antibacterial activity against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Pseudomonas aeruginosa, and Salmonella typhi (using microplate alamar blue assay; MABA), antifungal activity against Aspergillus niger, A. fumigatus, Candida albicans, C. glabarate, Fusarium lini, Microspurum canis, and Trichophyton rubrum (using agar tube dilution method), antileishmanial activity against Leishmania major and L. tropica using a 96-well serial dilution protocol, anti-inflammatory activity using the respiratory burst assay, cytotoxicity against HeLa (cervical cancer) and BJ (normal fibroblast) cells using MTT assay, insecticidal activity against Tribolium castaneum, Sitophilus oryzae, and Rhyzopertha dominica (using the contact toxicity method), larvicidal activity against Aedes aegypti, anti-DPPH• activity, and cytotoxicity against brine shrimp (Artemia salina) in a lethality assay. Eventually, the phytochemicals from the active fractions were studied by gas chromatography coupled with mass spectrometry (GC-MS). ResultsInterestingly, the DCM fraction was the most active, followed by the n-hexane fraction in the biological assays, including antibacterial (>80% inhibition), leishmanicidal (IC50 (L.major) = 29.4 μg/mL, and IC50 (L.tropica) = 30.0 μg/mL), anti-inflammatory (IC50 = 15.8 μg/mL), insecticidal (>80% inhibition), and larvicidal (100% inhibition of A. aegypti) assays. Further GC-MS analysis of the DCM and n-hexane fractions resulted in the characterization of 12 and 14 phytoconstituents, respectively, compared with the NIST library. Thymol and carvacrol were abundant in both fractions. To lesser quantities, the presence of monoterpenoids (p-cymen-8-ol, thymoquinone, 3-hydroxy-β-damascone, and 3-hydroxy-7,8-dihydro-β-ionol), phenylpropanoids (methoxyeugenol, elemicin, and 4-[(1E)-3-hydroxy-1-propenyl]-2-methoxyphanol, simple phenolics (salicylic acid and 4-methoxy-2,3,6-trimethyl-phenol), and a coumarin (6,7-dimethoxy-coumarin) were detected in the DCM fraction. On the other hand, besides a coumarin and monoterpenoids, the fatty acids (tetradecanoid acid, n-hexadecanoic acid, and linolenic acid) and a sesquiterpene (spathulenol) were observed in the n-hexane fraction. The EtOAc fraction scavenged the DPPH• radicals better than other fractions (IC50 = 41.4 μg/mL), while in brine shrimp lethality assay, the crude extract was more active than n-hexane and DCM fractions with LD50 = 385.20, 660.28, and 699.74 μg/mL, respectively. Surprisingly, the crude extract and fractions were ineffective against assayed fungal strains and tested cancer and non-cancer cell lines. ConclusionsOur findings showed that O. decumbens deserves to be a multi-bioactive medicinal plant, besides its ability for cereal protection against pests. To understand the principal mechanism of action, in silico, in vitro, and in vivo experiments may clarify the ambiguities and even figure out the synergistic behavior of the minor secondary metabolites.

Potential hemostatic compounds targeting urokinase plasminogen activator explored from three Euphorbiaceae species: Euphorbia maculata, Euphorbia humifusa, and Acalypha australis, with bio-affinity ultrafiltration UPLC-MS.

Numerous species of the Euphorbiaceae family, including Euphorbia maculata, Euphorbia humifusa, and Acalypha australis, have been used to manage bleeding disorders. However, few investigations have demonstrated their hemostatic potential, and their procoagulant compounds remain elusive. This study aimed to determine the most active procoagulant extracts from the three species' crude extract (CE) and fractions in order to screen out the active compounds and to analyze their possible mechanisms of action. An integrative approach, comprising prothrombin time and activated partial thromboplastin time evaluations and urokinase-type plasminogen activator (uPA) inhibitory assessment, followed by bio-affinity ultrafiltration paired with UPLC/QTOF-MS targeting uPA and docking simulations, was used. The extracts with highest procoagulant activity were the CE for both E. maculata (EMCE) and E. humifusa (EHCE) and the n-butanol fraction (NB) for A. australis (AANB). The most promising ligands, namely, isoquercetin, orientin, rutin, and brevifolin carboxylic acid, were selected from these lead extracts. All of these compounds exhibited pronounced specific binding values to the uPA target and showed tight intercalation with the crucial side chains forming the uPA active pocket, which may explain their mode of action. The activity validation substantiated their hemostatic effectivity in inhibiting uPA as they had better inhibition constant (Ki) values than the reference drug tranexamic acid. Collectively, the integrative strategy applied to these three species allowed the elucidation of the mechanisms underlying their therapeutic effects on bleeding disorders, resulting in the fast detection of four potential hemostatic compounds and their mode of action.