Fractionation Of Extract Research Articles

Aktivitas Penghambatan Enzim α-Amilase Ekstrak Etanol dan Fraksi Buah Belimbing Wuluh (Averrhoa bilimbi L.)

The enzyme α-amylase is one of the enzymes that plays a role in the process of degrading starch into maltose and glucose. Organic compounds such as polyphenols and flavonoids are inhibitors of α-amylase. The star fruit plant (Averrhoa bilimbi L.) has α-amylase enzyme inhibition activity and has the potential to be used in the therapy of type 2 diabetes mellitus. This study aims to determine whether ethanol extract and the fraction of star fruit (Averrhoa bilimbi L.) have α-amylase enzyme inhibition activity, inhibition against α-amylase enzyme and better inhibition activity. The test of α-amylase enzyme inhibition activity was carried out using 3 test groups, namely negative control, positive control (acarbose) and samples (ethanol extract and star fruit fraction) with concentrations of 12.5 ppm, 25 ppm, 50 ppm, 100 ppm and 200 ppm. The results of testing the inhibitory activity of α-amylase enzyme of ethanol extract and the fraction of star fruit (Averrhoa bilimbi L.) obtained IC50 values of 5.581 μg/mL, 76.725 μg/mL, 43.152 μg/mL and 55.447 μg/mL, respectively. Based on these results, it can be concluded that the ethanol extract of star fruit (Averrhoa bilimbi L.) at a concentration of 200 ppm has a very strong activity in inhibiting the α-amylase enzyme with an inhibition percentage of 85.217% and an IC50 value of 5.581 μg/mL.

Assessment of the Interaction of Acetylcholinesterase Binding with Bioactive Compounds from Coffee and Coffee Fractions Digested In Vitro in the Gastrointestinal Tract.

The aim of the study was to evaluate the degree of acetylcholinesterase (AChE) inhibition by green and light- and dark-roasted coffee extracts and their fractions after digestion in a simulated gastrointestinal tract. The analysis was carried out using isothermal titration calorimetry, molecular docking, and dynamics simulations. The results showed that 3-O-caffeoylquinic acid binds strongly to AChE through hydrogen interactions with the amino acids ARG289A, HIS440A, and PHE288A and hydrophobic interactions with TYR121A in the active site of the enzyme. The Robusta green coffee extract (ΔG = -35.87 kJ/mol) and dichlorogenic acid fraction (ΔG = -19-29 kJ/mol) showed the highest affinity. Dichlorogenic acids (3,4-O-dicaffeoylquinic acid, 4,5-O-dicaffeoylquinic acid, and 3,4-O-dicaffeoylquinic acid) have high affinity for AChE as single compounds (ΔG(ITC) = -48.99-55.36 kJ/mol, ΔG(LF/AD) = -43.38-45.38 kJ/mol). The concentration necessary to reduce AChE activity by 50% amounted to 0.22 μmol/μmol chlorogenic acids to the enzyme.

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.

Evaluation of the Hypoglycemic and Hypolipidemic Potential of Extract Fraction of Quercus baloot Griff Seeds in Alloxan-induced Diabetic Mice.

The discovery and development of new phytomedicines can be greatly aided by plants because of their tremendous therapeutic benefits, efficiency, cost-effectiveness, lack of side effects, and cheaper therapies. In this regard, Quercus baloot, generally known as oak, is used in folkloric medicine for treating and preventing various human disorders, including diabetes. For this purpose, the present study aimed to evaluate crude methanolic extract and various fractions of Quercus baloot for antihyperlipidemic and antihyperglycemic potential followed by the analysis of active compounds. The hypoglycemic and hypolipidemic activity was evaluated in Swiss male Albino mice by administering an oral dose of 150-300 mg/kg of Q. baloot extracts in alloxan induced diabetic mice for 14 days. The results revealed that crude methanolic extract at a dose of 300 mg/kg exhibited a significant reduction in the blood glucose level (198.50 ± 1.99 mg/dl) at day 14 and the same treatment significantly increased the body weight (31.26 ± 0.27 g) at day 14 in comparison to the control group. Moreover, the biochemical parameters were investigated which presented an increase in high-density lipids (HDL) (30.33 ± 0.33 mg/dl), whereas low-density lipids (LDL) showed a significant decrease (105.66 ± 0.26 mg/dl). Additionally, triglyceride levels 104.83 ± 0.70 mg/dl, and total cholesterol 185.50 ± 0.76 mg/dl are significantly decreased. In serum biochemical analysis creatinine and hepatic enzyme markers, like serum glutamate pyruvate transaminase (32.00 ± 0.36 U/mg), serum glutamate oxaloacetate transaminase (34.33 ± 0.61 U/mg), and alkaline phosphatase (157.00 ± 0.73 U/mg), were significantly reduced by the crude methanolic extract at a dose of 300 mg/kg as compared to the control group. The antioxidant enzymes like Superoxide dismutase (4.57 ± 0.011), peroxidases dismutase (6.53 ± 0.014, and catalase (8.38 ± 0.014) at a dosage of 300 mg/kg of methanolic extract exhibited a significant increase. The histopathological study of the diabetic heart, liver, and pancreas showed substantial restoration of damaged tissues in the methanolic extract 150 and 300 mg/kg treated group, which supports the effectiveness of Q. baloot seeds. The gas chromatography-mass spectrometry analysis of methanolic extract identified 10 antidiabetic active compounds in the Q. baloot seeds, validating the antihyperglycemic activity. Thus, methanolic crude extract at the doses 150 and 300 mg/kg of Q. baloot showed significant antihyperlipidemic and antihyperglycemic activities, which validate the folkloric utilization of Q. baloot as a remedy in diabetes. In conclusion, the 300 mg/kg methanolic extract of Q. baloot has notable hypoglycemic and hypolipidemic potential, supporting the plant's traditional medicinal usage in the treatment of diabetes and its complications. Further studies are needed for the purification, characterization, and structural clarification of bioactive compounds.

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.

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<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<0.05) reductions in liver enzymes (ALT, AST and ALP), total and conjugated bilirubin of the treated infected mice and also decreased significantly (p<0.05) total protein and albumin levels of the treated mice relative to control. The leaf extract and fractions further improved significantly (p<0.05) the levels of oxidative stress markers enzymes and molecules (CAT, GPx, GST, SOD) of the treated infected mice with no significant (p>0.05) effect on GSH. The MDA levels in the livers of the treated infected mice were significantly (p<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.

Antiplasmodial activities of n-hexane and water fractions of Dennettia tripetala leaf extract in Wistar rats

Many people die from malaria, a global health crisis. Most global malaria cases and deaths occur in sub-Saharan Africa. The rising resistance of Plasmodium spp. to antimalarial medications is hindering efforts to eradicate the illness. This study tested Dennettia tripetala leaf extract, N-hexane, and water fractions for antimalarial activity in rats. We used standard methods to screen Dennettia tripetala solvent (N-hexane and water) for phytochemical components. We tested the antimalarial potential of rats using Peters' 4-day suppressive, Rane's curative, and prophylactic tests. After conducting an exhaustive investigation, it was discovered that both fractions contained varying amounts of flavonoids, alkaloids, terpenoids, and saponins. There was a significant amount of antimalarial activity demonstrated by both fractions across all test models (P<0.05). Compared to the water fraction, Dennettia tripetala's n-hexane fraction was highly chemosuppressive and curative. Dennettia tripetala n-hexane has chemoprophylactic, curative, and chemosuppressive effects. The curative effect was 62.3% to 72.4%, and the chemosuppressive effect was 51.2% to 88.5%. Chemoprophylactic activity was 32.1%–61.2%. At 250–1500 mg/kg, Combretum nigricans butanol suppressed 40.3%, 54.1%, and 69.1%, respectively. Therapeutic effects were 26.2%, 36.9%, and 34.5%, while chemoprophylaxis was 48.4%, 70.0%, and 87.4%. Both Dennettia tripetala solvent fractions have antimalarial activity, suggesting they may be effective in different malaria therapy stages.

Industrial SMB chromatography as an alternative to the conventional refining of raw cane sugar

Simulated moving bed (SMB) chromatography is a well-established and widespread unit operation in the beet sugar industry and has been practiced commercially for more than thirty years. SMB chromatography has proven to be very efficient at removing inorganic ash components and colorants from sugar liquors, particularly those colorants with relatively high molecular masses. Recent optimization of the separation using raw sugar with a color ranging from 2000 IU up to 4000 IU has focused on minimizing the capital and operating costs of a chromatography system to replace the various purification and decolorization steps in conventional raw cane sugar refining. Decolorization levels exceeding 89% have been reliably achieved in the extract fraction from chromatography, along with ash removals ranging from 92% to 97%. As only water is added to purify the sugar, the application of this technology, in combination with modern industrial heat pumps and mechanical vapor recompression, allows for a powerful CO2-neutral process, with all the associated benefits regarding permits and marketability. The potential of this technology includes integration into new refinery projects (either standalone or back-end) by replacing affination, carbonatation and ion exchange decolorization (with raw sugar colors of up to 5000 IU), to expansion of existing refinery sites for slip stream processing of raw sugar syrups or refinery run-off syrups after clarification, and to the introduction of new process steps, such as direct liquid sugar production from cane raw sugar.

In vitro and in vivo pharmacological evaluation of polygonum persicaria l.

The present study was designed to investigate the in vitro antioxidant, cytotoxic, anthelmintic and in vivo analgesic, anti-diarrheal, hypoglycemic and CNS anti-depressant properties of different solvent fractions of methanol extract of leaf and flower of Polygonum persicaria L., a notable medicinal plant in Bangladesh. The crude methanol extracts were partitioned separately to petroleum-ether, carbon tetrachloride, chloroform and aqueous fractions. In DPPH scavenging assay for antioxidant test, the aqueous fraction of flower extract showed maximum antioxidant activity with IC50 of 0.60 μg/ml. Most of the fractions revealed prominent lethality (LC50) against brine shrimp nauplii. The test samples also exhibited dose-dependent anthelmintic activity against Pheretima posthuma. The leaf extract and flower extract (200-and 400 mg/kg b.w.) can decrease the painful sensation in mice. Leaf extracts and flower extracts displayed 57.69 and 50% reduction of diarrhea induced by castor oil in mice, respectively. The plant samples exhibited hypoglycemic effect and CNS anti-depressant effect in mice. Based on the possibility of bioactivity, the plant can be thoroughly examined to determine its potential efficacy and to support its traditional uses. Bangladesh J. Bot. 53(3): 475-485, 2024 (September)

Antioxidant, membrane stabilizing and thrombolytic potentials of the leaves of erythrina fusca lour

Erythrina fusca (Family: Fabaceae) is famous for many traditional healing. In the present study, the leaves of E. fusca were collected, dried, powdered and then extracted with methanol. It was further partitioned into n-hexane, dichloromethane and aqueous soluble fractions. All the extracts were subjected to antioxidant, membrane stabilizing and thrombolytic assays. In case of free radical scavenging assay, the aqueous soluble fraction showed highest scavenging activity. In ferric reducing anti-oxidant power assay, the dicholoromethane extract showed highest reduction of ferric ion in a dose dependent manner. In total antioxidant assay, the methanol extract and dichloromethane extract showed stronger capacity as antioxidant. During the determination of total phenolic and flavonoid contents, the methanol extract and dichloromethane soluble fraction showed higher contents of phenolics and flavonoids, respectively. The erythrocyte membrane stabilizing assay revealed the capacities of methanol extract and its aqueous fraction to stabilize the RBC membrane in both hypotonic solution- and heat- induced hemolytic conditions. However, the clot lysis ability of the E. fusca leaf extracts was not so prominent. Bangladesh J. Bot. 53(3): 439-445, 2024 (September)

Gintonin-Enriched Panax ginseng Extract Fraction Sensitizes Renal Carcinoma Cells to TRAIL-Induced Apoptosis through DR4/5 Upregulation.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising chemotherapeutic agent because of its selective apoptotic action on cancer cells. However, resistance to TRAIL-induced apoptosis remains a challenge in many cancers. The gintonin-enriched Panax ginseng extract fraction (GEF) has diverse pharmacological benefits. We explored the combined efficacy of GEF and TRAIL in inducing apoptosis in human renal cell carcinoma (RCC) cells. The effect of GEF treatment on the viability, clonogenic potential, wound healing, and TRAIL-induced apoptotic signaling of RCC cells was studied in vitro. Our investigation revealed that GEF pre-treatment sensitized RCC cells to TRAIL-induced apoptosis, as evidenced by DNA fragmentation and cell proliferation, colony formation, and migration inhibition. This sensitization was linked to the upregulation of death receptors 4 and 5 and alterations in apoptotic protein expression, notably, the decreased expression of the Mu-2-related death-inducing gene, a novel anti-apoptotic protein. Our findings underscore the necessity of caspase activation for GEF/TRAIL-induced apoptosis using the pan-caspase inhibitor Z-VAD-FMK. This study demonstrates that GEF sensitizes human RCC cells to TRAIL-induced apoptosis by upregulating DR4/5 and modulating apoptotic protein expression. These findings suggest a promising strategy for overcoming TRAIL resistance in cancer therapy and highlight the potential of GEF as a valuable adjunct to TRAIL-based treatments.

Assessment of Piper longum L. (Piperaceae) leaves toxicity on the adults of Tribolium castaneum (Herbst, 1797) (Coleoptera: Tenebrionidae)

BackgroundNumerous insect pests attack stored grains causing both qualitative and quantitative losses. The most damaging pest that infests dry stored produce is the red flour beetle, Tribolium castaneum, a secondary pest of stored goods. This pest, especially in its adult stage, exhibits resistance to chemical insecticides, thereby rendering the traditional pesticides ineffective in controlling it. Phyto-derivatives, which are strong insecticides and also ecologically benign, have gained interest as non-chemical solutions for controlling this pest. Hence, the objective of this study was to investigate the potential of Piper longum leaf extract insecticidal action as an environmentally benign insecticide for the first time against the adults of T. castaneum. In this study, P. longum leaf ethanol extract was tested against the adults of T. castaneum by petri dish bioassay method. Ad hoc studies to verify significant mortality for the initial confirmation of adulticidal activity were conducted for 24 h at different dosages of 62.5, 125, 250, 500 and 1000 mg/L of P. longum leaf ethanol extract. Thereafter, dosages set at 10, 20, 30 and 40 mg/L for the fractions of P. longum leaf ethanol extract were conducted. Prior to this, the leaf extract of this plant was subjected to column chromatography for fractionation. The fractions tested for adulticidal activity were subjected to gas chromatography–mass spectroscopy.ResultsSignificant adulticidal action with 100% adult mortality was observed in ethanol extract of P. longum leaves. Among the fourteen fractions (F0–F13) obtained tested, only fractions, F5, F10 and F13, demonstrated adulticidal activity, and the remaining fractions displayed poor activity. One hundred per cent morality was noted in T. castaneum adults after 96 h at 40 mg/L in F5 and F10, and in F13 at 20 mg/L, and their respective LD50 values were 17.6, 26.6 and 10.0 mg/L. The fractions F5, F10 and F13 contained fatty acids, viz., hexadecanoic acid, dotriacontane and heptacosane in F5; tetradecanoic acid and nonadecanoic acid in F10; and octadecanoic acid, aspartame and tridecanoic acid in F13, revealed through gas chromatography–mass spectroscopy.ConclusionsThe results of the study showed that P. longum ethanol leaf extract revealed significant adulticidal activity and is a promising toxic agent to the adults of T. castaneum. The fatty acids in the ethanolic leaf extract fractions of P. longum could have caused toxicity to the adults of T. castaneum. According to the current literature survey, this is the first research report on the adulticidal activity of P. longum leaf extracts against the adults of T. castaneum.

Cytotoxic and antibacterial compounds from Schinus terebinthifolia Raddi fruits

Chromatographic procedures of extracts of Schinus terebinthifolia Raddi fruits afforded (Z)-masticadienoic (1) and 3β-masticadienolic (2) acids, tetrahydroamentoflavone (3), and 4-O-methyl gallic acid (4). Addicionally, the derivative 6-oxo masticadienoic acid (1a) was prepared by an allylic oxidation. The chemical structures of obtained compounds were elucidated by spectrometric data analyses. Furthermore, both the semi-synthetic derivative and the metabolites were subjected to in vitro cytotoxicity against the A549 human lung cancer cell line, as well as antimicrobial activity tests. Compounds 2 and 1a exhibited cytotoxicity towards A549 cells with IC50 values of 20.13 and 6.11 µM, respectively. In the tests against pathogens, the CHCl3 and EtOAc soluble fractions of MeOH extract along with the pure compounds, exhibited antibacterial activity against Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa. Except for 4-O-methyl gallic acid, the other pure compounds showed inhibitory microbial activities with MIC values ranging from 0.25 μg/mL to 25 μg/mL doses.

Discovery of Active Antibacterial Fractions of Different Plant Part Extracts of clove (Syzigium aromaticum) Against Streptococcus mutans

Nowadays, dental cavities caused by Streptococcus mutans are a major focus of research in Indonesia. While several antibiotics are available to combat this bacterium, concerns about antibiotic resistance have prompted researchers to explore natural remedies. Clove (Syzigium aromaticum) is a commonly studied natural remedy against dental cavities and S. mutans. Among the different parts of the clove plant, clove bud is the most widely used against dental cavities or S. mutans, and the potential of other clove parts has not been thoroughly explored. Identifying which parts of the clove plant have higher concentrations of active ingredients and exhibit the strongest antibacterial activity is important. Therefore, this study evaluated the antibacterial activity of three different parts, i.e., leaf, stems, and buds of the clove plant ethanolic extracts against S. mutans. The ethanolic extracts of clove leaf, stems, and buds were prepared using the maceration method with 70% ethanol, and their activity against S. mutans was tested using the disc diffusion method at three different concentrations (10%, 5%, 2.5% b/v). Fractionation was carried out using hexane and water to obtain two fractions: hexane and water fraction. These fractions were then subjected to antibacterial assays. The ethanolic leaf, stems, and bud extracts exhibited varying antibacterial activity levels. The best activity was observed with the 10% clove bud ethanolic extract, which produced an inhibition zone of 20.83 ± 0.77 mm. The leaf and stem extracts showed inhibition zones of 16.38 ± 3.84 mm and 17.95 ± 5.15 mm, respectively. Furthermore, the hexane-soluble fraction of the clove bud displayed the highest activity with an inhibition zone diameter of 23.7 ± 3.21 mm at 10%. This activity was twice as high as ampicillin, used as the positive control. In conclusion, clove bud remains the best source of antibacterial compounds against S. mutans. Fractionation of the bud extract using hexane can significantly enhance its activity. Further investigation should be conducted to optimize the effectiveness of this active fraction for use as an anti-dental caries treatment.

Aktivitas Antioksidan Fraksi Larut Etil Asetat dari Ekstrak Akar Tabar Kedayan (Aristolochia papilifolia Ding Hou)

Kedayan root is a medicinal plant that is traditionally used by the community as medicine, especially as an antidote to poison. This research aimed to determine the antioxidant activity of the ethyl acetate soluble fraction of Tabar Kedayan root extract (Aristolochia papillfolia Ding hou). The fractionation method used is solid-liquid extraction using ethyl acetate solvent to obtain a soluble fraction of ethyl acetate, then vacuum liquid chromatography fractionation using the eluent ratio n-hexane: ethyl acetate and obtain fraction A, fraction B, fraction C, fraction D, fraction E, fraction F. Testing of antioxidant activity using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method spectrophotometrically. The data obtained was analyzed using linear regression to determine IC50. The results of the C fraction test on the ethyl acetate soluble fraction of Tabar Kedayan roots (A. papillfolia) have antioxidant activity with an IC50 value of 139.11 ppm.

Altered oral microbiota of drug-resistant organism carriers exhibit impaired gram-negative pathogen inhibition.

The oral microbiome has been understudied as a reservoir for clinical pathogens, including drug-resistant strains. Understanding how alterations in microbiome functioning render this site vulnerable to colonization is essential, as multidrug-resistant organisms (MDRO) carriage is a major risk factor for developing serious infections. To advance our knowledge of oral MDRO carriage and protection against pathogen colonization conferred by native microbiota, we examined microbiomes from individuals colonized by MDROs (n=33) and non-colonized age-matched controls (n=30). Shotgun metagenomic analyses of oral swabs from study participants revealed significant differences in microbial communities with depletion of Streptococcus spp. among those colonized by multidrug-resistant gram-negative bacilli (RGNB), compared to non-carriers. We utilized metagenomic sequencing to characterize the oral resistome and find antimicrobial resistance genes are present in higher abundance among RNGB carriers versus non-carriers. High-throughput co-culture screening revealed oral bacteria isolated from MDRO non-carriers demonstrate greater inhibition of gram-negative pathogens, compared to isolates from carriers. Moreover, biosynthetic gene clusters from streptococci are found in higher abundance from non-carrier microbiomes, compared to RGNB carrier microbiomes. Bioactivity-guided fractionation of extracts from Streptococcus isolate SID2657 demonstrated evidence of strong E. coli and A. baumannii inhibition in a murine model of infection. Together, this provides evidence that oral microbiota shape this dynamic microbial community and may serve as an untapped source for much-needed antimicrobial small-molecules.

Antileishmanial, antitrypanosomal and anti-coronavirus activities of benzophenanthridine alkaloids and other specialized metabolites isolated from the root bark of Zanthoxylum zanthoxyloides (Lam.) B.Zepernick & Timler

Strong antileishmanial and antitrypanosomal activities were highlighted for the crude methanolic extract (IC50 = 0.61 and 2.15 μg/mL, respectively) of Zanthoxylum zanthoxyloides (Lam.) B.Zepernick & Timler root bark, as well as for its apolar partitions (cyclohexane: IC50 = 0.66 and 5.17 μg/mL, respectively and dichloromethane: IC50 = 0.07 and 0.22 μg/mL, respectively), with a good selectivity index (SI) towards WI-38 cells. In addition, cyclohexane and dichloromethane extracts exhibited a dose-dependent inhibition of human coronavirus HCoV-229E infection in hepatoma Huh-7 cells expressing or not the cellular protease TMPRSS2 (IC50 values of 5.29 μg/mL and 4.87 μg/mL, respectively). Fractionation of these active extracts led to the isolation of a new racemic benzophenanthridine alkaloid named zanthoxyloithrine (1), together with 13 known compounds. Their structures were elucidated by spectroscopic techniques including IR, UV, HR-MS, 1D and 2D NMR and electronic circular dichroism. In parallel, HR-ESI-MS/MS based dereplication and molecular networking analysis were performed to identify unpurified compounds in cyclohexane and dichloromethane extracts. Zanthoxyloithrine (1) showed strong antileishmanial (IC50 = 0.14 μM, SI = 52.0) and antitrypanosomal (IC50 = 0.36 μM, SI = 20.8) activities. In addition, compound (1) demonstrated a high antiviral activity against HCoV-229E with IC50 value of 6.70 μM in presence of TMPRRS2 and without significant toxicity on Huh-7 cells. Other purified benzo[c]phenanthridine alkaloids also showed anti-coronavirus and antiparasitic activities.

LC-MS/MS Analysis of crude Flavonoid Compounds from <i>Justicia secunda</i> from Democratic Republic of the Congo and evaluation of their antisickling Activity

<i>Justicia secunda</i> Vahl (Acanthaceae) is a tropical herbaceous plant traditionally used by indigenous peoples of the Democratic Republic of the Congo (DRC) to treat symptoms associated with sickle cell disease (SCD), a genetic condition with life-threatening complications. Given the lack of side-effect-free drugs, alternative herbal therapies that can reduce or reverse red blood cell sickling may serve as safe and effective treatments. The primary objective of this study is to conduct a phytochemical screening of crude flavonoids extracted from <i>J. secunda</i> using LC-ESI/MS/MS. <i>J. secunda</i> extracts were evaluated for their phytoconstituents and anti-sickling properties. The anti-sickling properties were assessed through reverse sickling estimation, polymerization and oxidation inhibition, and osmotic fragility assays. Fractionation of the methanol extract and chemical profiling were performed by flash chromatography and high-performance thin-layer chromatography analysis respectively. Characterization and identification of the crude flavonoids were done using chromatography and spectroscopy methods. Chemical screening of crude flavonoids revealed the presence of 12 flavonoid O-glycosides based on one flavonol (methoxy-kaempferol) and two flavones (luteolin and apigenin). The crude flavonoid extracts demonstrated maximum reversal of sickled red blood cells (91.9 ± 0.8%), significant inhibition of hemoglobin polymerization (77.8 ± 7.6%) and hemoglobin S auto-oxidation (96.3 ± 0.2), and provided maximum protection of red blood cells against hyperhemolysis. The results validate the traditional use of <i>J. secunda</i> and identify potential compounds for the development of novel anti-sickling agents.

Anti-arthritic studies of ethnomedicine Gaultheria trichophylla Royle extract and salicylate-rich fraction using complete Freud's adjuvant-induced rats: molecular docking and network pharmacology analysis.

Gaultheria trichophylla Royle is a traditional treatment for inflammatory conditions including arthritis. The objective was to evaluate the anti-arthritic activity of the extracts and salicylate-rich fractions through adjuvant-induced arthritis, histopathological analysis, radiological imaging, hematological, biochemical parameters along with using bioinformatic tools. In vivo anti-arthritic efficacy of the extract and SRF (at 100, 200, 300, and 150mg/kg doses) was assessed using healthy albino rats. Molecular docking of identified compounds along with network pharmacology analysis helped to determine the route of action of drug. Both the extract and SRF showed dose-dependent anti-arthritic activity by decreasing the joint diameter, increase in pain threshold and body weight compared with negative control group. Along with SRF (150mg/kg), EEGT (300 and 200mg/kg) shows significant (P < 0.01) anti-arthritic activity by lowering levels of WBC, platelets, serum C-reactive protein (CRP), and rheumatoid factor (RF) and raising levels of RBC and Hb. The modified biochemical measures (AST, ALT, ALP, and total protein level) further supported the anti-arthritic action. Histopathology and radiology study showed that EEGT (300 and 200mg/kg), SRF (150mg/kg) and diclofenac (10mg/kg) inhibited joint destruction. GCMS analysis showed the presence of methyl salicylate, sitosterol, calcifediol, and ergosta-5,22-dien-3-ol, acetate as important bioactive constituents. Moreover, as the significant node in the pharmacology network and docking against TNF-α, a classical therapeutic target in RA showed potential of G. trichophylla in treatment of RA. The results showed that G. trichophylla have effectively reduced the inflammation of the joints.

UPLC‐MS/MS and chemometrics analyses reveal chemical profile and anti‐inflammatory activity of Bienertia cycloptera Bunge fractions

AbstractIntroductionBienertia cycloptera is a species belonging to the Chenopodiaceae family. According to earlier reports, a unique research study on the phytochemistry and biological analysis of that species was conducted.ObjectiveThis study presents an integrated metabolomics investigation combined with multivariate analysis of various extractive fractions of B. cycloptera aerial parts. This study is the first attempt to explore the anti‐inflammatory metabolites from B. cycloptera, showing its significance as a valuable traditional medicine.MethodologyBy comparing retention times, quasi‐molecular ions, and MS/MS fragment ions with databases and literature references, metabolite annotation was accomplished using ultra performance liquid chromatography (UPLC)/triple quadrupole mass spectrometry (MS). Moreover, the effects of the studied samples on the gene expression of the four pro‐inflammatory cytokines (IL‐1β, IL‐6, TNF‐α, and INF‐γ) using polymerase chain reaction (PCR) and comparing their results by those caused by piroxicam were tested to determine their anti‐inflammatory efficacy.ResultsChemical profiling revealed diverse metabolites, with 62 chromatographic peaks identified across various chemical classes. UPLC‐MS/MS of different B. cycloptera fractions unveiled distinct chemical profiles. Results showed distinct chemical compositions in each fraction, with petroleum ether fraction enriched in sterols and fatty acids; methylene chloride fraction in alkaloids, sterols, and cardenolides; ethyl acetate fraction in alkaloids, flavonoids, cardenolides, and phenolic acids; and n‐butanol fraction in flavonoids, alkaloids, and phenolic acids. Multivariate data analysis illustrated clustering patterns among petroleum ether, methylene chloride, ethyl acetate, and n‐butanol fractions. OPLS‐DA models were constructed to discern inter‐class differences, identifying discriminatory metabolites. In vitro cytotoxicity and anti‐inflammatory assays demonstrated the safety and efficacy of B. cycloptera fractions, with significant downregulation of pro‐inflammatory markers. Further analysis revealed specific metabolites associated with anti‐inflammatory effects, such as p‐hydroxybenzoic acid, vanillic acid, tachioside, ferulic acid, staphylionoside D, humilixanthin, bergaptol, vulgaxanthin I, and portulacaxanthin III.ConclusionThe findings of this study provide valuable insights into the chemical composition and bioactivity of B. cycloptera fractions, suggesting their potential as therapeutic agents and warranting further investigation.