Differential Interleukin-2 Transcription Kinetics Render Mouse but Not Human T Cells Vulnerable to Splicing Inhibition Early after Activation

T cells are nodal players in the adaptive immune response against pathogens and malignant cells. Alternative splicing plays a crucial role in T cell activation, which is analyzed mainly at later time points upon stimulation. Here we have discovered a 2-h time window early after stimulation where optimal splicing efficiency or, more generally, gene expression efficiency is crucial for successful T cell activation. Reducing the splicing efficiency at 4 to 6 h poststimulation significantly impaired murine T cell activation, which was dependent on the expression dynamics of the Egr1-Nab2-interleukin-2 (IL-2) pathway. This time window overlaps the time of peak IL-2 de novo transcription, which, we suggest, represents a permissive time window in which decreased splicing (or transcription) efficiency reduces mature IL-2 production, thereby hampering murine T cell activation. Notably, the distinct expression kinetics of the Egr1-Nab2-IL-2 pathway between mouse and human render human T cells refractory to this vulnerability. We propose that the rational temporal modulation of splicing or transcription during peak de novo expression of key effectors can be used to fine-tune stimulation-dependent biological outcomes. Our data also show that critical consideration is required when extrapolating mouse data to the human system in basic and translational research

Southern Corn Rootworm (Coleoptera: Chrysomelidae) Adult Emergence and Population Growth Assessment After Selection With Vacuolar ATPase-A double-stranded RNA Over Multiple Generations

The southern corn rootworm, Diabrotica undecimpunctata howardi Barber (Coleoptera: Chrysomelidae), was exposed over multiple generations to vacuolar (v)ATPase-A double-stranded (ds)RNA, first as adults and later, as neonate larvae. During adult selection, high mortality and lower fecundity were observed in the RNAi-selected cages after beetles were exposed to sublethal dsRNA concentrations that varied between LC40 and LC75. During larval selection, a delay in adult emergence and effects on population growth parameters were observed after neonates were exposed to sublethal dsRNA concentrations that varied between LC50 and LC70. Some of the parameters measured for adult emergence such as time to reach maximum linear adult emergence, time elapsed before attaining linear emergence, termination point of the linear emergence, and total days of linear emergence increase, were significantly different between RNAi-selected and control colonies for at least one generation. Significant differences were also observed in population growth parameters such as growth rate, net reproductive rate, doubling time, and generation time. After seven generations of selection, there was no indication that resistance evolved. The sublethal effects caused by exposures of southern corn rootworm to dsRNAs can affect important life history traits and fitness especially through delays in adult emergence and reduction in population growth. Although changes in susceptibility did not occur, the observation of sublethal effects suggests important responses to potential selection pressure. Assuming resistance involves a recessive trait, random mating between susceptible and resistant individuals is an important factor that allows sustainable use of transgenic plants, and delays in adult emergence observed in our studies could potentially compromise this assumption

Reduced susceptibility of western corn rootworm (\u3ci\u3eDiabrotica virgifera virgifera\u3c/i\u3e LeConte) populations to Cry34/35Ab1‑expressing maize in northeast Nebraska

The western corn rootworm (WCR; Diabrotica virgifera virgifera LeConte) is a significant pest of maize (Zea mays L.) across the United States Corn Belt. Transgenic maize hybrids expressing insecticidal proteins derived from Bacillus thuringiensis (Bt) have been used to manage WCR since 2003. Widespread resistance to Cry3Bb1 (and associated cross-resistance to mCry3A and eCry3.1Ab) has placed increased selection pressure on Cry34/35Ab1 in single-protein and pyramided transgenic maize hybrids. Data on the susceptibility of Nebraska WCR populations to Cry34/35Ab1 has not been published since 2015 and plant-based bioassays conducted in 2017–2018 confirmed resistance to Cry3Bb1 + Cry34/35Ab1 maize, suggesting resistance to Cry34/35Ab1 has evolved in the Nebraska landscape. Therefore, plant-based bioassays were conducted on F1 progeny of WCR populations collected from northeast Nebraska in 2018 and 2019. Larval survival and development were used to classify resistance to Cry34/35Ab1 in each WCR population. Bioassays confirmed incomplete resistance to Cry34/35Ab1 maize in 21 of 30 WCR populations; 9 of 30 WCR populations remained susceptible to Cry34/35Ab1. Collectively, results indicate that northeast Nebraska WCR populations were in the initial stages of resistance evolution to Cry34/35Ab1 during 2018–2019. Appropriate resistance management strategies are needed to mitigate resistance and preserve efficacy of rootworm-active products containing Cry34/35Ab1

Isotropic-medium three-dimensional cloaks for acoustic and electromagnetic waves

We propose a generalization of the two-dimensional eikonal-limit cloak derived from a conformal transformation to three dimensions. The proposed cloak is a spherical shell composed of only isotropic media; it operates in the transmission mode and requires no mirror or ground plane. Unlike the well-known omnidirectional spherical cloaks, it may reduce visibility of an arbitrary object only for a very limited range of observation angles. In the short-wavelength limit, this cloaking structure restores not only the trajectories of incident rays, but also their phase, which is a necessary ingredient to complete invisibility. Both scalar-wave (acoustic) and transverse vector-wave (electromagnetic) versions are presented.Comment: 17 pages, 12 figure

Exploring Optimization for the Random-Field Ising Model

The push-relabel algorithm can be used to calculate rapidly the exact ground states for a given sample with a random-field Ising model (RFIM) Hamiltonian. Although the algorithm is guaranteed to terminate after a time polynomial in the number of spins, implementation details are important for practical performance. Empirical results for the timing in dimensions d=1,2, and 3 are used to determine the fastest among several implementations. Direct visualization of the auxiliary fields used by the algorithm provides insight into its operation and suggests how to optimize the algorithm. Recommendations are given for further study of the RFIM

Exploring optimization for the random-field Ising model

The push-relabel algorithm can be used to calculate rapidly the exact ground states for a given sample with a random-field Ising model (RFIM) Hamiltonian. Although the algorithm is guaranteed to terminate after a time polynomial in the number of spins, implementation details are important for practical performance. Empirical results for the timing in dimensions d=1,2, and 3 are used to determine the fastest among several implementations. Direct visualization of the auxiliary fields used by the algorithm provides insight into its operation and suggests how to optimize the algorithm. Recommendations are given for further study of the RFIM.Comment: 10 page

Fipronil metabolism, oxidative sulfone formation and toxicity among organophosphate- and carbamate-resistant and susceptible western corn rootworm populations

Fipronil toxicity and metabolism were studied in two insecticide-resistant, and one susceptible western corn rootworm (Diabrotica virgifera virgifera, LeConte) populations. Toxicity was evaluated by exposure to surface residues and by topical application. Surface residue bioassays indicated no differences in fipronil susceptibility among the three populations. Topical bioassays were used to study the relative toxicity of fipronil, fipronil. the mono-oxygenase inhibitor piperonyl butoxide, and fipronil\u27s oxidative sulfone metabolite in two populations (one resistant with elevated mono-oxygenase activity). Fipronil and fipronil-sulfone exhibited similar toxicity and application of piperonyl butoxide prior to fipronil resulted in marginal effects on toxicity. Metabolism of [14C]fipronil was evaluated in vivo and in vitro in the three rootworm populations. In vivo studies indicated the dominant pathway in all populations to be formation of the oxidative sulfone metabolite. Much lower quantities of polar metabolites were also identified. In vitro studies were performed using sub-cellular protein fractions (microsomal and cytosolic), and glutathione-agarose purified glutathione-Stransferase. Oxidative sulfone formation occurred almost exclusively in in vitro microsomal reactions and was increased in the resistant populations. Highly polar metabolites were formed exclusively in in vitro cytosolic reactions. In vitro reactions performed with purified, cytosolic glutathione-Stransferase (MW=27kDa) did not result in sulfone formation, although three additional polar metabolites not initially detectable in crude cytosolic reactions were detected. Metabolism results indicate both cytochromes P450 and glutathione-S-transferases are important to fipronil metabolism in the western corn rootworm and that toxic sulfone formation by P450 does not affect net toxicity