Vitamin D supplementation modulates autophagy in the pristane-induced lupus model.

INTRODUCTION/OBJECTIVES: Clinical evidence of skeletal muscle involvement is not uncommon in systemic lupus erythematosus (SLE). Because of the poor understanding of signaling pathways involved in SLE muscle wasting, the  aim of this study was to evaluate the effects of vitamin D supplementation on skeletal muscle in mice with pristane-induced lupus. METHODS: Balb/c mice with lupus-like disease induced by pristane injection were randomized into three groups: pristane-induced lupus (PIL; n = 10), pristane-induced lupus + vitamin D supplementation (PIL + VD; n = 10) and healthy controls (CO; n = 8). Physical function was evaluated on days 0, 60, 120 and 180. The tibialis anterior and gastrocnemius muscles were collected to evaluate myofiber cross-sectional area (CSA) and protein expression. RESULTS: The PIL + VD group showed lower muscle strength compared to the CO and PIL groups at different time points. PIL mice showed similar myofiber CSA compared to CO and PIL + VD groups. LC3-II expression was higher in PIL compared to CO and PIL + VD groups. MyoD expression was higher in PIL mice compared to PIL + VD, while myostatin expression was higher in PIL + VD than PIL group. Myogenin expression levels were decreased in the PIL + VD group compared with the CO group. The Akt, p62 and MuRF expressions and mobility assessment showed no significance. CONCLUSIONS: Changes in skeletal muscle in PIL model happen before CSA reduction, possibly due to autophagy degradation, and treatment with Vitamin D has a impact on physical function by decreasing muscle strength and time of fatigue.. Vitamin D supplementation has a potential role modulating physical parameters and signaling pathways in muscle during pristane-induced lupus model.

Ethnoveterinary for food-producing animals and related food safety issues: A comprehensive overview about terpenes.

Alternatives to the use of conventional veterinary drugs in food-producing animals have gained attention, such as the use of natural products (NPs), mainly to soften the risks to the animal, the environment, and consumer's health. Although NPs have consistent advantages over conventional drugs, they cannot be considered risk free under food safety matters. In this way, this document presents a comprehensive overview of the importance of considering both the pharmacological and toxicological properties of the constituents of a NP from plants intending the standardization and regulation of its use in food-producing animals. Terpenes are the most diverse class of natural substances present in NP of vegetal origin with a broad range of biological activities that can be explored in veterinary science; however, certain plants and terpenes also have significant toxic effects, a fact that can harm the health of animals and consequently generate economic losses and risks for humans. In this context, this review gathered scientific data of vegetal species of importance to ethnoveterinary for food-producing animals, which produce terpenes, its biological effects, and their implications on food safety issues for consumers. For this, more than 300 documents were selected from different online scientific databases. The present data and discussion may contribute to the rational commercial exploration of this class of NPs in veterinary medicine.

Synergistic effects of binary mixtures of linalool with pyrethroids against fall armyworm / Efeitos sinérgicos de misturas binárias de linalol com piretroides contra lagarta-do-cartucho

The present work aimed to determine the toxicity of linalool and evaluate the lethal and toxic effects of linalool associated with pyrethroids in binary mixtures to fall armyworm (Spodoptera frugiperda). The insects used in the experiment were obtained from stock breeding initiated from larvae collected from conventional corn plants, grown in an experimental area, in the city of Uberlândia, Minas Gerais. Also, it was obtained essential oil from a variety of Ocimum basilicum, with a high content of linalool (80%), found naturally, as a measure of comparison of different linalool (97.5%) assays. Dose-response bioassays with 3rd instar larvae were performed to determine lethal dose for 50% mortality (LD50) of linalool. Toxicity tests were also performed with O. basilicum essential oil and with pyrethroid insecticides: deltamethrin and its commercial product (Decis 25 EC, Bayer®). After this, combinations between different doses of these products were made and applied on 3rd instar larvae of Spodoptera frugiperda (Smith). Linalool presented high toxicity to S. frugiperda (LD50 = 0.177 µL a.i. µL-1). It was observed neurotoxic effects after the linalool application since the insects presented an aspect of confusion, followed by extreme agitation and finally death. All binary mixtures caused mortality higher than the products applied alone (deltamethrin and linalool) used at 100% LD50, except to 75% LD50 deltamethrin added to 25% LD50 linalool, whose mortality did not differ the products alone, in 24 hours. It was obtained over 90% larval mortality when linalool was combined with 25% LD50 of deltamethrin, in 24 and 48 hours after application, and over 80% of mortality when linalool was combined with 25% LD50 of Decis, only in 48 hours after application. We conclude that linalool is a potential insecticidal and can be associated with pyrethroids to control of S. frugiperda. Further studies are required in order to evaluate the synergistic combinations against field populations of S. frugiperda.

O objetivo deste trabalho foi determinar a toxicidade do linalol e avaliar os efeitos tóxicos e letais do linalol associado a piretroides em misturas binárias para lagarta do cartucho do milho (Spodoptera frugiperda). Os insetos utilizados no experimento foram obtidos de criação estoque iniciada a partir de larvas coletadas em plantas de milho convencional, cultivado em área experimental, no município de Uberlândia, Minas Gerais. Também foi obtido óleo essencial de uma variedade de Ocimum basilicum, com alto teor de linalol (80%), encontrado naturalmente, como medida de comparação para ensaios com linalol (97.5%). Os bioensaios do tipo dose-resposta com larvas de 3º instar foram realizados para determinar a dose letal do linalol para 50% de mortalidade da população (DL50). Também foram realizados testes de toxicidade com óleo essencial de Ocimum basilicum e com inseticidas piretroides: deltametrina e seu produto comercial (Decis 25 EC, Bayer®). Em seguida, foram realizadas combinações entre diferentes doses desses produtos e aplicadas em larvas de 3º instar de Spodoptera frugiperda (Smith). De acordo com os resultados, observou-se que o linalol apresentou alta toxicidade para S. frugiperda (DL50 = 0,177 µL a. i. µL-1). Foram observados efeitos neurotóxicos após a aplicação do linalol, uma vez que os insetos apresentaram um aspecto de confusão, seguido de extrema agitação e, por fim, morte. Todas as combinações binárias causaram mortalidade maior que os produtos aplicados isoladamente (deltametrina e linalol) utilizando-se 100% da DL50, exceto para 75% DL50 de deltametrina somada a 25% DL50 de linalol, cuja mortalidade não diferiu dos produtos isolados, em 24 horas após a aplicação. Foi obtida mais de 90% de mortalidade de larvas quando se combinou linalol com 25% da DL50 de deltametrina, em 24 e 48 horas após a aplicação, e mais de 80% de mortalidade quando se combinou linalol com 25% da DL50 do produto comercial, somente 48 horas após a aplicação. Concluímos que o linalol é um potencial inseticida e pode ser associado a piretroides no controle de S. frugiperda. Mais estudos são necessários em vista de avaliar as combinações sinérgicas contra populações de campo de S. frugiperda.

Anti-Estrogenic Activity of Guajadial Fraction, from Guava Leaves (Psidium guajava L.).

The research of natural products has allowed for the discovery of biologically relevant compounds inspired by plant secondary metabolites, which contributes to the development of many chemotherapeutic drugs used in cancer treatment. Psidium guajava leaves present a diverse phytochemical composition including flavonoids, phenolics, meroterpenoids, and triterpenes as the major bioactive constituents. Guajadial, a caryophyllene-based meroterpenoid, has been studied for potential anticancer effects tested in tumor cells and animal experimental models. Moreover, guajadial has been reported to have a mechanism of action similar to tamoxifen, suggesting this compound as a promisor phytoestrogen-based therapeutic agent. Herein, the anti-estrogenic action and anti-proliferative activity of guajadial is reported. The enriched guajadial fraction was obtained by sequential chromatographic techniques from the crude P. guajava dichloromethane extract showing promising anti-proliferative activity in vitro with selectivity for human breast cancer cell lines MCF-7 and MCF-7 BUS (Total Growth Inhibition = 5.59 and 2.27 µg·mL-1, respectively). Furthermore, evaluation of anti-estrogenic activity in vivo was performed demonstrating that guajadial enriched fraction inhibited the proliferative effect of estradiol on the uterus of pre-pubescent rats. These results suggest a relationship between anti-proliferative and anti-estrogenic activity of guajadial, which possibly acts in tumor inhibition through estrogen receptors due to the compounds structural similarity to tamoxifen.

Citral presents cytotoxic and genotoxic effects in human cultured cells.

Citral, 3,7-dimethyl-2,6-octadien-1-al, one of the main components of the essential oils obtained from several plants, is used as a food additive and as a fragrance for detergents, cosmetics and other toiletries. The literature shows disparity regarding citral genotoxicity. Thus, the main objective of our work was to evaluate the genotoxic effects of citral in human cell cultures, HepG2 and leukocytes. Cytotoxicity assays (trypan blue and MTT) showed citral toxic effects in HepG2 cells (with metabolizing liver enzymes), which contrasted with the absence of toxicity in leukocytes. After citral exposure, both cell types did not demonstrate clastogenic/aneugenic effects in the micronucleus test. However, for the comet assay, citral exposure lead to significant genotoxic effects in both HepG2 (even to citral low concentrations) and leukocytes. The use of citral must be viewed with caution due to its ability to induce DNA damages, especially after being metabolized by cells with active liver enzymes.

Botanical and synthetic pesticides alter the flower visitation rates of pollinator bees in Neotropical melon fields.

The ecological and economic contributions of pollinator bees to agricultural production have been threatened by the inappropriate and excessive use of pesticides. These pesticides are often applied in areas with ecological peculiarities (e.g., the Neotropical savannah-like region termed as Cerrado) that were not considered during the product development. Here, we conducted field experiments with melon (i.e., Cucumis melo L.) plants cultivated under Brazilian Cerrado conditions and evaluated the impacts of botanical (i.e., neem-based insecticide) and synthetic (i.e., the pyrethroid insecticide deltamethrin and the fungicides thiophanate-methyl and chlorothalonil) pesticides on the flower visitation rates of naturally occurring pollinator bees. Our results revealed that both honey bees (i.e., Apis mellifera L.) and non-Apis bees visited melon flowers and the intensity of bee visitation was moderately correlated with yield parameters (e.g., number of marketable fruits and fruit yield). Pesticide treatments differentially affected bee species. For instance, Plebeia sp. bees were not affected by any pesticide treatment, whereas both A. mellifera and Halictus sp. bees showed reduced visitation intensity after the application of deltamethrin or neem-based insecticides. Fungicide treatment alone did not influence the bee's visitation intensity. Deltamethrin-treated melon fields produced significantly lighter marketable fruits, and the melon yield was significantly lower in melon fields treated with the neem-based insecticide. Thus, our findings with such pollinator bees reinforce the idea that field applications of botanical pesticides may represent as risky as the applications of synthetic compounds, indicating that these alternative products should be submitted to risk assessments comparable to those required for synthetic products.

Neem oil based nanopesticide as an environmentally-friendly formulation for applications in sustainable agriculture: An ecotoxicological perspective.

Sustainable agriculture encourages practices that present low risks to the environment and human health. To this end, zein (corn protein) can be used to develop nanocarrier systems capable of improving the physicochemical properties of biopesticides, reducing their possible toxicity. Neem oil extracted from the Azadirachta indica tree contains many active ingredients including azadirachtin, which is the active ingredient in multiple commercially available biopesticides. In this study, we describe the preparation and characterization of neem oil-loaded zein nanoparticles, together with evaluation of their toxicity towards nontarget organisms, using Allium cepa, soil nitrogen cycle microbiota, and Caenorhabditis elegans aiming to achieve the safer by design strategy. The spherical nanoparticles showed an average diameter of 278 ±â€¯61.5 nm and a good stability during the experiments. In the toxicity assays with A. cepa, the neem oil-loaded zein nanoparticles mitigated the increase in the DNA relative damage index caused by the neem oil. Molecular genetic analysis of the soil nitrogen cycle microbiota revealed that neem oil-loaded zein nanoparticles did not change the number of genes which encode nitrogen-fixing enzymes and denitrifying enzymes. In C. elegans, the neem oil-loaded zein nanoparticles had no toxic effect, while neem oil interfered with pharyngeal pumping and GST-4 protein expression. These neem oil-loaded zein nanoparticles showed promising results in the toxicity studies, opening perspectives for its use in crop protection in organic agriculture.

Apis mellifera (Insecta: Hymenoptera) in the target of neonicotinoids: A one-way ticket? Bioinsecticides can be an alternative.

The recent decline of Apis mellifera populations around the world has been subject of intense research due to ecological and economic damages resulting from the loss of pollination services. The intensive use of insecticides from the neonicotinoids group is among the possible causal factors of this decline, including also sub-lethal effects. However, the use of synthetic insecticides has been increased on a global scale in the recent decades. In order to evaluate an alternative to the use of neonicotinoids, this work investigated the effects of a bioinsecticide and its major compound on A. mellifera (Apidae: Hymenoptera), one of the main pollinators of crop plants. For this, bees were exposed, by contact and ingestion, to the essential oil of Cymbopogon martinii (Poaceae: Poales), to geraniol (major compound) and the insecticide imidacloprid to evaluate the toxicity and behavioral effects as well as the locomotion changes and immune responses of bees treated with these compounds. In general, toxicity was greater through ingestion and the insecticide imidacloprid was more toxic to A. mellifera compared to the essential oil and its major compound. The individual and collective behaviors (i.e. trophallaxis, grooming, avoidance) as well as the immune responses of bees were not significantly affected by bioinsecticides. However, the locomotion response and flight orientation of the bees were significantly altered by insecticide when administered by ingestion. Our results highlight the potential of C. martinii essential oil and its major compound as a possible alternative to mitigate the harmful effects of neonicotinoids on bees.

Acute Toxicity and Sublethal Effects of Terpenoids and Essential Oils on the Predator Chrysoperla externa (Neuroptera: Chrysopidae).

The search for new safer insecticides has increased in recent agriculture. Botanical compounds such as terpenoids and plant essential oils with insecticidal activity could represent important tools in pest management, and their risk assessment against non-target organisms is necessary since they may serve as a precursor for the synthesis of new insecticide active ingredients. For this study, the acute toxicity and sublethal effects of seven terpenoids and three essential oils with recognized insecticidal activity were evaluated on the predator Chrysoperla externa (Hagen) (Neuroptera: Chrysopidae) in laboratory bioassays. Results indicate that these compounds feature relative selectivity to the predator C. externa; however, sublethal effects on reproduction were recorded for some compounds. The phenolic monoterpenoids carvacrol and thymol were more acutely toxic than other terpenoids screened, with LD50 <20,000 µg/g; however, they were less toxic than natural pyrethrins (toxicity standard) in these bioassays. Sublethal effects on fecundity and fertility were observed for R-(+)-limonene, while oregano oil only affected fecundity. The compounds evaluated here have potential to be used as insecticides and can serve as backbone for future synthesis of selective active ingredients; however, a complete risk assessment to C. externa and other non-target organisms is necessary for their incorporation in future crop protection paradigms.

Cytotoxic and genotoxic potential of geraniol in peripheral blood mononuclear cells and human hepatoma cell line (HepG2).

Geraniol is an acyclic monoterpene alcohol present in the essential oil of many aromatic plants and is one of the most frequently used molecules by the flavor and fragrance industries. The literature also reports its therapeutic potential, highlighting itself especially as a likely molecule for the development of drugs against cancer. In view of these considerations, this study was designed to evaluate the cytotoxic and genotoxic potential of geraniol, in an in vitro protocol, using two types of human cells: one without the ability to metabolize (peripheral blood mononuclear cells - PBMC), and the other with this capability (human hepatoma cell line - HepG2) through the comet assay and the micronucleus test. Four concentrations (10, 25, 50, and 100 µg/mL) were selected for the genotoxic assessment for PBMC and three (1.25, 2.5, and 5 µg/mL) for HepG2 cells based on cytotoxicity tests (MTT assay). Results showed that geraniol did not present genotoxic or clastogenic/aneugenic effects on both cell types under the conditions studied. However, caution is advised in the use of this substance by humans, since a significant reduction in viability of HepG2 and a marked decrease in cell viability on normal PBMC were verified.

Comparative toxicity of essential oil and blends of selected terpenes of Ocotea species from Pernambuco, Brazil, against Tetranychus urticae Koch

ABSTRACT Essential oils from the leaves of two species of the genus Ocotea that occur in the Atlantic Forest in the state of Pernambuco, Brazil, were analyzed using gas chromatography-mass spectrometry. The acaricidal activity of these oils as well as 11 selected components and blends were evaluated in fumigation and residual contact tests against the two-spotted spider mite (Tetranychus urticae). Sixty-seven constituents were identified, totaling 97.3 ± 0.3% and 97.8 ± 0.5% of the oils from O. duckei and O. glomerata, respectively. Sesquiterpene was the dominant class. The compounds β-caryophyllene (18.6 ± 0.1%) and aromadendrene (17.3 ± 0.6%) were the main constituents of the oils from O. duckei and O. glomerata, respectively. Acaricidal action varied depending on the method employed, species and chemical nature of the selected constituents. The mites were susceptible to the oils and chemical constituents using the fumigation method. The O. duckei oil was respectively 2.5-fold and 1.5-fold more toxic than the O. glomerata oil using the fumigation and residual contact methods. Among the selected constituents, β-caryophyllene was the most toxic, independently of the method employed. The individual toxicity of the selected compounds and their blends as well as the role of these constituents in the overall toxicity of the essential oils are also discussed.

Comparative toxicity of essential oil and blends of selected terpenes of Ocotea species from Pernambuco, Brazil, against Tetranychus urticae Koch.

Essential oils from the leaves of two species of the genus Ocotea that occur in the Atlantic Forest in the state of Pernambuco, Brazil, were analyzed using gas chromatography-mass spectrometry. The acaricidal activity of these oils as well as 11 selected components and blends were evaluated in fumigation and residual contact tests against the two-spotted spider mite (Tetranychus urticae). Sixty-seven constituents were identified, totaling 97.3 ± 0.3% and 97.8 ± 0.5% of the oils from O. duckei and O. glomerata, respectively. Sesquiterpene was the dominant class. The compounds ß-caryophyllene (18.6 ± 0.1%) and aromadendrene (17.3 ± 0.6%) were the main constituents of the oils from O. duckei and O. glomerata, respectively. Acaricidal action varied depending on the method employed, species and chemical nature of the selected constituents. The mites were susceptible to the oils and chemical constituents using the fumigation method. The O. duckei oil was respectively 2.5-fold and 1.5-fold more toxic than the O. glomerata oil using the fumigation and residual contact methods. Among the selected constituents, ß-caryophyllene was the most toxic, independently of the method employed. The individual toxicity of the selected compounds and their blends as well as the role of these constituents in the overall toxicity of the essential oils are also discussed.

Nanocapsules Containing Neem (Azadirachta Indica) Oil: Development, Characterization, And Toxicity Evaluation.

In this study, we prepared, characterized, and performed toxicity analyses of poly(ε-caprolactone) nanocapsules loaded with neem oil. Three formulations were prepared by the emulsion/solvent evaporation method. The nanocapsules showed a mean size distribution around 400 nm, with polydispersity below 0.2 and were stable for 120 days. Cytotoxicity and genotoxicity results showed an increase in toxicity of the oleic acid + neem formulations according to the amount of oleic acid used. The minimum inhibitory concentrations demonstrated that all the formulations containing neem oil were active. The nanocapsules containing neem oil did not affect the soil microbiota during 300 days of exposure compared to the control. Phytotoxicity studies indicated that NC_20 (200 mg of neem oil) did not affect the net photosynthesis and stomatal conductance of maize plants, whereas use of NC_10 (100:100 of neem:oleic acid) and NC_15 (150:50 of neem:oleic acid) led to negative effects on these physiological parameters. Hence, the use of oleic acid as a complement in the nanocapsules was not a good strategy, since the nanocapsules that only contained neem oil showed lower toxicity. These results demonstrate that evaluation of the toxicity of nanopesticides is essential for the development of environmentally friendly formulations intended for applications in agriculture.

Selectivity of plant extracts for Trichogramma pretiosum Riley (Hym.: Trichogrammatidae).

We evaluated the selectivity of three plant extracts with potential insecticidal effects for the parasitoid Trichogramma pretiosum Riley, which is commonly used in biological pest control. The plant extracts assayed were an acetone extract of Toona ciliata M. Roem., commercial neem oil, and a nanoencapsulated formulation of neem oil (NC40). The toxicity of the plant extracts to T. pretiosum was evaluated according to the recommendations of the International Organization for Biological Control- IOBC Working Group. We assessed the susceptibility of adults of the maternal and F1 generations and immature stages of T. pretiosum to the extracts. Females exposed to egg cards treated with commercial neem oil parasitized almost 70% fewer eggs than control eggs treated with water; and this extract was therefore classified as slightly harmful. When the eggs were offered to females 24h after treatment with neem oil and aqueous NC40, the parasitism rate also decreased, and the two extracts were classified as slightly harmful. Adult emergence was lower for parasitoids that fed on host eggs offered 24h after the treatment with the T. ciliata extract, which was considered slightly harmful. The emergence of T. pretiosum from eggs, larvae and pupae treated with the different plant extracts, did not decrease compared to development stages treated with the water control. The use of T. pretiosum, combined with the application of an ethanol extract of T. ciliata and a nanoencapsulated formulation of neem, appears to be feasible in view of these low toxicity indices.

Antifungal activity of plant-derived essential oils on Candida tropicalis planktonic and biofilms cells.

Dental prosthesis supports Candida species growth and may predispose the oral cavity to lesions. C. tropicalis has emerged as a colonizer of prosthesis and has shown resistance to clinically used antifungal agents, which has increased the search for new antifungals. This work describes the effectiveness of fifteen essential oils (EOs) against C. tropicalis The EOs were obtained by hydrodistillation and were chemically characterized by gas chromatography-mass spectrometry. The antifungal activities of the EOs were evaluated by the microdilution method and showed that Pelargonium graveolens (Geraniaceae) (PG-EO) was the most effective oil. Geraniol and linalool were the major constituents of PG-EO. The 2,3-Bis-(2-Methoxy-4-Nitro-5-Sulfophenyl)-2H-Tetrazolium-5-Carboxanilide (XTT) assay showed that all the clinical C. tropicalis strains formed viable biofilms. Scanning electron microscopy examination of the biofilms revealed a complex architecture with basal layer of yeast cells and an upper layer of filamentous cells. Treatments with PG-EO, linalool, and geraniol significantly reduced the number of viable biofilm cells and inhibited biofilm formation after exposure for 48 h. PG-EO, geraniol, and linalool were not toxic to normal human lung fibroblasts (GM07492A) at the concentrations they were active against C. tropicalis Together, our results indicated that C. tropicalis is susceptible to treatment with PG-EO, geraniol, and linalool, which could become options to prevent or treat this infection.

Morphological alterations in the synganglion and integument of Rhipicephalus sanguineus ticks exposed to aqueous extracts of neem leaves (Azadirachta indica A. JUSS).

Currently, the necessity of controlling infestation by ticks, especially by Rhipicephalus sanguineus, has led researchers and public health managers around the world to search for new and more efficient control methods. This way, we can highlight neem (Azadirachta indica A. Juss) leaf, bark, and seed extracts, which have been very effective on tick control, and moreover causing less damage to the environment and to the host. This study showed the potential of neem as a control method for R. sanguineus through morphological and morphometric evaluation of the integument and synganglion of females, in semiengorged stage. To attain this, routine techniques of optical microscopy, scanning electron microscopy and morphometry of the cuticle and subcuticle of the integument were applied. Expressive morphological alterations were observed in both organs, presenting a dose-dependent effect. Integument epithelial cells and nerve cells of the synganglion showed signs of cell vacuolation, dilated intercellular boundaries, and cellular disorganization, alterations not previously reported in studies with neem. In addition, variations in subcuticle thickness were also observed. In general, the effects of neem are multiple, and affect the morphology and physiology of target animals in various ways. The results presented in this work are the first evidence of its effects in the coating and nervous system of ticks, thus allowing an indication of neem aqueous extracts as a potential control method of the brown dog tick and opening new perspectives on acaricide use.

Toxicity of terpenes on fibroblast cells compared to their hemolytic potential and increase in erythrocyte membrane fluidity.

Terpenes are considered potent skin permeation enhancers with low toxicity. Electron paramagnetic resonance (EPR) spectroscopy of the spin label 5-doxyl stearic acid (5-DSA) was used to monitor the effect of sesquiterpene nerolidol and various monoterpenes on membrane fluidity in erythrocyte and fibroblast cells. In addition, the hemolytic levels and cytotoxic effects on cultured fibroblast cells were also measured to investigate possible relationships between the cellular irritation potentials of terpenes and the ability to modify membrane fluidity. All terpenes increased cell membrane fluidity with no significant differences between the monoterpenes, but the effect of sesquiterpene was significantly greater than that of the monoterpenes. The IC(50) values for the terpenes in the cytotoxicity assay indicated that 1,8-cineole showed lower cytotoxicity and α-terpineol and nerolidol showed higher cytotoxicity. The correlation between the hemolytic effect and the IC(50) values for fibroblast viability was low (R=0.61); however, in both tests, nerolidol was among the most aggressive of terpenes and 1,8-cineole was among the least aggressive. Obtaining information concerning the toxicity and potency of terpenes could aid in the design of topical formulations optimized to facilitate drug absorption for the treatment of many skin diseases.

Toxicity of Neem's oil, a potential biocide against the invasive mussel Limnoperna fortunei (Dunker 1857).

The golden mussel Limnoperna fortunei (Dunker 1857) is one of the most distributed Nuisance Invasive Species (NIS) in South America, and a threat of great concern for the industry of the area. In this study, we carried out toxicity tests made with a Neem's oil solution with L. fortunei larvae and benthonic adults (7, 13 and 19 ± 1 mm). Tests with non-target species (Daphnia magna, Lactuca sativa and Cnesterodon decemmculatus) were also made with the aim to evaluate the potential toxicity of the Neem's solution in the environment. The LC(100) of Neem's solution obtained for larvae was 500 µl/L, a value much higher than the one obtained for D. magna and C. decemmaculatus. Thus, we recommend that it should not be used in open waters. However, since the adults were killed in 72 h and the larvae in 24 h, this product can be used in closed systems, in man-made facilities.

Structure-activity relationships of larvicidal monoterpenes and derivatives against Aedes aegypti Linn.

In the search for larvicidal compounds against Aedes aegypti L. (Diptera: Culicidae), a collection of monoterpenes were selected and evaluated. R- and S-limonene exhibited the highest larvicidal potency (LC(50)=27 and 30 ppm, respectively), followed by γ-terpinene (LC(50)=56 ppm) and RS-carvone (LC(50)=118 ppm). Structural characteristics which may contribute to the understanding of the larvicidal activity of monoterpenes were empirically identified. The presence of heteroatoms in the basic hydrocarbon structure decreases larvicidal potency. Conjugated and exo double bonds appear to increase larvicidal potency. Replacement of double bonds by more reactive epoxides decreases the larvicidal potency. The presence of hydroxyls in the cyclic structure resulted in decreased potency, probably due to increased polarity indicanting that lipophilicity seems to play an important role in increasing the larvicidal potency in this set of compounds.

Three austin family compounds from Penicillium brasilianum exhibit selective blocking action on cockroach nicotinic acetylcholine receptors.

Austin (AT) and its derivatives (dehydroaustin (DAT) and acetoxydehydroaustin (ADAT)) produced by Penicillium brasilianum MG-11 exhibit toxicity to insects, yet their targets are unknown. Here, we used whole-cell patch-clamp electrophysiology to investigate the action of AT family compounds on cockroach acetylcholine (ACh), γ-aminobutyric acid (GABA) and l-glutamate receptors expressed in the American cockroach (Periplaneta americana) neuron. U-tube application of AT or its derivatives did not induce any current amplitudes, suggesting that they did not act as agonist of these three receptors. In the second step of experiments, they were bath-applied for 1min before co-application with the corresponding ligand. We found that AT and its derivatives had no effect on GABA and l-glutamate-induced currents, whereas they significantly reduced ACh- and epibatidine-induced currents, showing that these compounds acted as selective antagonists of nicotinic acetylcholine receptors (nAChRs) expressed in the cockroach neuron. Of the compounds, DAT showed the highest blocking potency for nAChRs, differentially attenuating the peak and slowly desensitizing current amplitude of ACh-induced responses with pIC(50) (=-logIC(50) (M)) values of 6.11 and 5.91, respectively. DAT reduced the maximum normalized response to ACh without a significant shift in EC(50), suggesting that the blocking action is not competitive with ACh.