Novel Histopathological and Molecular Effects of Natural Compound Pellitorine on Larval Midgut Epithelium and Anal Gills of Aedes aegypti

The yellow fever mosquito, Aedes aegypti, is a vector for transmitting dengue fever and yellow fever. In this study, we assessed the histopathological and molecular effects of pellitorine, an isobutylamide alkaloid, on the third instar of Ae. aegypti larvae. At 5 mg/l concentration of pellitorine, the whole body of the treated larvae became dark in color, particularly damaged thorax and abdominal regions. Pellitorine was targeted mainly on midgut epithelium and anal gills, indicating variably dramatic degenerative responses of the midgut through a sequential epithelial disorganization. The anterior and posterior midgut was entirely necrosed, bearing only gut lumen residues inside the peritrophic membranes. Pellitorine caused comprehensive damage of anal gill cells and branches of tracheole and debris was found in hemolymph of the anal gills. RT-PCR analysis indicates that the compound inhibited gene expression encoding V-type H⁺-ATPase and aquaporine 4 after treatment with 2.21 mg/l pellitorine. These results verify that pellitorine merits further study as a potential larvicide with a specific target site and a lead molecule for the control of mosquito populations.     

Epigenomics: novel aspect of genomic regulation

An International Symposium on Epigenomics took place at Yonsei University, Korea in December, 2006. The meeting brought to light new aspects of genome regulation by DNA and protein modification.     

Rescheduling Behavioral Subunits of a Fixed Action Pattern by Genetic Manipulation of Peptidergic Signaling

The ecdysis behavioral sequence in insects is a classic fixed action pattern (FAP) initiated by hormonal signaling. Ecdysis triggering hormones (ETHs) release the FAP through direct actions on the CNS. Here we present evidence implicating two groups of central ETH receptor (ETHR) neurons in scheduling the first two steps of the FAP: kinin (aka drosokinin, leucokinin) neurons regulate pre-ecdysis behavior and CAMB neurons (CCAP, AstCC, MIP, and Bursicon) initiate the switch to ecdysis behavior. Ablation of kinin neurons or altering levels of ETH receptor (ETHR) expression in these neurons modifies timing and intensity of pre-ecdysis behavior. Cell ablation or ETHR knockdown in CAMB neurons delays the switch to ecdysis, whereas overexpression of ETHR or expression of pertussis toxin in these neurons accelerates timing of the switch. Calcium dynamics in kinin neurons are temporally aligned with pre-ecdysis behavior, whereas activity of CAMB neurons coincides with the switch from pre-ecdysis to ecdysis behavior. Activation of CCAP or CAMB neurons through temperature-sensitive TRPM8 gating is sufficient to trigger ecdysis behavior. Our findings demonstrate that kinin and CAMB neurons are direct targets of ETH and play critical roles in scheduling successive behavioral steps in the ecdysis FAP. Moreover, temporal organization of the FAP is likely a function of ETH receptor density in target neurons.     

Vapor phase toxicity of plant essential oils to Cadra cautella (Lepidoptera: Pyralidae)

The toxicities of 44 plant essential oils against larvae of Cadra cautella (Walker) were examined using direct contact and vapor phase toxicity bioassays and compared with the lethal activity of chlorpyrifos-methyl, diazinon, dichlorvos, and fenthion, four widely used organophosphorus insecticides. Responses varied according to plant material used and exposure dose and time. In a filter paper contact toxicity bioassay, potent toxicity was produced from buchu leaf, niaouli, and rosemary oils at 2.4 mg/cm2 and armoise, cypress, galbanum, and mace oils at 4.7 mg/cm2. In vapor phase toxicity bioassays with larvae, cypress, galbanum, niaouli, and rosemary oils were much more effective in closed containers than in open containers, indicating that the lethal effects of these oils were largely because of action in the vapor phase. As judged by 24-h LC50 values, potent fumigant action was observed with niaouli oil (64.7 mg/liter air) and rosemary oil (64.6 mg liter/air). Cypress and galbanum oils exhibited weak fumigant activity. These essential oils were less active than dichlorvos (0.86 mg/liter air). Little or no fumigant action was observed with chlorpyrifos-methyl, diazinon, and fenthion. Essential oils described herein, particularly niaouli and rosemary oils, merit further study as potential larvicides for the control of C. cautella.     

Kolaviron inhibits dimethyl nitrosamine-induced liver injury by suppressing COX-2 and iNOS expression via NF-κB and AP-1

AimsKolaviron, a bioflavonoid isolated from the seeds of Garcinia kola has been reported to possess anti-inflammatory, antioxidant, antigenotoxic and hepatoprotective activities in model systems via multiple biochemical mechanisms. The present study investigated the possible molecular mechanisms underlying the hepatoprotective effects of kolaviron.Main methodsBiomarkers of hepatic oxidative injury, histological and immunohistochemical techniques were used. In addition, the protein expression levels of cyclooxygenase (COX-2) and inducible nitric oxide synthase (iNOS) were evaluated by western blotting while DNA-binding activities of nuclear factor kappa B (NF-κB) and activator protein-1 (AP-1) were determined by electrophoretic mobility shift assay.Key findingsKolaviron administered orally at doses of 100 and 200 mg/kg for 7 days significantly lowered the activities of serum transaminases and γ-glutamyl tranferase induced by single intraperitoneal administration of dimethyl nitrosamine (DMN) (20 mg/kg) and preserved the integrity of the hepatocytes. Also, kolaviron at both doses reduced the DMN induced elevated hepatic levels of malondialdehyde and reversed DMN mediated decrease in hepatic glutathione. The hepatoprotective effect of kolaviron was compared to that of curcumin, an established hepatoprotective agent. Kolaviron inhibited the DMN induced expression of COX-2 and iNOS. Immunohistochemical staining of rat liver verified the inhibitory effect of kolaviron on DMN-induced hepatic COX-2 expression. Furthermore, kolaviron abrogated DMN induced binding activity of NF-κB as well as AP-1.SignificanceThe ability of kolaviron to inhibit COX-2 and iNOS expression through down regulation of NF-κB and AP-1 DNA binding activities could be a mechanism for the hepatoprotective properties of kolaviron.     

Morphological and Molecular Characterization of the Causal Agent of Downy Mildew on Quinoa (<Emphasis Type="Italic">Chenopodium quinoa</Emphasis>)

Downy mildew is an economically important and widespread disease in quinoa (Chenopodium quinoa) growing areas. Although in many studies Peronospora farinosa is most commonly regarded as the causal agent of the disease, identification and classification of the pathogen remain still uncertain due to its taxonomic confusion. Thirty-six Peronospora isolates from quinoa with different geographic origins including Argentina, Bolivia, Denmark, Ecuador, and Peru were morphologically and molecularly compared with Peronospora species from other Chenopodium species. The morphology of three herbarium specimens was similar to that of P. variabilis, which originated from C. album, characterized by flexuous to curved ultimate branchlets and pedicellated conidia. Phylogenetic analysis based on ITS rDNA sequences also placed the quinoa pathogen within the same clade as P. variabilis. Within the ITS rDNA sequences of the quinoa pathogens, two base substitutions were found, which separated the majority of the Danish isolates from isolates from South America, but no sequence difference was found among the isolates from different cultivars of quinoa. The present results indicate that the pathogen responsible for the quinoa downy mildew is identical to Peronospora variabilis and that it should not be lumped with P. farinosa as claimed previously by most studies.