Characterization of a homolog of DEAD-box RNA helicases in Toxoplasma gondii as a marker of cytoplasmic mRNP stress granules

Toxoplasma gondii is an obligate intracellular protozoan which infects one-third of the human population. Due to its high infection prevalence, Toxoplasma offers an ideal system for the study of host–parasite interaction. Similar to other eukaryotes, Toxoplasma maintains levels and localization of cytoplasmic mRNAs throughout its life cycle as part of a gene regulation network to meet all cellular and biochemical requirements. More recently, it was reported that the presence of cytoplasmic mRNA granules could contribute to the parasite pathogenesis and viability. Here we identified a novel Toxoplasma DEAD-box RNA helicase, referred to as Toxoplasma gondiiHomolog of DOZI (TgHoDI), because of its high homology (81%) to Plasmodium DOZI. TgHoDI is the functional ortholog of yeast DHH1, and its function was authenticated by complementation studies in Δdhh1 yeast strain. We demonstrated that TgHoDI is a marker of cytoplasmic RNA stress granules, which assemble when the parasites experience cellular stresses and translational arrest.     

A Quantitative Comparison of the Behavior of Human Ventricular Cardiac Electrophysiology Models in Tissue

Numerical integration of mathematical models of heart cell electrophysiology provides an important computational tool for studying cardiac arrhythmias, but the abundance of available models complicates selecting an appropriate model. We study the behavior of two recently published models of human ventricular action potentials, the Grandi-Pasqualini-Bers (GPB) and the O''Hara-Virág-Varró-Rudy (OVVR) models, and compare the results with four previously published models and with available experimental and clinical data. We find the shapes and durations of action potentials and calcium transients differ between the GPB and OVVR models, as do the magnitudes and rate-dependent properties of transmembrane currents and the calcium transient. Differences also occur in the steady-state and S1–S2 action potential duration and conduction velocity restitution curves, including a maximum conduction velocity for the OVVR model roughly half that of the GPB model and well below clinical values. Between single cells and tissue, both models exhibit differences in properties, including maximum upstroke velocity, action potential amplitude, and minimum diastolic interval. Compared to experimental data, action potential durations for the GPB and OVVR models agree fairly well (although OVVR epicardial action potentials are shorter), but maximum slopes of steady-state restitution curves are smaller. Although studies show alternans in normal hearts, it occurs only in the OVVR model, and only for a narrow range of cycle lengths. We find initiated spiral waves do not progress to sustained breakup for either model. The dominant spiral wave period of the GPB model falls within clinically relevant values for ventricular tachycardia (VT), but for the OVVR model, the dominant period is longer than periods associated with VT. Our results should facilitate choosing a model to match properties of interest in human cardiac tissue and to replicate arrhythmia behavior more closely. Furthermore, by indicating areas where existing models disagree, our findings suggest avenues for further experimental work.     

A New Brain Drug Delivery Strategy: Focused Ultrasound-Enhanced Intranasal Drug Delivery

Central nervous system (CNS) diseases are difficult to treat because of the blood-brain barrier (BBB), which prevents most drugs from entering into the brain. Intranasal (IN) administration is a promising approach for drug delivery to the brain, bypassing the BBB; however, its application has been restricted to particularly potent substances and it does not offer localized delivery to specific brain sites. Focused ultrasound (FUS) in combination with microbubbles can deliver drugs to the brain at targeted locations. The present study proposed to combine these two different platform techniques (FUS+IN) for enhancing the delivery efficiency of intranasally administered drugs at a targeted location. After IN administration of 40 kDa fluorescently-labeled dextran as the model drug, FUS targeted at one region within the caudate putamen of mouse brains was applied in the presence of systemically administered microbubbles. To compare with the conventional FUS technique, in which intravenous (IV) drug injection is employed, FUS was also applied after IV injection of the same amount of dextran in another group of mice. Dextran delivery outcomes were evaluated using fluorescence imaging of brain slices. The results showed that FUS+IN enhanced drug delivery within the targeted region compared with that achieved by IN only. Despite the fact that the IN route has limited drug absorption across the nasal mucosa, the delivery efficiency of FUS+IN was not significantly different from that of FUS+IV. As a new drug delivery platform, the FUS+IN technique is potentially useful for treating CNS diseases.     

HRAS1 and LASS1 with APOE are associated with human longevity and healthy aging

The search for longevity‐determining genes in human has largely neglected the operation of genetic interactions. We have identified a novel combination of common variants of three genes that has a marked association with human lifespan and healthy aging. Subjects were recruited and stratified according to their genetically inferred ethnic affiliation to account for population structure. Haplotype analysis was performed in three candidate genes, and the haplotype combinations were tested for association with exceptional longevity. An HRAS1 haplotype enhanced the effect of an APOE haplotype on exceptional survival, and a LASS1 haplotype further augmented its magnitude. These results were replicated in a second population. A profile of healthy aging was developed using a deficit accumulation index, which showed that this combination of gene variants is associated with healthy aging. The variation in LASS1 is functional, causing enhanced expression of the gene, and it contributes to healthy aging and greater survival in the tenth decade of life. Thus, rare gene variants need not be invoked to explain complex traits such as aging; instead rare congruence of common gene variants readily fulfills this role. The interaction between the three genes described here suggests new models for cellular and molecular mechanisms underlying exceptional survival and healthy aging that involve lipotoxicity.     

Categories of resistance in St. Augustinegrass lines to southern chinch bug (Hemiptera: Blissidae)

The southern chinch bug, Blissus insularis Barber (Hemiptera: Blissidae), is the most important insect pest of St. Augustinegrass, Stenotaphrum secundatum (Walt.) Kuntze, in Florida and other Gulf Coast states. Resistance to southern chinch bug was identified previously in St. Augustinegrass lines 'FX-10' and NUF-76. Choice and no-choice tests and ovipositional and developmental studies were conducted to determine the categories of resistance in FX-10 and NUF-76 to southern chinch bug. When adult chinch bugs had a choice among attached stolons of three susceptible lines ('Floratam', 'Bitter Blue', and 'Palmetto') and the two resistant lines, chinch bugs were found significantly more often over a 5-d period on the susceptible lines. This result indicates the presence of antixenosis in the resistant lines FX-10 and NUF-76. In a no-choice study, chinch bugs produced less than half as many excretory spots on FX-10 as on the susceptible lines. Significantly fewer excretory spots produced by chinch bugs confined on NUF-76 accumulated by days 3 and 5 after release; however, on the first 2 d, the accumulative number of excretory spots was not significantly less than that found on susceptible lines. The no-choice study confirmed a high level of antixenosis in FX-10, a moderate level of antixenosis in NUF-76, and possible antibiosis in NUF-76. Ovipositional and developmental studies were conducted using only Floratam (a widely planted cultivar that was formerly resistant to B. insularis) and the two resistant lines. Adults released on Floratam produced 11 and 5 times more eggs and 18 and 9 times more offspring than adults on FX-10 and NUF-76, respectively. Plant anatomical and biochemical studies are required to investigate the exact cause of antixenosis in FX-10 and NUF-76 and possible antibiosis in NUF-76.     

Oral 18F-fluoro-2-deoxyglucose for primate PET studies without behavioral restraint: Demonstration of principle

We describe a method of orally administering ¹⁸F-fluoro-2-deoxyglucose (FDG) for positron emission tomography (PET) scans to determine local cerebral metabolic rates for glucose (LCMRGlc), normalized to that of whole brain, in fully conscious, non-restrained primates. Oral FDG-PET studies were performed in both non-restrained and chaired monkeys, and in one human where results could be compared with traditional intravenous FDG administration. The oral route of FDG administration gave images and whole brain-normalized PET LCMRGlc results comparable to those obtained by the intravenous route. This oral FDG-PET method may provide a useful means by which to obtain measures of LCMRGlcs for brain structures, relative to each other, in non-restrained, non-druggd primates in field and laboratory studies. This method might also have clinical applications for PET studies of children. Am. J. Primatol. 42:215–224, 1997. © 1997 Wiley-Liss, Inc.     

Changes in household food and drink purchases following restrictions on the advertisement of high fat,salt, and sugar products across the Transport for London network: A controlled interrupted time series analysis

BackgroundRestricting the advertisement of products with high fat, salt, and sugar (HFSS) content has been recommended as a policy tool to improve diet and tackle obesity, but the impact on HFSS purchasing is unknown. This study aimed to evaluate the impact of HFSS advertising restrictions, implemented across the London (UK) transport network in February 2019, on HFSS purchases.Methods and findingsOver 5 million take-home food and drink purchases were recorded by 1,970 households (London [intervention], n = 977; North of England [control], n = 993) randomly selected from the Kantar Fast Moving Consumer Goods panel. The intervention and control samples were similar in household characteristics but had small differences in main food shopper sex, socioeconomic position, and body mass index. Using a controlled interrupted time series design, we estimated average weekly household purchases of energy and nutrients from HFSS products in the post-intervention period (44 weeks) compared to a counterfactual constructed from the control and pre-intervention (36 weeks) series. Energy purchased from HFSS products was 6.7% (1,001.0 kcal, 95% CI 456.0 to 1,546.0) lower among intervention households compared to the counterfactual. Relative reductions in purchases of fat (57.9 g, 95% CI 22.1 to 93.7), saturated fat (26.4 g, 95% CI 12.4 to 40.4), and sugar (80.7 g, 95% CI 41.4 to 120.1) from HFSS products were also observed. Energy from chocolate and confectionery purchases was 19.4% (317.9 kcal, 95% CI 200.0 to 435.8) lower among intervention households than for the counterfactual, with corresponding relative reductions in fat (13.1 g, 95% CI 7.5 to 18.8), saturated fat (8.7 g, 95% CI 5.7 to 11.7), sugar (41.4 g, 95% CI 27.4 to 55.4), and salt (0.2 g, 95% CI 0.1 to 0.2) purchased from chocolate and confectionery. Relative reductions are in the context of secular increases in HFSS purchases in both the intervention and control areas, so the policy was associated with attenuated growth of HFSS purchases rather than absolute reduction in HFSS purchases. Study limitations include the lack of out-of-home purchases in our analyses and not being able to assess the sustainability of observed changes beyond 44 weeks.ConclusionsThis study finds an association between the implementation of restrictions on outdoor HFSS advertising and relative reductions in energy, sugar, and fat purchased from HFSS products. These findings provide support for policies that restrict HFSS advertising as a tool to reduce purchases of HFSS products.

Amy Yau and co-workers study food and drink purchases after the introduction of advertising restrictions in London, UK.     

Fluorescence Microscopy with Antisera Against Specific Cellular Structures: Double Photography Method for Cell Identification in Populations of Multiple Cell Types

Immunofluorescent staining techniques using antitubulin antibody have been difficult to apply to meiotic tissue (testis) because of the large number of cell types present. Such techniques customarily use a fluorescent dye to counterstain nuclei, and this counterstain is hard to distinguish because of the fluorescence of the antitublin. By counterstaining with dilute hematoxylin, we can photograph the same field using UV and then conventional illumination. This double photography allows us to identify precisely the many types of cells present, and it will be a useful tool for reexamining the staging of spermatogenesis.     

Survey traps for parasitoids, and coccinellid predators of the citrus blackfly, Aleurocanthus woglumi

The attractiveness of sticky traps of eight colors for two parasitoids Amitus hesperidum Silverstri and Prospaltella opulenta Silvestri, and seven species of coccinellid predators of the citrus blackfly, Aleurocanthus woglumi Ashby (Homoptera: Aleyrodidae) was evaluated in insectory and field tests.Yellow traps captured significantly more parasitoids and coccinellids than other colors tested; captures were greatest in traps placed in the lower half of citrus trees. These traps can be used to survey for and monitor the population trends of the citrus blackfly and its natural enemies.

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Résumé Nous avons évalué l'attraction de Amitus hesperidum Silvestri et Prospaltella opulenta Silvestri, parasitoïdes d'Aleurocanthus woglumi Ashby et de 7 espèces de Coccinellides prédatrices de cette espèce, par des pièges à glu de 8 couleurs différentes (jaune, orange, rouge, bleu, vert, noir, blanc et transparent).Les pièges jaunes capturent significativement plus de parasitoïdes et de coccinelles que les autres couleurs; les captures sont plus importantes avec des pièges placés dans la moitié inférieure des citronniers. Ces pièges peuvent être utilisés pour suivre et contrôler l'évolution des populations d'A. woglumi et de ses parasitoïdes et coccinelles prédatrices.

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Fla. Agr. Exp. Stn. J. Ser. No. 2271     

Biophysical Factors Influence Methane Fluxes in Subtropical Freshwater Wetlands Using Eddy Covariance Methods

Ecosystems - Wetlands are the largest natural source of methane (CH4); however, the contribution of subtropical wetlands to global CH4 budgets is still unclear due to difficulties in accurately...