Reservoir hosts experiencing food stress alter transmission dynamics for a zoonotic pathogen.

Food limitation is a universal stressor for wildlife populations and is increasingly exacerbated by human activities. Anthropogenic environmental change can significantly alter the availability and quality of food resources for reservoir hosts and impact host-pathogen interactions in the wild. The state of the host's nutritional reserves at the time of infection is a key factor influencing infection outcomes by altering host resistance. Combining experimental and model-based approaches, we investigate how an environmental stressor affects host resistance to West Nile virus (WNV). Using American robins (Turdus migratorius), a species considered a superspreader of WNV, we tested the effect of acute food deprivation immediately prior to infection on host viraemia. Here, we show that robins food deprived for 48 h prior to infection, developed higher virus titres and were infectious longer than robins fed normally. To gain an understanding about the epidemiological significance of food-stressed hosts, we developed an agent-based model that simulates transmission dynamics of WNV between an avian host and the mosquito vector. When simulating a nutritionally stressed host population, the mosquito infection rate rose significantly, reaching levels that represent an epidemiological risk. An understanding of the infection disease dynamics in wild populations is critical to predict and mitigate zoonotic disease outbreaks.

The vector ecology of introduced Culex quinquefasciatus populations, and implications for future risk of West Nile virus emergence in the Galápagos archipelago.

Culex quinquefasciatus Say (Diptera: Culicidae), an important vector of West Nile virus (WNV) in the U.S.A., was first detected on the Galápagos Islands (Ecuador) in the 1980s. However, little is known of its ecology, distribution or capacity for arbovirus transmission in the Galápagos. We characterize details of lifecycle (including gonotrophic period), temporal abundance, spatial distribution, vector competence and host-feeding behaviour. Culex quinquefasciatus was detected on five islands of the Galápagos during 2006-2011. A period of 7-14 days was required for egg-adult emergence; water salinity above 5 ppt was demonstrated to hinder larval development. Blood-meal analysis indicated feeding on reptiles, birds and mammals. Assessment of WNV vector competency of Galápagos C. quinquefasciatus showed a median infectious dose of 7.41 log10 plaque-forming units per millilitre and evidence of vertical transmission (minimal filial infection rate of 3.7 per 1000 progeny). The distribution of C. quinquefasciatus across the archipelago could be limited by salt intolerance, and its abundance constrained by high temperatures. Feeding behaviour indicates potential to act as a bridge vector for transmission of pathogens across multiple taxa. Vertical transmission is a potential persistence mechanism for WNV on Galápagos. Together, our results can be used for epidemiological assessments of WNV and target vector control, should this pathogen reach the Galápagos Islands.

Multi-omics analysis of antiviral interactions of Elizabethkingia anophelis and Zika virus.

The microbial communities residing in the mosquito midgut play a key role in determining the outcome of mosquito pathogen infection. Elizabethkingia anophelis, originally isolated from the midgut of Anopheles gambiae possess a broad-spectrum antiviral phenotype, yet a gap in knowledge regarding the mechanistic basis of its interaction with viruses exists. The current study aims to identify pathways and genetic factors linked to E. anophelis antiviral activity. The understanding of E. anophelis antiviral mechanism could lead to novel transmission barrier tools to prevent arboviral outbreaks. We utilized a non-targeted multi-omics approach, analyzing extracellular lipids, proteins, metabolites of culture supernatants coinfected with ZIKV and E. anophelis. We observed a significant decrease in arginine and phenylalanine levels, metabolites that are essential for viral replication and progression of viral infection. This study provides insights into the molecular basis of E. anophelis antiviral phenotype. The findings lay a foundation for in-depth mechanistic studies.

Lecanemab Clarity AD: Quality-of-Life Results from a Randomized, Double-Blind Phase 3 Trial in Early Alzheimer's Disease.

BACKGROUND: Lecanemab is a humanized IgG1 monoclonal antibody binding with high affinity to amyloid-beta protein protofibrils. In phase 3 development, lecanemab has been shown to reduce markers of amyloid in early Alzheimer's disease and reduce decline on clinical endpoints of cognition and function at 18 months. OBJECTIVES: To describe the health-related quality-of-life (HRQoL) results from Clarity AD which were exploratory outcomes in this trial. DESIGN: Clarity AD was an 18-month, multi-center, double-blind, phase 3 trial. SETTING: Early Alzheimer's disease. PARTICIPANTS: Individuals 50-90 years of age with a diagnosis of mild cognitive impairment or mild dementia due to Alzheimer's disease and positron emission tomography or cerebrospinal fluid evidence of cerebral amyloid accumulation. INTERVENTION: Placebo or lecanemab 10-mg/kg IV biweekly. MEASUREMENTS: HRQoL was measured at baseline and every 6 months using the European Quality of Life-5 Dimensions (EQ-5D-5L; by subject) and Quality of Life in AD (QOL-AD; by subject and proxy). Study partner burden was measured using the Zarit Burden Interview (ZBI). RESULTS: A total of 1795 participants were enrolled (lecanemab:898; placebo:897). At month 18, adjusted mean change from baseline in EQ-5D-5L and QOL-AD by subject showed 49% and 56% less decline, respectively. QOL-AD rated by study partner as proxy resulted in 23% less decline. ZBI adjusted mean change from baseline at 18 months resulted in 38% less increase of care partner burden. Individual HRQoL test items and dimensions also showed lecanemab benefit. CONCLUSIONS: Lecanemab was associated with a relative preservation of HRQoL and less increase in caregiver burden, with consistent benefits seen across different quality of life scales and within scale subdomains. These benefits provide valuable patient reported outcomes which, together with previously reported benefits of lecanemab across multiple measures of cognition, function, disease progression, and biomarkers, demonstrate that lecanemab treatment may offer meaningful benefits to patients, care partners, and society.

Fine-scale spatial and temporal population genetics of Aedes japonicus, a new US mosquito, reveal multiple introductions.

The newly introduced mosquito Aedes japonicus has expanded from its original range in Northeastern Asia to 29 US states (including Hawaii) plus Canada and northern Europe. Our objectives were to test an earlier hypothesis of multiple introductions of this species to the Northeastern US and evaluate putative temporal changes in genetic makeup. Using a panel of seven microsatellite loci, we confirmed the existence of two abundant genetic forms in specimens originally collected in 1999-2000 (F(ST) value based on microsatellite data = 0.26) that matches the disjunctive distribution of mitochondrial haplotypes. To examine the distribution of the two genetic 'types' across Pennsylvania we created a fine-scale genetic map of Ae. japonicus using 439 specimens collected from 54 Pennsylvania counties in 2002-2003. We also made direct comparisons between collections in 1999-2000 and new collections made in 2004-2005 obtained from the same areas in the northeastern US. We observed that the strong association between mtDNA haplotype and microsatellite signature seen in 1999-2000 had weakened significantly by 2002 across Pennsylvania, a trend continued to some extent in 2004-2005 in PA, NJ, and NY, indicating that once easily distinguishable separate introductions are merging. The two expanding genetic forms create a complex correlation between spatial and genetic distances. The existence of multiple introductions would be obscured without sampling early and across time with highly polymorphic molecular markers. Our results provide a high-resolution analysis of the spatial and temporal dynamics of a newly introduced disease vector and argue that successive introductions may be a common pattern for invasive mosquitoes.

Computerized tomography. An adjunct to early diagnosis in the cauda equina syndrome of ankylosing spondylitis.

This article describes a case of the cauda equina syndrome associated with ankylosing spondylitis. The typical myelographic features and the computerized tomography (CT) scan of the lumbar spine are included. The CT scan may aid in elucidating the pathogenesis of the disorder.

Topiramate placebo-controlled dose-ranging trial in refractory partial epilepsy using 200-, 400-, and 600-mg daily dosages. Topiramate YD Study Group.

We conducted a randomized double-blind comparison of three doses of the novel antiepileptic drug (AED) topiramate (200, 400, and 600 mg/day) and placebo as adjunctive therapy in patients with refractory partial onset epilepsy receiving one or two other AEDs at therapeutic concentrations. A total of 181 patients completed the 12-week baseline phase and were randomized to double-blind therapy. Median percent reductions from baseline in average monthly seizure rate, the principal efficacy evaluation, were 13% for placebo, 30% for topiramate 200 mg/day, 48% for topiramate 400 mg/day, and 45% for topiramate 600 mg/day. For the seizure rate comparison of active drug to placebo p values were: topiramate 200 mg/day, p = 0.051; topiramate 400 mg/day, p = 0.007; topiramate 600 mg/day, p < 0.001. Percent responders ( > or = 50% reduction in seizure rates) were 18% for placebo, 27% for topiramate 200 mg/day, 47% for topiramate 400 mg/day (p = 0.013), and 46% for topiramate 600 mg/day (p = 0.027). A significant (p = 0.003) reduction in secondarily generalized seizures compared with placebo treatment was also documented with topiramate. Topiramate plasma concentrations were closely related to dosage, and there were no significant interactions between topiramate and other AEDs. The minimal effective dose of topiramate in this study population was approximately 200 mg/day. Mild or moderate CNS symptoms were the primary treatment-emergent adverse events, but treatment-limiting adverse events occurred in only 9% of patients given topiramate compared with 7% given placebo. Results of this initial well-controlled study in patients indicate that topiramate is a very promising new AED.

Felbamate: a double-blind controlled trial in patients undergoing presurgical evaluation of partial seizures.

We studied the efficacy and safety of felbamate, an investigational antiepileptic drug, in a unique, double-blind, placebo-controlled trial. Sixty-four patients with refractory partial-onset seizures who completed a routine evaluation for epilepsy surgery met seizure frequency entry criteria. Each patient received felbamate or placebo in addition to the anticonvulsant regimen present at the conclusion of the presurgical evaluation. The treatment phase consisted of an 8-day inpatient period and a 21-day outpatient period. The efficacy variable was time to fourth seizure. The difference in time to fourth seizure was statistically significant (p = 0.028) in favor of felbamate. Eighty-eight percent of the patients in the placebo group had a fourth seizure during the treatment phase compared with 46% of the patients in the felbamate group (p = 0.001). Adverse experiences with felbamate were generally mild or moderate in severity. This trial demonstrated the ability of felbamate to quickly and safely reduce the occurrence of frequent partial-onset seizures and maintain effective seizure control following reductions in the dosages of standard antiepileptic drugs.

Binding of western equine encephalomyelitis virus to brush border fragments isolated from mesenteronal epithelial cells of mosquitoes.

Brush border fragments (BBF) were isolated from mesenteronal epithelial cells of mosquitoes that are either susceptible (WS Culex tarsalis) or refractory (WR Cx. tarsalis; Culex pipiens) to peroral infection by western equine encephalomyelitis (WEE) virus. The isolated BBF were combined with radiolabeled WEE virus in a binding assay to compare the amount of virus bound by BBF from susceptible and refractory mosquitoes. BBF and WEE virus were mixed in a microcentrifuge tube, incubated for 1 h and centrifuged at 27,000 X g-30 min to pellet WEE virus bound to BBF. Optimal binding occurred at pH 7.2, 20 degrees C and there was no requirement for divalent cations. BBF isolated from perorally susceptible mosquitoes (WS Cx. tarsalis) bound significantly greater amounts of radiolabeled WEE virus, compared to BBF isolated from refractory mosquitoes (WR Cx. tarsalis; Cx. pipiens), in all experiments. The binding of WEE virus to BBF from WS Cx. tarsalis appears to be specific, based on saturation and competitive binding studies; binding to BBF from WR Cx. tarsalis and Cx. pipiens is nonspecific. Scatchard analysis of the binding data for BBF from WS Cx. tarsalis yields an estimated 1.8-3.5 X 10(6) binding sites per mesenteronal epithelial cell with an affinity constant of (Ka) of 2.2 X 10(11) M-1.

Western equine encephalomyelitis virus: in vivo infection and morphogenesis in mosquito mesenteronal epithelial cells.

The infection and morphogenetic events associated with the replication of Western equine encephalomyelitis (WEE) virus within the mesenterons of Aedes dorsalis and three strains of Culex tarsalis are compared and contrasted. WEE virus apparently penetrates mesenteronal epithelial cells in vivo through membrane fusion. Profiles of apparent membrane fusion events were observed between virus particles and the microvillar surface of the mesenteron and naked nucleocapsids are observed intracellularly along the apical margin of the mesenteronal epithelial cell within 3 h of ingestion of the bloodmeal. Further, no viral particles were found in association with endocytotic nor lysosomal vacuoles during the initial phases of infection. In those strains of Cx. tarsalis that supported viral replication and in Ae. dorsalis, accumulations of nucleocapsids and maturation of WEE virus were evident along basolateral membranes of the mesenteron by 22-24 h after ingestion of the blood-meal. Maximal extracellular nascent virus occurred between 30-36 hrs. The Knights Landing strain of Cx. tarsalis revealed no subcellular morphological alteration in response to infection throughout the period of study. However, distinct morphological structures associated with the infection were observed in strains or species with enhanced susceptibility compared to Knights Landing (i.e., Cx. tarsalis WS-3 and Ae. dorsalis). In both, apical accumulations of nucleocapsids were apparent by 29 h post infection. These nucleocapsids were most often embedded in a rather amorphous matrix and occasionally in association with membrane profiles; presumably endoplasmic reticulum. Ae. dorsalis also demonstrated some alterations in response to WEE viral infection that were unique relative to Cx. tarsalis and some of these may be considered cytopathological. First, progeny virions were observed repeatedly within lysosomal figures. Second, extensive cytoplasmic vacuolization was noted and occasionally it appeared that these vacuolated cells were being sloughed off into the lumen of the mesenteron.

West Nile virus activity in the United States, 2001.

West Nile virus (WNV) first appeared in the naive environment of the Western Hemisphere in 1999 in New York. Genetic analysis determined that the virus was introduced into the United States from the Mediterranean Basin. This review discusses the spread of the virus in 2001 from the initial focus in Queens, New York, to widespread activity in the eastern and midwestern United States. It concentrates on viral ecology, epizootiology, pathology, prediction, and prevention. Research questions to further our understanding of the transmission cycle of WNV are discussed, including host-preference studies, molecular confirmation of implicated mosquito vectors, and survival of WNV in the temperate environment of the United States. Comparisons are drawn with two other arboviruses enzootic in the United States, eastern equine encephalitis, and St. Louis encephalitis viruses. Although not recently introduced, these two viruses also demonstrated increased activity in the United States in 2001.

Cardiac dysfunction in a patient with familial hypokalemic periodic paralysis.

A 19-year-old white man with familial hypokalemic periodic paralysis developed evidence of cardiac dysfunction during a episode of flaccid paralysis. This consisted of elevated total creatine phosphokinase (CPK), an increased myocardial fraction of CPK (myocardial band), alteration in the lactic dehydrogenase isoenzyme pattern, severe bradycardia, and evidence of left ventricular dysfunction. These findings, in conjunction with selected cases from the literature, suggest the possibility that cardiomyopathy may be a heretofore unrecognized complication of this disorder.

West Nile virus infection in birds and mammals.

West Nile virus (WNV) was found throughout New York State in year 2000. The epicenter was located in New York City with a high level of activity in the immediately surrounding counties, including Rockland, Westchester, Nassau, and Suffolk. During 2000, WNV testing was performed by the Wadsworth Center on 3,687 dead birds, representing 153 species, 46 families, and 18 orders. There were 1,203 WNV-positive birds, representing 63 species, 30 families and 14 orders. The percentage of WNV-positive birds was 33% for all birds tested throughout the state, with no significant difference in infection rates in migratory versus resident birds, although significantly more resident birds were submitted for testing. The highest apparent mortality for the entire season was observed in American crows in Staten Island, a location that also showed the highest minimal infection rate in Culex pipiens complex mosquitoes. Studies examining tissue tropism of WNV in corvids and noncorvids from the epicenter and from remote locations indicated that the kidney was the most consistently infected tissue in birds, regardless of level of infection. The brain was the next most consistently positive tissue. The differences in infection among the tissues were most apparent when low levels of virus were present. Experimental mouse inoculation demonstrated a classical flavivirus infection pattern.

Genetic variation among isolates of western equine encephalomyelitis virus from California.

The mechanism for long-term maintenance of western equine encephalomyelitis (WEE) virus in California was investigated by studying genetic variation in the E2 portion of the genome of 55 strains of WEE virus isolated since 1938 from different locations in California. Four major lineages were evident: virus strains isolated from the Central Valley since 1993 and Los Angeles in 1991 formed lineage A; southern California strains isolated since 1978 and isolates from the Central Valley from 1978 to 1987 formed lineage B; northern California isolates from 1968 to 1971 formed lineage C; and early isolates from 1938 to 1961 formed a fourth lineage, D. The separation of strains from north and south of the Tehachapi and San Bernardino Mountains (i.e., the Central Valley and southern California, respectively) since 1991 indicates that there has been little recent movement of virus between the two regions and recent strains from these two locations appear to be evolving independently. However, within the Central Valley and within southern California, virus appears to circulate freely, perhaps by movement of birds or mosquito vectors. Although the current virus lineage in the Central Valley may have been introduced from an unknown source in 1991, introduction and establishment of new viral genotypes from outside California do not seem to occur regularly. It appears most likely that virus is maintained in separate geographic areas of California through local persistence in enzootic foci.

Quantitation of La Crosse virus in venereally infected Aedes triseriatus.

The replication of two strains of La Crosse (LAC) virus, prototype and an unpassaged field isolate, was followed in intrathoracically inoculated male and venereally infected female Aedes triseriatus mosquitoes. The venereal infection rate following induced mating was 13.6% of 44 females inseminated by males inoculated with prototype virus, and 26.3% of 38 females inseminated by males inoculated with an unpassaged field strain. The males consistently contained concentrations of virus in reproductive tracts equal to or greater than those in salivary glands (SG). In contrast, venereally infected females generally contained the highest concentrations of virus in SG. Prototype LAC virus was demonstrable in nearly all organs of the infected females by 10 days following mating; the unpassaged field strain was detectable in all organ systems tested by day 6. There appeared to be no significant differences in replication between the prototype and field isolate viruses.

Vector competence of mosquitoes as a marker to distinguish Central American and Mexican epizootic from enzootic strains of Venezuelan encephalitis virus.

Two epizootic strains of Venezuelan encephalitis (VE) virus from Central America and Mexico were transmitted by a colonized epizootic vector mosquito, Aedes taeniorhynchus, at higher rates than were two enzootic strains when the mosquitoes were infected by intrathroacic inoculation or feeding of virus. Differences in transmission rates also occurred with colonized Aedes aegypti, but were less marked. Following intrathoracic inoculation of A. taeniorhynchus or A. aegypti, epizootic strains grew to slightly higher concentrations in the mosquitoes than did enzootic strains. Intestinal thresholds of infection for A. taeniorhynchus were slightly lower for epizootic than for enzootic virus strains, but were essentially equal for A. aegypti. Only a small percentage of individual Culex pipiens quinquefasciatus mosquitoes supported growth of epizootic VE virus, and only 1 of 6 tested C. p. quinquefasciatus transmitted virus by bite. Thus, transmission and growth of virus in these Aedes mosquitoes distinguished between these epizootic and enzootic strains of VE virus.

Experimental St. Louis encephalitis virus infection of sloths and cormorants.

Experimental infection of 11 Bradypus variegatus and Choloepus hoffmanni sloths with St. Louis encephalitis (SLE) virus produced detectable viremias of seven to 27 (median 13) days duration and maximum titers of 2.7 to 6.5 (median 5.1) log10 median suckling mouse intracranial lethal doses (SMicLD50) per ml. Experimental SLE viremia onset was delayed and maximum titer depressed in two sloths concurrently infected with naturally acquired viruses. SLE viremias in four experimentally inoculated cormorants Phalacrocorax olivaceus were shorter, and of equal or lower titer, than in sloths. Colonized Culex pipiens quinquefasciatus mosquitoes were infected by feeding on sloths circulating at least 4.8 log10 SMicLD50 of SLE virus per ml, and subsequently transmitted the infection to mice and chicks. An uninoculated baby Bradypus became infected by contact transmission from its mother. The antibody response of sloths to SLE virus was slow, being undetectable until several weeks post-inoculation. However, both sloth species developed high and long-lasting neutralizing and hemagglutination-inhibition antibody titers. The complement-fixation antibody response in Bradypus was lower and slower to develop than in Choloepus. Sloths with naturally acquired SLE virus antibody did not become detectably viremic after experimental inoculation. Neither sloths nor cormorants become overly ill from SLE virus infection.

Effect of temperature of extrinsic incubation on the vector competence of Culex tarsalis for western equine encephalomyelitis virus.

Culex tarsalis was a less competent vector of western equine encephalomyelitis (WEE) virus after 2-3 weeks' extrinsic incubation at 32 degrees C than after incubation at 18 degrees or 25 degrees C. The high temperature itself was not directly detrimental to mosquito infection as all mosquitoes were initially infected, but subsequently some females were able to limit viral multiplication and/or dissemination. Elevated maintenance temperatures enhanced the expression of modulation, and elevated larval rearing temperatures selected for those females with this trait. This is the first report of an inverse relationship between temperature of extrinsic incubation within the range of 25 degrees-32 degrees C and vector competence of a mosquito for an arbovirus.