Phytochemical Analysis, Pharmacological Activities, Isolation and Characterization of Bioactive Compounds from the Roots of Sterculia urens Roxb.

The present study was intended to explore the pharmacological significance of the crude root extract of Sterculia urens Roxb. Further the bio-active compounds were isolated and characterized using chromatographic and spectroscopic techniques. Soxhlet extraction apparatus was utilized for isolation of the chemical constituents from the root using a series of solvents such as n-hexane, ethyl acetate, methanol and water. The pharmacological activities such as inhibition of DPPH radical, α-amylase enzyme activity, albumin denaturation along with antibacterial and thrombolytic activities. The isolation of purified bioactive constituents was carried using preparative HPLC technique and the purified compounds were characterized using spectroscopic techniques like NMR, IR and mass. Among the crude root extracts, methanolic extract shows high DPPH radical scavenging activity with IC50 concentration of 26.74 ± 0.08 μg/mL. The IC50 concentrations in α-amylase enzyme inhibition activity was 263.96 ± 0.90, 127.73 ± 1.23 and 223.54 ± 4.76 μg/mL, respectively for ethyl acetate, methanol and water extracts, respectively. The methanolic extract shows high albumin denaturation inhibition assay than other extracts with IC50 concentration as 137.09 ± 0.20 μg/mL, which is very close to standard ascorbic acid. The methanolic extract also shows high % clot lysis than other extracts and results were comparable with 100 μL of streptokinase standard. The preparative HPLC followed by spectral analysis confirm that two known alkaloids (Sterculinine I & II) and three known flavonoids (gossypetin, apigenin and 6-hydroxyluteolin) were purified and characterized from the root methanol of Sterculia urens Roxb. The purified and identified compounds were reported for the first time in Sterculia urens Roxb.

Influence of Prolonged Serotonin and Ergovaline Pre-Exposure on Vasoconstriction Ex Vivo

Ergot alkaloid mycotoxins interfere in many functions associated with serotonergic neurotransmitters. Therefore, the objective was to evaluate whether the association of serotonin (5-hydroxytryptamine, 5-HT) and ergot alkaloids during a 24 h pre-incubation could affect the vascular contractile response to ergot alkaloids. To evaluate the effects of 24 h exposure to 5-HT and ergot alkaloids (ergovaline, ERV), two assays were conducted. The first assay determined the half-maximal inhibitory concentration (IC50) following the 24 h pre-exposure period, while the second assay evaluated the effect of IC50 concentrations of 5-HT and ERV either individually or in combination. There was an interaction between previous exposure to 5-HT and ERV. Previous exposure to 5-HT at the IC50 concentration of 7.57 × 10−7 M reduced the contractile response by more than 50% of control, while the exposure to ERV at IC50 dose of 1.57 × 10−10 M tended to decrease (p = 0.081) vessel contractility with a response higher than 50% of control. The 24 h previous exposure to both 5-HT and ERV did not potentiate the inhibitory response of blood vessels in comparison with incubation with each compound alone. These results suggest receptor competition between 5-HT and ERV. More studies are necessary to determine the potential of 5-HT to treat toxicosis caused by ergot alkaloids.

Synthesis, Crystal Structure, and Biological Evaluation of Novel 5-Hydroxymethylpyrimidines

Pyrimidine displays a wide array of bioactivities, and thence, it is still considered a potent unit of new drug research. Its derivative, 5-hydroxymethylpyrimidine, can be found as a scaffold of nontypical nitrogen bases in DNA and as a core of some natural bioactive compounds. In this study, we obtained a series of 5-hydroxymethylpyrimidines that vary in the 4-position by the reduction of proper esters. All compounds were characterized by spectroscopic analysis, and single-crystal X-ray diffraction was performed for some of them. Biological investigations estimated cytotoxic properties against normal (RPTEC) and cancer (HeLa, HepaRG, Caco-2, AGS, A172) cell lines. It was found that the derivatives with an aliphatic amino group at the 4-position are generally less toxic to normal cells than those with a benzylsulfanyl group. Moreover, compounds with bulky constituents exhibit better anticancer properties, though at a moderate level. The specific compounds were chosen due to their most promising IC50 concentration for in silico study. Furthermore, antimicrobial activity tests were performed against six strains of bacteria and one fungus. They demonstrated that only derivatives with at least three carbon chain amino groups at the 4-position have weak antibacterial properties, and only the derivative with 4-benzylsulfanyl constituent exhibits any antifungal action.

An Assessment of InP/ZnS as Potential Anti-Cancer Therapy: Quantum Dot Treatment Increases Apoptosis in HeLa Cells

InP/ZnS quantum dots (QDs) are an emerging option in QD technologies for uses of fluorescent imaging as well as targeted drug and anticancer therapies based on their customizable properties. In this study we explored effects of InP/ZnS when treated with HeLa cervical cancer cells. We employed XTT viability assays, reactive oxygen species (ROS) analysis, and apoptosis analysis to better understand cytotoxicity extents at different concentrations of InP/ZnS. In addition, we compared the transcriptome profile from the QD-treated HeLa cells with that of untreated HeLa cells to identify changes to the transcriptome in response to the QD. RT-qPCR assay was performed to confirm the findings of transcriptome analysis, and the QD mode of action was illustrated. Our study determined both IC50 concentration of 69 µg/mL and MIC concentration of 167 µg/mL of InP/ZnS. It was observed via XTT assay that cell viability was decreased significantly at the MIC. Production of superoxide, measured by ROS assay with flow cytometry, was decreased, whereas levels of nitrogen radicals increased. Using analysis of apoptosis, we found that induced cell death in the QD-treated samples was shown to be significantly increased when compared to untreated cells. We conclude InP/ZnS QD to decrease cell viability by inducing stress via ROS levels, apoptosis induction, and alteration of transcriptome.

Protective Effects of Gold Nanoparticles Against Malathion-Induced Cytotoxicity in Caco-2 Cells

Malathion is an organophosphorus insecticide widely used in agriculture, residential area, and public health programs with a known mechanism of toxicity of inhibition of acetylcholinesterase and induction of oxidative stress. Gold nanoparticles (AuNPs) represent stable and easily synthesized nanoparticles with extensive use in consumer products and medicine. Due to the antioxidant property of AuNPs, it is possible that AuNPs may prevent malathion-induced oxidative damage. In this study, the cytotoxicity of malathion and AuNPs (10 and 20 nm) were measured separately in Caco-2 cells. Then the protective effects of AuNPs were evaluated by measuring the oxidative stress (lipid peroxidation level and glutathione content) and acetylcholinesterase activity. The calculated IC50s values at 48 hr were 326.8±0.32, 43.09±0.65, and 41.46±0.24 µg/ml for malathion, AuNPs 10 and 20 nm, respectively. Then, the lowest concentration of AuNPs (1 µg/ml) and IC50 concentration of malathion (326.8 µg/ml) were selected to evaluate the effects of pretreatment of Caco-2 cells with AuNPs before exposure to malathion were evaluated. Interestingly, the results showed remarkably significant protective effects of AuNPs by attenuation the different parameters of oxidative stress and cytotoxicity induced by malathion in cells (P<0.001). It is the first report showing the protective effects of AuNPs against malathion-induced cytotoxicity in the Caco-2 cell line.

Self-Assembly of pH-Labile Polymer Nanoparticles for Paclitaxel Prodrug Delivery: Formulation, Characterization, and Evaluation

The efficacy of paclitaxel (PTX) is limited due to its poor solubility, poor bioavailability, and acquired drug resistance mechanisms. Designing paclitaxel prodrugs can improve its anticancer activity and enable formulation of nanoparticles. Overall, the aim of this work is to improve the potency of paclitaxel with prodrug synthesis, nanoparticle formation, and synergistic formulation with lapatinib. Specifically, we improve potency of paclitaxel by conjugating it to α-tocopherol (vitamin E) to produce a hydrophobic prodrug (Pro); this increase in potency is indicated by the 8-fold decrease in half maximal inhibitory concentration (IC50) concentration in ovarian cancer cell line, OVCA-432, used as a model system. The efficacy of the paclitaxel prodrug was further enhanced by encapsulation into pH-labile nanoparticles using Flash NanoPrecipitation (FNP), a rapid, polymer directed self-assembly method. There was an 1100-fold decrease in IC50 concentration upon formulating the prodrug into nanoparticles. Notably, the prodrug formulations were 5-fold more potent than paclitaxel nanoparticles. Finally, the cytotoxic effects were further enhanced by co-encapsulating the prodrug with lapatinib (LAP). Formulating the drug combination resulted in synergistic interactions as indicated by the combination index (CI) of 0.51. Overall, these results demonstrate this prodrug combined with nanoparticle formulation and combination therapy is a promising approach for enhancing paclitaxel potency.

Comparison the effects of Sambucus ebulus leaf and fruit extracts on Leishmania major in-vitro

Background;: Leishmaniasis is one of the major diseases caused by the intracellular parasite of Leishmania. It has become one of the most dangerous health problems today. Our aim of the present study is to compare the effects of Sambucus ebulus leaf and fruit extracts on Leishmania major in vitro. Methods: In this study, we used MTT, promastigote and amastigote assay to evaluate the effect of different concentrations of the extract on parasite and we compared their effects. The flow cytometry technique was also used to detect the apoptotic effect of the extracts on promastigotes. Results: According to MTT experiment IC50 concentration of leaf and fruit extracts on parasite was 157 μg/ml and 265 μg/ml, respectively. After analysis by flow cytometry, leaf and fruit extracts also showed apoptosis effect. Leaf and fruit extract caused 40.2 and 2.67 percent apoptosis. Conclusion: Based on the above assessment, we determined that the S. ebulus leaf extract has a more toxic effect on promastigotes and amstigotes than its fruit extract and maybe in the future that be used as a drug candidate.

Anti-dermatophytes Activity of Origanum compactum Essential Oil at Three Developmental Stages

The main aim of this study is to determine the chemical compounds of Origanum compactum essential oils (OCEO) at three phenological stages (vegetative, flowering, and post-flowering) and to evaluate their antifungal activity against three dermatophytes fungal strains: Trichophyton violaceum, Trichophyton tonsurans, and Trichophyton mentagrophytes using direct contact method. The main compounds of OCEO are carvacrol, thymol, p-cymene, and γ-terpinene. Oregano EOs showed important antifungal activities with some variability between EOs and fungal testing. At a concentration of 0.75% (v/v), the best inhibition values showed with OCEO at vegetative stage against Trichophyton mentagrophytes (95.17%), Trichophyton tonsurans (92.47%), and Trichophyton violaceum (91.41%). The OCEO at vegetative stage also showed the best IC50 (concentration of an inhibitor where the response is reduced by half) values of 52.86, 0.56, and 0.57% (v/v) against Trichophyton mentagrophytes, Trichophyton tonsurans, and Trichophyton violaceum, respectively. The findings reveal that OCEO is a good source of anti-dermatophytes agents.

MAP2K6-FP Enhances the Sensitiveness of Paclitaxel for Ovarian Cancer via Inducing Autophagy

BackgroundPaclitaxel is recommended as a first-line chemotherapeutic agent against ovarian cancer, but drug resistance becomes a major limitation. The key molecule or mechanism associated with paclitaxel resistance in ovarian cancer still remains unclear. Recent studies have revealed an association between autophagy and drug resistance.MethodsWe previously synthesized a MAPK kinase-recombinant fusion protein, MAP2K6-FP, that contains 3 domains: a protein transduction domain TAT, a human ovarian cancer HO8910 cell-specific binding peptide, and a potential antitumor effector domain MKK6(E). In this study, we investigated the effect of MAP2K6-FP on HO8910 cells treated with paclitaxel.ResultsThe IC50 (concentration by which 50% cell growth was inhibited) was 20 μM for paclitaxel alone, 1.5 μg/mL for MAP2K6-FP alone, and 0.3 μg/mL for MAP2K6-FP and 15 μM for paclitaxel if combined, respectively. In addition, immunohistochemistry assay demonstrated that tumor tissues from ovarian cancer patients showed higher expression of LC-3, the autophagy-related protein, compared with normal ovarian tissues. MAP2K6-FP (0, 2.5, 5, 10, 20, and 40 μg/mL) dose-dependently increased the LC-3 expression in HO8910 cells. Immunofluorescence assay showed that paclitaxel alone increased the expression of LC-3 in HO8910 cells, which was further enhanced by the combination with MAP2K6-FP. Downregulation of LC-3 expression using LC-3 small interfering RNA inhibited the cytotoxicity effect of MAP2K6-FP. Furthermore, either MAP2K6-FP alone or in combination with paclitaxel increased the ratio of expressions of Beclin-1/Bcl-2, another autophagy-related markers, compared with paclitaxel alone.ConclusionsMAP2K6-FP enhanced the sensitiveness of paclitaxel for ovarian cancer via inducing autophagy.

In vitro mycelial sensitivity of Macrophomina phaseolina to fungicides

Black root rot, caused by Macrophomina phaseolina (Tass.) Goid., is the most common root disease in soybean fields. This study aimed to determine the in vitro mycelial sensitivity, measured by the IC50 (concentration to inhibit 50% of the fungus mycelial growth) of a M. phaseolina isolate obtained from soybean, to different fungicides (thiram, iprodione, carbendazim, pyraclostrobin, fluquinconazol, tolyfluanid, metalaxyl and penflufen + trifloxystrobin), at six concentrations (0.01 mg L-1, 0.10 mg L-1, 1.00 mg L-1, 10.00 mg L-1, 20.00 mg L-1 and 40.00 mg L-1 of the active ingredient). The 0.00 mg L-1 concentration represented the control, without fungicide addition. The mycelial growth evaluation was performed with the aid of a digital pachymeter, by measuring the colonies diameter, when the fungus growth in the control treatment reached the Petri dish edge. The experimental design was completely randomized, with four replications. Concerning the fungitoxicity of active ingredients, a variation from non-toxic to highly fungitoxic was observed to the M. phaseolina isolate, with IC50 values ranging from 0.23 mg L-1 to > 40.00 mg L-1, being carbendazim the most efficient one (IC50 = 0.23 mg L-1). The fungus showed insensitivity to the active ingredients of fluquinconazole, metalaxyl, thiram and tolyfluanid.