Active Fraction Research Articles

Antihyperglycemic Effects of Active Fraction of Ethanol Extract from Castanopsis costata Leaves and Channa striata Extract on Rats

Anti-hyperglycemic experiments were conducted on male Wistar rats divided into six groups: Group 1 (negative control), Group 2 (active fraction of Castanopsis costata ethanol extract at 200 mg/kg body weight), Group 3 (Channa striata extract at 250 mg/kg body weight), Group 4 (active fraction of Castanopsis costata ethanol extract at 50 mg/kg body weight + Channa striata extract at 250 mg/kg body weight), Group 5 (active fraction of Castanopsis costata ethanol extract at 100 mg/kg body weight + Channa striata extract at 250 mg/kg body weight), and Group 6 (active fraction of Castanopsis costata ethanol extract at 200 mg/kg body weight + Channa striata extract at 250 mg/kg body weight). The most notable reduction in blood glucose levels was observed in Group 4, with an average decrease of -55.40%, followed by Group 6 (-55.19%), Group 5 (-52.80%), Group 2 (-52.01%), and Group 3 (-52.88%). These findings indicate that combining the active fraction of Castanopsis costata ethanol extract with Channa striata extract effectively lowers blood glucose levels in Wistar rats. Keywords: Antihyperglycemia, Active fraction of ethanol extract, Blood glucose level

Sub-chronic toxicity of the active fraction of a modified Huang-Lian-Jie-Du Decoction

A traditional Chinese herbal medicine formula named Huang-Lian-Jie-Du Decoction (HLJDD) has been used to cure various inflammatory diseases with a long history. However, one component of HLJDD Gardeniae fructus has remarkable liver and kidney toxicities. Therefore, it was altered with Dictamni cortex to form a modified HLJDD (MHLJDD). In this study, we aimed to evaluate the sub-chronic toxicity of the active fraction of MHLJDD (MHLJDD-F) in rats. Adult rats of both sexes were intragastrically administered with vehicle or MHLJDD-F (at the dose of 170, 340, and 680 mg/kg/day) once daily for 90 days. Half of the rats from each group were kept for an additional 30-day period to observe the drug withdrawal effect. The signs of toxicity and mortality of the rats were observed, and the body weight and food consumption were recorded. Blood was collected for hematological and biochemical analyses and major organs were weighed and harvested for histopathological examinations. The results revealed that no systemic toxicity of MHLJDD-F was found during the experiments. Organ coefficients and pathological alterations of major organs were comparable to the control rats. The no-observed adverse effect level (NOAEL) of MHLJDD-F was found up to 680 mg/kg/day. All these results demonstrated that long-term oral administration of MHLJDD-F did not cause significant toxicity, which is worthy to be widely applied as a new herbal medicine in pre-clinical and clinical studies.

Myrtus communis leaf compounds as novel inhibitors of quorum sensing-regulated virulence factors and biofilm formation: In vitro and in silico investigations

Antibiotic resistance of the Gram-negative bacterium Pseudomonas aeruginosa and its ability to form biofilm through the Quorum Sensing (QS) mechanism are important challenges in the control of infections caused by this pathogen. The extract of Myrtus communis (myrtle) showed strong anti-QS effect on Chromobacterium. violaceum 6267 by inhibiting 80 % of the production of violacein pigment at a sub-MIC concentration of 1/8 (31.25 μg/mL). In addition, the extract exhibited an inhibitory effect on virulence factors of P. aeruginosa PAO1 at half MIC (125 μg/mL), significantly reducing the formation of biofilms (72.02 %), the swarming activity (75 %), and the production of protease (61.83 %) and pyocyanin (97 %). The active fraction also downregulated the expression of selected regulatory genes involved in the biofilm formation and QS in the P. aeruginosa PAO1 strain. These genes included the autoinducer synthase genes (lasI and rhlI), the genes involved in the expression of their corresponding receptors (lasR and rhlR), and the pqsA genes. The analysis of the active fraction by HPLC/UV/MS and NMR allowed the identification of three phenolic compounds, 3,5-di-O-galloylquinic acid, myricetin 3-O-α-l-rhamnopyranoside (myricitrin), and myricetin 3-O-(2″-O-galloyl)-ß-d-galactopyranoside. In silico studies showed that 3,5-di-O-galloylquinic acid, with an affinity score of −9.20 kcal/mol, had the highest affinity to the active site of the CviR protein (3QP8), a QS receptor from C. violaceum. Additionally, myricetin 3-O-α-l-rhamnopyranoside (myricitrin) and myricetin 3-O-(2″-O-galloyl)-ß-d-galactopyranoside interact to a lesser extent with 3QP8. In conclusion, this study contributed significantly to the discovery of new QS inhibitors from M. communis leaves against resistant Gram-negative pathogens.

Network pharmacology, molecular docking study, and in vivo validation of the wound healing activity of the Red Sea soft coral Paralemnalia thyrsoides (Ehrenberg 1834) ethanolic extract and bioactive metabolites

BackgroundWounds are a major health issue on a global scale, putting a great deal of financial, commercial, and social strain on healthcare organizations, patients, and individuals. So, this study aims to investigate the in vitro antioxidant activity of Paralemnalia thyrsoides soft coral total ethanolic extract. Also, bio-guided in vivo wound healing validation enhanced by the evaluation of related gene expression of Paralemnalia thyrsoides total extract, derived fractions, and three known metabolites was done. Furthermore, utilizing network pharmacology, we identified ten hub target genes associated with wound healing, including AKT1(RAC-alpha serine/threonine–protein kinase), IL6 (interleukin-6), MAPK3 (mitogen-activated protein kinase 3), MMP9 (matrix metalloproteinase 9), and APP (amyloid P protein precursor). We conducted molecular docking to assess how the three compounds interact with these hub genes and inflammatory cytokines (IL-1β (interleukin-1 beta), TGF-β (transforming growth factor-beta), TNF-α (tumor necrosis factor-alpha), and NF-KB (nuclear factor-kappa B) linked to wound healing.ResultsIn vitro antioxidant activity of the total ethanolic extract of Paralemnalia thyrsoides revealed potent scavenging activity against H2O2 with IC50 of 178.62 μg/mL. Additionally, the bio-guided scheme of the in vivo wound healing assay leads to the most active fraction, petroleum ether, with a healing percentage of 85% ± 4. Several chromatographic procedures upon petroleum ether fraction led to the isolation of three known compounds with significant in vivo wound healing potential which are recognized as triacontan-1-ol (1), 24-methylcholesterol (2) 6α-acetyl-7α-acetate-1(10)-α-13-nornardosine [C16H24O4] (3). Noteworthy downregulation in Cox-2 (Cyclooxygenase-2), Cox-1 (Cyclooxygenase-1), IL-1β, TGF-β, TNF-α, NF-KB, and INF-ϒ (interferon-ϒ) relative genes expression and upregulation in TGF-β, and IL-10 (interleukin-10) relative genes expression proved that compounds (3), (2), and (1) were, respectively, significant. The in silico study suggests that both C16H24O4 and 24-methyl cholesterol have potential in wound healing, possibly involving the regulation of RAC-alpha serine/threonine-protein kinase (AKT1).ConclusionOur study highlights the antioxidant and wound-healing potential of Paralemnalia thyrsoides soft coral that can be attributed to its diverse chemical metabolites. The in vivo and in silico findings highlighted that P. thyrsoides can be an effective remedy for wound restoration with the need for extensive future detailed clinical studies to prove these results.

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

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

The combination of three CD4-induced antibodies targeting highly conserved Env regions with a small CD4-mimetic achieves potent ADCC activity.

The majority of naturally-elicited antibodies against the HIV-1 envelope glycoproteins (Env) are non-neutralizing (nnAbs), because they are unable to recognize the Env timer in its native "closed" conformation. Nevertheless, it has been shown that nnAbs have the potential to eliminate HIV-1-infected cells by Antibody-Dependent Cellular Cytotoxicity (ADCC) provided that Env is present on the cell surface in its "open" conformation. This is because most nnAbs recognize epitopes that become accessible only after Env interaction with CD4 and the exposure of epitopes that are normally occluded in the closed trimer. HIV-1 limits this vulnerability by downregulating CD4 from the surface of infected cells, thus preventing a premature encounter of Env with CD4. Small CD4-mimetics (CD4mc) sensitize HIV-1-infected cells to ADCC by opening the Env glycoprotein and exposing CD4-induced (CD4i) epitopes. There are two families of CD4i nnAbs, termed anti-cluster A and anti-CoRBS Abs, which are known to mediate ADCC in the presence of CD4mc. Here, we performed Fab competition experiments and found that anti-gp41 cluster I antibodies comprise a major fraction of the plasma ADCC activity in people living with HIV (PLWH). Moreover, addition of gp41 cluster I antibodies to cluster A and CoRBS antibodies greatly enhanced ADCC mediated cell killing in the presence of a potent indoline CD4mc, CJF-III-288. This cocktail outperformed broadly-neutralizing antibodies and even showed activity against HIV-1 infected monocyte-derived macrophages. Thus, combining CD4i antibodies with different specificities achieves maximal ADCC activity, which may be of utility in HIV cure strategies.

Identification and anti-inflammatory activity of flavonoids from Cardiospermum halicacabum in murine RAW 264.7 macrophage-A bioassay-guided approach

Cardiospermum halicacabum Linn. (CH), known as balloon vine, is used for food and medicinal purposes, including inflammation treatment. In Tamil cuisine, CH stems and leaves feature in curries, crepes, sauces, and soups. While CH's anti-inflammatory effects are documented, its therapeutic compounds are unclear. This study aims to isolate anti-inflammatory constituents from CH using a bioassay-guided approach. CH aerial part was extracted with three different solvents and evaluated for their anti-inflammatory activity. The active ethanol-water extract (CH-EW) was subjected to solvent-solvent partitioning to get five fractions. Among fractions, the methanol fraction (CH-EW-ML) showed potent anti-inflammatory as well as antioxidant activity along with high flavonoid content. Further fractionation of the active methanol fraction resulted in the isolation of two flavonoids, apigenin-7-O-β-D-glucuronide (AAG) and diosmetin-7-O-β-D-glucuronide (DMG). DMG was isolated from CH for the first time. The compounds were evaluated for their nitric oxide (NO) inhibition activity in the RAW 264.7 cell line. DMG showed the most potent activity (IC50 = 16.72 ± 1.01 μM). Further studies showed that DMG can inhibit the production of reactive oxygen species (ROS) and proinflammatory cytokines as well as upregulate the expression of anti-inflammatory cytokines. This study represents the first evidence that DMG can inhibit NO, ROS, TNF-α, and IL-6 while upregulating IL-10. Our findings suggest that incorporating CH infusion into the diet may alleviate inflammation, with DMG being a promising therapeutic component for its anti-inflammatory effects.

Anticancer activity of the ethylacetate fraction of Vernonia amygdalina Delile towards overexpression of HER-2 breast cancer cell lines

Vernonia amygdalina Delile, coming from the Asteraceae tribe, contains active compounds that can treat breast cancer. This study examines the anticancer activity of Vernonia amygdalina Delile leaves, an active fraction of MCF-7/HER-2 breast cancer cells. Thin-layer chromatography determines the phytochemical screening. Cytotoxic and proliferation analyses were determined using the MTT method for the MCF-7/HER-2 breast cancer cell line. The cell cycle and apoptosis profiles were examined using flow cytometry. The results can be summarized in this study: Vernonia amygdalina Delile fraction contains a flavonoid with great potential for pharmacological activities, for instance, inhibiting the growth of cancer cells. The ethyl acetate fraction was more potent and cytotoxic on MCF-7/HER-2 cells (IC50 = 66 μg/mL) than extract ethanol (IC50 = 130 μg/mL). The ethylacetate fraction of Vernonia amygdalina Delile has been proven to inhibit cell proliferation by decreasing cell viability with an IC50 of 66 μg/mL concentration incubated for 24, 48, and 72 hrs with cell viability values of 64.46%, 61.67%, and 53.89%, respectively, compared to the ethanol extract IC50 of 130 μg/mL concentration with cell viability values of 56.0%, 50.19%, and 58.67%, respectively, which induced apoptosis and inhibited the cell cycle in the G2-M phase. To conclude, the ethyl acetate fraction of Vernonia amygdalina Delile can be used as an anticancer against the MCF-7/HER-2 cell line with an IC50 of 66 μg/mL, inhibiting cell proliferation and the cell cycle and inducing apoptosis activity in the MCF-7/HER-2 cell.

Antimalarial Studies and Phytochemical Profiling of Laportea aestuans (L.) Chew Extractives

Background and objectives This study validated the antimalarial efficacy of Laportea aestuans extract using chemosuppressive and curative models, identified its most active fraction, and performed chemical profiling of the active fraction. Methodology The aerial part of L. aestuans (LA) was extracted with 80% methanol and assayed for its chemosuppressive and curative antimalarial activities through the oral administration of the extract (100, 200, 400 and 800 mg/kg) to groups consisting of five mice each. The extract was thereafter suspended in water and partitioned with dichloromethane to afford the dichloromethane (DLA) and aqueous (ALA) fractions. The fractions (10, 20, 40, and 80 mg/kg) were thereafter tested for their antimalarial activity followed by GC-MS analysis of the most active fraction. Results The toxicity level was found to be above 2000 mg/kg. The activity recorded for the four-day chemosuppressive antimalarial tests showed a dose-dependent activity up to 400 mg/kg. The activity was significantly lower than that of chloroquine. The antimalarial activity of the extract at the lowest dose of 100 mg/kg was 42%. In the curative test, the extract of LA gave a dose-dependent activity that was significantly higher than the negative control. The DLA and ALA fractions gave a comparable curative activity across all doses. At 10 mg/kg, ALA and DLA gave percentage clearance of 81.25 ± 1.84 and 87.45 ± 1.35, respectively, which were significantly higher than that of the negative control (0.00 ± 0.00) and significantly lower than that of the positive control (94.93 ± 0.26). The chemical profiling revealed the presence of non-polar and medium-polar constituents in the dichloromethane fraction. Conclusion The good antimalarial activity elicited by LA extractives supports its usage in ethnomedicine as an antimalarial agent.

Comparing the impact of insufficient physical activity on cardiovascular disease in survivors of childhood Hodgkin lymphoma and sibling controls: A report from the Childhood Cancer Survivor Study (CCSS).

10024 Background: Survivors of childhood Hodgkin lymphoma are at increased risk of cardiovascular disease (CVD) from anthracycline and radiation exposures. Whether the treatment-induced risk can be mitigated with physical activity is not known. We aimed to compare the impact of insufficient physical activity on CVD in Hodgkin lymphoma survivors and controls. Methods: Participants in the Childhood Cancer Survivor Study (CCSS) reported physical activity, which was translated into metabolic equivalent of task hours/week (MET) and categorized as 0, 1-8, or ≥9 METs. We estimated rate ratios (RR) for new onset (incident) congestive heart failure and any CVD (including heart attack, congestive heart failure, arrhythmia, and valvular disease requiring interventions) during follow-up, accounting for age, sex, race/ethnicity, smoking, risky drinking, overweight/obesity, and treatment exposures (survivors only). Then we calculated population attributable fractions (PAF) for insufficient physical activity (< 9 METs). The attributable relative rate (ARR) was calculated by multiplying the PAF in survivors with the RR for CVD incidence in survivors compared to controls, resulting in an ARR for survivors that was compared to the PAF in controls. Results: In 2,357 Hodgkin survivors [50% female; [mean (SD)] 14.3 (4.1) years at diagnosis; 31.6 (5.8) years at start of follow-up, and 17.8 (7.6) years from diagnosis] and 3,949 sibling controls, 233 survivors and 19 controls developed congestive heart failure. 462 survivors and 95 controls developed any CVD. The PAF for congestive heart failure attributable to insufficient physical activity was larger in controls (38.0%) than survivors (22.3%). However, with RR 20.3 (95% CI 12.6-32.5) for congestive heart failure for survivors compared to controls, the ARR in survivors (452.1%) revealed a 12 times higher incidence of congestive heart failure attributable to insufficient physical activity in Hodgkin survivors than in controls (452.1%/38.0%). The ARR indicates that 4.5 of the 20-fold higher risk of congestive heart failure in survivors vs. siblings is attributable to insufficient physical activity in the survivors. For any CVD, the PAFs were comparable in survivors (13.7%) and controls (11.7%). With RR 10.2 (95% CI 8.2-12.7) for any CVD for survivors compared to controls, the ARR for any CVD in survivors (139.7%) was also 12 times higher than the incidence attributable to insufficient physical activity in controls (139.7%/11.7%). Conclusions: Our results suggest that the magnitude of treatment-induced CVD risk that can be mitigated by physical activity is considerable. This offers a modifiable target and provides rationale for exercise interventions aimed at reducing CVD in survivors at high risk.

Understanding Solar Activity after the Maunder Minimum: Sunspot Records by Rost and Alischer

The Maunder Minimum was a period with significantly reduced solar activity between 1645 and 1715, approximately. The transition between the low solar activity in the Maunder Minimum and the subsequent “normal” regime of solar activity was gradual. However, there are discrepancies in the solar activity level from sunspot number indices and solar activity proxies in that period. Among the contemporaneous observers, Johann L. Rost and Sebastian Alischer were two key sunspot observers to understand the solar activity in this transition just after the Maunder Minimum. We have revised all their sunspot records, counting the number of groups and individual sunspots to derive reliable data for the solar activity level for the period 1716–1726. We found significant misinterpretations of the sunspot group counting assigned to these astronomers in the existing group number databases. Our new group sunspot counting significantly reduces the number of groups for Rost and Alischer’s observations compared to entries in existing databases. Furthermore, our sunspot number estimates (obtained from the active day fraction methodology) of the maximum amplitude of Solar Cycles −3 and −4 are significantly lower than the amplitudes according to the official sunspot number, but they are compatible with sunspot number values obtained from solar activity proxies such as radioisotopes. Our result would imply that solar activity after the Maunder Minimum recovered more gradually and with a lower intensity than previously considered.

Further Polycyclic Quinones of Micromonospora sp.

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

Discovery of Biofilm Inhibitors from the Microbiota of Marine Egg Masses.

Biofilms commonly develop in immunocompromised patients, which leads to persistent infections that are difficult to treat. In the biofilm state, bacteria are protected against both antibiotics and the host's immune system; currently, there are no therapeutics that target biofilms. In this study, we screened a chemical fraction library representing the natural product capacity of the microbiota of marine egg masses, namely, the moon snail egg collars. This led to the identification of active fractions targeting both Pseudomonas aeruginosa and Staphylococcus aureus biofilms. Subsequent analysis revealed that a subset of these fractions were capable of eradicating preformed biofilms, all against S.aureus. Bioassay-guided isolation led us to identify pseudochelin A, a known siderophore, as a S.aureus biofilm inhibitor with an IC50 of 88.5 μM. Mass spectrometry-based metabolomic analyses revealed widespread production of pseudochelin A among fractions possessing S.aureus antibiofilm properties. In addition, a key biosynthetic gene involved in producing pseudochelin A was detected on 30% of the moon snail egg collars and pseudochelin A is capable of inhibiting the formation of biofilms (IC50 50.6 μM) produced by ecologically relevant bacterial strains. We propose that pseudochelin A may have a role in shaping the microbiome or protecting the egg collars from microbiofouling.

Cymbopogon proximus phytochemicals induce S-phase arrest in A549 lung cancer cell lines via CDK2/cyclin A2 inhibition: gas chromatography-mass spectrometry and molecular docking analyses.

Cymbopogon proximus comprises several phytoconstituent classes that are reported to possess anticancer activity; however, studies on the anticancer potentials of theplant are lacking. C.proximus was extracted using solvents with increasing polarity. In-vitro cytotoxic activity of C.proximus extracts was examined against liver (HepG2), lung (A549), prostate (PC3), and bone (MG63) cell lines using MTT assay in comparison to doxorubicin. Flow cytometry was used to analyze the cell cycle for identification of the phase of inhibition. Chemical composition of the most active fraction was examined using the GC/MS technique. Molecular docking was used to explore the mechanism of cytotoxicity against A549, and the results were confirmed by Western blot analysis. Petroleum ether fraction was the highly effective fraction against A549 with IC50=14.02±2.79. GC/MS analysis of Pet.Eth led to the identification of nine compounds in unsaponifiable matter and 27 components in the saponifiable fraction. Di-N-octyl phthalate, 3-β-hydroxylean-11.13(18)-dien-30-oic acid methyl ester, elemol hydrocarbons, linoelaidic acid and linoleic acid demonstrated the lowest docking binding scores and similar binding modes against CDK2 as compared to that attained by the native ligand R-Roscovitine "CDK2 ATP inhibitor". Western blot analysis demonstrated that CDK2/cyclinA2 protein expression has been suppressed in A549 cell lines by Pet.Eth fraction.

#879 Impacts on ionized magnesium and total magnesium by the conversion to higher dialysate magnesium concentration

Abstract Background and Aims Hypermagnesemia is detrimental to hemodialysis patients whose renal function is compromised. However, recent observational study suggested the lower mortality rate was observed in the patients with mild hypermagnesemia. Even though the serum magnesium concentration (tMg) of the patients were frequently higher than the reference range of the tMg, one study reported 90% of the cases were within the reference range of the ionized magnesium (iMg) which is a physiologically active fraction of magnesium (Mg) in the blood. This is due to the higher serum phosphorous level in the hemodialysis patients, and negative correlation between iMg and the serum phosphorous, and anion gap. Current guideline recommends Mg level should be managed to be 3.0 mg/dL at pre-dialysis blood test. However, not much are known what iMg level is optimal. This study aimed to assess the impacts on iMg and tMg level of the patients by the conversion of the routinely-used dialysate containing 1.0 mEq/L (0.5 mmol/L) Mg to higher dialysate Mg concentration. Method This longitudinal study was conducted at Shingashi Renal Centre, Fujimino Renal Centre, and Kawagoe-minami Renal Centre. Out of 289 stable maintenance hemodialysis patients agreed to participate in the study, 42 were excluded due to death, changing hospital, or hospitalization. A total of 247 patients who are on regular hemodialysis treatment at least 6 months with commercially available dialysate mixture TA1 (Mg concentration: 1.0 mEq/L (0.5 mmol/L)) were included for the analysis. Dialysate Mg concentration was then increased to 1.25 mEq/L (0.625 mmol/L) by using a commercially available dialysate mixture TA5. Blood tests were carried out on the day of the first hemodialysis session of the week (Monday or Tuesday) before dialysate Mg concentration conversion and at 1, 4, 12, and 24 weeks. The iMg concentrations were analyzed using the ion-selective electrode on blood gas analyzer StatProfile Prime Plus (Nova Biomedical, Waltham, MA, USA). The tMg concentrations were determined by using Xylidylblue. Results Mean age of the 247 enrolled patients was 70.3ion-selective electrode on blood gas analyzer StatProfile Prime Plus (Nova Bidecreased significantly (TA1-pre:2.58±0.36mg/dL→TA1-post:2.18±0.16mg/dL, TA5-pre (24wk): 2.74±0.35mg/dL→TA5-post: 2.44±0.15mg/dL). After conversion to higher Mg dialysate TA5, post-dialysis tMg increased in comparison with TA1. After conversion to the higher Mg dialysate, iMg increased as well significantly (TA1-pre:0.64±0.11mmol/L→TA1-post:0.49±0.05mmol/L, TA5-pre:0.70±0.11mmol/L→0.60±0.04mmol/L) . However, iMg of TA1-post was close to the lower limit of the reference range of iMg (Fig. 1). Conclusion Hemodialysis patients’ iMg level became manageable within reference range by the conversion of the dialysate from TA1 to TA5 and the clinical advantage of iMg monitoring with hemodialysis patients were demonstrated. A slight increase in dialysate Mg concentration from 1.0 mEq/L to 1.25 mEq/L increased the iMg/tMg ratio, and iMg was within the normal range even when the pre-dialysis reference value of 3 mg/dL was not reached.

In Vivo Antidiarrheal Potential of the Leaf Extract of Maytenus addat (Loes.) Sebsebe and Its Major Compound.

Diarrhea continues to be one of the top causes of death in children under the age of five, particularly in developing nations. In Ethiopian traditional medicine, a variety of medicinal plants are used to treat diarrhea. One of these plants is Maytenus addat (Loes.) Sebsebe (fam. Celastraceae), which is endemic to the Afromontane forests, especially along forest margins, of Ethiopia. The air-dried powdered leaves of M. addat were macerated with 80% methanol to yield a crude extract. Additionally, the powdered plant material underwent sequential solvent extraction using chloroform, methanol, and water to obtain solvent fractions. The 80% methanol leaf extract, solvent fractions, and an isolated compound from M. addat were evaluated for their antidiarrheal activity using castor oil-induced diarrheal model, anti-enteropooling test, and charcoal meal test in mice. The results showed that the 80% methanolic leaf extract significantly reduced the onset of diarrhea, the weight of feces, and the frequency of defecation in all the tested doses. The methanol and water fractions of the hydroalcoholic extract also exhibited dose-dependent antidiarrheal activity, with the methanol fraction showing the highest activity at 400 mg/kg dose. Subsequently, the most active methanol fraction was subjected to C-18 solid phase extraction, resulting in the isolation of a 3-hydroxyflavone, identified as quercetin by ESI-qToF-MS, 1H, and 13C-NMR spectroscopic techniques. Quercetin demonstrated a strong antidiarrheal activity in a dose-dependent manner. Thus, the present study provided evidence that the leaves of M. addat possess genuine antidiarrheal activity upholding the traditional medicinal use of the plant for the treatment of diarrhea. The findings also suggest that quercetin is responsible, in full or in part, for the activity of the plant.

Anti-inflammatory and antinociceptive effects, toxicity, and phytochemical analysis of Beaumontia grandiflora Wall.

Preliminary pharmacological studies revealed that the EtOAc fraction (BGEA) might be the main active fraction with anti-inflammatory and antinociceptive effects in Beaumontia grandiflora Wall. Further assays on BGEA at doses of 200, 400, and 800 mg/kg using four animal models showed that it could inhibit the xylene-induced ear edema, carrageenan-induced paw edema, and acetic acid-induced writhing and prolong the latency time in the hot-plate test. ELISA analysis revealed that the anti-inflammatory activity of BGEA might be associated with the decrease of TNF-α, IL-1β, and IL-6 levels and the increase of the IL-10 level. The acute toxicity test showed that except for the n-BuOH fraction, the LD50 values of the extract and other three fractions were higher than 2000 mg/kg bw. Finally, 14 compounds were identified from BGEA by LC-MS. This research provides some basis for the folk use of B. grandiflora in the treatment of inflammation and pain-related diseases.

Exploring Southern Ecuador's Traditional Medicine: Biological Screening of Plant Extracts and Metabolites.

Ecuador stands as a nation inheriting a profound ancestral legacy in the utilization of medicinal plants, reflective of the rich biodiversity embraced by various ethnic groups. Despite this heritage, many of these therapeutic resources remain insufficiently explored concerning their toxicity and potential pharmacological effects. This study focused on a comprehensive evaluation of cytotoxicity and the potential subcellular targets within various extracts and nine isolated metabolites from carefully selected medicinal plants. Assessing their impact on the breast cancer cell line (MCF7), we subsequently examined the most active fractions for effects on the cell cycle, microtubule network, centrosome duplication, γH2AX foci, and E-cadherin. The investigated crude extracts and isolated compounds from Ecuadorian medicinal plants demonstrated cytotoxic effects, influencing diverse cellular pathways. These findings lend credence to the traditional uses of Ecuadorian medicinal plants, which have served diverse therapeutic purposes. Moreover, they beckon the exploration of the specific chemicals, whether in isolation or combination, responsible for these observed activities.

Short-term effects of irrigation and nitrogen management on paddy soil carbon pools under deep placement of basal fertilizer nitrogen

Active soil organic carbon (SOC) fractions are major driving factors of soil fertility. Understanding the effects of water and fertilizer management on changes in active SOC fractions helps improve soil quality and maintain high agricultural productivity. We conducted a 3-year field experiment in Northeast China. In this experiment, natural soil (CKT) was used as a blank, and two irrigation regimes were established: conventional flooded irrigation (FI) and controlled irrigation (CI). Four nitrogen application levels were set for both irrigation regimes under deep placement of basal fertilizer N: Nd0 (0 kg ha–1), Nd (110 kg ha–1), Nd1 (99 kg ha–1), and Nd2 (88 kg ha–1). After 3 years, at similar N fertilizer application rate, the rice yield, total organic carbon (TOC), and active SOC fraction content of CI were higher under CI than FI. The growth rate of rice yield was 3.8% − 8.63% under CI than FI. Under CI, the rice yield, active SOC fractions contents and carbon pool management index (CPMI) did not decrease with decreasing N application rate but instead reached the highest level in the CNd1 treatment. Overall, CI with Nd1 treatment appears to be the best practice for improving soil fertility and crop productivity in Northeast China.

Evaluation and isolation of anti-cancer compounds from the endophytic fungus Penicillium funiculosum isolated from Persicaria salicifolia (Brouss. Ex Willd.) seeds growing in Egypt

Background The endophytic fungus Penicillium is an important source of natural bioactive products. Persicaria salicifolia (Brouss. ex Willd) (family Polygonaceae) is a widely distributed plant on the Nile River and was reported to have several biological activities, such as antioxidative, antibacterial, and anti-inflammatory effects. Objective This study aimed to explore the potential activities (cytotoxic, anti-oxidant, and anti-microbial) of the endophytic fungus Penicillium funiculosum isolated from Persicaria salicifolia seeds growing in Egypt. Materials and methods The endophytic fungus Penicillium funiculosum was isolated from the seeds of the Persicaria salicifolia plant. The fungi were grown on Basmati rice as a solid media for the enhancement of pure fungi production. The collected fungi were extracted with ethyl acetate and fractionated using n-hexane and methanol. All fractions were examined for their cytotoxic, antioxidant, and antimicrobial activities. The secondary metabolites were isolated from the active fractions through column chromatography, and the isolated compounds were identified by spectroscopic technique. Molecular docking analysis was applied to the isolated compounds. Results and conclusion Ethyl acetate extract (Pf-2) of P. funiculosum was proved to have a highly potent antioxidant (IC50 37.5±0.70 μg/ml) and cytotoxic effects. It was affected on hepatic cancer (HepG-2), human colon carcinoma (HCT-116), lung carcinoma (A-549), and human breast cancer (MCF-7) cells with IC50 values (μg/ml) 4.26±0.2, 6.66±0.9, 9.36±0.3, and 9.41±0.7, respectively. Pf 2 was subjected to fractionation, resulting in four fractions (Pf 2–1 to Pf 2–4). The most potent cytotoxic fraction, Pf 2–2, was further fractionated into six sub-fractions: Pf 2–2 A to Pf 2–2 F. Pf 2–2 A possessed the most potent cytotoxic activity. The ethyl acetate extract (Pf 2) also had antimicrobial activity against gram-positive Bacillus subtilis and gram-negative bacteria (E. coli). Four compounds were isolated from Pf 2–2 A and identified by spectroscopic methods: NMR (1H and 13C) and Mass as (1) (9E, 11E, 13Z, 15Z)-tetracosa-9, 11, 13, 15-tetraenoic acid, (2) 3-(1Z, 3E-hexa-1, 3-dienyl)-4b-methyl-tetradecahydrophenanthrene, (3) mannitol, (4) d-Cerebroside A-glucose. It is the first time to report the isolation of compound (2) from the genus Penicillium and compounds (1, 3, and 4) from the species P. funiculosum. Compound 2 was the most potent cytotoxic one. Molecular docking was studied for polar compounds 1, 3, and 4 using the COX-2 enzyme, which indicated that compound 4 was the most potent anti-inflammatory one.