EVALUTION OF MODIFIED AUTOCIDAL GRAVID OVITRAPS FOR CONTROL OF AEDES AEGYPTI IN ST. AUGUSTINE, FLORIDA.

Aedes aegypti is an anthropophilic mosquito that vectors dengue, chikungunya, Zika, and yellow fever viruses. The US Center for Disease Control and Prevention (CDC)'s autocidal gravid ovitraps (AGOs) may facilitate the control of container-inhabiting Aedes mosquitoes and curb arbovirus outbreaks by taking advantage of oviposition-seeking behavior using pesticide-free technology. The AGOs, manufactured by SpringStar Inc., were tested during the summer of 2018 in St. Augustine, FL. A total of 1,718 AGOs were deployed for study in 3 different 40-acre (∼18.2 ha) plots at a density of 5-7 AGOs per house and a coverage of >90% for all AGO test sites. The AGOs were modified using tap water instead of infusion water to reduce the capture of nontarget organisms. Each intervention and reference area was monitored weekly using BioGents Sentinel traps and Sentinel AGOs. Generalized linear mixed models showed that changes to Aedes mosquito populations were more seasonal than treatment driven. Homeowners expressed positivity about traps and believed the traps were both effective and had directly contributed to increased quality of life.

Evaluation of Modified Autocidal Gravid Ovitraps for Control of Aedes Aegypti in St. Augustine, Florida.

Aedes aegypti is an anthropophilic mosquito that vectors dengue, chikungunya, Zika, and yellow fever viruses. The US Center for Disease Control and Prevention (CDC)'s autocidal gravid ovitraps (AGOs) may facilitate the control of container-inhabiting Aedes mosquitoes and curb arbovirus outbreaks by taking advantage of oviposition-seeking behavior using pesticide-free technology. The AGOs, manufactured by SpringStar Inc., were tested during the summer of 2018 in St. Augustine, FL. A total of 1,718 AGOs were deployed for study in 3 different 40-acre (∼18.2 ha) plots at a density of 5-7 AGOs per house and a coverage of >90% for all AGO test sites. The AGOs were modified using tap water instead of infusion water to reduce the capture of nontarget organisms. Each intervention and reference area was monitored weekly using BioGents Sentinel traps and Sentinel AGOs. Generalized linear mixed models showed that changes to Aedes mosquito populations were more seasonal than treatment driven. Homeowners expressed positivity about traps and believed the traps were both effective and had directly contributed to increased quality of life.

Multisite test-retest reliability and compatibility of brain metrics derived from FreeSurfer versions 7.1, 6.0, and 5.3.

Automatic neuroimaging processing tools provide convenient and systematic methods for extracting features from brain magnetic resonance imaging scans. One tool, FreeSurfer, provides an easy-to-use pipeline to extract cortical and subcortical morphometric measures. There have been over 25 stable releases of FreeSurfer, with different versions used across published works. The reliability and compatibility of regional morphometric metrics derived from the most recent version releases have yet to be empirically assessed. Here, we used test-retest data from three public data sets to determine within-version reliability and between-version compatibility across 42 regional outputs from FreeSurfer versions 7.1, 6.0, and 5.3. Cortical thickness from v7.1 was less compatible with that of older versions, particularly along the cingulate gyrus, where the lowest version compatibility was observed (intraclass correlation coefficient 0.37-0.61). Surface area of the temporal pole, frontal pole, and medial orbitofrontal cortex, also showed low to moderate version compatibility. We confirm low compatibility between v6.0 and v5.3 of pallidum and putamen volumes, while those from v7.1 were compatible with v6.0. Replication in an independent sample showed largely similar results for measures of surface area and subcortical volumes, but had lower overall regional thickness reliability and compatibility. Batch effect correction may adjust for some inter-version effects when most sites are run with one version, but results vary when more sites are run with different versions. Age associations in a quality controlled independent sample (N = 106) revealed version differences in results of downstream statistical analysis. We provide a reference to highlight the regional metrics that may yield recent version-related inconsistencies in published findings. An interactive viewer is provided at http://data.brainescience.org/Freesurfer_Reliability/.

Genetic map of regional sulcal morphology in the human brain from UK biobank data.

Genetic associations with macroscopic brain structure can provide insights into brain function and disease. However, specific associations with measures of local brain folding are largely under-explored. Here, we conducted large-scale genome- and exome-wide associations of regional cortical sulcal measures derived from magnetic resonance imaging scans of 40,169 individuals in UK Biobank. We discovered 388 regional brain folding associations across 77 genetic loci, with genes in associated loci enriched for expression in the cerebral cortex, neuronal development processes, and differential regulation during early brain development. We integrated brain eQTLs to refine genes for various loci, implicated several genes involved in neurodevelopmental disorders, and highlighted global genetic correlations with neuropsychiatric phenotypes. We provide an interactive 3D visualisation of our summary associations, emphasising added resolution of regional analyses. Our results offer new insights into the genetic architecture of brain folding and provide a resource for future studies of sulcal morphology in health and disease.

Effects of ApoE4 and ApoE2 genotypes on subcortical magnetic susceptibility and microstructure in 27,535 participants from the UK Biobank.

Disrupted iron homeostasis is associated with several neurodegenerative diseases, including Alzheimer's disease (AD), and may be partially modulated by genetic risk factors. Here we evaluated whether subcortical iron deposition is associated with ApoE genotype, which substantially affects risk for late-onset AD. We evaluated differences in subcortical quantitative susceptibility mapping (QSM), a type of MRI sensitive to cerebral iron deposition, between either ApoE4 (E3E4+E4E4) or ApoE2 (E2E3+E2E2) carriers and E3 homozygotes (E3E3) in 27,535 participants from the UK Biobank (age: 45-82 years). We found that ApoE4 carriers had higher hippocampal (d=0.036; p=0.012) and amygdalar (d=0.035; p=0.013) magnetic susceptibility, particularly individuals aged 65 years or older, while those carrying ApoE2 (which protects against AD) had higher QSM only in the hippocampus (d=0.05; p=0.006), particularly those under age 65. Secondary diffusion MRI microstructural associations in these regions revealed greater diffusivity and less diffusion restriction in E4 carriers, however no differences were detected in E2 carriers. Disease risk conferred by ApoE4 may be linked with higher subcortical iron burden in conjunction with inflammation or neuronal loss in aging individuals, while ApoE2 associations may not necessarily reflect unhealthy iron deposits earlier in life.

Modelling distributions of Aedes aegypti and Aedes albopictus using climate, host density and interspecies competition.

Florida faces the challenge of repeated introduction and autochthonous transmission of arboviruses transmitted by Aedes aegypti and Aedes albopictus. Empirically-based predictive models of the spatial distribution of these species would aid surveillance and vector control efforts. To predict the occurrence and abundance of these species, we fit a mixed-effects zero-inflated negative binomial regression to a mosquito surveillance dataset with records from more than 200,000 trap days, representative of 53% of the land area and ranging from 2004 to 2018 in Florida. We found an asymmetrical competitive interaction between adult populations of Aedes aegypti and Aedes albopictus for the sampled sites. Wind speed was negatively associated with the occurrence and abundance of both vectors. Our model predictions show high accuracy (72.9% to 94.5%) in validation tests leaving out a random 10% subset of sites and data since 2017, suggesting a potential for predicting the distribution of the two Aedes vectors.

Sunshine versus gold: The effect of population age on genetic structure of an invasive mosquito.

The genetic diversity and structure of invasive species are affected by the time since invasion, but it is not well understood how. We compare likely the oldest populations of Aedes aegypti in continental North America with some of the newest to illuminate the range of genetic diversity and structure that can be found within the invasive range of this important disease vector. Aedes aegypti populations in Florida have probably persisted since the 1600-1700s, while populations in southern California derive from new invasions that occurred in the last 10 years. For this comparison, we genotyped 1,193 individuals from 28 sites at 12 highly variable microsatellites and a subset of these individuals at 23,961 single nucleotide polymorphisms (SNPs). This is the largest sample analyzed for genetic structure for either region, and it doubles the number of southern California populations previously analyzed. As predicted, the older populations (Florida) showed fewer indicators of recent founder effect and bottlenecks; in particular, these populations have dramatically higher genetic diversity and lower genetic structure. Geographic distance and driving distance were not good predictors of genetic distance in either region, especially southern California. Additionally, southern California had higher levels of genetic differentiation than any comparably sized documented region throughout the worldwide distribution of the species. Although population age and demographic history are likely driving these differences, differences in climate and transportation practices could also play a role.

Toward Engineering Biosystems With Emergent Collective Functions.

Many complex behaviors in biological systems emerge from large populations of interacting molecules or cells, generating functions that go beyond the capabilities of the individual parts. Such collective phenomena are of great interest to bioengineers due to their robustness and scalability. However, engineering emergent collective functions is difficult because they arise as a consequence of complex multi-level feedback, which often spans many length-scales. Here, we present a perspective on how some of these challenges could be overcome by using multi-agent modeling as a design framework within synthetic biology. Using case studies covering the construction of synthetic ecologies to biological computation and synthetic cellularity, we show how multi-agent modeling can capture the core features of complex multi-scale systems and provide novel insights into the underlying mechanisms which guide emergent functionalities across scales. The ability to unravel design rules underpinning these behaviors offers a means to take synthetic biology beyond single molecules or cells and toward the creation of systems with functions that can only emerge from collectives at multiple scales.

Diel activity patterns of adult female mosquitoes (Diptera: Culicidae) determined by a novel rotated trap in northeastern Florida, U.S.A.

A novel rotator trap was evaluated to determine the diel activity patterns and physiological state of adult female mosquitoes in St. Augustine, FL, U.S.A. Culex nigripalpus were most active from 19:00-21:00, followed by 1:00-3:00, based on collections from the novel rotator trap. Furthermore, analysis of the physiological state of female mosquitoes collected by the novel rotator trap suggested that non-parous (nulliparous) host-seeking mosquitoes were more frequently active in the 21:00-23:00 and 09:00-19:00 time frames. Parous host-seeking mosquitoes were more frequently collected from 19:00-21:00 and 1:00-3:00. A low abundance of gravid females was collected by the rotator trap, so analysis of their activity periods was inconclusive. These results indicate that the novel rotated trap could be used to detect the diel activity patterns of adult mosquitoes in mosquito control programs and more testing should be conducted in the future.

Autocidal Gravid Ovitrap Incorporation with Attractants for Control of Gravid and Host-Seeking Aedes aegypti (Diptera: Culicidae).

Aedes aegypti Linnaeus (Diptera: Culicidae) is a primary vector of multiple arboviruses including Zika, dengue, chikungunya, and yellow fever. After incorporating BG lure or BG lure + octenol, the CDC's Autocidal Gravid Ovitrap (AGO) was more effective at collecting and controlling host-seeking female Ae. aegypti than gravid female mosquitoes. The addition of octenol to the AGO did not increase the number of female mosquitoes captured by the AGO, compared with the AGO alone. The AGO baited with the BG lure captured a significant number of host-seeking female Ae. aegypti. This finding indicates that the combination of AGO with BG lure could enable the trap to control female Ae. aegypti mosquitoes representing either host-seeking or gravid physiological states.

Automated Visualization of Genetic Designs Using DNAplotlib.

Visualization of complex genetic systems can help efficiently communicate important design features and clearly illustrate overall structures. To aid in the creation of such diagrams, standards such as the Synthetic Biology Open Language Visual (SBOLv) have been established to ensure that specific symbols and shapes convey the same meaning for genetic parts across the field. Here, we describe several ways that the computational tool DNAplotlib can be used to automate the generation of SBOLv standard-compliant diagrams covering simple genetic designs to large libraries of genetic constructs.

Characterization of Carbonic Anhydrase 9 in the Alimentary Canal of Aedes aegypti and Its Relationship to Homologous Mosquito Carbonic Anhydrases.

In the mosquito midgut, luminal pH regulation and cellular ion transport processes are important for the digestion of food and maintenance of cellular homeostasis. pH regulation in the mosquito gut is affected by the vectorial movement of the principal ions including bicarbonate/carbonate and protons. As in all metazoans, mosquitoes employ the product of aerobic metabolism carbon dioxide in its bicarbonate/carbonate form as one of the major buffers of cellular and extracellular pH. The conversion of metabolic carbon dioxide to bicarbonate/carbonate is accomplished by a family of enzymes encoded by the carbonic anhydrase gene family. This study characterizes Aedes aegypti carbonic anhydrases using bioinformatic, molecular, and immunohistochemical methods. Our analyses show that there are fourteen Aedes aegypti carbonic anhydrase genes, two of which are expressed as splice variants. The carbonic anhydrases were classified as either integral membrane, peripheral membrane, mitochondrial, secreted, or soluble cytoplasmic proteins. Using polymerase chain reaction and Western blotting, one of the carbonic anhydrases, Aedes aegypti carbonic anhydrase 9, was analyzed and found in each life stage, male/female pupae, male/female adults, and in the female posterior midgut. Next, carbonic anhydrase 9 was analyzed in larvae and adults using confocal microscopy and was detected in the midgut regions. According to our analyses, carbonic anhydrase 9 is a soluble cytoplasmic enzyme found in the alimentary canal of larvae and adults and is expressed throughout the life cycle of the mosquito. Based on previous physiological analyses of adults and larvae, it appears AeCA9 is one of the major carbonic anhydrases involved in producing bicarbonate/carbonate which is involved in pH regulation and ion transport processes in the alimentary canal. Detailed understanding of the molecular bases of ion homeostasis in mosquitoes will provide targets for novel mosquito control strategies into the new millennium.

Accuracy of a dynamic surgical guidance probe for screw insertion in the cervical spine: a cadaveric study.

STUDY DESIGN: A fresh frozen cadaver study was conducted. OBJECTIVE: To report the cortical breach rate using the dynamic surgical guidance (DSG) probe versus traditional freehand technique for cervical lateral mass, cervical pedicle and cervical laminar screws. METHODS: Nine male fresh frozen cadaveric torsos were utilized for this study. Each investigator was assigned three specimens that were randomized by fixation point, side and order of technique for establishing a screw pilot hole. The technique for screw hole preparation utilized was either a DSG probe in the "on" mode or in the "off" mode using a freehand technique popularized by Lenke et al. Levels instrumented included C1 lateral mass, C2 pedicle screws and lamina screws, and C6-T1 pedicle screws. Fluoroscopy and other navigational assistance were not used for screw hole preparation or screw insertion. All specimens were CT imaged following insertion of all screws. A senior radiologist evaluated all scans and determined that a misplaced screw was a breach of ≥2 mm. RESULTS: A total of 104 drillings were performed, 52 with DSG and 52 without DSG There were 68 total pedicle drillings, 34 in each group. There were 18 drillings in the lamina and lateral mass. There was no significant difference between surgeons or between the left and right side. All breaches were in the pedicle, and none in the lamina or lateral mass. The breach rate for PG "on" was 6/68 = 8.96% (95% CI 3.69, 19.12%). The breach rate for PG "off" was 20/68 = 29.41% (95% CI 19.30, 41.87%). Of the 20 pedicle breaches in the non-DSG group, 7 were lateral and superior, 8 were lateral, 4 medial and 1 inferior. Of the six pedicle breaches in the DSG group, two were lateral/superior, two were lateral and two were medial in the pedicle. CONCLUSIONS: The dynamic surgical guidance probe is a safe tool to assist the surgeon with screw placement in the cervical spine. Additionally, the DSG potentially avoids the cumulative risks associated with fluoroscopy and provides real-time feedback to the surgeon allowing correction at the time of breach. Level of evidence Level IV.

Effects of environmental and clinical interferents on the host capture efficiency of immobilized bacteriophages.

Bacteriophage-functionalized surfaces are a new class of advanced functional material and have been demonstrated to be applicable for use as antimicrobial surfaces in medical applications (e.g., indwelling medical devices or wound dressings) or as biosensors for bacterial capture and detection. However, the complex composition of many real life samples (e.g., blood, natural waters, etc.) can potentially interfere with the interaction of phage and its bacterial host, leading to a decline in the efficiency of the phage-functionalized surface. In this study, the bacterial capture efficiency of two model phage-functionalized surfaces was assessed in the presence of potential environmental and biomedical interferents. The two phage-bacteria systems used in this study are PRD1 with Salmonella Typhimurium and T4 with Escherichia coli. The potential interferents tested included humic and fulvic acids, natural groundwater, colloidal latex microspheres, host extracellular polymeric substances (EPS), albumin, fibrinogen, and human serum. EPS and human serum decreased the host capture efficiency for immobilized PRD1 and T4, and also impaired the infectivity of the nonimmobilized (planktonic) phage. Interestingly, humic and fulvic acids reduced the capture efficiency of T4-functionalized surfaces, even though they did not lead to inactivation of the suspended virions. Neither humic nor fulvic acids affected the capture efficiency of PRD1. These findings demonstrate the inadequacy of traditional phage selection methods (i.e., infectivity of suspended phage toward its host in clean buffer) for designing advanced functional materials and further highlight the importance of taking into account the environmental conditions in which the immobilized phage is expected to function.

The impact of protein concentration on mannitol and sodium chloride crystallinity and polymorphism upon lyophilization.

The effect of protein concentration, in the range of 0-26 mg/mL for two Fc-fusion proteins, on the crystallinity and polymorphism of mannitol and sodium chloride in a lyophilized model formulation was examined. Mannitol hydrate levels were quantified based on moisture data and correlated to the X-ray diffraction peak area. In all formulation conditions, sodium chloride did not crystallize in samples with >44% total amorphous content. As protein concentration increased through the range of 1-5 mg/mL prior to lyophilization, beta-mannitol decreased in amount, becoming undetectable at protein concentrations above 5 mg/mL. Conversely, delta-mannitol increased as a function of protein concentration, reaching a maximum level at approximately 5 mg/mL protein. Above 10 mg/mL protein, mannitol crystallization was increasingly inhibited. Sucrose control vials showed higher levels of mannitol hydrate than either model protein. Both proteins behaved comparably with respect to mannitol crystallinity and polymorphism despite significant differences in molecular weight. Because of the differences between protein and sucrose control samples, protein concentration must be taken into consideration when assessing the lyophilization of mannitol containing solutions.

Insignificant change in Antarctic snowfall since the International Geophysical Year.

Antarctic snowfall exhibits substantial variability over a range of time scales, with consequent impacts on global sea level and the mass balance of the ice sheets. To assess how snowfall has affected the thickness of the ice sheets in Antarctica and to provide an extended perspective, we derived a 50-year time series of snowfall accumulation over the continent by combining model simulations and observations primarily from ice cores. There has been no statistically significant change in snowfall since the 1950s, indicating that Antarctic precipitation is not mitigating global sea level rise as expected, despite recent winter warming of the overlying atmosphere.

Synthesis and characterization of water-soluble hydroxybutenyl cyclomaltooligosaccharides (cyclodextrins).

We have examined the synthesis of hydroxybutenyl cyclomaltooligosaccharides (cyclodextrins) and the ability of these cyclodextrin ethers to form guest-host complexes with guest molecules. The hydroxybutenyl cyclodextrin ethers were prepared by a base-catalyzed reaction of 3,4-epoxy-1-butene with the parent cyclodextrins in an aqueous medium. Reaction byproducts were removed by nanofiltration before the hydroxybutenyl cyclodextrins were isolated by co-evaporation of water-EtOH. Hydroxybutenyl cyclodextrins containing no unsubstituted parent cyclodextrin typically have a degree of substitution of 2-4 and a molar substitution of 4-7. These hydroxybutenyl cyclodextrins are randomly substituted, amorphous solids. The hydroxybutenyl cyclodextrin ethers were found to be highly water soluble. Complexes of HBen-beta-CD with glibenclamide and ibuprofen were prepared and isolated. In both cases, the guest content of the complexes was large, and a significant increase in the solubility of the free drug was observed. Dissolution of the complexes in pH 1.4 water was very rapid, and significant increases in the solubility of the free drugs were observed. Significantly, after reaching equilibrium concentration, a decrease in the drug concentration over time was not observed.