Median Effective Concentration Research Articles

Pickering emulsion-enhanced Vibrio fischeri assay for ecotoxicity assessment of highly hydrophobic polycyclic aromatic hydrocarbons

Accurate ecotoxicity assessment of contaminated soil is critical to public health, and the luminescent bacteria (Vibrio fischeri) method is the most commonly used. Hydrophobic compounds such as polycyclic aromatic hydrocarbons (PAHs) in soil cannot be in contact with luminescent bacteria due to their low water solubility so that the luminescence inhibitory effect cannot be observed. The underestimated biological toxicity makes the test unreliable and en-dangers public health and safety. The commonly adopted improved method of adding cosolvents has limited effect, it was only effective for low-hydrophobicity chemicals and could not be used for ecotoxicity evaluation of high-hydrophobicity chemicals. Therefore, we constructed Pickering emulsions using luminescent bacteria modified with n-dodecanol in which PAHs were dissolved in the oil phase, n-tetradecane. Then the luminescent bacteria could tightly adhere to the oil-water interface and contact PAHs. As a result, their bioluminescence was suppressed to varying degrees depending on the chemical species and concentrations. With no solubility limitation, highly hydrophobic PAHs could even completely inhibit bacterial bioluminescence, hence the toxicity information was accurately displayed and the median effect concentration (EC50) values could be calculated. This Pickering emulsion-based method was successfully applied for the accurate ecotoxicity evaluation of highly hydrophobic PAHs and soil samples contaminated with them, which all previous methods could not achieve. This method overcomes the problem of ecotoxicity evaluation of hydrophobic compounds, and has great potential for practical application, whether it is pure chemicals or various real samples from the ecological environment.

Sensitivity of Krasnodar Venturia inaequalis Populations to the Sterol Demethylation Inhibitor Difenoconazole.

Difenoconazole sensitivity was assessed in three populations of the apple scab agent Venturia inaequalis of the Krasnodar region. One of the populations was fungicide naïve population; its sample was collected in natural habitats of the eastern crabapple Malus orientalis in foothills of the Severskii raion. The two other populations were from commercial orchards of the apple variety Reinette Simirenko (the Krasnoarmeiskii and Dinskoi raions), where fungicide treatments with various agents, including those with difenoconazole as an active ingredient, were performed on a regular basis. Single-spore V. inaequalis isolates were isolated from fresh leaves with signs of the disease or fallen leaves with fungal fruiting bodies. The median effective concentration (EC50) was defined as the concentration that halves the colony growth and was estimated in a series of in vitro experiments with 120 isolates. Difenoconazole (Score EC, 250 mg/L, Syngenta) was used at six concentrations: 0.005, 0.01, 0.025, 0.05, 0.5, and 1 mg a.i./L. Mean EC50 values were 0.0078mg a.i./L in the natural population and 0.12 and 0.25 mg a.i./L in the orchard populations. Fungicide sensitivity was lower in both of the orchard populations; their resistance factors were estimated at 16 and 32. The proportion of sensitive and low-sensitive isolates differed between the two orchard populations. A discriminatory dose, or single-assessment concentration (SAC), of 0.01 mg a.i./L was proposed to simplify the laboratory monitoring of the difenoconazole sensitivity in V. inaequalis by using a test for relative growth (RG) of the mycelium. Comparable results were obtained with EC50 and RG at the discriminatory dose. The portion of isolates with RGs exceeding the threshold (RG > 70%) was 97% in one of the orchard populations. The results indicate that difenoconazole resistance develops in V. inaequalis populations from commercial orchards of the Krasnodar region.

The median effective concentration of propofol in combination with different doses of esketamine during gastrointestinal endoscopy in adults

We designed a four-arm randomized controlled trial to investigate the median effective concentration (EC50) of propofol in combination with different doses of esketamine inducing appropriate depth of anaesthesia during gastrointestinal endoscopy in adults. One hundred patients aged 18-65years planning for gastrointestinal endoscopy were divided into four groups randomly: esketamine 0, 0.15, 0.25 and 0.5mg/kg groups (n = 25). Propofol doses followed the Dixon and Massey up-and-down method with different starting between groups. The primary endpoint was the EC50 of propofol. Secondary outcomes included the cumulative dose of propofol, the duration of the procedure, recovery time, and adverse effects. The EC50 (median, 95% confidence interval) of propofol was significantly less in the esketamine 0.5mg/kg group compared with the esketamine 0, 0.15, and 0.25mg/kg groups [1.34 (1.15, 1.54) vs. 3.48 (3.25, 3.71), 2.82 (2.58, 3.07), and 2.36 (2.11, 2.61), respectively; p < 0.001]. The total dose of propofol (mean ± SD) required for the whole procedure was significantly less in the esketamine 0.5mg/kg group compared with the esketamine 0, 0.15, and 0.25mg/kg groups [95.5 ± 43.1 vs. 277.4 ± 49.0, 207.8 ± 31.6, and 135.1 ± 27.7, respectively; p < 0.001]. The recovery time was significantly longer in esketamine 0 and 0.5mg/kg group compared with other two groups (p < 0.001). More patients in the esketamine 0.5mg/kg group experienced visual disturbance compared with the other groups (p = 0.016). Additionally, the incidence of hypotensionin the esketamine 0mg/kg group after inducation was higher compared with other groups (p < 0.001). In summary, the administration of esketamine significantly and dose-dependently reduced the dose of propofol required to accomplish procedures.

In Vitro Assessment of Berberine against Ichthyophthirius multifiliis in Goldfish.

Ichthyophthirius multifiliis is a pathogenic ciliate parasite, which infects almost all freshwater teleost fish and leads to significant economic losses. The present study aimed to evaluate the acute toxicity of berberine to the free-living stages of I. multifiliis, that is, theronts and tomonts. Our results indicated that 99.30% of I. multifiliis theronts were killed by a concentration of 15 mg/L berberine during the 4 h exposure time, while berberine had no effect on protomonts. Nevertheless, berberine at a concentration of 5 mg/L could effectively reduce the release of theronts from tomonts treated for 4 h. Additionally, according to the transmission electron microscopy results, berberine at 15 mg/L could strongly change the shape of protomonts, destroy their organelles, and significantly decrease the number of ribosomes. The median lethal concentration (LC50) of berberine for goldfish at 96 h was 528.44 mg/L, which was almost 67 times the median effective concentration (EC50) of berberine for killing theronts (7.86 mg/L). The results demonstrated that berberine could be an effective and safe potential parasiticide for killing I. multifiliis.

Novel 1,3,4-oxadiazole sulfonate/carboxylate flavonoid derivatives: synthesis and biological activity.

With the long-term use of traditional bactericides and antiviral agents, drug resistance has become increasingly prominent, resulting in impaired crop growth and yields. Based on this, the introduction of small molecular active groups into natural products has become the direction of research for green pesticides. In this study, novel 1,3,4-oxadiazole sulfonate/carboxylate flavonoid derivatives were explored. Among them, D4 exhibited good inhibitory effects on plant bacteria. It is worth mentioning that D4 (15 μg ml-1 ) exhibited an excellent median effective concentration (EC50 ) value against Xanthomonas oryzae pv. oryzae (Xoo), which was better than bismerthiazol (73 μg ml-1 ) and thiodiazole copper (100 μg ml-1 ). The EC50 for D4 was much lower than the two positive controls (bismerthiazol, thiodiazole copper), making D4 more potent in this assay of bacterial growth inhibition. In addition, mechanism research using scanning electron microscopy revealed that D4 could cause deformation or rupture of the cell membranes of Xoo and Pseudomonas syringae pv. actinidiae. Moreover, D4 exhibited the best EC50 for in vivo curative (132 μg ml-1 ) and protective (101 μg ml-1 ) activities against tobacco mosaic virus, which were more effective than ningnanmycin. Microscale thermophoresis data suggested that D4 [dissociation constant(Kd ) = 0.038 ± 0.011 μmol L-1 ] exhibited a stronger binding capacity than the control agent ningnanmycin (Kd = 4.707 ± 2.176 μmol L-1 ). The biological activity data and mode of action demonstrated that D4 had the best antibacterial and antiviral effects. Compound D4 discovered in the current work may be a very promising agricultural drug. © 2022 Society of Chemical Industry.

Interaction of the Fungal Metabolite Harzianic Acid with Rare-Earth Cations (Pr3+, Eu3+, Ho3+, Tm3+).

Rare-earth elements (REEs) are in all respect a class of new contaminants that may have toxic effects on organisms and microorganisms and information on their interactions with natural ligands should be of value to predict and control their diffusion in natural environments. In the current study, we investigate interactions of tripositive cations of praseodymium, europium, holmium, and thulium with harzianic acid (H2L), a secondary metabolite produced by selected strains of fungi belonging to the Trichoderma genus. We applied the same techniques and workflow previously employed in an analogous study concerning lanthanum, neodymium, samarium, and gadolinium tripositive cations. Therefore, in the current study, HPLC-ESI-HRMS experiments, circular dichroism (CD), and UV-Vis spectrophotometric absorption data, as well as accurate pH measurements, were applied to characterize bonding interactions between harzianic acid and Pr3+, Eu3+, Ho3+, and Tm3+ cations. Problems connected to the low solubility of harzianic acid in water were overcome by employing a 0.1 M NaClO4/(CH3OH + H2O 50/50 w/w) mixed solvent. For Pr3+, Ho3+, and Tm3+, only the mono complexes PrL+, HoL+, and TmL+ were detected and their formation constant determined. Eu3+ forms almost exclusively the bis complex for which the corresponding formation constant is reported; under our experimental conditions, the mono complex EuL+ is irrelevant. Combining the results of the present and previous studies, a picture of interactions of harzianic acid with rare-earth cations extending over 8 of the 17 REEs can be composed. In order to complement chemical information with toxicological information, a battery of bioassays was applied to evaluate the effects of praseodymium, europium, holmium, and thulium tripositive cations on a suite of bioindicators including Aliivibrio fischeri (Gram-negative bacterium), Raphidocelis subcapitata (green alga), and Daphnia magna (microcrustacean), and median effective concentration (EC50) values of Pr3+, Eu3+, Ho3+, and Tm3+ for the tested species were assessed.

Novel botanical active component derivatives containing carboxamide and 1,3,4-Thiadiazole thioether moieties: Design, synthesis, and inhibitory activity.

In this study, using the botanical active components of carvacrol, thymol, guaiacol, and sesamol as the lead structures, 19 novel botanical active component derivatives containing carboxamide and 1,3,4-thiadiazole thioether moieties (5a−5s) were synthesized and structurally characterized by 1H NMR, 13C NMR, and HRMS. The antibacterial bioassay results in vitro showed that compound 2-(2-methoxyphenoxy)-N-(5-(methylthio)-1,3,4-thiadiazol-2-yl)acetamide (5k) revealed excellent inhibitory activities against Xanthomonas axonopodis pv. citri (Xac) and Xanthomonas oryzae pv. oryzicolaby (Xoc), with the median effective concentration (EC50) values of 22 and 15 μg/ml, respectively, which were even better than those of thiodiazole copper and bismerthiazol. Meanwhile, all the target compounds revealed lower in vitro inhibitory effects on Mucor bainieri (M. bainieri), Mucor fragilis (M. fragilis), and Trichoderma atroviride (T. atroviride), than carbendazim.

Toxicity of tyre wear particle leachates to marine phytoplankton

Tyre wear particles (TWP) are some of the dominant sources of microplastics in the aquatic environment. Once TWP enter aquatic systems, they can leach certain plastic additives that can be potentially toxic to biota. However, little is known about the impact of TWP lixiviates on marine phytoplankton, the base of marine food webs. This study aims to determine the acute toxic effect of leachates derived from TWP on three phytoplankton species: the cryptophyte Rhodomonas salina, the diatom Thalassiosira weissflogii and the dinoflagellate Heterocapsa steinii, using the median effect concentration (EC50) for specific growth rate as endpoint. Leachates were obtained by incubating 1 g L−1 of < 250 µm TWP in artificial seawater for 3 days. Each phytoplankton species was exposed to leachates at five different concentrations, and cell concentrations were measured every 24 h over 3 days. Leachates from TWP were toxic to marine phytoplankton. The dinoflagellate H. steinii was the most sensitive species, with 72-h EC50 of 23% leachate concentration, whereas R. salina and T. weissflogii exhibited EC50 values of 64% and 73%, respectively. Our results suggest that TWP leachates have a negative effect on phytoplankton growth, although more field data on the concentration of TWPs and their leachates is needed to fully evaluate the environmental impact of TWP.

A Bioavailability Model to Predict the Impact of pH and Dissolved Organic Carbon on Ciprofloxacin Ecotoxicity to the Cyanobacterium Microcystis aeruginosa.

Ciprofloxacin (CIP) is a pseudopersistent antibiotic detected in freshwater worldwide. As an ionizable chemical, its fate in freshwater is influenced by water chemistry factors such as pH, hardness, and dissolved organic carbon (DOC) content. We investigated the effect of pH, DOC, and Ca2+ levels on the toxicity of CIP to Microcystis aeruginosa and developed a bioavailability model on the basis of these experimental results. We found that the zwitterion (CIP+/- ) is the most bioavailable species of CIP to M. aeruginosa, whereas DOC is the most dominant factor reducing CIP toxicity, possibly via binding of both CIP+/- and CIP+ to DOC. pH likely also regulates CIP-DOC binding indirectly through its influence on CIP speciation. In addition, higher tolerance to CIP by M. aeruginosa was observed at pH < 7.2, but the underlying mechanism is yet unclear. Calcium was identified as an insignificant factor in CIP bioavailability. When parameterized with the data obtained from toxicity experiments, our bioavailability model is able to provide accurate predictions of CIP toxicity because the observed and predicted total median effective concentrations deviated by <28% from each other. Our model predicts that changes in pH and DOC conditions can affect CIP toxicity by up to 10-fold, suggesting that CIP in many natural environments is likely less toxic than in standard laboratory toxicity experiments. Environ Toxicol Chem 2022;41:2835-2847. © 2022 SETAC.

Epigallocatechin Gallate Stabilized by Cyclodextrin Inactivates Influenza Virus and Human Coronavirus 229E.

Natural products are attractive antiviral agents because they are environment-friendly and mostly harmless. Epigallocatechin gallate (EGCg), a type of catechin, is a well-known natural antiviral agent that can inhibit various viruses. However, EGCg easily oxidizes and loses its physiological activity. Although this problem can be overcome by combining EGCg with cyclodextrin (CD-EGCg), which makes it stable in water at high concentrations, the antiviral effect of this compound remains unclear. Here, we show that in Madin–Darby canine kidney (MDCK) and MRC-5 cells, CD-EGCg is cytotoxic for 50% of cells at 85.61 and 65.34 ppm, respectively. Furthermore, CD-EGCg mainly shows its antiviral effect during the adsorption step for all four influenza virus strains (median effect concentration (EC50) was 0.93 to 2.78 ppm). Its antiviral effect post-adsorption is less intense, and no inhibitory effect is observed on influenza viruses pre-adsorption. Moreover, human coronavirus 229E (HCoV-229E) was inhibited at the adsorption step in short contact (EC50 = 2.5 ppm) and long contact conditions (EC50 = 0.5 ppm) by mixing CD-EGCg with HCoV-229E. These results suggest that CD-EGCg effectively inhibits various viruses that require an adsorption step, and is an effective tool for preventing infection.

Synthesis of Hinokitiol Sulfonate Derivatives and Their Anti-Oomycete and Nematicidal Activities.

In order to explore novel natural product-based anti-oomycete and nematicidal agents, sixteen unreported 2-sulfonyloxyhinokitiol derivatives were prepared using the principle of active splicing, and structurally confirmed by proton nuclear magnetic resonance (1 H-NMR), carbon-13 nuclear magnetic resonance (13 C-NMR), high-resolution mass spectrometry (HRMS), and melting point. Moreover, we evaluated the title compounds as anti-oomycete and nematicidal agents against two serious agricultural pests of Phytophthora capsici and Meloidogyne incongnita. Among the sixteen hinokitiol esters tested: (1) Compounds 3a and 3m exhibited the most potent anti-oomycete activity compared to zoxamide against P. capsici, and the median effective concentration (EC50 ) values of 3a, 3m, and zoxamide were 18.64, 21.11, and 23.15 mg/L, respectively; Further studies showed that the existence of seven membered ring and carbonyl group was the necessary condition for the high anti-oomycete activity of hinokitiol. (2) Compounds 3n and 3p exhibited more promising nematicidal activity than hinokitiol, and the median lethal concentration (LC50 ) values of 3n, 3p and 1 against M. incongnita were 0.2111, 0.2079, and 0.3933 mg/L, respectively. This result will pave the way for further modification of hinokitiol to develop potential new fungicides and nematicides.

A first attempt to evaluate the toxicity to Phaeodactylum tricornutum Bohlin exposed to rare earth elements

The increasing use and demand of rare earth elements in many emerging technologies is leading to a potentially higher input to the marine environment. This study compared for the first time the effect of lanthanum (La), cerium (Ce), neodymium (Nd), samarium (Sm), europium (Eu), gadolinium (Gd), dysprosium (Dy), and erbium (Er) to the microalga Phaeodactylum tricornutum Bohlin. The algal growth inhibition was investigated after 72 h of exposure. The median effect concentrations (EC50) ranged from 0.98 mg/L to 13.21 mg/L and elements were ranked as follows: Gd &amp;gt; Ce &amp;gt; Er &amp;gt; La &amp;gt; Eu &amp;gt; Nd &amp;gt; Dy &amp;gt; Sm. The comparison of predicted no effect concentrations (PNEC) for hazard and risk assessment with measured environmental concentrations showed that ecological risks deriving from REEs could be present, but limited to specific environments like estuarine waters. The results support evidence of actions to manage the REE impact in seawater environments, looking to improve the monitoring tailored to the different and dynamic nature of ecosystems.

Collembola are Among the Most Pesticide-Sensitive Soil Fauna Groups: A Meta-Analysis.

Pesticides are a major concern because of their deleterious impacts on biodiversity and on the ecological functions provided by living organisms. Although earthworms are well studied, smaller-sized organisms, such as Collembola, also contribute to the agroecosystem functioning, and their sensitivity to pesticides makes them good bioindicators of soil quality. Using data from 21 publications, we performed a meta-analysis to compare the pesticide sensitivity of Collembola with other soil invertebrate groups and discuss the relevance of including tests on representatives of this microarthropods group in European regulation tests. We defined a paired observation as the median lethal concentration or the median effect concentration values for both Collembola species and another soil fauna group (Acari, enchytraeids, earthworms, isopods, and nematodes) under a unique combination of author, year, substance, and type of soil (61 and 57 paired observations for reproduction and lethal effects). In some studies, paired comparisons were available for several groups of soil fauna. We demonstrated that Collembola are among the most sensitive soil fauna groups to a variety of pesticides, notably for effects on reproduction, mostly compared with earthworms and enchytraeids. Because there are several modes of exposure and explaining factors, we suggest moving from a single-species study to a food-chain approach integrating different taxonomic groups. Differences between soil fauna groups in sensitivity or response to pesticides could have effects on soil communities and also on soil functions. Environ Toxicol Chem 2022;41:2333-2341. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

The median effective analgesic concentration of ropivacaine in ultrasound-guided interscalene brachial plexus block after arthroscopic rotator cuff repair.

Background: The median effective analgesic concentration (MEAC) of ropivacaine in interscalene brachial plexus block (ISBPB) for postoperative analgesia after arthroscopic rotator cuff repair (ARCR) has not been determined. Therefore, this study aimed to evaluate the MEAC after ARCR using 10 ml ropivacaine. Method: This study was conducted on 40 patients with American Society of Anesthesiologists grade I or II who had selective ARCR. The 10 ml ropivacaine was administered for determined, with an initial concentration of 0.3% using up-and-down sequential allocation. After successful or unsuccessful postoperative analgesia, the concentration of ropivacaine was decreased or increased by 0.05% in the next patient. We defined successful postoperative analgesia as a visual analog scale score of <4 at rest within the initial 8 h after ISBPB. The analytic techniques of linear, linear-logarithmic, exponential regressions and centered isotonic regression were used for calculating MEAC. The secondary outcomes was sufentanil consumption, time to 1st rescue analgesic, onset time of sensory block and motor block. Results: The concentration of ropivacaine administered ranged from 0.1% to 0.35%. The MEAC from the four different methods (linear, linear-logarithmic, exponential regressions and centered isotonic regression) were 0.207% (95% CI, 0.168–0.355%), 0.182% (95% CI, 0.165–0.353%), 0.196% (95% CI, 0.154–0.356%), and 0.163%, respectively. Of the four models, exponential regression had the least residual standard error (0.0990). Conclusion: The MEAC derived from the four statistical models for 10 ml ropivacaine in ultrasound-guided ISBPB for postoperative analgesia was distributed within a narrow range of 0.163%–0.207%. The exponential regression model calculated by the goodness-of-fit test at a concentration of 0.196% best fits the study data. Clinical Trial Registration: http://www.chictr.org.cn/showproj.aspx?proj=127449, identifier ChiCTR2100047978

Synthesis and antifungal activity evaluation of novel pyridine derivatives as potential succinate dehydrogenase inhibitors

Two series of novel pyridine derivatives (1a−l, 2a−l) were synthesized and characterized by IR, 1H NMR, 13C NMR, and HRMS. The crystal structures of compounds 1 h and 2l were characterized by single crystal X-ray diffraction and crystallized in the monoclinic and orthorhombic system, respectively. The in vitro antifungal activity of synthesized compounds were determined against five plant pathogenic fungi namely Gibberella zeae, Helminthosporium maydis, Rhizoctonia solani, Botrytis cinerea, and Sclerotinia sclerotiorum and most of the target compounds revealed significant antifungal activity at 20 mg/L. Of these, the inhibitory rate and the median effect concentrations (EC50) of compounds 1 m, 2i, 2o, and 2p were more close to fluopyram against B. cinerea. Meanwhile, the half inhibitory concentrations (IC50) of compounds 1 m, 2i, 2o, and 2p against succinate dehydrogenase (SDH) and their score in molecular docking were also more close to fluopyram, indicating the stronger antifungal activities of the four compounds were well associated with the affinities of these compounds in interacting with SDH. Therefore, we concluded that compounds 1 m, 2i, 2o, and 2p might be potential succinate dehydrogenase inhibitors (SDHIs), which was been reported for the first time.

In Vitro and in Vivo Studies of the Antiparasitic Effect of Aspirin Against Dactylogyrus Extensus (Monogenea) Invasion in Carp (Cyprinus carpio)

Abstract Aspirin is one of the most widely used medications globally as an analgesic and antipyretic drug. Aspirin use as an antiparasitic against fish parasites has never been tested. The parasite Dactylogyrus is considered a serious threat to freshwater aquaculture in relation to considerable losses. The present study is aimed to evaluate the antiparasitic effect of aspirin against Dactylogyrus extensus on Cyprinus carpio and alteration in gill histopathology. In vitro, aspirin exhibited antiparasitic activity with median effective concentration (EC50) values at one and six minutes of 8.137 g/L−1 and 1.629 g/L−1, were assessed for the first time. In vivo antiparasitic efficacy of aspirin to D. extensus was found to be 46.49%. Severe lesion profile was observed in histopathological evaluations of the gills.

Design, synthesis, and biological evaluation of novel berberine derivatives against phytopathogenic fungi.

The abuse of chemical fungicides not only leads to toxic residues and resistance in plant pathogenic fungi, but also causes environmental pollution and side effects on in humans and animals. Based on the antifungal activities of berberine, seven different types of berberine derivatives (A1-G1) were synthesized, and their antifungal activities against six plant pathogenic fungi were evaluated (Rhizoctonia solani, Botrytis cinerea, Fusarium graminearum, Phytophthora capsici, Sclerotinia sclerotiorum, and Magnaporthe oryzae). The results for antifungal activities in vitro showed that berberine derivative E1 displayed good antifungal activity against R. solani with a median effective concentration (EC50 ) of 1.77 μg ml-1 , and berberine derivatives F1 and G1 demonstrated broad-spectrum antifungal activities with EC50 values ranging from 4.43 to 42.23 μg ml-1 against six plant pathogenic fungi. Berberine derivatives (E2-E29, F2-F18, and G2-G9) were further synthesized to investigate the structure-activity relationship (SAR), and compound E20 displayed significant antifungal activity against R. solani with an EC50 value of 0.065 μg ml-1 . Preliminary mechanism studies showed that E20 could cause mycelial shrinkage, cell membrane damage, mitochondrial abnormalities and the accumulation of harmful reactive oxygen species, resulting in cell death in R. solani. Moreover, in vivo experimental results showed that the protective effect of E20 was 97.31% at 5μg ml-1 , which was better than that of the positive control thifluzamide (50.13% at 5μg ml-1 ). Berberine derivative E20 merits further development as a new drug candidate with selective and excellent antifungal activity against R. solani. © 2022 Society of Chemical Industry.

Functional analysis reveals ionotropic GABA receptor subunit RDL is a target site of ivermectin and fluralaner in the yellow fever mosquito, Aedes aegypti.

Ionotropic γ-aminobutyric acid (iGABA) receptors are involved in various physiological activities in insects, including sleep, olfactory memory, movement, and resistance to viruses. Ivermectin and fluralaner can disturb the insect nervous system by binding to iGABA receptors, and are therefore an effective means for controlling insect pests. However, the molecular mechanisms underlying the insecticidal effect of both the compounds on Aedes. aegypti remain unexplored. In this study, we investigated the spatiotemporal expression profile of Ae. aegypti RDL (Ae-RDL), a subunit of iGABA receptor. RDL dsRNA suppressed the expression of Ae-RDL mRNA in Ae. aegypti larvae and adult by 60% and 50.67%, resepectly. However, the physiology of Ae. aegypti larvae was not significantly affected. The mortality of Ae. aegypti larvae and adult females subjected to Ae-RDL knockdown significantly decreased after exposure to ivermectin and fluralaner. Additionally, Ae-RDL was cloned into Xenopus laevis oocytes and characterized using the two-electrode voltage-clamp method. The inward current was induced by GABA binding to the functional Ae-RDL homomeric receptors at a median effective concentration (EC50 ) of 100.4 ± 59.95 μM (n > 3). The significant inhibitory effect of ivermectin and fluralaner on inward current indicated that both insecticides exerted a significant antagonistic effect on Ae-RDL. However, ivermectin also showed strong agonistic as well as weak activation effects on Ae-RDL. These contrasting effects of ivermectin on Ae-RDL depended on ivermectin concentration. Our study revealed that Ae-RDL subunit is a target of ivermectin and fluralaner, providing new insights into the insecticidal mechanism of both compounds in Ae. aegypti. © 2022 Society of Chemical Industry.

Developmental Toxic Effects of Thiram on Developing Zebrafish (Danio rerio) Embryos.

Thiram, an oxidized dimer of dithiocarbamate, has fungicidal and ectoparasiticidal roles. This study aimed to determine the effects of thiram on the development of zebrafish (ZF) embryos. The developmental toxicity test was performed in accordance with the OECD 236 test guidelines, and ZF embryos were subjected to several thiram concentrations and a DMSO (0.01%) control. Subsequently, embryo mortalities and developmental anomalies were evaluated at different hours post fertilization (hpf). Thiram was highly toxic to ZF, with calculated median lethal concentrations (LC50) of thiram at 48 and 96 h as 13.10 ± 2.17 and 8.87 ± 2.09 μg/L, respectively. Thiram-treated embryos/larvae exhibited a variety of deformities, such as abnormal somites, reduced eye pigment, abnormal tail shape, yolk sac edema, hatching defects, and curved spines, with a median effective concentration (EC50) of 3.88 ± 1.23, 5.04 ± 1.82, 6.23 ± 0.92, 5.24 ± 2.22, 1.39 ± 0.25, and 2.60 ± 0.82 μg/L, respectively. Teratogenic index (TI) values ranged from 1.42 to 6.66 for the scored deformities. At 48 hpf, the average heartbeat of the control group was 177.20 ± 5.63 per minute, while the highest thiram-treated group (40 μg/L) was 99.50 ± 18.12 per minute. In addition, cardiac-related issues, such as pericardial edema and abnormal blood flow, were observed in thiram-treated ZF embryos. Overall, these findings suggest that thiram is teratogenic to ZF.

Therapeutic Efficacy of Arnica in Hamsters with Cutaneous Leishmaniasis Caused by Leishmania braziliensis and L. tropica

Leishmaniasis may occur in three different clinical forms, namely, visceral, mucocutaneous and cutaneous, which are caused by different species of trypanosomatid protozoans of the genus Leishmania. Pentavalent antimonials are the leading treatment for cutaneous leishmaniasis despite the hepatic, renal, and cardiac toxicity. In addition, the response of some Leishmania species to pentavalent antimonials is increasingly poorer, and therefore new and more potent therapeutic alternatives are needed. Arnica montana L., Asteraceae, is a traditional medicinal plant of Europe and preparations of its flowers are commonly used externally to treat disorders of the musculoskeletal system as well as superficial inflammatory conditions. Previous studies have shown that Arnica tincture (AT), an ethanolic extract prepared from the flowerheads of Arnica montana as well as isolated Arnica sesquiterpene lactones (STLs) have antileishmanial activity in vitro against L. donovani and L. infantum, as well as in vivo against L. braziliensis. In this work, we studied the in vitro cytotoxicity and antileishmanial activity of AT and STLs against both L. braziliensis and L. tropica. The in vivo therapeutic effect of AT was studied in hamsters with cutaneous Leishmaniasis (CL) caused by experimental infection with L. braziliensis and L. tropica. Furthermore, various semisolid Arnica preparations were also evaluated against L. braziliensis. The STLs and the AT possess a very high in vitro activity against both Leishmania species with median effective concentrations (EC50) ranging from 1.9 to 5.9 μg/mL. The AT was not cytotoxic for human tissue macrophages, skin fibroblasts, and hepatic cells. The therapeutic response of hamsters infected with L. braziliensis to the topical treatment with AT was 87.5% at a dose of 19.2 μg STL/2× day/60 d, 72.7% at doses of 19.2 μg STL/1× d/60 d and 67% at a dose of 38.4 μg STL/2× d/60 d. In turn, the therapeutic response in hamsters infected with L. tropica was 100% when treated at a dose of 19.2 μg STL/2× day/60 d and 71% at a dose of 38.4 μg STL/2× d/60 d. On the other hand, the effectiveness of treatment with glucantime administered intralesionally at a dose of 200 mg/every three days for 30 days was 62.5% for L. braziliensis and 37.5% for L. tropica infection. These results are promising and encourage the implementation of clinical trials with AT in CL patients as a first step to using AT as a drug against CL.