Fatally impaired glucose digestion by propylene glycol in Aedes aegypti (Diptera: Culicidae) and co-formulation with terpenoids for enhancing attractive toxic sugar baits.

Propylene glycol (PG) demonstrates greater efficacy than other sugar polyols. However, the attributes it confers for toxicity and possible co-formulation with other ingredients are unknown. To evaluate this, α-glucosidase and glucose oxidase reactions were performed in Aedes aegypti (L.) (Diptera: Culicidae) to categorize if PG behaves similarly to prior studied sugar alcohols. A combination of no-choice and choice assays was used to determine effective ratios of PG and sucrose, competitiveness against a control of 10% sucrose, and whether mosquitoes recovered from PG consumption. The final trials included ß-cyclodextrin encapsulated cinnamon leaf oil, clove stem oil, patchouli oil, garlic oil, cedarwood oil, and papaya seed oil formulated with 5% sucrose + 5% PG. PG functioned as a linear competitive inhibitor of α-glucosidase. The efficacy of PG was synergized by co-ingestion with equivalent ratios of sucrose. Unlike the high diuretic response to other sugar alcohols, PG resulted in diminished excretion regardless of being co-formulated with sucrose or terpenoids. PG is not especially competitive against unadulterated sugar meals but is likewise not clearly repellent. Although mosquitoes did not recover from ingestion of the glycol meals, there was no indication that mortality would continue to accumulate once the treatments were removed. Of the terpenoids tested, cinnamon and patchouli caused ~50% or less mortality; garlic, cedarwood, and clove caused 80-90% mortality; and papaya seed caused 100% mortality, exceeding all other test groups and the formulation blank. PG is a useful supporting ingredient in attractive toxic sugar bait formulations with flexibility in formulation.

A CASE REPORT ON PRODUCT ROTATION TO MANAGE SEVERE LYSINIBACILLUS SPHAERICUS RESISTANCE IN CULEX PIPIENS FROM SALT LAKE CITY, UTAH.

The Salt Lake City Mosquito Abatement District (SLCMAD) detected a 20,000-fold resistance to Lysinibacillus sphaericus (Lsph) in Culex pipiens occurring in catch basins of Salt Lake City during 2016. In response, SLCMAD suspended use of Lsph and rotated use of spinosyn and s-methoprene products for the next three years. At the end of the third year, Lsph was evaluated again and efficacy similar to susceptible colony strains. During the second year of Lsph use, technicians observed lack of control of larvae at some urban sites. Bioassays performed during 2021 showed recurrence of some resistance to Lsph to varying degrees across SLCMAD urban areas. The rapidity with which resistant phenotypes reemerged clarifies that SLCMAD cannot in the near future rely on repeated use of Lsph, even after suspending use for three years and using within-season product rotations. Prior reports in other research groups have found long-term selection to Lsph, as is the case at SLCMAD, to not regress in spite of halting use of the products. However, our findings offer some optimism that regression may be relatively quick. More operational review is needed, and future work should characterize resistance alleles in field populations. Collectively, there is a lack of concrete data supporting the prevailing assumptions from adjacent industries that were adopted into mosquito abatement. We provide this short note as additional guidance for mosquito and vector control districts weighing options to remediate Lsph resistance.

Diuresis and α-glucosidase inhibition by erythritol in Aedes aegypti (Diptera: Culicidae) and viability for efficacy against mosquitoes.

BACKGROUND: Sugar alcohols, such as erythritol, are low-impact candidates for attractive toxic sugar baits (ATSB) to kill mosquitoes. To determine whether erythritol has a viable future in ATSB formulations, a suite of assays was conducted to diagnose toxicity mechanisms and starvation effects on mortality in Aedes aegypti (L.) as a model system. METHODS: We measured general carbohydrate load, glucosidase levels, and free glucose in intoxicated adult mosquitoes to observe whether sugar digestion was impaired. We assayed the effects of sugar combinations with erythritol on larvae and adults. To measure erythritol effects when mosquitoes were not resource-deprived, additional assays manipulated the prior starvation status. RESULTS: Up to 50,000 ppm of erythritol in water had no effect on larvae within 72 h, but an ammonia spike indicated diuresis in larvae as early as 4 h (F8,44 = 22.50, P < 0.0001) after sucrose/erythritol combinations were added. Adult consumption of erythritol was diuretic regardless of the sugar pairing, while sucrose and erythritol together generated above 80% mortality (F2,273 = 33.30, P < 0.0001) alongside triple the normal excretion (F5,78 = 26.80, P < 0.0004). Glucose and fructose paired individually with erythritol had less mortality, but still double the fecal excretion. When ingesting erythritol-laced meals, less sugar was detected in mosquitoes as compared to after sucrose meals (χ2 = 12.54, df = 1, P = 0.0004). CONCLUSIONS: Data showed that erythritol is a linear competitive inhibitor of α-glucosidase, marking it as a novel class of insecticide in the current research climate. However, the efficacy on larvae was null and not persistent in adult mosquitoes when compared across various starvation levels. Despite significant diuresis, the combined effects from erythritol are not acute enough for vector control programs considering ATSB against mosquitoes.

Do it yourself: 3D-printed miniature CDC trap for adult mosquito (Diptera: Culicidae) surveillance.

The central component of mosquito and vector surveillance programs globally is the adult mosquito trap, which is intended to collect host-seeking mosquitoes. The miniature CDC trap is a widely distributed trap style in part due to its relative affordability and compact nature. Despite already being a simple trap, in-house production methods, such as 3D printing, could improve the accessibility of the CDC trap by eliminating some of the supply chain variables. We present here several trials with the Salt Lake City (SLC) trap, a three-dimensional (3D) printed trap design. Functional assessments were made on secondary components and found no statistically significant differences when comparing CO2 line height (above vs. below fan), battery types (sealed lead acid vs. USB battery pack), and trap body collection shape (funnel body vs. simple/straight body). The SLC trap was compared directly to a commercial equivalent, the ABC trap, with comparative assessment on species diversity and evenness in collections and found to be statistically equivalent on all metrics. Methods also detail an accompanying optional transport system for a pressurized CO2/regulator set-up, should a practitioner elect not to use dry ice. Our final design is presented here with the publicly published stereolithography (STL) files and a detailed outline of the transport container system. Alternative models are available for in-house manufacture of mosquito traps, and we contribute these designs in an effort to stimulate further growth in vector surveillance.

Do It Yourself: A Modernized Gravid Trap Design for Mosquito Surveillance.

Gravid traps have become a common and frequently essential surveillance tool for parous Culex spp. vectors of West Nile virus and other encephalitis-causing pathogens. The recent closing of BioQuip Products Inc., an entomological supply company, has jeopardized the commercial availability of gravid traps. The Salt Lake City Mosquito Abatement District presents herein a template for making your own gravid trap, but with some modernizations for quieter fans and longer lasting, light weight, lithium battery packs. At the time of writing, the materials cost for the fan ($14 USD), toolbox ($13), cables ($9), ABS pipe ($2.50), aluminum brackets ($10), catch container with lid ($9), trap net ($10), USB battery pack ($35) and the negligible amount of 3D-printed filament ($2), is approximately half the cost (not including labor) of the formerly available commercial model. Additionally, performance validation in the laboratory (t4,9 = 0.1191, P < 0.9109) and within two field sites (χ2 = 0.107, P < 0.744) demonstrated no significant differences in collections of gravid Culex pipiens. We do not present an overhaul of the previous gravid trap blueprint, but the quality-of-life updates to the trap design, the feasibility of in-house manufacture, and the mirrored collection efficacy to the commercial model can allow improved maintenance of gravid trap surveillance networks without a commercial supplier.

Evaluation of An Aerial Application of Duet Hd® Against Aedes Dorsalis and Culex Tarsalis in Rural Habitats of the Great Salt Lake, Utah.

The Salt Lake City Mosquito Abatement District (SLCMAD) has been conducting aerial applications using an organophosphate insecticide against adult mosquitoes for several decades. In order to evaluate a potential rotation product, aerial applications of Duet HD™, a pyrethroid, were conducted under operational conditions against wild populations of Aedes dorsalis and Culex tarsalis and against colony strains of Cx. pipiens and Cx. quinquefasciatus. The erratic wind patterns of the greater Salt Lake area did not prevent sufficient droplet deposition flux at 9 monitoring locations spread across a 5,120-acre (2,072 ha) spray block within rural habitats. Three separate aerial application trials showed great efficacy against Ae. dorsalis. In contrast, Cx. tarsalis exhibited inconsistent treatment-associated mortalities, suggesting the presence of less susceptible or resistant field populations as a result of spillover from agricultural or residential pyrethroid usage. Bottle bioassays to diagnose pyrethroid resistance using field-collected Cx. tarsalis indicated that some populations of this species, especially those closest to urban edges, failed to show adequate mortality in resistance assays. Despite challenging weather conditions, Duet HD worked reasonably well against susceptible mosquito species, and it may provide a crucial role as an alternative for organophosphate applications within specific and sensitive areas. However, its area-wide adoption into control applications by the SLCMAD could be problematic due to reduced impacts on the most important arboviral vector species, Cx. tarsalis, in this area. This study demonstrates the importance of testing mosquito control products under different operational environments and against potentially resistant mosquito populations by municipal mosquito control districts.

Do it yourself: fabricating and evaluating a mosquito (Diptera: Culicidae) blood-feeding device to replace a commercial option.

Tools for rearing hematophagous insects, such as mosquitoes (Diptera: Culicidae), in an insectary are essential for research and operational evaluations in vector biology and control. There is an abundance of low-cost options for practitioners without conventional infrastructure. However, few midrange options exist that provide a balance of efficiency and low material waste. We present here a reproducible design for an electrically powered blood-feeding device that offers long-term reusability, low material waste, and customizability for different species or experiments. The limitation is the requirement for electricity, but the gain is a simple, low-skill device that can be modified as needed. To validate the design, assessments of feeding angle and blood-feeding success were compared between the Salt Lake City Mosquito Abatement District artificial membrane feeder (SLAM) and a commercial system (Hemotek). Engorgement in Aedes aegypti (80-90%), Culex pipiens (50-80%), and Culex tarsalis (30-75%) was similar between the 2 units, resulting in nearly identical fecundity outcomes between devices. Additionally, 45° angles were more successful, generally, than presenting the feeders flat or vertical to the mosquitoes (df3,48, P = 1.014 × 10-15). This angle is simple to present with the SLAM device. Materials for in-house reproduction of the SLAM system are now widely available in regions with access to e-commerce and shipped goods. This results in a device schematic that should fit well into a relatively modular, do-it-yourself paradigm where the practitioner needs only to assemble some materials without complex engineering. This article provides schematics, cost comparison, and validation of the in-house-made SLAM system.

Enhancing toxic sugar meals against Aedes aegypti (Diptera: Culicidae) by adulterating with erythritol in combination with other active ingredients.

Attractive toxic sugar baits (ATSBs) are an underexploited method for mosquito control. For ATSBs to be more widely accepted, demonstrably effective ingredients need to be verified. We investigated erythritol as a toxic additive in sugar meals against Aedes aegypti (L.) for potential future use in ATSBs. Erythritol is a sugar alcohol that is commonly used as a sugar substitute, while also being toxic to mosquitoes. Our studies tested formulations of erythritol, sucrose, and blends of both. Secondary investigations included combinations with the active ingredients Bacillus thuringiensis israelensis, spinosyn, and boric acid. Adult Ae. aegypti were separated into test groups and provided various combinations. Formulations containing erythritol, with or without another toxicant, exhibited 90% mortality within 72 h of observation (P = 0.03192). Additionally, erythritol appeared more effective when combined with sucrose in a 1:1 ratio (5% concentration each). This combination showed a 24% and 85% increase in mortality when combined with boric acid and Bti, respectively, at 48 h compared with equivalent groups containing only 10% sucrose. Erythritol appears to kill adult mosquitoes, even in relatively low concentrations, without another toxicant being required. However, erythritol also effectively enhances kill of main ingredient toxicants such as boric acid and Bti, showing a supporting role. The low concentration of erythritol needed to provide significant kill, its ability to fill in as both a sugar base and toxicant, and its ability to be safely handled by humans makes erythritol a strong candidate for use as a supporting ingredient in future bait formulations.

Infrared light sensors permit rapid recording of wingbeat frequency and bioacoustic species identification of mosquitoes.

Recognition and classification of mosquitoes is a critical component of vector-borne disease management. Vector surveillance, based on wingbeat frequency and other parameters, is becoming increasingly important in the development of automated identification systems, but inconsistent data quality and results frequently emerge from different techniques and data processing methods which have not been standardized on wingbeat collection of numerous species. We developed a simple method to detect and record mosquito wingbeat by multi-dimensional optical sensors and collected 21,825 wingbeat files from 29 North American mosquito species. In pairwise comparisons, wingbeat frequency of twenty six species overlapped with at least one other species. No significant differences were observed in wingbeat frequencies between and within individuals of Culex quinquefasciatus over time. This work demonstrates the potential utility of quantifying mosquito wingbeat frequency by infrared light sensors as a component of an automated mosquito identification system. Due to species overlap, wingbeat frequency will need to integrate with other parameters to accurately delineate species in support of efficient mosquito surveillance, an important component of disease intervention.

No Evidence That Salt Water Ingestion Kills Adult Mosquitoes (Diptera: Culicidae).

Various products and insecticides are available that purport to reduce wild populations of adult mosquitoes. Recently, several manufacturers and general public comments on the internet have promoted devices that claim that ingestion of salt will significantly reduce populations of wild mosquitoes to near zero; there are no known scientific efficacy data that support these claims. We tested the survival of nine mosquito species of pest and public health importance across four adult diets: Water Only, Sugar Water Only (8.00%), Salt Water Only (1.03%), and Sugar + Salt Water. Species included the following: Aedes aegypti (L.), Aedes albopictus (Skuse), Aedes dorsalis (Meigen), Aedes notoscriptus (Skuse), Aedes vigilax (Skuse), Anopheles quadrimaculatus (Say), Culex pipiens (L.), Culex quinquefasciatus (Say), and Culex tarsalis (Coquillett). Male and female mosquitoes were placed in cages and allowed to feed on liquid diets under controlled environmental conditions for 1 wk. For seven of the nine species, adult survival was significantly higher in the presence (Sugar Water, Sugar + Salt Water) versus the absence (Water Only, Salt Only) of sugar, with no indication that salt had any effect on survival. Anopheles quadrimaculatus showed intermediate survival in Sugar + Salt to either Sugar Only or no sugar diets, whereas Aedes dorsalis showed low survival in Salt Only versus other diets. Based on our data and coupled with the fact that mosquitoes have physiological and behavioral adaptations that allow them to avoid or process excess salt (as found in blood meals), we conclude that there is no scientific foundation for salt-based control methods of mosquitoes.

Trap Comparison for Surveillance of the Western Tree Hole Mosquito (Diptera: Culicidae).

The western tree hole mosquito, Aedes sierrensis (Ludlow), is a common nuisance mosquito and vector of Dirofilaria immitis (Leidy), the etiologic agent of dog heartworm, in western North America. Here, we compare weekly mosquito collections made with Mosquito Magnet (MM) traps, Biogents Sentinel (BGS) traps, and Biogents Bowl (BGS Bowl) traps set in Salt Lake City, UT, from the start of June to mid-August 2017. We found the number of mosquitoes decreased with rainfall and temperature independently of trap type. The highest number of mosquitoes were caught by BGS traps baited with carbon dioxide (CO2) and BG lure, which collected 62% (n = 422) of all mosquitoes, followed by the MM at 31% (n = 213), and both the BGS and BG Bowl with BG lure had 3.5% (n = 24) each. Aedes sierrensis females were caught weekly at similar densities (mean ±â€…SD) in BGS with CO2 and lure (1.17 ±â€…2.93) and the MM (1.17 ±â€…2.66) traps during the study period. Given that BGS with CO2 and lure traps have several operational advantages over MM traps, including a quicker setup, smaller size, and lower cost, we consider BGS with CO2 and lure traps as the best suited surveillance tool to detect and remove Ae. sierrensis in the western United States and similar settings throughout North America.

High Resistance to Bacillus sphaericus and Susceptibility to Other Common Pesticides in Culex pipiens (Diptera: Culicidae) from Salt Lake City, UT.

Biorational mosquito larvicides based on microbial organisms and insect growth regulators (IGRs) have played a vital role in integrated mosquito control, particularly since the invasion of West Nile virus to the United States in 1999. Products that are formulated with technical powder of the bacterium, Bacillus sphaericus Neide (recently Lysinibacillus sphaericus Meyer and Neide), are among the ones that have been extensively applied to combat Culex and other mosquito species. Due to the simplicity of the binary toxins, resistance to this pesticide in laboratory and field populations of Culex pipiens L. complex has occurred globally since 1994. A Cx. pipiens population with a high level of resistance to B. sphaericus (VectoLex WDG) was identified in Salt Lake City, UT, in September 2016. The resistance ratios in this population were 20,780.0- and 23,926.9-fold at LC50 and LC90, respectively, when compared with a susceptible population of a laboratory reference colony of the same species. This B. sphaericus-resistant population remained mostly susceptible to other commonly used pesticides to control arthropods of public health and urban significance, including ones based on microbial organisms (Bacillus thuringiensis subsp. israelensis, spinosad, spinetoram, abamectin), IGRs (pyriproxyfen, methoprene, diflubenzuron, novaluron), organophosphate (temephos), neonicotinoid (imidacloprid), phenylpyrazole (fipronil), oxadiazine (indoxacarb), and pyrethroid (permethrin). Results are discussed according to the modes of action of the pesticides tested, and suggestions are made to manage B. sphaericus-resistant mosquito populations.

Profiles of Amino Acids and Acylcarnitines Related with Insecticide Exposure in Culex quinquefasciatus (Say).

Culex quinquefasciatus Say is a vector of many pathogens of humans, and both domestic and wild animals. Personal protection, reduction of larval habitats, and chemical control are the best ways to reduce mosquito bites and, therefore, the transmission of mosquito-borne pathogens. Currently, to reduce the risk of transmission, the pyrethroids, and other insecticide groups have been extensively used to control both larvae and adult mosquitoes. In this context, amino acids and acylcarnitines have never been associated with insecticide exposure and or insecticide resistance. It has been suggested that changes in acylcarnitines and amino acids profiles could be a powerful diagnostic tool for metabolic alterations. Monitoring these changes could help to better understand the mechanisms involved in insecticide resistance, complementing the strategies for managing this phenomenon in the integrated resistance management. The purpose of the study was to determine the amino acids and acylcarnitines profiles in larvae of Cx. quinquefasciatus after the exposure to different insecticides. Bioassays were performed on Cx. quinquefasciatus larvae exposed to the diagnostic doses (DD) of the insecticides chlorpyrifos (0.001 µg/mL), temephos (0.002 µg/mL) and permethrin (0.01 µg/mL). In each sample, we analyzed the profile of 12 amino acids and 31 acylcarnitines by LC-MS/MS. A t-test was used to determine statistically significant differences between groups and corrections of q-values. Results indicates three changes, the amino acids arginine (ARG), free carnitine (C0) and acetyl-carnitine (C2) that could be involved in energy production and insecticide detoxification. We confirmed that concentrations of amino acids and acylcarnitines in Cx. quinquefasciatus vary with respect to different insecticides. The information generated contributes to understand the possible mechanisms and metabolic changes occurring during insecticide exposure.

Reemergence of St. Louis Encephalitis Virus, California, 2015.

St. Louis encephalitis virus infection was detected in summer 2015 in southern California after an 11-year absence, concomitant with an Arizona outbreak. Sequence comparisons showed close identity of California and Arizona isolates with 2005 Argentine isolates, suggesting introduction from South America and underscoring the value of continued arbovirus surveillance.

Endovascular treatment of vasculogenic erectile dysfunction.

The treatment of erectile dysfunction (ED) has been a fascination involving multiple medical specialities over the past century with urologic, cardiac and surgical experts all contributing knowledge toward this multifactorial disease. With the well-described association between ED and cardiovascular disease, angiography has been utilized to identify vasculogenic impotence. Given the success of endovascular drug-eluting stent (DES) placement for the treatment of coronary artery disease, there has been interest in using this same technology for the treatment of vasculogenic ED. For men with inflow stenosis, DES placement to bypass arterial lesions has recently been reported with a high technical success rate. Comparatively, endovascular embolization as an approach to correct veno-occlusive dysfunction has produced astonishing procedural success rates as well. However, after a thorough literature review, arterial intervention is only recommended for younger patients with isolated vascular injuries, typically from previous traumatic experiences. Short-term functional outcomes are less than optimal with long-term results yet to be determined. In conclusion, the hope for a minimally invasive approach to ED persists but additional investigation is required prior to universal endorsement.

Detection of eastern equine encephalomyelitis virus RNA in North American snakes.

The role of non-avian vertebrates in the ecology of eastern equine encephalomyelitis virus (EEEV) is unresolved, but mounting evidence supports a potential role for snakes in the EEEV transmission cycle, especially as over-wintering hosts. To determine rates of exposure and infection, we examined serum samples from wild snakes at a focus of EEEV in Alabama for viral RNA using quantitative reverse transcription polymerase chain reaction. Two species of vipers, the copperhead (Agkistrodon contortrix) and the cottonmouth (Agkistrodon piscivorus), were found to be positive for EEEV RNA using this assay. Prevalence of EEEV RNA was more frequent in seropositive snakes than seronegative snakes. Positivity for the quantitative reverse transcription polymerase chain reaction in cottonmouths peaked in April and September. Body size and sex ratios were not significantly different between infected and uninfected snakes. These results support the hypothesis that snakes are involved in the ecology of EEEV in North America, possibly as over-wintering hosts for the virus.

Winter biology of wetland mosquitoes at a focus of eastern equine encephalomyelitis virus transmission in Alabama, USA.

At temperate latitudes, vectors and pathogens must possess biological mechanisms for coping with cold temperatures and surviving from one transmission season to the next. Mosquitoes that overwinter in the adult stage have been proposed as winter maintenance hosts for certain arboviruses. In the cases of West Nile virus (family Flaviviridae, genus Flavivirus) and St. Louis encephalitis virus (family Flaviviridae, genus Flavivirus), discovery of infected overwintering females lends support to this hypothesis, but for other arboviruses, in particular Eastern equine encephalomyelitis virus (family Togaviridae, genus Alphavirus, EEEV), overwintering of the virus in mosquito hosts as not been demonstrated. In the current study, we collected overwintering mosquitoes from a focus of EEEV transmission in the southeastern United States to determine whether mosquitoes serve as winter maintenance hosts for EEEV and to document overwintering biologies of suspected vectors. No virus was detected via reverse transcription-polymerase chain reaction of > 500 female mosquitoes collected during three winters. Investigation into the winter biologies indicated that Anopheles punctipennis (Say), Culex erraticus (Dyar & Knab), Culex peccator Dyar & Knab, and Uranotaenia sapphirina (Osten Sacken) overwinter as females. Females of these species were collected from hollow trees and emergence traps placed over ground holes. Southern magnolia, Magnolia grandiflora L., trees were preferred overwintering sites of culicine mosquitoes. Emergence from underground overwintering sites peaked in mid-March, when air temperatures reached 18-22 degrees C, and the first blood-engorged females of Cx. erraticus and Cx. peccator were collected during this same period. Blood-fed Culex territans Walker females were collected as early as mid-February. This work provides insight into the overwintering biologies of suspected virus vectors at a site of active EEEV transmission and provides limited evidence against the hypothesis that EEEV persists through intertransmission periods in overwintering mosquitoes.

Phylogenetic analysis of eastern equine encephalitis virus isolates from Florida.

Florida has the highest degree of endemicity for eastern equine encephalitis virus (EEEV) of any state in the United States and is the only state with year-round transmission of EEEV. To further understand the viral population dynamics in Florida, the genome sequence of six EEEV isolates from central Florida were determined. These data were used to identify the most polymorphic regions of the EEEV genome from viruses isolated in Florida. The sequence of these polymorphic regions was then determined for 18 additional Florida isolates collected in four geographically distinct regions over a 20-year period. Phylogenetic analyses of these data suggested a rough temporal association of the Florida isolates, but no clustering by region or by source of the isolate. Some clustering of northeastern isolates with Florida isolates was seen, providing support for the hypothesis that Florida serves as a reservoir for the periodic introduction of EEEV into the northeastern United States.

Glomerular-specific protein kinase C-ß-induced insulin receptor substrate-1 dysfunction and insulin resistance in rat models of diabetes and obesity.

Insulin resistance has been associated with the progression of chronic kidney disease in both diabetes and obesity. In order to determine the cellular mechanisms contributing to this, we characterized insulin signaling in renal tubules and glomeruli during diabetic and insulin-resistant states using streptozotocin-diabetic and Zucker fatty-insulin-resistant rats. Compared with nondiabetic and Zucker lean rats, the insulin-induced phosphorylation of insulin receptor substrate-1 (IRS1), Akt, endothelial nitric oxide synthase, and glycogen synthase kinase 3α were selectively inhibited in the glomeruli but not in the renal tubules of both respective models. Protein, but not mRNA levels of IRS1, was decreased only in the glomeruli of streptozotocin-diabetic rats likely due to increased ubiquitination. Treatment with the protein kinase C-ß inhibitor, ruboxistaurin, enhanced insulin actions and elevated IRS1 expression. In glomerular endothelial cells, high glucose inhibited the phosphorylation of Akt, endothelial nitric oxide synthase, and glycogen synthase kinase 3α; decreased IRS1 protein expression and increased its association with ubiquitin. Overexpression of IRS1 or the addition of ruboxistaurin reversed the inhibitory effects of high glucose. Thus, loss of insulin's effect on endothelial nitric oxide synthase and glycogen synthase kinase 3α activation may contribute to the glomerulopathy observed in diabetes and obesity.

Isolation of genotype V St. Louis encephalitis virus in Florida.

We isolated and characterized St. Louis encephalitis virus (SLEV) from cloacal swabs of naturally exposed adult sentinel chickens in 2006. Phylogenetic analysis of SLEV strains isolated in Florida indicated that Brazilian SLEV circulated in 1972 and 2006; lineages were VA and VB.