Repellency and Larvicidal Activity of Essential oils from Xylopia laevigata, Xylopia frutescens, Lippia pedunculosa, and Their Individual Compounds against Aedes aegypti Linnaeus.

In order to find new alternatives for vector control and personal protection, we evaluated the larvicidal and repellent activity of essentials oils from plants found in the Northeast of Brazil against Aedes aegypti Linnaeus mosquitoes. The plants tested include Xylopia laevigata, Xylopia frutescens, and Lippia pedunculosa and their major compounds, piperitenone oxide, and (R)-limonene. The essential oil of L. pedunculosa and its major volatile compounds were shown to be toxic for Ae. aegypti larvae with a LC50 lower than 60 ppm. The essential oil of plants from the Xylopia genus, on the other hand, showed no activity against Ae. aegypti, proving to be toxic to mosquito larvae only when concentrations were higher than 1000 ppm. All plants tested provided some degree of protection against mosquitoes landing, but only the essential oil of L. pedunculosa and the volatile compound piperitenone oxide suppressed 100% of mosquitoes landing on human skin, in concentrations lower than 1%. Among the plants studied, the essential oil of L. pedunculosa and its volatiles compounds have shown the potential for the development of safe alternative for mosquito larvae control and protection against Ae. aegypti mosquito bites.

Detection and Distribution of V1016Ikdr Mutation in the Voltage-Gated Sodium Channel Gene in Aedes aegypti (Diptera: Culicidae) Populations From Sergipe State, Northeast Brazil.

Aedes aegypti (L.) resistance to pyrethroids was recorded in Brazil few years after its introduction as the adulticide in the National Dengue Control Program campaigns. Altered susceptibility to pyrethroids had been reported in the state of Sergipe, northeast Brazil, through biological assays, even before its use against Ae. aegypti in the state. Metabolic and target-site resistance mechanisms were also revealed in samples from Aracaju, the capital of Sergipe. Herein, we investigated the presence and distribution of the kdr mutation V1016Ikdr in Ae. aegypti populations from different municipalities of the state. Aedes aegypti eggs were collected from seven municipalities located in areas showing different climatic types and infestation levels. Approximately 20 Ae. aegypti females from each municipality (total of 135 subjects) were individually submitted to allele-specific polymerase chain reaction (AS-PCR) for the 1016 site of the voltage-gated sodium channel (NaV). The V1016Ikdr mutation was found in subjects from all the municipalities under study with a high frequency of heterozygotes in several locations. Homozygous recessive subjects (resistant kdr genotype) were found only in one municipality. The results suggest a wide distribution of the V1016Ikdr mutation in the northeast Brazil, which indicates urgent need for monitoring the effectiveness of the pyrethroids currently used for vector control.

An environmentally safe larvicide against Aedes aegypti based on in situ gelling nanostructured surfactant systems containing an essential oil.

HYPOTHESIS: Liquid crystalline precursors, which are in situ gelling nanostructured surfactant systems, can undergo phase transition in aqueous solution and become more structured aggregates, controlling release of larvicides and acting as biotechnology alternatives for dengue control. Such systems can contain bioactive substances as Citrus sinensis essential oil (CSEO) which exhibits biological activity against Aedes aegypti (Ae. aegypti) larvae. EXPERIMENTS: The formulations were composed by fixed concentration of CSEO stabilized by Polyoxypropylene (5) Polyoxyethylene (20) Cetyl Ether (PPG-5 CETETH-20): oleic acid (OA) 2:1, increasing water content. The phase diagram was established and systems structure was evaluated by polarized light microscopy (PLM), small angle X-ray scattering (SAXS) and rheology. Median lethal concentration was determined against Ae. aegypti larvae. FINDINGS: The phase diagram exhibited four regions: liquid crystal (LC), emulsion, microemulsion (ME) and phase separation. The PLM and SAXS distinguished microemulsions, lamellar and hexagonal LC structures. Flow and oscillatory tests showed that increasing water content increases elasticity from Newtonian to non-newtonian behavior confirming the in situ gelation behavior. The larvicidal activity of formulations indicates that these nanostructured systems improved the oil solubility in aqueous medium and in addition are potential environmental larvicide against Ae. aegypti larvae.

Composition and acaricidal activity of Lippia sidoides essential oil against two-spotted spider mite (Tetranychus urticae Koch).

The essential oils from accessions of Lippia sidoides Cham. (Verbenaceae) were characterized by GC and GC/MS and investigated for their acaricidal activity against the two-spotted spider mite (Tetranychus urticae Koch). Twenty-nine compounds were identified with potential acaricidal activity. Glass receptacles were used as test chambers. For each dose and exposure time combination, three replicates were used. Each replicate consisted of 30 adult females of T. urticae, 10 mites in each leaf disk of Canavalia ensiformis placed in a Petri dish. Increasing amounts of oil or terpene were applied on a blotting paper strip, fixed on the inner surface of the glass recipient cover, corresponding to 2, 4, 6, 8, and 10 microL/L of air, respectively. Exposure periods were 24, 48, and 72 h. Data obtained in these experiments were submitted to probit analysis. The essential oil of L. sidoides, thymol and carvacrol exhibited potent acaricidal activity against T. urticae.

Effects of essential oils on Aedes aegypti larvae: alternatives to environmentally safe insecticides.

The essential oils from leaves of Hyptis fruticosa (Lamiaceae) Salzm., H. pectinata (Lamiaceae) Poit., and Lippia gracilis (Verbenaceae) HBK were investigated for their larvicidal activity against Aedes aegypti and analyzed by GC/MS. Fifty-nine compounds, representing 91.28-98.39% of the essential oils, have been identified. A standard solution was used to make 20 mL solutions ranging from 30 to 2000 ppm. Twenty larvae between third and fourth stages were added to the essential oil solution. A mortality count was conducted 24 h after treatment. Essential oils LC50 and their confidence limits at 95% probability were calculated by the methods of Reed-Muench and Pizzi, respectively. The essential oil of Lippia gracilis showed potent insecticidal effect against Aedes aegypti larvae, the vector of dengue fever. Carvacrol and caryophyllene oxide were the main responsible for the activity of L. gracilis and H. pectinata. Minor compounds are probably acting synergistically to achieve H. fruticosa activity.

Antinociceptive and antiedematogenic properties and acute toxicity of Tabebuia avellanedae Lor. ex Griseb. inner bark aqueous extract.

BACKGROUND: Tabebuia avellanedae is a tree from the Bignoniaceae family. Commonly know as "pau d'arco" in Brazil, its inner bark is used as analgesic, anti-inflammatory, antineoplasic and diuretic at the Brazilian northeast. A validation of the plant usage has not been previously performed. RESULTS: Antinociceptive and antiedematogenic effects of Tabebuia avellanedae Lor. ex Griseb. inner bark were measured by nociceptive experimental models in mice. A rat paw edema test induced by carrageenan (1%) was also performed in rats to access the plant's antiedematogenic effect. The inner bark aqueous extract, administered via oral in three different concentration, namely 100, 200 and 400 mg/Kg, reduced the nociception produced by acetic acid (0.6% in water, i.p.) by 49.9%, 63.7% and 43.8%, respectively. The aqueous extract (200 and 400 mg/Kg, p.o.) reduced formalin (1%) effects only at the second phase of the experiment by 49.3% and 53.7%, respectively. Naloxone (5 mg/Kg, i.p.) was not able to revert the extract effect, however caffeine (10 mg/Kg, i.p.) reverted its effect by 19.8% at the second phase of the formalin test. The aqueous extract (200 mg/Kg, p.o.) inhibited edema by 12.9% when we used the rat paw edema model. The acute toxicity was low in mice. CONCLUSION: The T. avellanedae inner bark aqueous extract presented antinociceptive and antiedematogenic activities at the used models, with a possible antinociceptive effect associated to the adenosine system.

In vitro selection of mutations in the human immunodeficiency virus type 1 reverse transcriptase that decrease susceptibility to (-)-beta-D-dioxolane-guanosine and suppress resistance to 3'-azido-3'-deoxythymidine.

Human immunodeficiency virus type 1 (HIV-1) isolates resistant to (-)-beta-D-dioxolane-guanosine (DXG), a potent and selective nucleoside analog HIV-1 reverse transcriptase (RT) inhibitor, were selected by serial passage of HIV-1(LAI) in increasing drug concentrations (maximum concentration, 30 microM). Two independent selection experiments were performed. Viral isolates for which the DXG median effective concentrations (EC(50)s) increased 7.3- and 12.2-fold were isolated after 13 and 14 passages, respectively. Cloning and DNA sequencing of the RT region from the first resistant isolate identified a K65R mutation (AAA to AGA) in 10 of 10 clones. The role of this mutation in DXG resistance was confirmed by site-specific mutagenesis of HIV-1(LAI). The K65R mutation also conferred greater than threefold cross-resistance to 2',3'-dideoxycytidine, 2', 3'-dideoxyinosine, 2',3'-dideoxy-3'-thiacytidine, 9-(2-phosphonylmethoxyethyl)adenine, 2-amino-6-chloropurine dioxolane, dioxolanyl-5-fluorocytosine, and diaminopurine dioxolane but had only marginal effects on 3'-azido-3'-deoxthymidine (AZT) susceptibility. However, when introduced into a genetic background for AZT resistance (D67N, K70R, T215Y, T219Q), the K65R mutation reversed the AZT resistance. DNA sequencing of RT clones derived from the second resistant isolate identified the L74V mutation, previously reported to cause ddI resistance. The L74V mutation also decreased the AZT resistance when the mutation was introduced into a genetic background for AZT resistance (D67N, K70R, T215Y, T219Q) but to a lesser degree than the K65R mutation did. These findings indicate that DXG and certain 2',3'-dideoxy compounds (e.g., ddI) can select for the same resistance mutations and thus may not be optimal for use in combination. However, the combination of AZT with DXG or its orally bioavailable prodrug (-)-beta-D-2, 6-diaminopurine-dioxolane should be explored because of the suppressive effects of the K65R and L74V mutations on AZT resistance.

Synthesis and antiviral activities of enantiomeric 1-[2-(hydroxymethyl) cyclopropyl] methyl nucleosides.

Cyclopropyl carbocyclic nucleosides have been synthesized from the key intermediate 2 which was converted to the mesylated cyclopropyl methyl alcohol 3. Condensation of compound 3 with various purine and pyrimidine bases gave the desired nucleosides. All synthesized nucleosides were evaluated for antiviral activity and cellular toxicity. Among them adenine 22 and guanine 23 derivatives showed moderate antiviral activity against HIV-1 and HBV. None of the other compounds showed any significant antiviral activities against HIV-1, HBV, HSV-1 and HSV-2 in vitro up to 100 microM.

Synthesis of 2',3'-dideoxy-2'-fluoro-L-threo-pentofuranosyl nucleosides as potential antiviral agents.

A series of 2',3'-dideoxy-2'-fluoro-L-threo-pentofuranosyl nucleosides has been synthesized as potential antiviral agents. The synthesized compounds were evaluated against HIV-1, HBV, HSV-1, and HSV-2. Among the synthesized analogues, only the cytosine derivative showed moderate antiviral activity against HIV and HBV.