Efficacy of various larvicides against Aedes aegypti immatures in the laboratory.

We conducted a laboratory study to evaluate the efficacy of control agents against small larvae, large larvae, and pupae of Aedes aegypti to determine an appropriate larvicide regime to employ in emergency dengue control programs. The control agents included Bacillus thuringiensis var. israelensis (Bti), pyriproxyfen (an insect growth regulator), a larvicidal oil, Aquatain AMF (polydimethylsiloxane, a monomolecular film), and temephos at the recommend application dosages and rates. Our results showed that Bti, pyriproxyfen, and temephos were efficacious (100% mortality) against larvae, irrespective of the instar stage, but not against pupae of Ae. aegypti (1.5-7.8% mortality). Aquatain AMF, on the other hand, was very effective at controlling the pupal stage (100% mortality), but had limited efficacy against small larvae (38.0% mortality) and large larvae (78.0% mortality). The larvicidal oil was effective against all immature stages (93.3-100% mortality). Therefore, we concluded that for effectively interrupting the dengue transmission cycle, larvicides that kill the pupal stage (Aquatain AMF or larvicidal oil) should be included in an emergency dengue control program in addition to Bti, pyriproxyfen, or temephos.

A dengue vector surveillance by human population-stratified ovitrap survey for Aedes (Diptera: Culicidae) adult and egg collections in high dengue-risk areas of Taiwan.

Aedes aegypti L. is the primary dengue vector in southern Taiwan. This article is the first report on a large-scale surveillance program to study the spatial-temporal distribution of the local Ae. aegytpi population using ovitraps stratified according to the human population in high dengue-risk areas. The sampling program was conducted for 1 yr and was based on weekly collections of eggs and adults in Kaohsiung City. In total, 10,380 ovitraps were placed in 5,190 households. Paired ovitraps, one indoors and one outdoors were used per 400 people. Three treatments in these ovitraps (paddle-shaped wooden sticks, sticky plastic, or both) were assigned by stratified random sampling to two areas (i.e., metropolitan or rural, respectively). We found that the sticky plastic alone had a higher sensitivity for detecting the occurrence of indigenous dengue cases than other treatments with time lags of up to 14 wk. The wooden paddle alone detected the oviposition of Ae. aegypti throughout the year in this study area. Furthermore, significantly more Ae. aegypti females were collected indoors than outdoors. Therefore, our survey identified the whole year oviposition activity, spatial-temporal distribution of the local Ae. aegypti population and a 14 wk lag correlation with dengue incidence to plan an effectively proactive control.

Screening of dengue virus in field-caught Aedes aegypti and Aedes albopictus (Diptera: Culicidae) by one-step SYBR green-based reverse transcriptase-polymerase chain reaction assay during 2004-2007 in Southern Taiwan.

We carried out virological surveillance of dengue virus (DENV) in field-caught Aedes mosquitoes during 2004-2007 to estimate the monthly prevalence of infected females in dengue high-risk areas of Taiwan. A total of 92,892 Aedes aegypti (43,133 females and 49,759 males) and 79,315 Aedes albopictus (57,319 females and 21,996 males) adults were collected, grouped into 25,654 pools, and processed for virus detection using a one-step SYBR Green-based real-time reverse transcriptase-polymerase chain reaction assay. DENVs were periodically and sympatrically detected in Ae. aegypti females in accordance with major dengue outbreaks and the corresponding dengue serotypes. Only 0.2% of 7628 pools of Ae. aegypti females were positive for DENVs. This resulted in an overall estimated infection rate (maximum likelihood estimation) of 0.970 per 1000 mosquitoes (95% confidence interval [CI] = 0.53-1.65). The total monthly infection rates ranged from 0.50 to 2.23 per 1000 mosquitoes (95% CI = 0.03-10.71). When sampling areas were scaled down to the city level, monthly infection rates increased to 0.73-12.59 (95% CI = 0.06-59.19). Monthly infection rates over all sampling areas and at the city level increased significantly by month. All positive pools were collected in July (one pool), August (two pools), September (one pool), October (three pools), November (four pools), and December (one pool). All four virus serotypes were detected in mosquitoes, which were consistent with dengue serotypes infecting humans in 2004 (DENV-4), 2005 and 2006 (DENV-2 and DENV-3), and 2007 (DENV-1). Our results provide supporting evidence that, in general, DENV infection rates were low in local Aedes mosquito population during 2004-2007 and that transovarial transmission may not be occurring or is occurring at much lower rates than evidenced in some endemic countries.

Drug-drug interactions of silymarin on the perspective of pharmacokinetics.

Silymarin, which is extracted from the milk thistle (Silybum marianum), has been used for centuries for treating hepatic disorders and its hepatoprotective effects have been known for hundreds of years. Silymarin is a mixture of polyphenoic flavonoids, which include silibinin (silybin A and silybin B), isosilyin A and B, silychristin A and B, silydianin and other phenol compounds. The pharmacokinetics of silibinin shows fast absorption and elimination. Silymarin undergoes phase I and phase II metabolism, especially phase II conjugation reactions, it undergoes multiple conjugation reactions, and is primarily excreted into bile and urine. Silymarin has a good safety profile, but little is known regarding its potential for drug interaction. Silymarin has limited effect on the pharmacokinetics of several drugs in vivo; despite silymarin decreasing the activity of cytochrome P-450 (CYPs) enzymes, UDP-glucuronosyltransferase (UGT) enzyme, and reducing P-glycoprotein (P-gp) transport. Health-care practitioners should caution patients against co-administration of silymarin and pharmaceutical drugs.

Hepatobiliary excretion of silibinin in normal and liver cirrhotic rats.

Silibinin is the main biologically active flavonolignan extracted from the seeds and fruits of milk thistle and has potential efficacy in the treatment of liver disease. The aim of the present study was to examine the hepatobiliary excretion of silibinin and its effect on dimethylnitrosamine (DMN)-induced liver cirrhosis. The experiments were divided into five groups: 10, 30, and 50 mg/kg silibinin alone, 30 mg/kg silibinin coadministered with cyclosporin A (CsA), and 50 mg/kg silibinin with liver cirrhosis induced by DMN. The data indicated that silibinin had dose-related pharmacokinetics in the dose ranges of 10 to 50 mg/kg. All of the unconjugated or total (unconjugated + conjugated) silibinin concentrations in the bile were significantly higher than those in plasma at the sampling time points at each dose, suggesting active hepatobiliary excretion. When coadministered with CsA, the area under the concentration versus time curve (AUC) in bile was significantly decreased. This result suggested that the active silibinin efflux might be partially inhibited by P-glycoprotein. In the DMN-induced liver cirrhotic rats, the AUC of plasma unconjugated silibinin was reduced by 53%; however, total silibinin was increased by 182%. These results together suggest that the phase II conjugative reaction of silibinin was blocked by treatment with DNM.

Analysis of silibinin in rat plasma and bile for hepatobiliary excretion and oral bioavailability application.

Silibinin is an herbal ingredient isolated from milk thistle. The aim of this study was to develop a simple liquid chromatographic system to assay silibinin in plasma and bile for pharmacokinetic study. Silibinin was given oral and intravenously. The plasma sample (25 microL) was vortex-mixed with 50 microL of internal standard solution (naringenin 10 microg/mL in acetonitrile) to achieve protein precipitation. Silibinin in the rat plasma and bile was separated using a reversed-phase C18 column (250 mm x 4.6 mm, 5 microm) with a mobile phase of acetonitrile -10 mM monosodium phosphate (pH 5.45 adjusted with orthophosphoric acid) (50:50, v/v) and the flow-rate of 1 mL/min. The UV detection wavelength was 288 nm. The concentration-response relationship from the present method indicated linearity over a concentration range of 0.5-100 microg/mL. Intra- and inter-assay precision and accuracy of silibinin fell well within the predefined limits of acceptability (<15%). An ultrafiltration method was used in this experiment and the protein binding of silibinin was 70.3+/-4.6%. After silibinin administration in rats, the disposition of silibinin in the plasma and bile fluid was due to rapid distribution and equilibration between the blood and hepatobiliary system, and the bile levels of unconjugated silibinin and total silibinin were greater than those in the plasma. The oral bioavailability of silibinin in rats was estimated to be 0.73%.

Effect of silibinin on the pharmacokinetics of pyrazinamide and pyrazinoic acid in rats.

Pyrazinamide (PZA) is widely used in combination with other drugs in chemotherapy for tuberculosis. However, the dose-related liver injury is the main adverse effect of PZA and its metabolite [pyrazinoic acid (PA)]. Silibinin is the main flavonoid extracted from milk thistle (Silybum marianum), and it displays hepatoprotective properties. This study investigates the pharmacokinetics of PZA and PA and their interaction with silibinin in rats. The parallel study design was divided into six groups: PZA alone, PZA + long-term silibinin exposure, PZA + concomitant short-term silibinin exposure, PA alone, PA + long-term silibinin exposure, and PA + concomitant short-term silibinin exposure groups. The results indicate that the distribution ratio of PZA from bile to blood [area under the curve (AUC)(bile)/AUC(blood)] in the PZA + long-term silibinin exposure and PZA + concomitant short-term silibinin exposure groups was also not significantly different when compared with the PZA alone group. However, the bile-to-blood distribution ratio of PA was significantly decreased in the PA + long-term silibinin exposure and the PA + concomitant short-term silibinin exposure groups. On PZA administration, the blood, but not bile, levels of PA were markedly increased in the PZA + long-term silibinin exposure and PZA + concomitant short-term silibinin exposure groups, but the bile-to-blood ratio of PA was decreased. These results suggest that the excretion pathway of PA may be blocked by silibinin through xanthine oxidase and hepatobiliary excretion.

Regulation of hepatobiliary excretion of sinomenine by P-glycoprotein in Sprague-Dawley rats.

Sinomenine, an herbal ingredient isolated from Sinomenium acutum, is used for the amelioration of arthritis. Using microdialysis and a specially constructed hepato-duodenal shunt probe, the present study investigated the pharmacokinetics of sinomenine in rat blood and bile and the effects of P-glycoprotein modulation and cytochrome P450 inhibition. The results indicated that the pharmacokinetics of sinomenine in rat blood appeared to be dose dependent in the 3 to 30 mg/kg range. The disposition of sinomenine in the bile exhibited a slow elimination phase, reaching a peak concentration in 20-40 min following intravenous administration. The area under the concentration versus time curves (AUC's) for sinomenine in the bile were significantly greater than those in the blood at dosages of 3, 10, and 30 mg/kg with the blood-to-bile distribution ratios (k = AUC(bile) / AUC(blood)) being 3.85 +/- 0.29 and 3.52 +/- 0.28 at 10 and 30 mg/kg, respectively, indicating active hepatobiliary excretion. Coadministration with 20 mg/kg of cyclosporin A 10 min prior to sinomenine administration resulted in a significant reduction of the bile AUC's for the dosages of 10 and 30 mg/kg., resulting in the bile/blood distribution ratio being significantly reduced to 0.47 +/- 0.05 and 0.49 +/- 0.05, respectively. On the other hand, proadifen treatment increased both the blood and bile AUC's, resulting in insignificant effects on the blood-to-bile distribution ratios. In conclusion, our results indicated that sinomenine underwent active hepatobiliary elimination which may be regulated by the P-glycoprotein and that P-450 was likely involved in its metabolism.