Ethyl Acetate Fraction Research Articles

Dual Functionality of Papaya Leaf Extracts: Anti-Coronavirus Activity and Anti-Inflammation Mechanism.

Papaya leaves have been used as food and traditional herbs for the treatment of cancer, diabetes, asthma, and virus infections, but the active principle has not been understood. We hypothesized that the anti-inflammatory activity could be the predominant underlying principle. To test this, we extracted papaya leaf juice with different organic solvents and found that the ethyl acetate (EA) fraction showed the most outstanding anti-inflammatory activity by suppressing the production of nitric oxide (NO, IC50 = 24.94 ± 2.4 μg/mL) and the expression of pro-inflammatory enzymes, such as inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX-2), and cytokines including interleukins (IL-1β and IL-6), and a tumor necrosis factor (TNF-α) in lipopolysaccharide (LPS)-induced RAW 264.7 cells. Transcriptomic analysis and Western blot results revealed its anti-inflammatory mechanisms were through the MAPK signaling pathway by inhibiting the phosphorylation of ERK1/2, JNKs, and p38 and the prevention of the cell surface expression of TLR4. Furthermore, we discovered that the EA fraction could inhibit the replication of alpha-coronavirus (HCoV-229E) and beta-coronavirus (HCoV-OC43 and SARS-CoV-2) and might be able to prevent cytokine storms caused by the coronavirus infection. From HPLC-QTOF-MS data, we found that the predominant phytochemicals that existed in the EA fraction were quercetin and kaempferol glycosides and carpaine. Counter-intuitively, further fractionation resulted in a loss of activity, suggesting that the synergistic effect of different components in the EA fraction contribute to the overall potent activity. Taken together, our results provide preliminary evidence for papaya leaf as a potential anti-inflammatory and anti-coronavirus agent, warranting further study for its use for human health promotion.

Cleistocalyx nervosum var. paniala mitigates oxidative stress and inflammation induced by PM10 soluble extract in trophoblast cells via miR-146a-5p

Air pollution poses a significant global concern, notably impacting pregnancy outcomes through mechanisms such as DNA damage, oxidative stress, inflammation, and altered miRNA expression, all of which can adversely affect trophoblast functions. Cleistocalyx nervosum var. paniala, known for its abundance of anthocyanins with diverse biological activities including anti-mutagenic, antioxidant, and anti-inflammatory properties, is the focus of this study examining its effect on Particulate Matter 10 (PM10) soluble extract-induced trophoblast cell dysfunction via miRNA expression. The study involved the extraction of C. nervosum fruit using 70% ethanol, followed by fractionation with hexane, dichloromethane, and ethyl acetate. Subsequent testing for total phenolics, flavonoids, anthocyanins, and antioxidant activity revealed the ethyl acetate fraction (CN-EtOAcF) as possessing the highest phenolic and anthocyanin content along with potent antioxidant activity, prompting its selection for further investigation. In vitro studies on HTR-8/SVneo cells demonstrated that 5–10 µg/mL PM10 soluble extract exposure inhibited cell proliferation, migration, invasion, and induced apoptosis. However, pretreatment with 20–80 µg/mL CN-EtOAcF followed by 5 µg/mL PM10 soluble extract exposure exhibited protective effects against PM10 soluble extract-induced damage, including inflammation inhibition and intracellular ROS suppression. Notably, CN-EtOAcF down-regulated PM10-induced miR-146a-5p expression, with SOX5 identified as a potential target. Overall, CN-EtOAcF demonstrated the potential to protect against PM10-induced harm in trophoblast cells, suggesting its possible application in future therapeutic approaches.

Phytochemical and antimicrobial evaluation of Miconia burchellii Triana (Melastomataceae).

Four novel triterpenoid saponins, 1α,3β,23-trihydroxy-olean-12-en-28-oic acid 28-O-β-glycopyranoside ester (1), 1α,3β,23-trihydroxy-urs-12-en-28-oic acid 28-O-β-glycopyranoside ester (2), 3-O-β-galloyl-1α,23-dihydroxy-olean-12-en-28-oic acid 28-O-β-glycopyranoside ester (3) and 3-O-β-galloyl-1α,23-dihydroxy-urs-12-en-28-oic acid 28-O-β-glycopyranoside ester (4) and two new pentacyclic triterpenoids, 3-O-β-trans-p-coumaroyl-1α-hydroxy-urs-12-en-28-oic acid (5) and 3-O-β-cis-p-coumaroyl-1α-hydroxy-urs-12-en-28-oic acid (6), together with six known compounds were isolated from the ethanolic extracts of Miconia burchellii leaves and stem bark. Their structures were established by HR-MS, 1D and 2D NMR and comparison with literature data. Additionally, the antimicrobial activity of the leaves and stem bark extracts and fractions was evaluated using the micro broth dilution technique. The crude extract, the ethyl acetate and methanol fractions of the stem bark were the most active against the tested bacteria, showing growth inhibition of both Gram-positive (Staphylococcus aureus and Staphylococcus epidermidis) and Gram-negative (Escherichia coli and Pseudomonas aeruginosa). To the best of our knowledge, this is the first study to report on the antimicrobial activity of M. burchellii.

Antimalarial potentials of Solanum dasyphyllum fractions against Plasmodium berghei infected mice

BackgroundThe leaves of Solanum dasyphyllum are commonly utilized in the management of fever in traditional medicine, yet scientific evidence supporting their antimalarial effectiveness is lacking. ObjectiveThis study aimed to assess the antioxidant properties of solvent fractions derived from these leaves, alongside investigating antimalarial potential against Plasmodium berghei in mice. MethodsAntioxidant activity was evaluated through total phenolic content determination, DPPH radical scavenging, and Ferric ion reducing antioxidant power assay. The fractions were tested against Plasmodium berghei (NK65) in 4-day suppressive, prophylaxis, and curative models, with mice orally administered 1 % Tween 80 (control), Solanum dasyphyllum hexane fraction (SdHXF), Solanum dasyphyllum dichloromethane fraction (SdDMF), Solanum dasyphyllum ethyl-acetate fraction (SdEAF), and Solanum dasyphyllum aqueous fraction (SdAQF) (each at 125 mg/kg), and chloroquine (10 mg/kg). IC50 values were determined via linear regression analysis, and antimalarial study results were statistically analyzed using one-way ANOVA followed by Duncan multiple range tests, considering significance at P value < 0.05. ResultsThe SdHXF (60.4 %), SdDMF (56.8 %), SdEAF (86.4 %), and SdAQF (94.8 %) exhibited notable percentage chemosuppression compared to the control in the 4-day suppressive test. In the prophylaxis test, SdEAF (80.6 %) and SdAQF (80.0 %) demonstrated similar percentage chemosuppression. In the curative test, the mean survival time significantly increased in SdHXF (30.2 ± 1.3 days), SdDMF (30.8 ± 3.2 days), SdEAF (31.4 ± 2.3 days), and SdAQF (30.1 ± 2.0 days) compared to the control (10.3 ± 0.1 days). The antioxidant activity of the fractions increased with concentration; however, it was lower than that of the positive controls (gallic acid and ascorbic acid). ConclusionThese results suggest that the fractions possess potent antiplasmodial activity against Plasmodium berghei, particularly in the mid-polar fractions, indicating the potential of the plant as a source of novel antimalarial agents.

Hypoglycemic and antioxidant activities of Jasminum officinale L. with identification and characterization of phytocompounds

The utilization of plant-derived chemicals with anti-diabetic properties is widely promoted for its advantageous tactics in managing diabetes, as they are cost-effective and have minimal or no adverse effects. Therefore, this work investigates the medicinal plant Jasminum officinale L. leaves by extraction and bio-guided fractionation. The ethyl acetate fraction showed a higher yield of 36.4 %. A phytochemical test on Jasminum officinale confirmed flavonoids, saponins, phenols, and tannins. The highest total phenol and flavonoid contents in the ethyl acetate fraction of J. officinale are 103.01 ± 1.1 mg GAE/g and 80.29 ± 1.03 mg QUE/ value found in methanol crude extract. Furthermore, HPTLC analysis of the ethyl acetate fraction detected the existence of flavonoids (kaempferol) and phenols (gallic acid, quercetin, and rutin). The compounds detected at the greatest concentrations in the LC-M/MS analysis of the ethyl acetate fraction were cirsiliol, kaempferol, and 2-tridecanone. Additionally, J. officinale (IC50 33.845 ± 1.09 μg/mL) demonstrated the highest DPPH scavenging activity in EAF like that of ascorbic acid (IC50 22.27 ± 0.96 μg/mL). Also, in the FRAP assay, the IC50 of this fraction is 15.14 ± 0.25 μM Fe equivalents. In the range of alpha-amylase deactivating action, from 13.25 % to 74.51 %, and IC50 value (47.40 ± 0.29 μg/mL) was significantly higher in the ethyl acetate fraction of J. officinale leaf extract. Moreover, J. officinale leaf extract had a substantially higher retention of glucose level (23.92 ± 0.85 % to 87.21 ± 0.6 %), significantly higher anti-inflammatory activity with the lowest IC50 value (66.00 ± 1.84), and lipid peroxidation (IC50 value 34.67 ± 1.69) by utilizing egg yolk as a substrate for lipids. Overall, the study revealed that J. officinale has considerable anti-diabetic characteristics. However, further comprehensive research is necessary to ascertain the medicinal purposes of J. officinale and its chemical components, pharmacological effects, and clinical uses.

Stevia By-product Fraction has Antioxidant Capacity and has been Evaluated for its Antihyperglycemic and Antilipid Potential

Introduction/Objective: The ethanolic pretreatment of stevia leaves is considered ecologically favorable and increases its yield, purity level, and sensory profile of sweeteners. We investigated the by-product generated so that we can provide a viable and applicable destination to effectively contribute to the production and industrial chain of stevia sweeteners. Recently, an antioxidant and antidiabetic fraction was obtained through stevia methanolic extraction. However, this process is costly, has a low yield, and is non-green. In this study, we got an antioxidant fraction of stevia from its pretreatment by-product, the ethanolic extract, characterized its bioactive compounds – mainly phenolic acids and flavonoids – and evaluated its antidiabetic and antilipid capacity. Methods: The ethanolic extract was fractionated, then the ethyl acetate fraction was microencapsulated. Physicochemical, Mass Spectrometry (UHPLC/MS-MS), scanning electron microscopy, microencapsulation efficiency, and antioxidant capacity analyses were carried out. The antioxidant capacity was evaluated. Antihyperglycemic capacity was assessed by inhibition of α-glucosidase and α-amylase. For antilipid potential, inhibition of pancreatic lipase was measured. Results: Antioxidant potential was observed mainly in the fraction. Microencapsulation improved the stability of the fraction. Mass spectrometry allowed comparison with phenolic and flavonoid compounds from the methanolic and ethanolic fractions of the by-product. Many compounds were coincident, which may explain the similar antioxidant capacity found. The samples showed more than 90% inhibition of alpha-glucosidase, which may indicate potential antidiabetic activity. All samples showed inhibition of the pancreatic lipase enzyme higher than 80%, reaching 91.05% for the free fraction. Conclusion: This study represented the recovery of a by-product and showed that this antioxidant fraction deserves further investigation for its bioactive potential, especially antioxidant, antidiabetic, and anti-lipid.

Antioxidant and Antifungal Activities and Characterization of Phenolic Compounds Using Ultra-High Performance Liquid Chromatography and Mass Spectrometry (UPLC-MS) of Aqueous Extracts and Fractions from Verbesina sphaerocephala Stems.

The Verbesina gender represents the second most diverse group from the Asteraceae family in Mexico; Verbesina sphaerocephala is one of the most distributed species along the Mexican territory. This species has been poorly studied, reporting the presence of some bioactive compounds with antioxidant and antibacterial activity. In this study, phenolic and flavonoid contents and composition, antioxidant and antifungal activities of aqueous extracts of the stem of V. sphaerocephala and its fractions were determined. The results showed that the highest antifungal activity against Botrytis cinerea was shown by the aqueous extract (IC50: 0.10 mg/mL) and the ethyl acetate fraction (IC50: 14.8 mg/mL). In addition, the aqueous extract and the ethyl acetate fraction exhibited the highest phenolic (21.40 and 21.26 mg gallic acid equivalents per gram of dry extract, respectively) and flavonoid contents (11.53 and 3.71 mg rutin equivalents, respectively) and high antioxidant activity determined by the Total Antioxidant Capacity (20.62 and 40.21 mg ascorbic acid equivalents per gram of dry extract, respectively), Ferric Reducing Power (74.76 and 129.57 mg gallic acid equivalents per gram of dry extract, respectively), DPPH (IC50: 12.38 and 7.36 mg/mL, respectively), and ABTS (IC50: 5.60 and 7.76 mg/mL, respectively) methods. Twelve phenolic compounds were detected in the aqueous extract using UPLC-MS analysis, of which the major ones were protocatechuic, vanillic, and hydroxybenzoic acid, while in the ethyl acetate fraction, the presence of 18 phenolic compounds were identified, of which the majority were vanillin, rutin, and hydroxybenzoic acid. The results of this research demonstrate that the aqueous extract of V. sphaerocephala stems has phenolic compounds with antifungal and antioxidant activity.

Evaluation of inhibitory potential of the flavonoid-rich fraction of Myrica esculenta Buch.-Ham. ex D. Don against DSS-induced colonic inflammation in mice

ObjectiveMyrica esculenta (family Myricaceae), a valuable plant species in China and India, has been traditionally used by many tribes and local communities to manage gut disorders. Scientific validation of its anti-ulcerative colitis activity was aimed. MethodsThe ethyl acetate fraction of Myrica esculenta (MeEa) was prepared and evaluated for its potency against DSS-induced ulcerative colitis (UC) in mice at 200 and 400 mg/kg BW oral dose. The effective dose of MeEa was determined through its effect on DSS-induced UC and was further analyzed through its effects on disease activity index (DAI), colon length, colon weight/length ratio, spleen weight, serum and colon tissue cytokine level, cell count and hemoglobin content. Further, the effect was determined through histopathology and FITC-dextran-induced membrane permeability assay. ResultsBetween the two doses, MeEa at 400 mg/kg BW was found to be the most effective dose in terms of reduced DAI scores, protected colon length from shortening, decreased colon weight/length ratio, reduced spleen weight, decreased pro-inflammatory cytokine level and stabilized the anti-inflammatory cytokine level in serum and colon tissue. MeEa 400 reduced cell counts and increased hemoglobin content and platelet count. MeEa 400 also prevented the colon by protecting epithelial cells and crypts. MeEa 400 provided significant protection from intestinal leakage and reduced FITC dextran level in serum. DiscussionMeEa 400 possesses significant anti-inflammatory potential and acts via attenuation of DSS-induced UC and inhibition of DAI scores. It reduces pro-inflammatory cytokines and stabilizes anti-inflammatory cytokine levels, reduces cell count, and protects epithelial tissue and crypts in the colon, as well as intestinal membrane leakage that occurred due to FITC-dextran administration in mice.

Phenolic Contents and Antioxidant Properties of Bauhinia rufescens, Ocimum basilicum and Salvadora persica, Used as Medicinal Plants in Chad

The plants Bauhinia rufescens, Ocimum basilicum and Salvadora persica are well known in traditional African medicine, and particularly in traditional Chadian medicine. They are commonly used to treat infectious diseases, inflammatory diseases, fevers, gastroenteritis and other medical conditions. The aim of this study was to perform a phytochemical screening to determine the antioxidant properties of different extracts and fractions from the three plants. Ethanolic extracts and solvent fractions were prepared and analyzed for total phenolic content (TPC), total flavonoid content (TFC) and total tannin content (TTC). LC-MS and an online screening HPLC-ABTS system identified phytochemicals with antioxidant activities. DPPH and ABTS reduction methods were used to test the extracts and fractions for their antioxidant potential. The results showed that the TPC of O. basilicum was higher than that of B. rufescens, ranging from 64.70 ± 5.2 to 411.16 ± 8.11 mgGAE/g DW. B. rufescens extracts and fractions, on the other hand, showed higher TFC, ranging from 69.5 ± 5.3 to 408.26 ± 8.42 mgQE/g DW, and higher TTC, ranging from 4.57 ± 2.45 to 62.19 ± 4.7 mgTAE/g DW. The maximum TPC, TFC and TTC in both plants were recorded in the ethyl acetate fractions. S. persica extracts and fractions showed a very low quantity of TPC, TFC and TTC. Based on LC-MS and HPLC-ABTS analysis, rosmarinic acid was identified as the major component in the extracts and all fractions of O. basilicum, and epicatechin, procyanidin B and quercetin were found in B. rufescens. S. persica did not exhibit specific substances with antioxidant activity and was therefore not considered for further assays. DPPH and ABTS results showed that ethyl acetate fractions of B. rufescens and O. basilicum have the strongest antioxidant activities. This study indicates that B. rufescens and O. basilicum are good sources of phytochemicals with antioxidant properties, suitable for medicinal use in Chadian communities. Additionally, the antioxidant-rich extracts from these plants hold significant potential for cosmetic development, enhancing skin health and protecting against oxidative-stress-induced damage.

Functional Properties and Components of Koenigia alpina Extract.

Koenigia alpina (All.) T.M.Schust. & Reveal (alpine knotweed) is a perennial herb belonging to the Polygonaceae family. Several studies have examined Polygonaceae species' potential applications as cosmeceutical materials; however, the potential of K. alpina as a cosmeceutical has not yet been studied. Hydrogen peroxide (H2O2) and lipopolysaccharide were used to induce an inflammatory response in RAW 264.7 cells. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) radicals and H2O2 were used to evaluate the free-radical scavenging activity of K. alpina extract and its protective effect against reactive oxygen species (ROS)-induced cell damage. The whitening, antiaging, and cell proliferation/migration effects of the extracts were evaluated via tyrosinase inhibition, collagenase/elastase inhibition, and wound healing assays, respectively. The anti-inflammatory effect was confirmed by evaluating nitric oxide (NO) production in RAW 264.7 cells. High-performance liquid chromatography (HPLC), UV, and MS/MS were used to determine the main components of the extract and fractions. The ethyl acetate (EA) fraction and its aglycone fraction showed very high free-radical scavenging activities (47.5 and 47.1µg/mL, respectively). The extract/fractions also showed significant tyrosinase inhibition (IC50 = 0.38mg/mL in EA fraction), collagenase inhibition (IC50 = 0.21mg/mL in EA fraction), and elastase inhibition (IC50 = 0.57mg/mL in aglycone fraction). NO production in lipopolysaccharide-induced RAW 264.7 cells was inhibited by the extract/fractions. The extract also promoted the closure of scratch wounds in HaCaT cells. The K. alpina extract/fractions contained cardamonin, quercetin, and quercitrin. K. alpina extracts/fractions showed antioxidant, antiaging, whitening, and anti-inflammatory activities, suggesting they may have potential as antiaging cosmeceuticals.

Phytochemical profiling and anticancer potential of Cymbopogon citratus extract

Objective: To evaluate the anticancer potential of Cymbopogon citratus extract. Methods: GC-MS analysis was used to identify phytocomponents in the methanolic extract of Cymbopogon citratus. A fractionation method was employed to isolate and assess the bioactivity of different fractions and their cytotoxic activities against cancer cell lines HCT116, LoVo, Caco-2, and HT-29 were investigated. A dual staining method with acridine orange and ethidium bromide was used to assess the effect of the extract on cell apoptosis. Additionally, the expression levels of Bax and TP53 were quantified using realtime PCR in Caco-2 cells treated with the ethyl acetate fraction of Cymbopogon citratus extract. A protein array was employed to profile key pro- and anti-apoptotic proteins in Caco-2 cells. Moreover, molecular docking studies were conducted to investigate the interactions between key compounds of Cymbopogon citratus extract and specific apoptosis-related protein domains (PDB IDs: 7wql and 4bkx). Results: A significant growth inhibition was observed in Caco-2 cells treated with Cymbopogon citratus extract. Among the seven fractions of the plant extract, the ethyl acetate fraction showed the highest cytotoxicity against Caco-2 cells with an IC50 value of (6.16 ± 0.01) μg/mL. The immunofluorescence assay showed that the ethyl acetate fraction could induce apoptosis of Caco-2 cells. Moreover, the fraction upregulated the gene expressions of Bax and TP53 in a dose-dependent manner. The docking analysis demonstrated the interaction of five compounds isolated from the ethyl acetate fraction with key proteins in Caco-2 cells, indicating their anticancer properties. Conclusions: Cymbopogon citratus extract shows anticancer activity against Caco-2 cells by inducing apoptosis. It may be a promising candidate for the treatment of colon cancer, which needs further investigation.

OPTIMISATION METHOD FOR OBTAINING A BIOLOGICALLY ACTIVE SUBSTANCES FROM THE PLANT PETROSIMONIA BRACHIATA

Kazakhstan is rich in medicinal plants, so it needs to be studied. Ten species of the Petrosimonia belonging to the Chenopodiaceae family are growing in Kazakhstan. These plants have been adapted to harsh climatic conditions in the country’s salty, saline, and desert areas. Petrosimonia plants are a new species that has not been fully studied. Chinese scientists studied Petrosimonia sibirica species and found that the plant contains alkaloids, steroids, terpenes, flavonoids, and phenolic acids and has antibacterial activity (Wen, 2015; Ying, 2016). The qualitative and quantitative compositions of biologically active substances in P. sibirica, P. Glaucescens, P. triandra, and P. brachiata growing in Kazakhstan were studied. To prepare phytopreparations from P. Sibirica, P. Glaucescens and P. triandra plants were extracted using the classical maceration method, and individual representatives of biologically active compounds were isolated (Toktarbek, 2021; Seitimova, 2022). In this study, a phytochemical study of P. brachiata was conducted. The method for obtaining biologically active complexes with high efficiency was optimised by using supercritical fluid and ultrasonic extraction methods. These methods have a short extraction time, can be carried out at room temperature, are cost effective, and are a modern green chemical approach. Using supercritical fluid CO2 extraction (180 bar, 2 hours), the plant was purified from lipophilic substances. Further, a biologically active complex was obtained by pouring 70% ethanol-water solvent into the plant raw material and performing ultrasound extraction. The obtained complex was analysed by thin-layer chromatography using different organic solvent systems. Based on our analysis, steroids, terpenes, phenolic acids, and flavonoid glycosides were found in the extract. Hexane and ethyl acetate fractions were obtained as a result of liquid-liquid extraction of the extract using organic solvents. Stigmasterol 3-O-β-D-galactopyranoside, isovanillic acid, quercetin 3-O-β-D-glucopyranoside, and isorhamnetin 3-O-α-L-rhamnopyranoside were isolated by washing the hexane and ethyl acetate fractions in a silica gel column with an organic solvent system. The isolated compounds will be tested for biological activity.

Isolation and Characterization of Cytotoxic Compounds from Detarium microcarpum Guill. and Perr. Stem Bark.

Globally, about 8.2 million cancer-related deaths are recorded annually. Sadly, most of the deaths result from the toxicity of most chemotherapeutic agents. Hence, there are growing demands for chemotherapeutic agents with high specificity and selectivity. This study was designed to assess the cytotoxic potential of Detarium microcarpum and isolate cytotoxic compounds with better selectivity profiles. Detarium microcarpum Stem bark (DMS) was collected and authenticated at the Forest Herbarium Ibadan (FHI), and a voucher (FHI-111954) was issued. Dried DMS was pulverized and extracted into 70% methanol. The extract was partitioned into hexane, dichloromethane, and ethyl acetate fractions. The cytotoxicities of the extract, fractions, and isolated compounds were determined. The cytotoxicity of the isolated compounds was tested against different cell lines, including human breast (AU565 and MDA MB231), oral adenosquamous (CAL27), and cervical (HeLa) cancer cells, as well as healthy (3T3) non-cancer cells. Methyl gallate, eriodictyol, quercetin, quebrachitol, catechin, catechin gallate, and gallic acid, isolated from dichloromethane and ethyl acetate fractions, displayed weak cytotoxicity against breast (AU565 and MDAMD- 231) and cervical (HeLa) cancer cell lines. Interestingly, all the compounds, except gallic acid (48.91±4.51% inhibition), displayed potent cytotoxicity on oral cancer cells. Methyl gallate and quercetin displayed the highest activity, with IC50 values of 89.57±1.98μM and 78.19±1.49μM, respectively. Interestingly, all the compounds were not toxic to healthy non-cancer (3T3) cells. The compounds displayed anticancer activity specific to oral cancer cells and were highly selective for cancer cells without causing significant toxicity to healthy non-cancer cells.

Nutmeg (Myristica fragrans) Extracts Accelerate Oral Mucosal Wound Healing In Vivo

Background: Oral mucosal pain and sores are common concerns in dental care, often resulting from surgeries, infections, trauma, or autoimmune disorders. Conventional treatments like NSAIDs offer relief but carry adverse side effects. Nutmeg (Myristica fragrans) has been traditionally used for its analgesic, anti-inflammatory, and antimicrobial properties. However, its potential in oral mucosal wound healing remains underexplored. Methods: This study utilized male Wistar rats (Rattus norvegicus) to assess the wound healing effects of nutmeg extracts and fractions. Nutmeg was extracted using maceration, and the fractions were obtained through liquid-liquid extraction. Rats were divided into control and treatment groups, and topical nutmeg treatments were applied daily to incised gingival mucosa. Wound closure time was monitored daily, and healing rates were analyzed. Results: The ethyl acetate fraction of nutmeg exhibited the fastest wound healing, achieving an average healing rate of 88.79%, closely matching the positive control (94.65%). Other fractions also demonstrated efficacy, with the aqueous fraction achieving 82.46% and the ethanol fraction 75.54%. The hexane fraction showed the slowest healing at 63.67%. Statistical analysis revealed significant differences between the groups (p &lt; 0.05). Conclusion: Nutmeg extracts, particularly the ethyl acetate fraction, significantly accelerated wound healing in the oral mucosa of Wistar rats, highlighting its potential as a natural wound-healing agent. Further research is needed to isolate the bioactive compounds responsible for these effects and to explore their clinical applications in dental care.

Effect of Justicia insularis Leaf Extract and Fractions on Oxidative Stress Markers, Liver Function Parameters and Liver Histology of Plasmodium berghei -Infected Mice

Justicia insularis (Family-Acanthaceae) is used in Ibibio ethnomedicine to treat malaria. The leaf extract and fractions of J. insularis were investigated for antioxidative stress and hepatoprotective activities in Plasmodium berghei-infected mice. The leaf extract (100-300 mg/kg, p.o.) exerted significant (p&lt;0.05) antimalarial activity against P. berghei infection in curative test with ethyl acetate fraction demonstrating the highest activity. The extract/fractions treatment caused significant (p&lt;0.05) reductions in liver enzymes (ALT, AST and ALP), total and conjugated bilirubin of the treated infected mice and also decreased significantly (p&lt;0.05) total protein and albumin levels of the treated mice relative to control. The leaf extract and fractions further improved significantly (p&lt;0.05) the levels of oxidative stress markers enzymes and molecules (CAT, GPx, GST, SOD) of the treated infected mice with no significant (p&gt;0.05) effect on GSH. The MDA levels in the livers of the treated infected mice were significantly (p&lt;0.05) reduced relative to control. Histology of liver sections revealed absence or significant reductions in pathological features in infected mice treated with leaf extract (100 mg/kg), DCM and ethyl acetate fractions compared to untreated infected mice. These results suggest that the leaf extract/fractions of Justicia insularis possess antioxidative stress and hepatoprotective potentials, which is an added advantage to its antimalarial property.

The Inhibitory Potentials of the Root Extract and Fractions of Brenania brieyi (De Wild) (Rubiaceae) on Lactate Dehydrogenase, Phospholipase A2, and α- Amylase in Plasmodium falciparum - Infected Persons

Medicinal plants have been used as an alternative in malaria chemotherapy. This study aimed to determine the in vitro inhibitory potentials of the extract and fractions of Brenania brieyi on Lactate Dehydrogenase (LdH), Phospholipase A2 (PLA2), and α-Amylase enzyme extracted from Plasmodium falciparum. Cold maceration and Liquid-Liquid partitioning were used to obtain the extract and fractions respectively, using different solvent of varying polarity. Phytochemical analysis was done using Gas Chromatography-Flame Ionization detector (GCFID) method to identify and quantify different phytoconstituents. In vitro enzyme assay (α-Amylase, LdH and PLA2) were performed using standard procedure. The extract contained Alkaloids such as Spartein (5.05 µg/ml), Dihydrocytisine (23.12 µg/ml), Aphyllidine (5.25 µg/ml), Ribalinidine (2.65 µg/ml), Epihedrine (1.91 µg/ml), Saponins such as sapogenin (5.70 µg/ml), Tannins (16.25 µg/ml), Flavonoids such as Kaempferol (4.59 µg/ml), Catechin (5.50 µg/ml), Proanthocyanidin (9.81 µg/ml), Anthocyanin (3.22 µg/ml), Narigenin (3.62 µg/ml), Flavonones (8.73 µg/ml), Steroids (8.58 µg/ml), Polyphenols such as Reservatrol (20.68 µg/ml) and Glycosides such as cardiac glycoside (4.42 µg/ml), Cyanogenic glycosides (17.72 µg/ml) while Terpenoids are absent. The percentage inhibition of the enzyme (PLA2, LdH and α-amylase) at 100 mg/ml of extract showed the highest inhibition (37 %, 0 % and 38.33 %) when compared to controls, while ethyl acetate fraction has the highest inhibition (98 %, 34 % and 92 %) at P ˂ 0.05 at 100 mg/kg respectively. This study justifies the use of ethnomedicinal remedies (B. brieyi) in treating malaria in infected individuals. The study also suggests that the extract and fractions may act through the inhibition of plasmodial enzymes in infected persons.

Cancer Chemotherapeutic Effect of Vernonia Amygdalina Delile on Glioblastoma Brain Cancer Cell.

This study is targeted at assessing the chemotherapy factor of the ethanol extract of Vernonia amygdalina Delile (VAD). U87 glioblastoma cells were treated with extract and a fraction of Vernonia amygdalina Delile (VAD) was harvested from the herbarium. Cytotoxicity was evaluated to determine the IC50 through microscopic observation followed by an MTT assay. Subsequently, flow cytometry with a FACS type was employed to conduct cell cycle and apoptosis analyses. Annexin V/PI and PI markers were used to assess apoptosis and cell cycle progression. The ethanol extract and ethyl acetate fraction of VAD showed promising effects as cancer chemotherapy in glioblastoma cells. The IC50 values for the extract and fraction were notably low, at 37.65 µg/ml and 10.12 µg/ml, respectively, for U87 cells. Analysis of apoptosis using FACS revealed a more pronounced apoptotic effect of the 15 µg/ml fraction of VAD on both early and late apoptosis compared to the 75 µg/ml extract of VAD. Although some differences in cell cycle properties were observed, there were no significant differences in cell cycle analysis between the extract and fraction. These findings underscore the efficacy of VAD's ethanol extract and ethyl acetate fraction as chemotherapeutic agents against U87 cancer cells. The low IC50 values and significant induction of early apoptosis highlight the cytotoxic effects of these treatments on U87 cells.

Bioassay-guided isolation and structure elucidation of anti-mycobacterium tuberculosis compounds from Galatella grimmii (Regel & Schmalh.) Sennikov

BackgroundGalatella is a genus in the family Asteraceae, represented by 35-45 species. Considering the high effectiveness of the ethyl acetate (EtOAc) fraction of G. grimmii against Mycobacterium tuberculosis (MIC = 0.5 µg/mL), a bioassay-directed fractionation of this extract was carried out.MethodsThe methanolic extract of the aerial parts of G. grimmii was obtained using maceration, then it was suspended in water and partitioned with petroleum ether, dichloromethane (CH2Cl2), EtOAc, and n-butanol (n-BuOH), successively. The most potent fraction (EtOAc), was selected for further isolation by Sephadex LH–20 and semi-preparative HPLC to obtain active compounds.ResultsFractionation of the EtOAc solvent fraction resulted in the characterization of five compounds, among them, compounds 1 and 2 showed the highest anti-mycobacterial effects with MICs of 0.062 and 1.00 µg/mL against H37Rv M. tuberculosis, respectively, which were higher than those of rifampin (MIC of 1.25 µg/mL) and isoniazid (MIC of 0.31 µg/mL), as positive controls. Also, compound 1 ​​inhibited all tested strains of drug-resistant Mycobacterium (MDR and XDR). Notably, the isolated compounds have been reported for the first time from G. grimmii.ConclusionDue to the potent anti-mycobacterial effect of isolated compounds from G. grimmii, this study could pave the way for developing a novel class of natural anti-tuberculosis compounds.

Phytochemical Characterization and Synergistic Antibacterial Effects of Colebrookea Oppositifolia Essential Oil as Adjuvants to Modern Antibiotics in Combating Drug Resistance.

The global threat of multi-drug-resistant bacteria has severely limited the options available for effective antibiotics. This study focuses on the antimicrobial activity and phytochemical characterization of C. oppositifolia extracts, aiming to identify novel plant-based therapeutic agents. C. oppositifolia specimens-leaves and inflorescence. Specimens were cleaned, sterilized, dried, and ground into a fine powder. Extracts were obtained using methanol and petroleum ether via a Soxhlet apparatus, followed by fractionation with chloroform, n-butanol, and ethyl acetate. Volatile oil was extracted through hydro distillation using a Clevenger apparatus. Phytochemical analysis was conducted to identify bioactive compounds. Biophysical techniques, including UV-visible spectrophotometry, TLC, HPLC, GC-MS, FTIR, and NMR, were employed for characterization. Antimicrobial activity was tested against S. aureus ATCC25922 and E. coli ATCC25922 using agar well and disc diffusion methods, and synergistic effects were assessed with erythromycin and amoxicillin. Methanol extract exhibited bacteriostatic activity with inhibition zones of 13.0 ± 0.2 mm for both S. aureus and E. coli. Petroleum ether, chloroform, n-butanol, and ethyl acetate fractions showed varying inhibition zones. Erythromycin demonstrated bactericidal activity, which was enhanced synergistically when combined with methanol extract and volatile oil, increasing inhibition zones against S. aureus. Phytochemical analysis identified phenols, flavonoids, tannins, coumarins, alkaloids, terpenoids, saponins, and glycosides. FTIR analysis revealed functional groups such as amines, aldehydes, nitriles, alkenes, and sulfones. GC-MS identified 24 compounds, with α-pinene, caryophyllene, and carene as major components. NMR spectra indicated no complex formation between oils and antibiotics, suggesting the compounds act as synergists. The C. oppositifolia extracts possess significant antimicrobial activity and synergistic potential, particularly against S. aureus. The presence of various bioactive compounds suggests a promising role in developing new plant-based therapeutics.

Potensi Aktivitas Antimalaria Residu Hidrodestilasi Kulit Kayu Manis (Cinnamomum Burmanii) Melalui Penghambatan Polimerisasi Heme

Cinnamon (Cinnamomum burmanii) is a herbal plant from Indonesia which is widely used in cooking, cosmetics and medicine. Cinnamon bark isolate is thought to contain compounds with the structure of isoquinoline which are known to be used as antimalarials. The aim of this study was to examine the potential antimalarial activity of extracts and fractions of cinnamon bark hydrodistillation residue based on the % inhibition and IC50 value. These results will also be compared with extracts and fractions of cinnamon bark without hydrodistillation as controls. The test method is carried out in vitro by heme polymerization inhibitor. The test results showed that the n-hexane fraction, DCM fraction did not have inhibitory activity, while the ethyl acetate fraction had better potential than other extracts and fractions, where the ethyl acetate fraction after hydrodistillation (residue) had an IC50 value of 71.84 µg/mL, while the ethyl acetate fraction without hydrodistillation (control) had an IC50 value of 120.05 µg/mL. This result is better than hydroxychloroquine as positive control which has an IC50 value of 184.98 µg/mL. These results indicate that the ethyl acetate fraction from the hydrodistillation residue of cinnamon bark can be utilized and has potential as an antimalarial.