Myo-Inositol Limits Kainic Acid-Induced Epileptogenesis in Rats.

Epilepsy is a severe neurological disease characterized by spontaneous recurrent seizures (SRS). A complex pathophysiological process referred to as epileptogenesis transforms a normal brain into an epileptic one. Prevention of epileptogenesis is a subject of intensive research. Currently, there are no clinically approved drugs that can act as preventive medication. Our previous studies have revealed highly promising antiepileptogenic properties of a compound-myo-inositol (MI) and the present research broadens previous results and demonstrates the long-term disease-modifying effect of this drug, as well as the amelioration of cognitive comorbidities. For the first time, we show that long-term treatment with MI: (i) decreases the frequency and duration of electrographic SRS in the hippocampus; (ii) has an ameliorating effect on spatial learning and memory deficit associated with epileptogenesis, and (iii) attenuates cell loss in the hippocampus. MI treatment also alters the expression of the glial fibrillary acidic protein, LRRC8A subunit of volume-regulated anion channels, and protein tyrosine phosphatase receptor type R, all expected to counteract the epileptogenesis. All these effects are still present even 4 weeks after MI treatment ceased. This suggests that MI may exert multiple actions on various epileptogenesis-associated changes in the brain and, therefore, could be considered as a candidate target for prevention of epileptogenesis.

Meta-Analysis of the Field Efficacy of Seed- and Soil-Applied Nematicides on Meloidogyne incognita and Rotylenchulus reniformis Across the U.S. Cotton Belt.

Meta-analysis was used to compare yield protection and nematode suppression provided by two seed-applied and two soil-applied nematicides against Meloidogyne incognita and Rotylenchulus reniformis on cotton across 3 years and several trial locations in the U.S. Cotton Belt. Nematicides consisted of thiodicarb- and fluopyram-treated seed, aldicarb and fluopyram applied in furrow, and combinations of the seed treatments and soil-applied fluopyram. The nematicides had no effect on nematode reproduction or root infection but had a significant impact on seed cotton yield response ([Formula: see text]), with an average increase of 176 and 197 kg/ha relative to the nontreated control in M. incognita and R. reniformis infested fields, respectively. However, because of significant variation in yield protection and nematode suppression by nematicides, five or six moderator variables (cultivar resistance [M. incognita only], nematode infestation level, nematicide treatment, application method, trial location, and growing season) were used depending on nematode species. In M. incognita-infested fields, greater yield protection was observed with nematicides applied in furrow and with seed-applied + in-furrow than with solo seed-applied nematicide applications. Most notable of these in-furrow nematicides were aldicarb and fluopyram (>131 g/ha) with or without a seed-applied nematicide compared with thiodicarb. In R. reniformis-infested fields, moderator variables provided no further explanation of the variation in yield response produced by nematicides. Furthermore, moderator variables provided little explanation of the variation in nematode suppression by nematicides in M. incognita- and R. reniformis-infested fields. The limited explanation by the moderator variables on the field efficacy of nematicides in M. incognita- and R. reniformis-infested fields demonstrates the difficulty of managing these pathogens with nonfumigant nematicides across the U.S. Cotton Belt.

Levamisole, a cocaine cutting agent, induces acute and subchronic systemic alterations in Wistar rats.

The use of the anthelmintic levamisole as a cocaine adulterant has been increasing worldwide. Complications caused by this association include systemic vasculitis, agranulocytosis, neutropenia, tissue necrosis, pulmonary hemorrhage, and renal injury. Data about toxicity of levamisole are scarce, therefore the aim of this study was to evaluate the acute and subchronic toxic effects of levamisole in rats. Male Wistar rats received saline or levamisole by intraperitoneal route at the doses of 12, 24 and 36 mg/kg in the acute toxicity test; and at 3, 6 and 12 mg/kg in the subchronic toxicity test. Toxicity was evaluated using behavioral, cognitive, renal, hematological, biochemical and histopathological parameters. Acute administration of levamisole caused behavioral and histopathological alterations. Subchronic administration caused behavioral, cognitive and hematological alterations (p < 0.0001 and p < 0.05, respectively), impairment of liver and kidney functions (p < 0.05), and changes of antioxidant defenses (p ≤ 0.0001). Both administrations produced toxic effects of clinical relevance, which make levamisole a dangerous cutting agent. Furthermore, the knowledge of these effects can contribute to the correct diagnosis and treatment of cocaine dependents with unusual systemic alterations.

N-alkyl isatin derivatives: Synthesis, nematicidal evaluation and protein target identifications for their mode of action.

Meloidogyne incognita is an economically dominant pathogen infesting a wide range of crops curbing their growth and productivity. Deregistration of frontline nematicides has necessitated exploration of innovative and novel class of structurally diverse nematicides with streamlined activity. In this context, N-alkylated derivatives of isatin known for their remarkable biological profile were synthesized, characterized and evaluated in vitro for their antinemic character followed by in silico studies for their mode of action and toxicological studies for their fitness as agrochemical. The antinemic evaluation was carried by egg hatch inhibition and juvenile mortality and its effect on egg hatching. Compounds 1 and 2a exhibited nematicidal potential and significantly decreased egg hatching and increased juvenile mortality. For egg hatch inhibition LC50 and LC95 values for 1 were found to be 0.125 and 1.380 mg/ml and for compound 2a, 0.457 and 8.511 mg/ml respectively. For juvenile mortality LC50 and LC95 values for 1 were found to be 0.109 and 0.776 mg/ml and for 2a, 0.190 and 1.380 mg/ml respectively. For insights into the mode of action of the synthesized molecules, in silico studies for the targeted effects were conducted which revealed novel interaction with pathogenic protein - Aspartyl protease. Computational studies on the drug-ability and potential toxicity of the selected compounds revealed they belonged to class IV and are safe. With good reasons, our compounds hold value for their exploration in agrochemical industry and thus, this study identifies a new scaffold with useful level of nematicidal activity for its use in agriculture industry.

Evaluation of Nematicidal Activity of Streptomyces yatensis KRA-28 against Meloidogyne incognita.

The root-knot nematode (Meloidogyne incognita) is an important pathogen in crop cultivation, however, few methods are available to control this parasitic roundworm. In this study, the nematicidal effects of approximately 30 Streptomyces strains isolated from soil samples of Mt. Naejang (Korea) were tested against Meloidogyne incognita, and the culture broth of the strains KRA- 24 and KRA-28 exhibited approximately 75% and 85% insecticidal activity, respectively, in in vitro assays. In in vivo pot experiments, these strains reduced the number of nematodes in the soil and the number of egg masses in the roots of red peppers. The two strains also survived in the presence of insecticidal agents (0.1 to 3.0%) such as fosthiazate, ethoprophos and terbufos when they were used in parallel. The mixture of KRA-24 or KRA-28 culture broth and fosthiazate exhibited nematicidal effects that were similar to those observed when KRA-24 or KRA-28 were used alone. Our results clearly suggest that the Streptomyces strains KRA-24 and KRA-28 should be promoted as a biocontrol agent against Meloidogyne incognita.

Up and down-regulation of mRNA in the cytotoxicity and genotoxicity of Plumbagin in HepG2/C3A.

Studies that evaluated the mechanisms of action of Plumbagin (PLB) and its toxicity may contribute to future therapeutic applications of this compound. We investigate biomarker important in the mechanisms of action correlate the expression of mRNA with the cytotoxic and genotoxic effects of PLB on HepG2/C3A. In the analysis of cytotoxicity, PLB decreased cell viability and membrane integrity at concentrations ≥ 15µM. Xenobiotic-metabolizing system showed strong mRNA induction of CYP1A1, CYP1A2, and CYP3A4, suggesting extensive metabolization. PLB induced apoptosis and an increase in the mRNA expression of genes BBC3, CASP3, and CASP8. At a concentration of 15µM, there was a reduction in the expression of PARP1 mRNA and an increase in the expression of BECN1 mRNA, suggesting that PLB may also induce cell death by autophagy. PLB induced an arrest at the G2/M phase due to DNA damage, as observed in the comet assay. This damage is associated with the increased mRNA expression of genes p21, GADD45A, and H2AFX and with changes in the expression of proteins H2AX, p21, p53, Chk1, and Chk2. These results allow a better understanding of the cellular action of PLB and of its toxicity, thereby contributing to the development of PLB-based drugs, with markers of mRNA expression possibly playing a role as indicators for monitoring toxicity in human cells.

Toxicities of 4,5-Dihydroisoxazoles Against Root-Knot Nematodes and in Silico Studies of Their Modes of Action.

The present work sought to contribute to the development of new nematicides. Benzaldehydes were initially converted to nitrile oxides that underwent 1,3-dipolar cycloaddition reactions with methyl acrylate to generate 4,5-dihydroisoxazoles. In in vitro tests, methyl 3-phenyl-4,5-dihydroisoxazole-5-carboxylate (1) and methyl 3-(4-chlorophenyl)-4,5-dihydroisoxazole-5-carboxylate (4) increased the mortality of Meloidogyne exigua and Meloidogyne incognita second-stage juveniles (J2). Compounds 1 and 4 presented necessary concentrations of 398 and 501 µg mL-1, respectively, to kill 50% of M. incognita J2 (LC50 values), while the value for carbofuran (positive control) was 168 µg mL-1. In in vivo tests, compounds 1 and 4 reduced the number of M. incognita galls in tomato roots by 70 and 40%, respectively, and the number of eggs by 89 and 44%. Using an in silico approach, we showed that compounds 1 and 4 were toxic to the nematodes by binding to the allosteric binding sites of the agonist-binding domains of the nematode nicotinic acetylcholine receptors. These results opened up possibilities for further investigations aimed at developing novel commercial nematicides.

Dimethyl disulfide (DMDS) as an effective soil fumigant against nematodes in China.

Root-knot nematode is an important soil pest in horticulture crops and constrains the protected cultivation development after methyl bromide (MB) was phased out in China. Dimethyl disulfide (DMDS) exhibits excellent efficacy against nematodes. Laboratory experiments and field trials were set up to clarify DMDS dose, efficacy, and yield. A dose-response experiment using three methods showed that DMDS presented high efficacy against the nematode Meloidogyne incongnita. The LC50 values of direct fumigation activity in the dessicator method were 0.086 and 0.070 mg L-1 for DMDS and 1,3-D, 29.865 and 18.851 mg L-1 for DMDS and 1,3-D of direct contact activity in the small tube method, 6.438 and 3.061 mg L-1 for DMDS and 1,3-D of soil fumigation activity in the soil fumigation method, respectively. The field trials indicated that DMDS showed an excellent efficacy of 80%-94% on root-knot nematode applied at 10-100 g m-2 on tomato in Tongzhou, Beijing. The crop yields showed no significant difference after applying 10-80 g m-2 DMDS. Results indicate that DMDS applied at 10 g m-2 for controlling root-knot nematode in Beijing is cost effective. In conclusion, DMDS is an excellent soil fumigant that can be used for controlling root-knot nematode and can be an potential novel alternative to MB in China.

The bioactivity of new chitin oligosaccharide dithiocarbamate derivatives evaluated against nematode disease (Meloidogyne incognita).

Plant-parasitic nematodes cause substantial crop losses annually; however, current nematicides are environmentally unfriendly and highly toxic to nontarget organisms. The development of green efficient nematicides from multifunctional natural bioactive substances such as chitin oligosaccharide (COS) is promising. In this paper, COS dithiocarbamate derivatives (COSDTC, COSDTA, COSDTB) were synthesized to increase nematicidal activity (against Meloidogyne incognita), and their structures were characterized by FTIR, NMR, TGA/DTG and elemental analysis. Furthermore, the nematicidal activities, egg hatching inhibitory activities, plant growth adjustment abilities, cytotoxicity and phytotoxicity of the derivatives were evaluated. The primary mechanism was assessed by heavy metal ion absorption and GSH-binding assays. The results showed COS dithiocarbamate derivatives could possess multiple efficacies, including high nematicidal activities and egg hatching inhibitory activities, plant growth regulating effects, low cell toxicities and phytotoxicities. Additionally, it was inferred that nematicidal activity may be correlated with GSH-binding activity but not heavy metal ion complexation. COS modification has immense potential for controlling plant-parasitic nematodes.

Fenbendazole induces apoptosis of porcine uterine luminal epithelial and trophoblast cells during early pregnancy.

Fenbendazole, is an effective benzimidazole anthelmintic that prevents parasite infection in both human and veterinary health care. Although the well-known and effect of benzimidazole was recently shown to have a broad spectrum of biological abilities, such as anticancer and anti-inflammation activities, the mechanism of benzimidazole's antiproliferative effect via cell signaling pathways and its role in preimplantation has not been studied. Therefore, the purpose of this study was to determine the effects of fenbendazole on porcine trophectoderm and luminal epithelial cells. First, we investigated cell viability in response to a low dose of fenbendazole, which highly inhibited cell proliferation. In addition, we investigated apoptotic molecules in the mitochondria, imbalanced intracellular calcium homeostasis, and the expression of some genes involved in apoptosis to explain the decrease in proliferation. Finally, we examined the intracellular mechanisms of fenbendazole by measuring the extracellular signal-regulated kinase, PI3K/AKT, and c-Jun N-terminal kinase signaling proteins by western blot analysis. Our findings suggest that fenbendazole functions as an effective anti-proliferative molecule that induces critical apoptosis in the porcine trophectoderm and uterine luminal epithelial cells by disrupting the mitochondria membrane potential during early pregnancy.

In vitro efficacies of solubility-improved mebendazole derivatives against Echinococcus multilocularis.

Recently, we introduced an epoxy group to mebendazole by a reaction with epichlorohydrin and obtained two isoforms, mebendazole C1 (M-C1) and mebendazole C2 (M-C2). The in vitro effects of mebendazole derivatives at different concentrations on Echinococcus multilocularis protoscoleces and metacestodes as well as cytotoxicity in rat hepatoma (RH) cells were examined. The results demonstrated that the solubility of the two derivatives was greatly improved compared to mebendazole. The mortality of protoscoleces in vitro reached to 70-80% after 7 days of exposure to mebendazole or M-C2, and M-C2 showed higher parasiticidal effects than mebendazole (P > 0.05). The parasiticidal effect of M-C1 was low, even at a concentration of 30 µm. The percentage of damaged metacestodes that were treated with mebendazole and M-C2 in vitro at different concentrations were similar, and M-C1 exhibited insignificant effects on metacestodes. Significant morphological changes on protoscoleces and metacestodes were observed after treatment with mebendazole and M-C2. In addition, the introduction of an epoxy group to mebendazole also reduced its cytotoxicity in RH cells. Our results demonstrate that the introduction of an epoxy group not only improved the solubility of mebendazole, but also increased its parasiticidal effects on E. multilocularis and reduced its cytotoxicity in RH cells.

Assessment of the effects of oxamyl on the bacterial community of an agricultural soil exhibiting enhanced biodegradation.

Modern agricultural practices largely rely on pesticides to protect crops against various pests and to ensure high yields. Following their application to crops a large amount of pesticides ends up in soil where they may affect non-target organisms, among which microorganisms. We assessed the effects of the carbamate nematicide oxamyl on the whole bacterial diversity of an agricultural soil exhibiting enhanced biodegradation of oxamyl through 16S rRNA amplicon next generation sequencing (NGS) and on the oxamyl-degrading bacterial community through cehA q-PCR analysis and 14C-oxamyl mineralization assays. Oxamyl was rapidly mineralized by the indigenous microorganisms reaching >70% within a month. Concomitantly, a significant increase in the number of oxamyl-degrading microorganisms was observed. NGS analysis of the total (DNA) and active (RNA) bacterial community showed no changes in α-diversity indices in response to oxamyl exposure. Analysis of the ß-diversity revealed significant changes in the composition of the soil bacterial community after 13 and 30 days of oxamyl exposure only when the active fraction of the bacterial community was considered. These changes were associated with seven OTUs related to Proteobacteria (5), Acidobacteria (1) and Actinobacteria (1). The relative abundance of the dominant bacterial phyla were not affected by oxamyl, except of Bacteroidetes and Gemmatimonadetes which decreased after 13 and 30 days of oxamyl exposure respectively. To conclude, oxamyl induced changes in the abundance of oxamyl-degrading microorganisms and on the diversity of the soil bacterial community. The latter became evident only upon RNA-based NGS analysis emphasizing the utility of such approaches when the effects of pesticides on the soil microbial community are explored.

In vitro nematicidal activity of natural and semisynthetic cadinenes from Heterotheca inuloides against the plant-parasitic nematode Nacobbus aberrans (Tylenchida: Pratylenchidae).

BACKGROUND: Nacobbus aberrans (Tylenchida: Pratylenchidae) is one of the main plant-parasitic nematodes species that affects crops in Mexico, generating substantial economic losses. Traditionally, the control of the nematodes is carried out using chemical products; however, research efforts are presently focused on the search for new methods for the control of this pest. Natural products derived from plants are an alternative for the control of populations of plant-parasitic nematodes. The genus Heterotheca (Asteraceae) is characterized by containing sesquiterpenes with cadinane skeleton, and some species of this genus exert nematicidal activity. RESULTS: We determined the effects of selected Heterotheca inuloides plant metabolites and some semisynthetic derivatives on the hatching of eggs isolated from the gelatinous matrix and infective second-stage juveniles (J2) of the false root-knot nematode N. aberrans using an in vitro experimental model. Among the evaluated compounds, nematodes were more susceptible to hydroxylated and quinone compounds, whereas the remaining compounds showed moderate or no activity. The presence of the hydroxyl group is essential for nematicidal potential, with changes at the hydroxyl group modifying the nematicidal activity. CONCLUSION: Flowers of Heterotheca inuloides contain bioactive compounds that showed nematicidal activity against N. aberrans. Here we report the nematicidal activities of cadinenes isolated from the flowers of H. inuloides and their semisynthetic derivatives against the false root-knot nematode N. aberrans. © 2018 Society of Chemical Industry.

Flubendazole induces mitotic catastrophe and apoptosis in melanoma cells.

Flubendazole (FLU) is a widely used anthelmintic drug belonging to benzimidazole group. Recently, several studies have been published demonstrating its potential to inhibit growth of various tumor cells including those derived from colorectal cancer, breast cancer or leukemia via several mechanisms. In the present study we have investigated cytotoxic effects of FLU on malignant melanoma using A-375, BOWES and RPMI-7951 cell lines representing diverse melanoma molecular types. In all three cell lines, FLU inhibited cell growth and proliferation and disrupted microtubule structure and function which was accompanied by dramatic changes in cellular morphology. In addition, FLU-treated cells accumulated at the G2/M phase of cell cycle and displayed the features of mitotic catastrophe characterized by formation of giant cells with multiple nuclei, abnormal spindles and subsequent apoptotic demise. Although this endpoint was observed in all treated melanoma lines, our analyses showed different activated biochemical signaling in particular cells, thus suggesting a promising treatment potential of FLU in malignant melanoma warranting its further testing.

Effects of the plant volatile trans­2-hexenal on the dispersal ability, nutrient metabolism and enzymatic activities of Bursaphelenchus xylophilus.

Bursaphelenchus xylophilus causes pine wilt disease (PWD), which severely damages pine species. The plant volatile trans­2-hexenal has strong activity against nematodes, although the precise mechanism of this inhibitory action remains unclear. In this paper, the fumigant effects of the LC10 and LC30 of trans­2-hexenal on B. xylophilus were demonstrated. The trans­2-hexenal treatments significantly inhibited the dispersal ability of nematodes. The results also indicated that trans­2-hexenal affects the metabolism of nutrients and the activity of digestive enzymes. Among detoxifying enzymes, after treatment with trans­2-hexenal, glutathione S-transferase activity increased significantly and general esterase activity decreased significantly. Based on these results, trans­2-hexenal disturbs the normal physiological and biochemical activities of this nematode. These results provide valuable insight into the nematicidal mechanisms of trans­2-hexenal.

Assessing Psychological Toxicity and Patient-Reported Distress as the Sixth Vital Sign in Cancer Care and Clinical Trials.

As the number of available cancer therapies continues to grow, there is increasing interest in their impact on cancer patients' lived experiences. Screening for distress is one way to measure psychological dimensions of cancer patients' experiences, and doing so is increasingly part of standard operations at major cancer centers across the US. To date, however, most clinical trials have not adequately captured patients' experiences as part of their outcome assessments, so clinicians lack data needed to guide their responses to psychological features of patients' illness experiences. As distress becomes the "sixth vital sign" in routine cancer care, we argue that clinical trials should assess patients' experiences in the same way that they robustly screen for adverse events and toxicities. New interventions are needed to address distress.

Toxicity of the bionematicide 1,4-naphthoquinone on non-target soil organisms.

The main goal of the present study was to evaluate the ecotoxicological effects of 1,4-naphthoquinone (1,4-NTQ), a natural-origin compound presenting nematicidal activity, that can be obtained from walnut husk, in plants and soil invertebrates, including non-target soil nematode communities. This research was part of an ongoing project that aims to develop environmentally-friendly nematicides obtained from agricultural residues. The battery of ISO tests included emergence and growth of corn (Zea mays) and rape (Brassica napus); avoidance with the earthworm Eisenia andrei and the collembolan Folsomia candida; and reproduction with the previous species plus the enchytraeid Enchytraeus crypticus. A novel soil nematode community assay was also performed. ISO tests and nematode assays were conducted using a natural uncontaminated soil that was spiked with a range of 1,4-NTQ concentrations. Toxicity of 1,4-NTQ was found for all test-species and the most sensitive were F. candida and E. andrei. After 7 days of exposure to 1,4-NTQ, nematode abundance decreased along the concentration gradient, and a partial recovery was observed after 14 days (1,4-NTQ <48 mg kg-1 soil). The number of nematode families consistently decreased in both periods. Overall, results indicate that a 1,4-NTQ concentration of <20 mg kg-1 could be environmentally safe but preliminary data suggest that it might be ineffective for the target-nematodes, root-knot nematodes, Meloidogyne spp., and root-lesion nematodes, Pratylenchus spp. In addition, if higher dosages of 1,4-NTQ bionematicide are necessary, the potential recovery of non-target organisms under real field scenarios also needs to be assessed.

Mixture toxicity of flubendazole and fenbendazole to Daphnia magna.

Nowadays, residual amounts of many pharmaceuticals can be found in various environmental compartments including surface and ground waters, soils and sediments as well as biota. Even though they undergo degradability, their environmental discharge is relatively continuous, thus they may be regarded as quasi-persistent contaminants, and are also frequently regarded as emerging organic pollutants. Benzimidazoles, especially flubendazole (FLU) and fenbendazole (FEN), represent two anthelmintic drugs belonging to this group. Although their presence in environmental matrices has been reported, there is relatively little data concerning their (eco)toxicological impact. Furthermore, no data is available on their mixture toxicity. FLU and FEN have been found to have a strong impact on an environmentally important non-target organism - Daphnia magna. Moreover, these compounds are usually present in the environment as a part of pharmaceutical mixtures. Therefore, there is a need to evaluate their mixture toxicity, which was the main aim of this study. Single substance toxicity tests were carried out in parallel with mixture studies of FLU and FEN, with the application of two well established concepts of Concentration Addition (CA) and Independent Action (IA). As a result, both models (CA and IA) were found to underestimate the toxicity of mixtures, however CA yielded more accurate predictions.

A nematicidal tannin from Punica granatum L. rind and its physiological effect on pine wood nematode (Bursaphelenchus xylophilus).

The ethanol extract of Punica granatum L. rind was tested to show significant nematicidal activity against pine wood nematode. Three nematicidal compounds were obtained from the ethanol extract by bioassay-guided fractionation and identified as punicalagin 1, punicalin 2, and corilagin 3 by mass and nuclear magnetic resonance spectral data analysis. Punicalagin 1 was most active against PWN among the purified compounds with the LC50 value of 307.08µM in 72h. According to the enzyme assays in vitro, punicalagin 1 could inhibit the activity of acetylcholinesterase, amylase and cellulase from PWN with IC50 value of 0.60mM, 0.96mM and 1.24mM, respectively. The morphological structures of PWNs treated by punicalagin 1 were greatly changed. These physiological effects of punicalagin 1 on PWN may helpful to elucidate its nematicidal mechanism.