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Dittrichia viscosa L., is a perennial plant belonging to the Asteraceae family, and this study was performed to investigate the chemical composition of its extract, using gas chromatography–mass spectrometry (GC–MS). Total phenol (TPC), flavonoid (TFC), and tannins contents (TTC), were quantified using colorimetric methods in two extracts (EtOH and ACE). The antioxidant activity was evaluated using DPPH scavenging, phosphomolebdenum test (TAC) and ferric reducing power assay (FRAP). The antimicrobial activity was determined against six nosocomial pathogens: Bacillus subtilis, Staphylococcus aureus, Salmonella enterica, Escherichia coli, Candida albicans, Aspergillus niger, using disc diffusion method and microdilution assay. The ACE and EtOH extracts had similar TPC: 151.18 ± 1.57 and 127.09 ± 15,81 mg GAE/ g DW. TFC & TTC recorded were also closely matched. The chemical composition revealed the presence of 18 phytochemical compounds with a total of 99.91%, where trimethylsilyl-meso-inositol (20.54%) was the major compound, followed by 5(4H)-Thebenidinone (16.80%). Both extracts showed high radical scavenging activity with an IC50 equal to 12.54 ± 0.2 μg/mL for EtOH, and 7.84 ± 0.1 μg/mL for ACE in DPPH test. In the FRAP test, we recorded an EC50 of 6.37 ± 0,012 mg/mL for EtOH, and 6 ± 0.022 mg/mL for ACE. The ACE presented higher antioxidant capacity (253.52 ± 2.98 mg AAE/g) compared to EtOH (189.14 ± 4,86 mg AAE/g) in the TAC assay. The higher inhibition zone was observed on B. subtilus (13 ± 0.1 mm) for EtOH, and the ACE was more effective on S. enterica (13.3 ± 0.08 mm). All the microbial strains were sensitive for both extracts, with MICs ranging from 0.93 mg/mL to 15 mg/mL.

The present work is carried out to evaluate biological activities of tubers extracts of Bunium mauritanicum. The methanol extract yield had higher (7.81%) tha n that of the aqueous extract (6.79%). The quantitative analysis of total phenols and flavonoid revealed that the highest concentration was recorded for the methanoic fraction with 89,442 ± 5,951μg EAG mg and 4.031 ± 0.141 μg EQ mg of extract respectively. In addition, the aqueous extract of tubers represents the most important antioxidant activity with an IC 50 of 0.14 mg ml against 2,2 diphenyl 1 picryl hydrazyl (DPPH Wherea s, the two extracts of the tubers have low reducing capacities compared to standard with EC 50 equal to 0.048 mg/ ml for the methanolic extract, 0.018 mg ml for t he aqueous extract and 0.009 mg/ ml for ascorbic acid. The methano l ic and aqueous extracts of Bunium mauritanicum reacted positively at least on one of the bacterial strains studied.

Dengue virus poses a serious worldwide health threat with up to 400 million infections occurring annually in over 100 countries. Currently, there are no specific therapeutics available, and the only licensed vaccine encompasses risk of hospitalization in immunologically naive individuals. In the current work for the first time we are reporting dengue virus type 2 (DENV2) inhibitory activity of newly designed 3-chlorobenzyl linked 1,9-diazaspiro[5.5]undecane derivatives. Compounds with substitutions featuring 2-methylbenzyl (SPO-6, EC50 = 11.43 ± 0.87 μM), 4-bromobenzyl (SPO-7, EC50 = 14.15 ± 0.50 µM), and 4-cyanobenzyl (SPO-13, EC50 = 20.77 ± 1.92 µM) groups and Ribavirin (IC50=50.9±18 µM, J. Gen. Virol.2006,87, 1947-1952) were found to be potent against DENV2 in a cell-based assay. Docking calculations identified NS5-methyltransferase as the most probable target for this series of compounds. Molecular dynamics simulations of NS5-methyltransferase reproduced the experimental binding affinity findings by yielding ΔGbind values that predicted SPO-6 to possess the most favorable binding energy of -27.2 ± 3.9 kcal/mol compared to SPO-7, SPO-13, and Ribavirin with -22.5 ± 4.7, -22.5 ± 4.6, and -20.0 ± 4.6 kcal/mol, respectively. In addition, based on Lipinski’s rule of 5 SPO-6 was found to be nontoxic and possessed a better score of drug likeness (5.87) in comparison to the standard drug ribavirin (1.72).

Peach scab is a fungal disease caused by Venturia carpophila that can significantly reduce peach yield and quality. Fungicide application is the main control measure for peach scab worldwide. To better understand the fungicide-resistance status and devise suitable management strategies, the sensitivity of 135 single-spore V. carpophila isolates to the commonly used fungicides carbendazim, iprodione, propiconazole, azoxystrobin, and boscalid were determined using a microtiter plate test method. Results showed that the mean effective concentrations to cause inhibitions by 50% (EC50) of tested isolates to iprodione, propiconazole, azoxystrobin, and boscalid were 16.287, 0.165, 0.570, and 0.136 µg/ml, respectively. The EC50 values of V. carpophila isolates to four fungicides displayed unimodal frequency distributions, indicating no resistance occurred to these fungicides. On the contrary, bimodal frequency distribution was observed for carbendazim, indicating that V. carpophila developed resistance to carbendazim. Resistance was widely detected from all 14 provinces studied. Molecular analysis showed that the point mutation E198K of the TUB2 gene determined high resistance, whereas E198G conferred moderate resistance. Moderate and high resistances were stable, and the resistant isolates did not show significant fitness penalties. On the contrary, some resistant isolates showed better competitiveness under certain stresses. This is the first report to detect the sensitivity of V. carpophila to fungicides, which enables future monitoring of fungicide resistance and provides basic information to allow the design of suitable peach scab management strategies.

In this study, the polyphenol content and the antioxidant and antimicrobial activities of hydroethanolic (MVE) and hydroacetonic (MVA) leaf extracts of Marrubium vulgare L. were examined. The results indicated that the total phenolic content was higher in MVA (112.09 ± 4.77 mg GAE/DW) compared to MVE extract (98.77 ± 1.68 mg GAE/DW). The total flavonoid content was also higher in MVA extract (21.08 ± 0.38 mg QE/g DW) compared to MVE (17.65 ± 0.73 mg QE/g DW). Analysis of the chemical composition revealed the presence of 13 compounds with a total of 96.14%, with the major compound being malic acid (22.57%). Both extracts possess a good total antioxidant activity. DPPH and FRAP assays indicated that the MVE extract possesses a better antioxidant activity, with IC50 = 52.04 µg/mL ± 0.2 and EC50 of 4.51 ± 0.5 mg/mL, compared to MVA extract (IC50 = 60.57 ± 0.6 µg/mL and EC50 of 6.43 ± 0.0411 mg/mL). Moreover, both extracts exhibited strong antimicrobial activity against certain nosocomial strains as indicted by the MIC values, which ranged between 0.93 mg/mL and 10 mg/mL. Taken together, these results reveal the importance of M. vulgare as a natural antioxidant with important antimicrobial activity.

Porcine reproductive and respiratory syndrome virus (PRRSV) continues to cause significant economic loss worldwide and remains a serious threat to the pork industry. Currently, vaccination strategies provide limited protection against PRRSV infection, and consequently, new antiviral strategies are urgently required. Andrographolide (Andro) and its derivative potassium dehydrographolide succinate (PDS) have been used clinically in China and other Asian countries as therapies for inflammation-related diseases, including bacterial and viral infections, for decades. Here, we demonstrate that Andro and PDS exhibit robust activity against PRRSV replication in Marc-145 cells and primary porcine alveolar macrophages (PAMs). The two compounds exhibited broad-spectrum inhibitory activities in vitro against clinically circulating type 2 PRRSV GD-HD, XH-GD, and NADC30-like HNhx strains in China. The EC(50) values of Andro against three tested PRRSV strain infections in Marc-145 cells ranged from 11.7 to 15.3 μmol/L, with selectivity indexes ranging from 8.3 to 10.8, while the EC(50) values of PDS ranged from 57.1 to 85.4 μmol/L, with selectivity indexes ranging from 344 to 515. Mechanistically, the anti-PRRSV activity of the two compounds is closely associated with their potent suppression on NF-κB activation and enhanced oxidative stress induced by PRRSV infection. Further mechanistic investigations revealed that PDS, but not Andro, is able to directly interact with PRRSV particles. Taken together, our findings suggest that Andro and PDS are promising PRRSV inhibitors in vitro and deserves further in vivo studies in swine. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12250-021-00455-y.

Studies on the tissue distribution of the New Synthetic Opioid U-47700 and its main metabolite N-desmethyl-U-47700 revealed about 6-fold higher metabolite concentrations in pig brain as compared to the parent compound. To better assess the toxic potential of this drug, the aim of this study was to assess the in vitro μ-opioid receptor (MOR) activation potential of the main metabolites of U-47700, N-desmethyl-U-47700 and N,N-bisdesmethyl-U-47700, using a live cell-based reporter assay based on NanoLuc Binary Technology®. Cells stably expressing human MOR and β-arrestin 2 (βarr2), each fused via a flexible linker to two complementary inactive subunits of the nanoluciferase, were seeded on poly-D-lysine-coated 96-well plates and treated with N-desmethyl-U-47700, N,N-bisdesmethyl-U-47700, U-47700, or hydromorphone as reference standard. MOR activation results in functional complementation of the nanoluciferase, which can be assessed via luminescence monitoring. The potency of the metabolites is lower than that of U-47700 (EC50 of 186 nM for U-47700, to 3770 nM for N-desmethyl-U-47700 and >5 μM for N,N-bisdesmethyl-U-47700). The maximal efficacy (Emax ) observed (relative to hydromorphone, set arbitrarily at 100%) decreased from 183% to 127% and 39.2% for U-47700, N-desmethyl-U-47700 and N,N-bisdesmethyl-U-47700, respectively. Thus, the loss of one or two methyl groups reduced the MOR activation potential, which was more pronounced if both methyl groups were removed. It is thus anticipated that the impact on MOR exerted by the higher metabolite concentration in brain has only little - if any - relevance for the strong toxic effects of U-47700.

Background Treatment and control of schistosomiasis rely on a single drug, praziquantel. New orally active antischistosomals featuring novel molecular scaffolds are urgently needed to prevent the emergence of resistance. Methods We screened 400 drug-like compounds contained in the open-access Pandemic Response Box (PRB) against newly transformed schistosomula (NTS) at a concentration of 10 µM scoring death, changes in motility, and morphological alterations. Compounds displaying an activity ≥66% at 72 h underwent testing against adult Schistosoma mansoni in vitro. Fast-acting (≥66% at 24 h), nontoxic drugs focusing on late-stage and approved drugs were investigated in the patent S. mansoni mouse model. Results We identified 26 hits active against NTS, of which 17 elicited ≥66% activity against adult S. mansoni following 24 h of drug exposure. The highest activity against adult S. mansoni was observed with MMV1581558 (EC50 value of 0.18 ± 0.01 µM) and nitazoxanide (0.47 ± 0.07 µM). Of the five compounds tested in vivo, MMV1581558 and the approved drug ozanimod reduced average worm burden versus controls by 42 % and 36 %, respectively, after a single oral dose of 200 mg/kg bodyweight in mice harboring a chronic S. mansoni infection. Conclusion MMV1581558 discovered from screening the PRB represents a novel antischistosomal scaffold with high in vitro antischistosomal activity amenable to chemical modification for drug development.

Three new compounds (1‒3), including a phenylpropanoid (1), a bistetrahydrofuran lignin (2), and a lignan dimer (3), and seven known lignans (4‒10), were isolated from an EtOH extract of the aerial parts of Triadica rotundifolia. The structures of the new compounds were elucidated by comprehensive spectroscopic analyses. New compounds 1−3 were evaluated for their antineuroinflammatory and antioxidant activities. Research on the antineuroinflammatory activity suggested that compounds 2 and 3 had significant inhibitory effects on NO production in LPS-induced BV-2 cells, with the IC50 values of 15.7 and 5.8 μM, respectively. Meanwhile, compounds 2 and 3 exhibited better DPPH free radical scavenging activities with EC50 values of 2.0 and 4.3 μM, respectively, than the positive control vitamin C (EC50 = 14.3 μM).

INTRODUCTION: Numerous efforts in natural product drug development are reported for the treatment of Coronavirus. Based on the literature, among these natural plants Artemisia annua L. shows some promise for the treatment of SARS-CoV-2. OBJECTIVE: The main objective of our study was to determine artemisinin content by liquid chromatography electrospray ionisation tandem mass spectrometry (LC-ESI-MS/MS), to investigate the in vitro biological activity of artemisinin from the A. annua plants grown in Turkey with various extracted methods, to elaborate in silico activity against SARS-CoV-2 using molecular modelling. METHODOLOGY: Twenty-one different extractions were applied. Direct and sequential extractions studies were compared with ultrasonic assisted maceration, Soxhlet, and ultra-rapid determined artemisinin active molecules by LC-ESI-MS/MS methods. The inhibition of spike protein and main protease (3CL) enzyme activity of SARS-CoV-2 virus was assessed by time resolved fluorescence energy transfer (TR-FRET) assay. RESULTS: Artemisinin content in the range 0.062-0.066%. Artemisinin showed significant inhibition of 3CL protease activity but not Spike/ACE-2 binding. The 50% effective concentration (EC50 ) of artemisinin against SARS-CoV-2 Spike pseudovirus was found greater than 50 µM (EC45 ) in HEK293T cell line whereas the cell viability was 94% of the control (P < 0.01). The immunosuppressive effects of artemisinin on TNF-α production on both pseudovirus and lipopolysaccharide (LPS)-induced THP-1 cells were found significant in a dose dependent manner. CONCLUSION: Further studies of these extracts for COVID-19 treatment will shed light to seek alternative treatment options. Moreover, these natural extracts can be used as an additional treatment option with medicines, as well as prophylactic use can be very beneficial for patients.

Fil: Martino, Renzo. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de la Química y Metabolismo del Fármaco; Argentina Fil: Martino, Renzo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Barreiro Arcos, María Laura. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciónes Biomédicas; Argentina Fil: Barreiro Arcos, María Laura. Consejo de Investigaciones Científicas y Técnica; Argentina Fil: Peralta, Ignacio. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de la Química y Metabolismo del Fármaco; Argentina Fil: Peralta, Ignacio. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Marrassini, Carla. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de la Química y Metabolismo del Fármaco; Argentina Fil: Marrassini, Carla. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Saint Martin, Elina Malén. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de la Química y Metabolismo del Fármaco; Argentina Fil: Saint Martin, Elina Malén. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Cogoi, Laura. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de la Química y Metabolismo del Fármaco; Argentina Fil: Cogoi, Laura. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Cremaschi, Graciela A. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciónes Biomédicas; Argentina Fil: Cremaschi, Graciela A. Consejo de Investigaciones Científicas y Técnica; Argentina Fil: Alonso, María Rosario. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de la Química y Metabolismo del Fármaco; Argentina Fil: Alonso, María Rosario. Consejo de Investigaciones Científicas y Técnica; Argentina Fil: Anesini, Claudia. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de la Química y Metabolismo del Fármaco; Argentina Fil: Anesini, Claudia. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Instituto de la Química y Metabolismo del Fármaco; Argentina Abstract: Most of the deaths from skin cancer are caused by melanoma, a malignancy in which STAT3 plays a crucial role. The inhibition of STAT3 is considered a potential target to induce cell death, tumor regression and metastasis inhibition. The objective of this work was to evaluate the activity of the aqueous extract of Larrea divaricata (Aq), a fraction rich in polyphenols (EA),and the isolated compound quercetin-3-methyl ether (Q3ME) on B16F10 melanoma cells. The effects of Aq, EA and Q3ME were assessed on B16F10 cells by determining the proliferation, viability, apoptosis induction and the expression and phosphorylation of STAT3. The phytochemical composition of the extracts was determined by High Performance Liquid Chromatography. Aq, EA and Q3ME presented antiproliferative activity on B6F10 cells through p-STAT3 inhibition and early and late apoptosis induction (EC50 EA= ≤0.1 μg/ml; Aq= 316±30 μg/ml; Q3ME=

We have investigated the interaction of α1 - and α2 -adrenoceptor subtypes in producing isometric contractions to NA in mouse whole spleen. The α1 -adrenoceptor antagonist prazosin (10-8 M) or the α2 -adrenoceptor antagonist yohimbine (10-6 M) alone produced only small shifts in NA potency in wild type (WT) mice, but the combination produced a large shift in NA potency. In spleen from α1A/D -KO mice, the effects of prazosin and the combination of prazosin and yohimbine were similar to their effects in WT mice. Hence, in α1A/D -KO mice, in which the only α1 -adrenoceptor present is the α1B -adrenoceptor, prazosin still antagonized contractions to NA. The α1A -adrenoceptor antagonist RS100329 (3 × 10-9 M) produced significant shifts in the effects of higher concentrations of NA (EC50 and EC75 levels) and the α1D -adrenoceptor antagonist BMY7378 (3 × 10-8 M) produced significant shifts in the effects of lower concentrations of NA (EC25 and EC50 levels). The effects of BMY7378 and RS00329 demonstrate α1D -adrenoceptor and α1A -adrenoceptor components and suggest that the α1B -adrenoceptor interacts with an α1D -adrenoceptor, and to a lesser extent an α1A -adrenoceptor, at low, and an α1A -adrenoceptor at high, NA concentrations. This study demonstrates the complex interaction between α1 - and α2 -adrenoceptor subtypes in producing contractions of mouse spleen and may have general implications for α-adrenoceptor mediated control of smooth muscle.

Plant extracts such as essential oils are a common yet valuable source of antioxidants for application in pharmaceutical and food preservative. In this study, the phytochemical profile of Baeckea frutescens (BF) and Leptospermum javanicum (LJ) essential oils were determined using GC-FID and GC/MS analyses. This resulted in the identification of 13 components (82.03%) in the essential oil of BF, the major compounds were p-cymene (1, 31.06%), α-pinene (2, 24.53%), 1,8-cineole (3, 11.49%), and γ-terpinene (4, 8.31%). Whilst, 32 components (84.62%) were detected in the essential oil of LJ, in which sabinene (5, 25.83%) and α-pinene (2, 12.17%) were the major components. The antioxidant activity of the essential oils were evaluated using a wide array of radical scavenging assays, such as DPPH radical scavenging, reducing power, metal chelating, and β-carotene bleaching assays. From the results, both BF and LJ essential oils exhibited appreciable antioxidant activity in all assays (EC50 0.29–1.39 mg/mL), except in DPPH radical scavenging assay (EC50 > 50 mg/mL). The essential oil of BF had a higher overall antioxidant capacity than LJ. With reference to the phytochemical profile, compounds 1–5 are potent antioxidants, which may substantially contribute to the antioxidant capacity of the essential oils.

Background and Aims: Melissa officinalis L. (lemon balm) is a perennial herb. Melissae folium and their preparations have been used for their sedative, spasmolytic and antibacterial actions. The study was aimed to investigate the qualities and also to compare the antioxidant activity potentials of the drug samples available in herbal markets and pharmacies in Turkey Methods: The percentages of the loss on drying and total ash were determined by gravimetric method and the percentage of total hydroxycinnamic derivatives was calculated by a spectrophotometric method according to European Pharmacopoeia. Drug samples were investigated for their potentials to scavenge the DPPH radical by using an in vitro method. Results: The percentages of the loss on drying were found to be between 8.51-16.53 %; whereas total ash amounts were determined between 9.41-11.33 %. The percentage of total hydroxycinnamic derivatives was found in the range of 4.45-12.97 %. The extracts of the samples were found to have DPPH radical scavenging activity with EC50 values ranging from 10.60 to 19.10 mu g/ml. Conclusion: In the assays for total ash and quantification of total hydroxy cinnamic derivatives all of the examined commercial samples were found to be compatible with standards in European Pharmacopoeia. Among the tested samples; a sample sold in pharmacy seems to have the best quality when its compared with the standards in European Pharmacopoeia., Ege University Re-search Fund [14-ECZ-009], This study was supported by Ege University Re-search Fund with the project number 14-ECZ-009.

Enlightened by the available structural biology information, a novel series of dihydrothiopyrano[4,3-d]pyrimidine derivatives were rationally designed via scaffold hopping and molecular hybridization strategies. Notably, compound 20a yielded exceptionally potent antiviral activities (EC50 = 4.44-54.5 nM) against various HIV-1 strains and improved resistance profiles (RF = 0.5-5.6) compared to etravirine and rilpivirine. Meanwhile, 20a exhibited reduced cytotoxicity (CC50 = 284 μM) and higher SI values (SI = 5210-63992). Molecular dynamics simulations were performed to rationalize the distinct resistance profiles. Besides, 20a displayed better solubility (sol. = 12.8 μg/mL) and no significant inhibition of the main CYP enzymes. Furthermore, 20a was characterized for prominent metabolic stability and in vivo safety properties. Most importantly, the hERG inhibition profile of 20a (IC50 = 19.84 μM) was a remarkable improvement. Overall, 20a possesses huge potential to serve as a promising drug candidate due to its excellent potency, low toxicity, and favorable drug-like properties.

This study aimed to determine in vitro anthelmintic activity of plant extracts of eleven plant species used traditionally in South Africa to treat various disorders including symptoms related to nematode infections, and to isolate bioactive compounds from the most active plant extract. Crude plant extracts were tested on different life-cycle stages of Haemonchus contortus. The cytotoxicity of the most active extracts, fractions and compounds was evaluated on Vero cells and the most potent extract, fractions and compounds were tested for their ability to kill the parasitic H. contortus and the free-living nematode Caenorhabditis elegans. Typha capensis acetone extract had the strongest egg hatching inhibitory effect with an EC50 of 184.94 μg/mL, and this extract also halted larval development of H. contortus with an EC50 of 83.30 μg/mL compared to the positive control (albendazole) with an EC50 of 2.66 μg/mL. Typha capensis crude extract and its butanol fraction had promising anthelmintic activity against both parasitic H. contortus and free-living C. elegans. Two compounds isolated from T. capensis, namely, isorhamnetin-3-O-β-d-glucoside and isorhamnetin 3-O-rutinoside, had antioxidant activity with IC50 values of 3.16 μg/mL and 0.96 μg/mL respectively, and good anthelmintic activity against H. contortus with IC50 values of 55.61 μg/mL and 145.17 μg/mL respectively. Identification of bioactive compounds from the T. capensis crude extract supports development of this extract as a complementary or alternative treatment against haemonchosis. However, further research is necessary to confirm the anthelmintic efficacy of the plant, including in vivo studies.

To discover novel antibacterial agents, a series of novel 1,3,4-thiadiazole derivatives containing an amide moiety were designed and synthesized, and their antibacterial activities were tested. Compound 30 was designed and synthesized according to the CoMFA model. Compound 30 exhibited higher antibacterial activities against Xanthomonas oryzae pv. oryzicola and Xanthomonas oryzae pv. oryzae, with EC50 values of 2.1 and 1.8 mg/L, respectively, which were superior to those of thiodiazole copper (99.6 and 92.5 mg/L). The protective and curative activities of compound 30 against rice bacterial leaf blight were 51.3 and 46.1%, respectively, which were better than those of thiodiazole copper (37.8 and 38.5%). The protective and curative activities of compound 30 against rice bacterial leaf streak were 45.9 and 40.5%, respectively, which were better than those of thiodiazole copper (36.2 and 31.1%). In addition, the protective activity of compound 30 against rice bacterial leaf streak was related to increased activities of related defense enzymes and upregulated the differentially expressed proteins of the glycolysis/gluconeogenesis pathway.

BackgroundEnterococcus faecalis commonly produce aminoglycoside-modifying enzymes (AMEs) and are implicated in polymicrobial infections.ObjectivesTo determine if AME-producing E. faecalis is capable of protecting Enterobacteriaceae and Pseudomonas aeruginosa from gentamicin exposure.MethodsTwo Klebsiella pneumoniae isolates, two Escherichia coli isolates, and two Pseudomonas aeruginosa isolates were investigated in monoculture time–kill experiments, and each Gram-negative organism was also evaluated during co-culture with either AME-producing or AME-deficient E. faecalis. A pharmacokinetic/pharmacodynamics analysis that utilized Log Ratio Areas and a Hill-type mathematical model was used to determine if the maximal killing or potency of gentamicin against the Gram-negative organisms was altered by the presence of the E. faecalis.ResultsThe maximal killing and potency of gentamicin was the same during monoculture and co-culture experiments for both K. pneumoniae isolates and one E. coli isolate (P > 0.05). In contrast, the maximal killing of gentamicin was attenuated against one E. coli isolate and both P. aeruginosa isolates during co-culture with E. faecalis (P ConclusionsThe AME-producing E. faecalis did not provide a consistent protective effect from aminoglycosides for the Gram-negative pathogens.

The aim of the presented work was to evaluate polar and non-polar extracts and fatty oil, isolated from pumpkin fruit (Cucurbita pepo L.), as a potential food supplements or raw material for cosmetic and pharmaceutical preparations. The extracts were isolated from the pumpkin peel, mesocarp and pulp and fatty oil from the seeds. The highest yield of total extractive matter and highest β-carotene content was determined in peel extracts at concentrations of 5.45 g/100gf.p.m and 136.22 μgg−1f.p.m, respectively. The presence of phenolic acids, flavonoids and carotenoids in polar and non-polar extracts was confirmed by UHPLC-DAD-ESI-MS and HPLC-DAD analysis, respectively. The yield of fatty oil in the seeds was 34.53%, while TLC and HPLC analysis showed the highest percentage of triglycerides (99.3%) and methyl esters (99.7%) in fatty oil and fatty acids methyl esters, respectively. Non-polar extracts of peel showed the highest antioxidant activity estimated by DPPH test (EC50 = 0.041 mgcm−3), fully comparable to activity of synthetic BHT antioxidant (EC50 = 0.021 mgcm−3). High linear correlation factor (0.9969) of EC50 values on β-carotene concentration suggests that β-carotene is the most responsible for significant antioxidant activity of non-polar extracts. Investigated isolates showed no antimicrobial activity against Lt. monocytogenes, St. aureus, E. coli, P. vulgaris, C. albicans, P. aeruginosa, B. cereus, K. pneumoniae and B. luteus under the examined conditions. Bearing in mind the results presented, the C. pepo isolates can be used as an alternative to synthetic antioxidants and a rich source of phytochemicals in food and pharmaceutical industry.

In-vitro antioxidant activity of therapeutically important plant Thymus serpyllum L. grown at different altitudes viz. foothill areas, lower Himalayan and higher Himalayan regions were evaluated against various radicals such as DPPH, ABTS and reducing power assays. The antioxidant constituents like total phenols, flavonoids and total tannins were also evaluated in the present study. The results revealed that the alcoholic, as well as the aqueous extracts of the plant, showed significant antioxidant potential against all the radicals. Extracts obtained from different altitude cultivation showed variation in IC50 values. The alcoholic and aqueous extracts prepared from the plants grown at foothill areas (Haldwani) showed the highest DPPH (IC50: ALC-0.566 mg/ml; AQ- 0.778 mg/ml), ABTS (IC50: ALC- 0.484 mg/ml; AQ- 0.533 mg/ml) and reducing power (EC50: ALC- 0.29 mg/ml; 0.42 mg/ml) activities, followed by high altitude cultivated plants, (Auli), with moderate antioxidant activity. The extracts from mid-altitude cultivation (Pithoragarh) exhibited the least antioxidant potential. The results showed that the amount of total phenolics and flavonoids were significantly correlated to the antioxidant activity. Higher the value of phenolics (TS3: 12.63mg CE/g> TS1: 11.51 mg CE/g> TS2: 10.70 mg CE/g) and flavonoids (TS3: 9.30 mg QE/g> TS1: 9.07mg QE/g> TS2: 6.59mg QE/g) in the extracts, greater was the antioxidant activity. Therefore, T. serpyllum grown in foothill areas was more beneficial in preparing various herbal formulations.

Five new triterpene acids, mesonaic acids A-C (1-3), 2α,3α,19α-trihydroxy-24-nor-4(23),12-oleanadien-28-oic acid (4), and 3α,19α,22α-trihydroxy-2-oxo-12-ursen-28-oic acid (5), and 10 known triterpene acid compounds (6-15) were isolated from a methanolic extract of Mesona procumbens. Triterpenes 1-3 possess unusual hexacyclic skeletons with a 13α,27-cyclopropane ring. Regarding their anti-inflammatory activity, compounds 1-3, 6, and 7 inhibited NO production with EC50 values lower than 15 μM, which were better than that of the positive control quercetin. Compounds 1-3, 6, and 7 markedly decreased levels of the inducible iNOS and COX-2 proteins in macrophages by inhibiting the activation of NF-κB through interference with the MAPK signaling pathway. Based on these data, compounds 1-3, 6, and 7 have great potential as NO inhibitors.

Plants are important sources of chemical compounds with various applications. These are usually associated with the secondary metabolism of plants which is tightly linked to the interactions with the surrounding environment. These compounds usually accumulate in low amounts, sometimes leading to the overexploitation of the source plant. Hence, development of alternative platforms for the production of these compounds represents a more sustainable solution. Among such platforms, hairy roots (HR) cultures are often used for their ability to produce the same secondary metabolites as the original plant. Cynara cardunculus L. or cardoon has been used in folk medicine to treat various diseases. Distinct parts of the plant have been studied and found to produce secondary metabolites with health promoting properties. In this work, HR cultures from cardoon were characterized regarding their secondary metabolites production and potential biological activities. Methanolic extracts contained compounds that belong, almost exclusively, to the hydroxycinnamic acid group. These extracts showed maximum total phenolic content and antioxidant activity of 46 mg GAE/g DW and 98 μmolTE/g DW, respectively, at day 35 of the growth curve. This extract also showed antiproliferative activity on colorectal cancer cells (EC50 1.16 ± 0.07 mg/mL). This work demonstrates the potential of cardoon HR as alternative sources for valuable hydroxycinnamic acid compounds with important biological activities. Hairy root cultures from cardoon produced mainly hydroxycinnamic acid compounds, which showed antioxidant activity and antiproliferative properties on colorectal cancer cell lines.

A novel series of peptidomimetic aldehydes was designed and synthesized to target 3C protease (3Cpro) of enterovirus 71 (EV71). Most of the compounds exhibited high antiviral activity, and among them, compound 18p demonstrated potent enzyme inhibitory activity and broad-spectrum antiviral activity on a panel of enteroviruses and rhinoviruses. The crystal structure of EV71 3Cpro in complex with 18p determined at a resolution of 1.2 Å revealed that 18p covalently linked to the catalytic Cys147 with an aldehyde group. In addition, these compounds also exhibited good inhibitory activity against the 3CLpro and the replication of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), especially compound 18p (IC50 = 0.034 μM, EC50 = 0.29 μM). According to our previous work, these compounds have no reasons for concern regarding acute toxicity. Compared with AG7088, compound 18p also exhibited good pharmacokinetic properties and more potent anticoronavirus activity, making it an excellent lead for further development. ispartof: JOURNAL OF MEDICINAL CHEMISTRY vol:65 issue:4 pages:2794-2808 ispartof: location:United States status: published

In completed research, ferrous gluconate -a food additive- used to preserve black color to prevent discoloration during storage in ripe black olives, and Allium cepa L. species. A. cepa L. roots were treated with different doses of ferrous gluconate. The effective concentration EC50 (0.068 g/l) was determined. A. cepa root tips were treated with EC50/2 (0.034 g/l), EC50 (0.068 g/l), 2XEC50 (0.136 g/l) dose for 24, 48, 72 hours, and afterward, the root tips were prepared for observation under the light microscope according to the method of preparing mitotic preparation. Chromosomal abnormality index (CAI) and genotoxic effect of ferrous gluconate in A. cepa root tip cells were determined. Repeated measurement ANOVA and TUKEY multiple comparison tests were used to investigate the effect of time and dose together on genotoxicity. C-mitosis, polyploidy, polar shifting in anaphase, polar shifting in telophase, equatorial plate shifting, laggard chromosome was observed by microscope. The highest CAI (70.16±4.85) was observed at 72h for 2XEC50 dose. Chromosomal aberration is also observed in control group. While the most common chromosomal aberration is determined as C-mitosis; The least observed chromosomal aberration is determined as polyploidy. Research results revealed that ferrous gluconate has a genotoxic effect on the root tip of A. cepa.

Leishmaniasis and toxoplasmosis are parasitic protozoal diseases that pose serious health concerns, especially for immunocompromised people. Leishmania major and Toxoplasma gondii are endemic in Saudi Arabia and are particularly common in the Qassim Region. The present work was conducted to evaluate the in vitro antileishmanial and antitoxoplasmal activity of methanolic extracts and phytochemical fractions from two plants, Euphorpia retusa and Pulicaria undulata, which are ethnobotanical agents used to treat parasitic infection. Whole E. retusa and P. undulata plants were extracted with methanol and fractionated using petroleum ether, chloroform, ethyl acetate, n-butanol, and water and then were tested in vitro against L. major promastigote and the amastigote stages of T. gondii; the cytotoxicity of the extracts was tested against Vero cell line. The methanolic extracts of E. retusa and P. undulata exhibited promising antitoxoplasmal activity against T. gondii with EC50 values 5.6 and 12.7 μg mL-1, respectively. The chloroform fraction of P. undulata was the most potent, exhibiting an EC50 of 1.4 μg mL-1 and SI value of 12.1. It was also the most active fraction against both L. major promastigotes and amastigotes, exhibiting an EC50 of 3.9 and 3.8 μg mL-1 and SI values 4.4 and 4.5, respectively. The chloroform fraction from P. undulata is a very good candidate for the isolation of active antitoxoplasmal and antileishmanial ingredients; therefore, further phytochemical analysis for active compound isolation is highly recommended.

Exposure of nontarget organisms to therapeutic agents can cause distinct toxic effects, even at low concentrations. Paracetamol is a painkiller drug, widely used in human and veterinary therapies, being frequently found in the aquatic compartment in considerable amounts. Its toxicity has already been established for some species, but its full ecotoxicological potential is still not sufficiently described. To characterize the ecotoxicity of paracetamol, the present study evaluated several parameters, such as acute immobilization (EC50 calculation), biochemical alterations, and behavioral effects, in two species of freshwater microcrustaceans of the genus Daphnia (D. magna and D. longispina). To increase the relevance of the data obtained, animals were exposed to levels of paracetamol similar to those already reported to occur in the wild. Data showed antioxidant responses in both species, namely an increase of catalase and GSTs activities in D. magna. On the contrary, effects of paracetamol on D. longispina included only an impairment of GSTs activity. Despite the absence of anticholinesterasic effects, behavioral modifications were also observed. This set of data indicates that realistic levels of paracetamol may trigger the activation of the antioxidant defense system of freshwater crustaceans, causing changes in behavioral traits (increase in swimming time, but with a reduction in swimming distance) of unknown etiology that are likely to affect normal life traits of wild populations. published

Nanotoxicology is argued and considered one of the emerging topics. In this study, polyaniline (PANI)/2-acrylamido-2-methylpropanesulfonic acid (AMPSA) capped silver nanoparticles (NPs)/graphene oxide (GO) quantum dots (QDs) nanocomposite (PANI/Ag (AMPSA)/GO QDs NC) as a nanoadsorbent has a potential for removal of toxic hexavalent chromium (Cr(VI)) ions from water. The acute toxicity of this NC was evaluated on Artemia salina and freshwater Ostracods (Cypridopsis vidua) larvae for 48 h. The measurements were made at 24 and 48 h with 3 repetitions. The 50% effective concentration (EC50) values of the NC were determined after the exposure of these organisms. According to the results of the optical microscope, it was found that both experimental organisms intake the NC. In the toxicity results of Ostracods, the NC had a highly toxic effect only at 250 mg/L after 48 h and the EC50 value was 157.6 ± 6.4 mg/L. For Artemia salina individuals, it was noted that they were less sensitive than the Ostracods and EC50 value was 476 ± 25.1 mg/L after 48 h. These results indicated that PANI/Ag (AMPSA)/GO QDs NC has low toxicity towards both investigated organisms.

There has been increasing concern over the toxic effects of microplastics (MP), nanoplastics (NP), and copper (Cu) on microalgae. However, the combined toxicity of the metal in the presence of polystyrene (PS) MP/NP on microalgae has not been well studied, particularly after long-term exposure (i.e., longer than 4 days). The primary aim of the present study was to investigate the effect of PS MP and NP on Cu toxicity on two freshwater microalgae, namely Chlorella sp. TJ6-5 and Pseudokirchneriella subcapitata NIES-35 after acute exposure for 4 days and up to 16 days. The results showed that both microalgae were sensitive to Cu, but tolerant to MP/NP. However, MP/NP increased the toxicity of Cu at EC50 in both microalgae, which was only noticeable in chronic exposure. Single and combined treatment of MP/NP and Cu induced higher oxidative stress and caused morphological and ultrastructural changes in both microalgae. The adsorption of Cu to MP and NP was low (0.23-14.9%), with most of the Cu present in free ionic form (81.6-105.8%). The findings on different sensitivity of microalgae to Cu in the presence of MP/NP may have significant implication as microalgae are likely to be exposed to a mixture of both MP/NP and Cu in the environment. For example, in air-blasting technology, MP and NP are used as abrasive medium to remove Cu-containing antifouling paints on hulls of ship and submerged surfaces. Wastewater treatment plants receive household wastes containing MP and NP, as well as stormwater runoffs and industrial wastes contaminated with heavy metals.

Several medicinal plants are used in the steep area of Algeria (Laghouat) for treatment of inflammation and diabetes. Furthermore, Hammada elegans Botsch. (Chenopodiaceae) a xerophytic plant popularly known as (Ajram) is widely spread perennial shrub in Laghouat region and it is traditionally used to treat inflammation and diabete. Then, the objective of this work is to study for the first time the in vivo anti-inflammatory, antidiabetic and acute toxicity effects of acetonic, methanolic and aqueous Hammada elegans Botsch extracts. The acute toxicity test was performed according to the OECD method using single increasing doses (50–1500 mg/kg bw). The anti-inflammatory effect is investigated in Wistar rats by using the rat paw edema assay. The antidiabetic activity was evaluated in vivo using three tests: short-term test (in non-diabetic rats), starch-induced hyperglycemia test (in non-diabetic rats) and long-term alloxan test (experimental diabetes). The acute toxicity results show no deaths in rats and no clinical signs of toxicity. The anti-inflammatory effects showed that all extracts significantly inhibit rat paw edema (EC50 less than 345.51 ± 0.29 mg/kg bw). Therefore, the acetonic extract (EC50 = 157.45 ± 0.33 mg/kg bw) had the more active anti-inflammatory activity than that of the standard inhibitor “Ibuprofen”. In addition, the evaluation of the antidiabetic activities by three tests shows that: in, in the short-term test, there was no important decrease in normal rats glucose rate, while in the starch-induced hyperglycemia test, the aqueous extract decreased significantly hyperglycemia (57.21 ± 1.24 mg AEAC / kg bw) compared to all tested extracts. While in the long-term test, the acetone extract significantly decreased hyperglycemia (9.18 ± 0.72 mg GEAC / kg bw) compared to all the tested extracts. Hammada elegans Botsch extracts seem to have therapeutic opportunities for the treatment of the inflammation and diabetes.

Copper (Cu) mining operations have a global footprint and have led to contamination of soils and Cu toxicity to plants. Understanding the controls of plant Cu uptake, including competition with other metals, such as zinc (Zn), is essential for improving plant growth in Cu-contaminated soils. The objective of this study was to evaluate the capacity of Zn to alleviate the toxicity of Cu in crops grown in Cu-contaminated soils. Lettuce was grown in 27 soils with ranges of total Cu and Zn concentrations of 82–1295 mg Cu kg−1 and 86–345 mg Zn kg−1 for a period of 21 days. Oat was grown in 21 soils with the same total Cu and Zn concentration ranges for a period of 62 days. Regression analyses were used to evaluate the impact of total soil Zn on plant growth in the Cu-contaminated soils. We show for the first time that Zn alleviates Cu toxicity in lettuce and oat grown in soils. Specifically, we observed a negative (toxicity) effect of total soil Cu and a positive (protective) effect of total soil Zn on shoot growth response for lettuce and oat. The effective concentration 50% (EC50) of Cu/Zn mass ratio was 7.0 ± 1.8 for lettuce shoot length and 5.9 ± 1.0 for oat shoot weight. These results indicate that the previously demonstrated efficacy of Zn in mitigating Cu phytotoxicity in hydroponic systems can extend to more complex soil systems. Further research should be done to evaluate specific Zn amendments for restoring vegetative growth in Cu-contaminated soils.

Background: Schistosomiasis has been identified as a major public health problem in tropical countries. The present study aimed to investigate the schistosomicidal effects of the methanolic extract of Argemone mexicana L. and its active component, berberine against Schistosoma mansoni on in-vitro experiments. Methods: S. mansoni adults were used. Various concentrations of the methanolic extract (10 - 200 µg/ml) and berberine (2.5 - 50 µM) were tested from 24 to 72 h. The viability of S. mansoni was confirmed with an invertoscope-microscope. Furthermore, cytotoxic (Hemolysis test), and antioxidant (DPPH radical scavenging assay) capacities were determined. Results: The viability tests on S. mansoni showed that A. mexicana at 50 μg/mL is lethal at 48 h and berberine at 10 μM is lethal at 24 h. The hemolytic activity at 1,000 μg/mL was 2.9% for A. mexicana and 90.2% for berberine. The antioxidant capacities shown by A. mexicana and berberine, were EC50 156.3 and 84.1 μg/mL, respectively. Conclusion: The extract of A. mexicana and berberine demonstrated high antischistosomal activities in low concentration and short exposure time on the in-vitro model.

Background Chikungunya fever is an endemic disease caused by the Chikungunya virus (CHIKV) to which there is no vaccine or effective antiviral drug treatment so far. Our study aimed to evaluate the potential anti-CHIKV activity of memantine hydrochloride (mtnH), a drug from the class of the aminoadamantanes approved for the treatment of Alzheimer´s disease, as a possible drug to be repurposed to the treatment of Chikungunya fever. Methods MtnH antiviral activity against CHIKV was determined by infecting BHK-21 cells with CHIKV-nanoluc, a virus carrying the marker nanoluciferase reporter, in the presence or absence of mtnH at concentrations ranging from 500 to 1.45 µM. The effective concentration of 50% inhibition (EC50) was calculated. Cell viability assay (determination of CC50) was also performed employing BHK-21 cells. Mutagenic assays were performed by the Salmonella Typhimurium/microsome assay (Ames test). Results MtnH presented a CC50 of 248.4 ± 31.9 µM and an EC50 of 32.4 ± 4 µM against CHIKV in vitro. The calculated selectivity index (SI) was 7.67. MtnH did not induce genetic mutation in Salmonella strains with or without an external metabolizing system. Conclusion With the data herein presented, it is possible to hypothesize mtnH as a viable candidate to be repurposed as an anti-CHIKV drug. Clinical assays are, therefore, encouraged due to the promising in vitro results. Graphic abstract The drug memantine hydrochloride is herein personified with a doubt: as a prior regulated drug against Alzheimer, could it follow the path against Chikungunya virus too?

The cell line RTgill-W1 was evaluated as an in vitro alternative model for acute fish whole-effluent toxicity (WET) testing. We determined the 50% effective concentration (EC50) that reduces the viability of RTgill-W1 cells for selected toxicants commonly found in effluent samples and correlated those values with the respective 50% lethal concentration (LC50) of freshwater (fathead minnow, Pimephales promelas) and marine (sheepshead minnow, Cyprinodon variegatus) fish species obtained from the literature. Excluding low water-soluble organics and the volatile sodium hypochlorite, significant correlations were measured for metal, metalloids, ammonia, and higher water-soluble organics between in vitro EC50 values and in vivo LC50 values for both species. Typically, toxicity studies with RTgill-W1 cells are conducted by adding salts to the exposure medium, which may affect the bioavailability of toxicants. Osmotic tolerance of RTgill-W1 cells was found between 150 and 450 mOsm/kg, which were set as the hypoosmotic and hyperosmotic limits. A subset of the toxicants were then reexamined in hypoosmotic and hyperosmotic media. Copper toxicity decreased in hyperosmotic medium, and nickel toxicity increased in hypoosmotic and hyperosmotic media. Linear alkylbenzene sulfonate toxicity was not affected by the medium osmolality. Overall, RTgill-W1 cells have shown potential for applications in measuring metal, metalloids, ammonia, and water-soluble organic chemicals in acute WET tests, as well as complementing current toxicity identification and reduction evaluation strategies. In the present study, RTgill-W1 cells have been established as a valid animal alternative for WET testing, and we show that through manipulation of medium osmotic ranges, sensitivity to nickel was enhanced. Environ Toxicol Chem 2021;40:1050-1061. © 2020 SETAC.

Anthracnose, caused by Colletotrichum spp., is the most significant field and post-harvest disease of mango worldwide and is mainly controlled through the use of systemic fungicides belonging to the methyl benzimidazole carbamate (MBC) class. In Mexico, this disease in mango has been associated with at least seven cryptic species belonging to the C. gloeosporioides complex. In this study, the distribution of sensitivity to MBC (benomyl and carbendazim) and prochloraz fungicides of 101 Colletotrichum spp. isolates obtained from 101 commercial orchards in Mexico’s most important mango-producing states was determined. All Colletotrichum spp. isolates were obtained from commercial mango orchards with applications of MBC fungicides and that had not been exposed to applications of prochloraz, or any other DMI (demethylation inhibitors) class fungicide. Isolates were evaluated using an in vitro test to determine the effective fungicide concentration at which 50% of mycelial growth is inhibited (EC50) for each isolate-fungicide combination. EC50 values for benomyl and carbendazim ranged from 0.0967 to 0.3995 µg mL−1 (median = 0.1923 µg mL−1) and from 0.0880 to 0.1703 µg mL−1 (median = 0.1171 µg mL−1), respectively, while those for prochloraz ranged from 0.0066 to 0.0813 µg mL−1 (median = 0.0253 µg mL−1). Our results revealed a wide distribution in Mexico of Colletotrichum spp. isolates from mango sensitive to MBC fungicides and prochloraz. Data obtained in this study will serve as a source of comparison of the evolution of the sensitivity of Colletotrichum spp. populations obtained from mango to MBC and DMI fungicides in the future.

To develop unique small-molecule inhibitors of hepatitis C virus (HCV), thiophen urea (TU) derivatives were synthesised and screened for HCV entry inhibitory activities. Among them, seven TU compounds exhibited portent anti-viral activities against genotypes 1/2 (EC50 < 30 nM) and subsequently, they were further investigated; based on the pharmacological, metabolic, pharmacokinetic, and safety profiles, J2H-1701 was selected as the optimised lead compound as an HCV entry inhibitor. J2H-1701 possesses effective multi-genotypic antiviral activity. The docking results suggested the potential interaction of J2H-1701 with the HCV E2 glycoprotein. These results suggest that J2H-1701 can be a potential candidate drug for the development of HCV entry inhibitors.

3D spheroid cultures are attractive candidates for application in in vitro drug-induced hepatotoxicity testing models to improve the reliability of biological information obtainable from a simple 2D culture model. Various 3D spheroid culture models exist for hepatotoxicity screening, but quantitative assays of spheroid response in situ are still challenging to achieve with the current 3D liver toxicity platforms. In this study, we developed a 3D printing-based HepG2 liver spheroid culture model for in situ quantitative evaluation and high-content monitoring of drug-induced hepatotoxicity. HepG2 liver spheroids grown in mini-fabricated hydrogel constructs using a 3D bioprinter were used to obtain the EC50 values and to measure the multi-parametric hepatotoxic effects, including mitochondrial permeability transition (MPT), cytosolic calcium levels, and apoptosis. Interestingly, the average fluorescence intensities of apoptotic and cell death markers, calculated for out-of-focus and in-focus spheroids, increased proportionally as a function of the drug concentration, allowing for the determination of the EC50 values. In addition, 3D HepG2 spheroids were more resistant to nefazodone-induced MPT than 2D HepG2 cells, indicating that the gelatin/alginate hydrogel culture system provides enhanced resistance to hepatotoxic drugs. The drug response of HepG2 liver spheroids was also found to be unrelated to the spheroid size. These results demonstrate that the present 3D cell-printing-based embedded HepG2 liver spheroid platform is a promising approach for screening and characterizing drug-induced hepatotoxicity.

The current study evaluated the antioxidant, α-amylase inhibitory and anti-Staphylococcus aureus activities of the fruiting bodies of Ganoderma lucidum. Methanolic extract of G. lucidum (MEGL) was investigated for its in vitro antioxidant activity using 1,1-diphenyl 2-picrylhydrazyl (DPPH) assay and EC50 value was determined. α-Amylase inhibitory and anti-S. aureus activities were carried out by conventional methods. The mushroom extract showed maximum free radical scavenging activity at a dose of 500 μg/mL with EC50 value of 22.1399 μg/mL. EC50 value for the inhibition of α-amylase activity was found 22.6011 μg/mL. G. lucidum extracts exhibited a concentration-dependent Anti-S. aureus activity. The vast differences of this inhibition between lower and higher doses were assessed significantly (p G. lucidum has moderate antioxidant, α-amylase inhibitory, and anti-S. aureus activities. Thus this mushroom may be the alternative plant source for the antioxidant, anti-diabetic and antibacterial agents and further in vivo studies and isolation of bioactive compounds are required.

Somatostatin receptor-4 (SST(4)) is highly expressed in brain regions affiliated with learning and memory. SST(4) agonist treatment may act to mitigate Alzheimer's disease (AD) pathology. An integrated approach to SST(4) agonist lead optimization is presented herein. High affinity and selective agonists with biological efficacy were identified through iterative cycles of a structure-based design strategy encompassing computational methods, chemistry, and preclinical pharmacology. 1,2,4-Triazole derivatives of our previously reported hit (4) showed enhanced SST(4) binding affinity, activity, and selectivity. Thirty-five compounds showed low nanomolar range SST(4) binding affinity, 12 having a K(i) < 1 nM. These compounds showed >500-fold affinity for SST(4) as compared to SST(2A). SST(4) activities were consistent with the respective SST(4) binding affinities (EC(50) < 10 nM for 34 compounds). Compound 208 (SST(4)K(i) = 0.7 nM; EC(50) = 2.5 nM; >600-fold selectivity over SST(2A)) display a favorable physiochemical profile, and was advanced to learning and memory behavior evaluations in the senescence accelerated mouse-prone 8 model of AD-related cognitive decline. Chronic administration enhanced learning with i.p. dosing (1 mg kg(−1)) compared to vehicle. Chronic administration enhanced memory with both i.p. (0.01, 0.1, 1 mg kg(−1)) and oral (0.01, 10 mg kg(−1)) dosing compared to vehicle. This study identified a novel series of SST(4) agonists with high affinity, selectivity, and biological activity that may be useful in the treatment of AD.

Purpose: To evaluate the in vivo anti-inflammatory effect and in vitro antioxidant activity of the areal part of Sedum sediforme (Jacq.) extracts.Methods: The plant was extracted with solvents of varying polarity (Methanol, chloroform and ethyl acetate, respectively) allowed its separation into three sub-fractions: crude extract, chloroform extract and ethyl acetate extract (CrE, ChE and EaE, respectively). Total polyphenol contents of the extracts were determined. The phorbolmyristate acetate (PMA) induced mice ear edema method was using to evaluate the anti-inflammatory activity. The in vitro scavenging activity was evaluated using enzymatic and non-enzymatic methods. Chelating ability of extracts was assessed using Fe2+–ferrozine complex.Results: The highest content of phenolics compound was in EaE. The administration of CrE (12.5 and 25 mg/kg) reduced ear edema induced by PMA (%I = 35.81 ± 3.18 % and 38.57 ± 2.80 %, respectively), the effect was comparable with that of diclofenac used as a reference drug (%I = 38.84 ± 1.87 %). The in vitro scavenging activity of S. sediforme extracts confirmed that the CrE has the highest enzymatic and non-enzymatic activity with a half maximal inhibitory concentration (IC50) = 0.063 ± 0.005 mg/mL and 0.178 ± 0.006 mg/mL, respectively. However, the ChE present an excellent chelating activity with a half-maximal effective concentration (EC50) of 0.397 ± 0.001 mg/mL.Conclusion: The results show that S. sediforme extracts have a strong antioxidant and antiinflammatory activities which lend some support their use in the traditional medicine. Keywords: Sedum sediforme, Anti-inflammatory, Superoxide scavenger, Cytochrome C, Metal chelating

As Alzheimer disease (AD) is a multifactorial condition, it should be tackled with drugs targeting multiple key pathways. A series of aliphatic (2–8) and aromatic (9–15) edaravone derivatives were synthesized, characterized, and evaluated as antioxidant agents using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2-2’-azino-bis-(3-ethylbenzothiazoline-6-sulfonate) (ABTS·+) assays, as well as acetylcholinesterase (AChE) inhibitors. In both antioxidant assays, even though the starting compound edaravone was more active, the best derivative was 5 with 50% effective concentration (EC50) of 0.0301 and 0.8106 mM respectively, followed by 3 (EC50 of 0.1920 mM and 3.5311 mM). In the AChE inhibition assay, the derivatives were not as active as the positive control galantamine, but a general better activity was shown from the aromatic compounds. The best results were for 10, with 41.9% of inhibition (concentration of 150 μg/mL), and 9 with 31.6%. Docking analysis of compound 10 showed hydrogen bonds with residues Ser200 and His440 in the AChE catalytic gorge. All synthesized derivatives 2–15 presented drug-like properties and are capable of crossing the blood–brain barrier and not be pumped out of it. These results indicate edaravone derivatives can function as scaffolds for AD drugs, though further derivatizations should be conducted to improve their antioxidant and AChE inhibition profiles.

Antibiotics are recently recognized as a group of emerging environmental contaminants that are frequently detected in various environmental matrixes. Relative root elongation (RRE) test is a rapid and effective strategy to evaluate the water/soil quality and the toxic effects of environmental contaminants on plants. In the present study, we examine the toxicity effect of ciprofloxacin (CIP), norfloxacin (NOR), and tetracycline (TET) to pakchoi individually and in combinations. Both independent action (IA) and concentration addition (CA) models are used for toxicity assessment. Results showed that the EC50 values of CIP, NOR, and TET are 193.59, 60.81, and 40.37 μM, respectively. Combinations of TET + CIP and TET + NOR caused more inhibitory effects on root elongation than those of CIP + NOR. Toxic Unit (TU) and Synergistic Ratio (SR) analysis showed that the relatively lower (higher) EC values are observed in the combinations with lower (higher) antibiotic concentrations, suggesting an effect of low-dose synergism and high-dose antagonism. The reliability of the simulation results from IA and CA models to predict that combined toxicity is highly dependent upon the results from the analysis of TU or SR.

The extract from leaves of Genipa americana L., Rubiaceae, has antichagasic effect. The major components of this extract, arabinogalactan-glycoconjugate fractions (PFI and PFII), have shown similar biological effects of the crude extract, becoming interesting to evaluate the antichagasic effect of PFI and PFII. This work aimed to access the trypanocidal effect of PFI and PFII against the benznidazole-resistant Trypanosoma cruzi Y strain and to investigate the mechanism of action. PFI and PFII were evaluated on epimastigote (24, 48, 72 h), trypomastigote (24 h), and amastigote (24 and 48 h) forms. The cytotoxicity on LLC-MK2 cells was made by MTT assay. Flow cytometry analysis with PE-annexin V (Ax), 7-AAD, and DCFH-DA staining and scanning electron microscopy (SEM) were performed in epimastigote forms for evaluation of the cell death pathway. PFI showed effect against epimastigote forms (EC50/24 h = 580 ± 0.17 μg/ml; EC50/48 h = 530 ± 0.13 μg/ml; EC50/72 h = 500 ± 0.14 μg/ml), while PFII did not show effect on any tested concentrations. In trypomastigotes, the PFI and PFII showed effect with EC50 values of 100 ± 0.09 and 23 ± 0.06 μg/ml respectively. PFI and PFII were also able to decrease amastigotes/100 cells parameter. PFI and PFII were not cytotoxic in LLC-MK2 cells at the highest tested concentration, resulting in selectivity index (SI) higher than 15 for PFI and higher than 65 for PFII. The increase in labeling of 7-AAD and DCFH-DA fluorescence on cytometry flow assays suggested necrosis as the cell death pathway of PFI in epimastigotes of T. cruzi, being confirmed by pores in the membrane observed on SEM. PFI and PFII are partially responsible by the trypanocidal effect of the crude extract of G. americana, with PFI showing the same action mechanism of crude extract and PFII being the most selective and potent fraction, presenting the biotechnological potential for antichagasic natural products.

The application of pesticides to get more agricultural products is increasing day by day. The use of a huge amount of pesticides raises public concerns about safety. Tetraconazole is a widely used and successful fungicide. Possible toxic, cytotoxic, and genotoxic effects of different doses of tetraconazole (1.00 mg/L, 5.00 mg/L, and 10.00 mg/L) were evaluated on the meristematic cells of Allium cepa L. root tips by means of physiological, cytogenetic, biochemical, and anatomical parameters. EC50 value for tetraconazole in terms of growth inhibition was calculated as 6.7 mg/L. Increasing doses of tetraconazole resulted in reduced germination ratio, root length, and weight gain. Total activities of superoxide dismutase (SOD) and catalase (CAT) enzymes as well as malondialdehyde (MDA) content were increased as a result of oxidative stress. As an evidence of genotoxicity, mitotic index (MI) level decreased, while scores for micronucleus (MN) and chromosomal aberrations (CAs) rose. In addition, various meristematic cell damages were detected in root tips of tetraconazole applied bulbs. As a result, the multiple toxic, cytotoxic, and genotoxic effects of tetraconazole fungicide were demonstrated through a wide range of parameters on A. cepa, which was found to be a versatile tool for testing hazardous pesticides.

The aim of this work was to evaluate the antioxidant and antibacterial activities of two marine algae, Cystoseira mediterranea and Padina pavonica, extracts. Total phenols (TPC), carotenoids, and phlorotannins contents of the extracts obtained by four extraction solvents were determined and compared. The highest TPC content was observed for aqueous extract of C. mediterranea with 37.09±0.46 mg GAE/g DE followed by ethanol extract of P. pavonica (24.28 ±0.99 mg GAE/g DE), which showed the highest phlorotannins content (1.18±0.18 mg PE/g DE), while its methanol extract held carotenoids content of 66.96 ±4.78 μg g–1 DE. Ethanol extract of C. mediterranea exhibited the best antioxidant activity with an EC50 of 58.3 ±1.16 μg ml . The antibacterial activity screening against MRSA and E. coli showed that ethanol extract of C. mediterranea towards a Methicillin resistant Staphyloccocus aureus (20.33±0.28 mm) and E. coli (15.66±0.57 mm) was more efficient with MICs about 80 mg ml–1 and 20 mg ml–1, respectively. Ethanol extract of C. mediterranea seems to have the highest potential for use in food industries.

[Image: see text] Bitter taste receptors (TAS2Rs) are recognized as being expressed on multiple cell types and organs, including human airway smooth muscle (HASM) cells, where agonists promote significant relaxation to constrictive stimuli. Thus, the HASM TAS2Rs have been targeted as novel bronchodilators for the treatment of asthma and other obstructive lung diseases. The TAS2R5 subtype, a dominant receptor on HASM, has few known agonists, all with reported low potency and efficacy. We screened multiple compounds by measuring [Ca(2+)](i) release in HASM (a consequence of receptor–G protein coupling) to establish structure–activity relationships and arrive at a potent agonist for TAS2R5. HASM physiological studies using magnetic twisting cytometry confirmed the relaxation effects of lead compounds. 1,10-Phenanthroline-5,6-dione had the greatest potency (EC(50) ≈ 120 nM), amounting to a >1000-fold improvement over the other compounds, and displayed maximal efficacy. These studies revealed critical structural requirements for favorable potencies and efficacies for a potential first-in-class bronchodilator targeting TAS2R5 of the airway.