The Plasmodium PI(4)K inhibitor KDU691 selectively inhibits dihydroartemisinin-pretreated Plasmodium falciparum ring-stage parasites.

Malaria control and elimination are threatened by the emergence and spread of resistance to artemisinin-based combination therapies (ACTs). Experimental evidence suggests that when an artemisinin (ART)-sensitive (K13 wild-type) Plasmodium falciparum strain is exposed to ART derivatives such as dihydroartemisinin (DHA), a small population of the early ring-stage parasites can survive drug treatment by entering cell cycle arrest or dormancy. After drug removal, these parasites can resume growth. Dormancy has been hypothesized to be an adaptive physiological mechanism that has been linked to recrudescence of parasites after monotherapy with ART and, possibly contributes to ART resistance. Here, we evaluate the in vitro drug sensitivity profile of normally-developing P. falciparum ring stages and DHA-pretreated dormant rings (DP-rings) using a panel of antimalarial drugs, including the Plasmodium phosphatidylinositol-4-OH kinase (PI4K)-specific inhibitor KDU691. We report that while KDU691 shows no activity against rings, it is highly inhibitory against DP-rings; a drug effect opposite to that of ART. Moreover, we provide evidence that KDU691 also kills DP-rings of P. falciparum ART-resistant strains expressing mutant K13.

Point source inoculation of mesocyclops (Copepoda: Cyclopidae) gives widespread control of Ochlerotatus and Aedes (Diptera: Culicidae) immatures in service manholes and pits in north Queensland, Australia.

This study details the novel application of predacious copepods, genus Mesocyclops, for control of Ochlerotatus tremulus (Theobald) group and Aedes aegypti (L.) mosquito larvae in subterranean habitats in north Queensland, Australia. During June 1997, 50 Mesocyclops sp. 1 were inoculated into one service manhole in South Townsville. Wet season rainfall and flooding in both 1998 and 2000 was responsible for the dispersal of copepods via the underground pipe system to 29 of 35 manholes over an area of 1.33 km2. Significant reductions in Aedes and Ochlerotatus larvae ensued. In these habitats, Mesocyclops and Metacyclops were able to survive dry periods, when substrate moisture content ranged from 13.8 to 79.9%. At the semiarid inland towns of Hughenden and Richmond, cracking clay soil prevents drainage of water from shallow service pits where Oc. tremulus immatures numbered from 292-18,460 per pit. Introduction of Mesocyclops copepods into these sites during May 1999 resulted in 100% control of Oc. tremulus for 18 mo. One uninoculated pit subsequently became positive for Mesocyclops with resultant control of mosquito larvae.

Epidemiological significanceof subterranean Aedes aegypti (Diptera: Culicidae) breeding sites to dengue virus infection in Charters Towers, 1993.

The objective of this study wasto determine the epidemiological significance of subterranean mosquito breeding sites to the 1993 outbreak of dengue fever (type 2) in the northern Queensland town of Charters Towers, Australia. In recent studies on subterranean mosquito breeding, containers such as wells and service manholes have been shown to be important breeding sites to Australia's only dengue vector, Aedes aegypti (L.). This study demonstrates a direct epidemiological association between subterranean breeding sites and dengue virus infection. The mean distance between residents seropositive for dengue 2 and the nearest subterranean container (113 m) was significantly less than for a randomly selected control (191 m), (F = 81.9; df = 1, 478; P < 0.001). Residents positive for dengue 2 antibodies was 2.47 (95% confidence interval 1.88-3.24) times higher for those living within 160 m of a well or service manhole, compared with those residing further away. These findings emphasize the importance of incluuding subterranean water containers in Ae. aegypti surveillance and control programs.

Laboratory evaluation of two native fishes from tropical North Queensland as biological control agents of subterranean Aedes aegypti.

The ability of 2 freshwater fishes, eastern rainbow fish Melanotaenia splendida splendida and fly-specked hardyhead Craterocephalus stercusmuscarum stercusmuscarum, native to North Queensland to prey on immature Aedes aegypti was evaluated under laboratory conditions. The predation efficiency of the 2 species was compared to the exotic guppy, Poecilia reticulata, which is commonly used as a biological control agent of mosquito larvae. Of the 3 fish species tested, M. s. splendida was shown to be the most promising agent for the biological control of Ae. aegypti that breed in wells. Melanotaenia s. splendida consumed significantly greater numbers of immature Ae. aegypti than P. reticulata, irrespective of developmental stage or light conditions. Unlike C s. stercusmuscarum, M. s. splendida could be handled, transported, and kept in captivity for extended periods with negligible mortality. However, M. s. splendida was also an efficient predator of Litoria caerulea tadpoles, a species of native frog found in wells during the dry season. This result may limit the usefulness of M. s. splendida as a biological control agent of well-breeding Ae. aegypti and suggests that predacious copepods, Mesocyclops spp., are more suitable. However, the use of M. s. splendida as a mosquito control agent in containers that are unlikely to support frog populations (e.g., aquaculture tanks and drinking troughs) should be given serious consideration.

Survival of Aedes aegypti (Diptera: Culicidae) eggs in surface and subterranean breeding sites during the northern Queensland dry season.

The effect of a protracted dry season on the viability of Ae. aegypti (L.) eggs was examined in Townsville, northern Queensland, Australia. Eggs were placed in several different surface and subterranean larval habitats; and after four dry season months, only 1-10% of eggs remained viable in the surface and subterranean sites, respectively. Low humidity and predation by Periplaneta americana (L.) were the major causes of egg mortality in eggs in surface sites. P. americana was the most significant cause of egg predation in subterranean breeding sites but fungi, especially Penicillium citrinum Thom, covered egg batches within 15 d. Mycotoxins produced by the spores of P. citrinum are believed to have killed embryonating eggs. The high mortality rate of Ae. aegypti eggs during the dry season suggests that this survival strategy is unlikely to contribute to rapid and successful recolonization of surface sites at the end of the wet season.

The importance of subterranean mosquito habitat to arbovirus vector control strategies in north Queensland, Australia.

In north Queensland, 14 localities were surveyed for mosquito larvae (third and fourth instar) during summer/autumn and winter from 1996 to 1999. Absolute population numbers in subterranean habitats, mainly service manholes and pits (97%) but also some wells, septic tanks, storm drains, and sumps, were expressed as a proportion of total numbers in these sites plus surface sites within a 100-m radius. When correction factors were applied to subterranean samples, the 472,477 larvae mainly of Aedes tremulus (Theobald) group, Aedes notoscriptus (Skuse), and Aedes aegypti (L.) comprised 78% of the total population. In relation to the proportion of the overall immature mosquito population from subterranean habitats (propsub), linear regression coefficients for minimum temperature, relative humidity, and Mesocyclops copepod prevalence were significant for winter data; but for summer, only relative humidity was significant. Linear regression coefficients for Mesocyclops prevalence approached significance (P = 0.061) in summer. When multiple linear regression was used to model propsub, 68% of the variation was accounted for by relative humidity and the prevalence of Mesocyclops. In the drier and cooler towns, increased use of subterranean sites during winter was caused by reduced availability of surface oviposition sites because of the dry season. In the wetter coastal towns, no such restrictions applied and ambient conditions remained more equitable all year round. Mesocyclops were surprisingly common, particularly in these coastal towns. Release of known numbers of Mesocyclops indicated that 3-sweep netting in service manholes was sensitive down to densities of one Mesocyclops per 10 liters, and overall recovery varied from 1 to 4%. In relation to control, service manholes represent a stable habitat for mosquito (7% positive overall) and Mesocyclops populations. If they remained wet, service manholes positive for mosquito immatures or Mesocyclops during summer/autumn had 96% and 85% chance, respectively, of being positive the following winter. Even allowing for the effect of drying, a mosquito-positive manhole had a 79% chance of remaining positive the following winter. In view of the importance of these sites as refuges from adverse ambient conditions, it is proposed that a winter control strategy using Mesocyclops presents a cost-effective control option to reduce the recolonization of surface sites when conditions become more suitable.

A sticky entry-exit trap for sampling mosquitoes in subterranean habitats.

This paper addresses the problems of sampling adult Aedes aegypti and other mosquitoes which utilize subterranean habitats such as wells and service manholes. The sticky pipe trap is a simple device with an adhesive paper insert that can be clipped to the undersides of service manholes to record the entry and exit of adult mosquitoes through the keyhole openings. This trap was 1st used successfully in Townsville, Charters Towers, and Saunders Beach in north Queensland, Australia, in dry seasons of 1996-97 to record usage by 5 species, mainly the Aedes tremulus group and Ae. aegypti, which together comprised 91% of the 1,140 adults collected. Both males and predominantly nulliparous females were recorded exiting manholes, whereas all freshwater-breeding species entering manholes were gravid, presumably seeking oviposition sites.

Calibrated funnel trap for quantifying mosquito (Diptera: Culicidae) abundance in wells.

Our article addresses the problems of sampling and quantifying the abundance of immature Aedes aegypti (L.) and other mosquitoes that use subterranean habitats such as wells and service manholes. Two versions of the funnel trap, Austrap and Vietrap, were extremely sensitive for detecting the presence of Ae. aegypti larvae in wells, and the modified version, the Vietrap, sampled on average 20.5% of 3rd and 4th instars when set overnight for 12 h. For both models, regression analyses between trap catch and absolute abundance indicated high coefficients of determination (R2 = 0.932 and 0.992) enabling accurate direct calibration. The effects of immature age, mosquito species, well diameter, and trap size on catch size were investigated. Because of its simplicity, efficacy, and inexpensiveness, the Vietrap will facilitate the comprehensive assessment of key larval habitats for Ae. aegypti, data critical for the development of effective control programs.

Skeletal anomalies and deformities in patients with deletions of 22q11.

Skeletal anomalies in patients with a 22q11.2 deletion are reported infrequently. We report the skeletal findings in 108 patients with a 22q11.2 deletion, of whom 37 (36%) had a skeletal anomaly. Twenty-two patients (20%) had anomalies of the limbs, 7 of the upper limb, including preaxial or postaxial polydactyly. An anomaly of the lower limb was found in 16 patients, including postaxial polydactyly, clubfoot, severely overfolded toes, and 2-3 toe cutaneous syndactyly. Chest films of 63 patients were examined; 30% of them had abnormal findings, most commonly supernumerary ribs (17%) or a "butterfly" vertebral body (11%). Hypoplastic vertebrae, hemivertebrae, and vertebral coronal clefts were also noted. Thus, skeletal anomalies are not uncommon in patients with a 22q11.2 deletion and may occur more frequently than recognized previously.

Surveillance of the mosquito Aedes aegypti and its biocontrol with the copepod Mesocyclops aspericornis in Australian wells and gold mines.

A survey of the dengue vector mosquito Aedes aegypti was undertaken using funnel traps to detect immature stages (larvae and pupae) in flooded disused mine shafts and wells in Charters Towers, Queensland, Northern Australia. The town has a history of dengue fever since 1885 when goldminers were the first recorded victims. During the latest dengue epidemic in 1993, 2% of the population had laboratory-confirmed dengue virus Type 2, despite source reduction of Ae.aegypti breeding-sites at ground level or above. This led to suspicions that dengue vector Ae.aegypti breeding-sites might be below ground level. When surveyed in March 1994, Ae.aegypti immatures were found in 9/10 wells and 1/6 mine shafts. The water in wells and mines had similar characteristics-except that turbidity was higher in the mines, which more often contained predators of mosquito immatures. The copepod Mesocyclops aspericornis was collected from water in 1/10 wells and 2/6 mine shafts. Laboratory predation trials resulted in 95.5-100% predation by 25 copepods/l on Ae.aegypti first-instar larvae up to 200 larvae/l. Five wells containing Ae.aegypti in the survey were inoculated with fifty indigenous M.aspericornis, and five wells (one positive and four negative in the survey) were left untreated as controls. Nine months later, in December 1994, Ae.aegypti had been eliminated from all five treated wells but all untreated control wells contained Ae.aegypti, except for one well that contained a natural population of M.aspericornis. The role of wells and mines as winter/ dry season refuges of Ae.aegypti in northern Australia is reviewed, and we recommend the use of M.aspericornis as a cost-effective, environmentally acceptable and persistent agent for the sustainable control of Ae.aegypti, especially in inaccessible breeding sites.

Contact lenses in the treatment of patients with overcorrected radial keratotomy.

Thirty-two eyes of 28 patients who underwent radial keratotomy had a hyperopic overcorrection after the surgical procedure. Anisometropia, with associated aniseikonia and depth perception problems were present in many of these patients. The patients were subsequently fit with contact lenses. Conventional methods of fitting contact lenses in these patients were unsuccessful. A trial lens technique is used with the initial lens having a base curve equal to the preoperative keratometry measurements and the power equal to the preoperative spherical equivalent. Superficial neovascularization developed within the radial incisions in 33% of the eyes fitted with soft contact lenses. Gas permeable contact lenses offered patients the best-corrected visual acuity as well as minimal complications. In addition, the anisometropia was minimized, and fluctuating vision was eliminated in cases in which it was a problem.